bluuebunny/crossref_metadata_2025 huggingface dataset:
PPD‐Specific Proliferative Response in Humans: I. Analysis of PPD‐Specific Proliferative Cells from Tuberculous Pleurisy Patients and Healthy Controls with Monoclonal Antibodies Specific for Human T Subsets
Authors: Okubo Yoshio , Kusama Shozo , Yano Akihiko
Published: November 2013

Peripheral mononuclear cells (PBL) from tuberculin reaction (TR)‐negative tuberculous pleurisy patients proliferated poorly with PPD, while the cells of pleural effusion from these patients showed a proliferative response to PPD as well as did the healthy control PBL. Surface antigens of peripheral blood and pleural effusion were examined by using monoclonal antibodies. The Leu 1‐positive cell population can be divided into four groups, namely (1) Leu 1+, Leu 2a+, Leu 3a+, (2) Leu 1+, Leu 2a+, Leu 3a−, (3) Leu 1+, Leu 2a−, Leu 3a+, and (4) Leu 1+, Leu 2a−, Leu 3a− cell populations. Results of analysis of surface antigens of PPD‐specific proliferative cells in peripheral blood and pleural effusion from tuberculous pleurisy patients as well as healthy controls indicate that the PPD‐specific proliferative response is mediated by Leu 1+, Leu 2a−, Leu 3a+ cells and Leu 1+, Leu 2a−, Leu 3a− cells.
Suppressive Effect by Cooperation of Splenic and Peritoneal Adherent Cells on Generation of Cytotoxic T Lymphocytes
Authors: Koga Yasuhiro , Taniguchi Kazuto , Kubo Chiharu , Nomoto Kikuo
Published: November 2013

When spleen cells primed in vivo against allogeneic lymphoid cells were used as responder cells in secondary mixed lymphocyte cultures, a high degree of cytotoxicity was generated even in the absence of splenic adherent cells. However, removal of adherent cells from such primed responder spleen cells reduced the cytotoxicity to some extent. On the other hand, when these responder cells were transferred into the peritoneal cavity of irradiated syngeneic mice together with antigenic cells, unseparated responder cells generated a lower degree of cytotoxicity than did adherent cell‐depleted responder cells. In an in vitro system, peritoneal adherent cells also suppressed the generation of cytotoxic T lymphocytes by unseparated responders; however they augmented the cytotoxic T lymphocyte generation by adherent cell‐depleted responders. These adherent cell populations with augmenting activity became inhibitory when they coexisted. The mechanism of this inhibitory action remains unclear.
Numerical Analysis of the Taxonomy of Nocardiae and Rhodococci
Authors: Tsukamura Michio
Published: November 2013

The genera Nocardia and Rhodococcus were clearly differentiated in the present study. Eleven characteristics were shown to be useful for differentiation between these two genera. Nocardia asteroides sunsu stricto previously defined by Tsukamura was divided into two taxa. One contained the type strain and was considered to retain the name Nocardia asteroides in a new sense. Another was named in the present study as Nocardia nova sp. nov. Tsukamura. The type strain of this species is ATCC 33726. The following seven characters were useful for differentiating N. nova from newly defined N. asteroides: 1) arylsulfatase activity after 14 days; 2) catalase activity (semiquantitative); 3) β‐esterase activity; 4) pyrazinamidase activity; 5) utilization of citrate as a sole source of carbon; 6) utilization of 2,3‐butylene glycol as a sole carbon source; and 7) resistance to 5‐fluorouracil (20 μg/ml).

The name Nocardia farcinica for Tsukamura's Kyoto‐I group should be rejected. This taxon has been named Nocardia paratuberculosis sp. nov. Tsukamura. The type strain is ATCC 23826.

Three new species of the genus Rhodococcus were proposed: Rhodococcus aichiensis sp. nov. Tsukamura (type strain, ATCC 33611); Rhodococcus chubuensis sp. nov. Tsukamura (type strain, ATCC 33609); Rhodococcus obuensis sp. nov. Tsukamura (type strain, ATCC 33610).
A Simple and Practical Method for Typing and Strain Differentiatin of Herpes Simplex Virus Using Infected Cell DNAs
Authors: Ueno Tetsuji , Suzuki Nobutaka , Sakaoka Hiroshi , Fujinaga Kei
Published: November 2013

A simple and practical method for typing and strain differentiation of herpes simplex virus (HSV) isolates, based upon analysis of the restriction endonuclease cleavage patterns of viral DNAs, was established by using unlabeled infected cell DNAs. The preparation of infected cell DNA is technically easier than that of purified viral DNA or of radiolabeled viral DNA. The method provides a powerful and practical tool for epidemiological and clinical studies of HSV infection, which can be performed in most diagnostic laboratories. In order to select suitable restriction endonucleases for the study of HSV isolates, the cleavage patterns of viral DNAs (strains MacIntyre, HF, UW‐268, and SAV) with 12 enzymes were analyzed. Several enzymes, Bam HI, Kpn I, Pst I, Sal I, Sst I, and Xho I, were found to be useful for both typing and strain differentiation. With this method, HSV isolates from different individuals and from the same individual were analyzed by digestion of their infected cell DNAs with Bam HI. Six isolates from epidemiologically unrelated individuals were readily typed and differentiated from each other. Three isolates from the same individual showed very similar patterns. However, there was a small degree of difference between the first two isolates and the third isolate.
Crystallization and Some Properties of Leukocidin from Pseudomonas aeruginosa
Authors: Hirayama Toshiya , Kato Iwao , Matsuda Futami , Noda Masatoshi
Published: November 2013

A leukocidin was isolated and purified from autolysates of Pseudomonas aeruginosa strain 158 by a combination of procedures such as column chromatography on DEAE‐Sephadex A‐50, gel filtration on Sephadex G‐100, and zone electrophoresis on pevikon. The purified preparation showed a single band in each experiment using electrophoreses in the presence or absence of sodium dodecyl sulfate (SDS), and the agar‐gel Ouchterlony immunodiffusion test. The purified pseudomonal leukocidin was crystallized by salting out with saturated ammonium sulfate at pH 7.0 in a needle‐leaf like form. The molecular weight of the leukocidin was estimated to be 42,500 by SDS‐polyacrylamide gel electrophoresis, 40,000 by gel filtration, and 44,700 (3.3 s20,w) by sucrose density gradient centrifugation. The isoelectric point of the leukocidin was estimated to be at pH 6.3 by isoelectrofocusing. Morphological studies of a leukocidin‐treated leukocyte showed that the formation of vacuoles of cytosolic granules and the swelling of the lobulated nuclei occurred prior to leukocyte enlargement.

In a slide adhesion test with rabbit polymorphonuclear leukocytes (1 × 106), the minimum cytotoxic dose for the destruction of all leukocytes was 13–20 ng of the crystallized toxin. Rabbit lymphocytes were one‐thirtieth as sensitive as rabbit leukocytes. Leukocidin did not act on rabbit erythrocytes or on platelets.
Abortive Infection of L Cells by Influenza B Virus: Defect in Bud Formation
Authors: Takeura Shigeki , Aoki Hiizu , Tsurumi Tatsuya , Nishiyama Yukihiro , Fujioka Hisashi , Yoshii Saiji , Maeno Koichiro
Published: November 2013

Host‐dependent restriction of influenza B virus replication in L cells was analysed in comparison with productive infection in MDCK or 1–5C‐4 cells. The synthesis and intracellular distribution of virus‐specific proteins and the production of cytoplasmic ribonucleoproteins in nonpermissive L cells were similar to those in permissive MDCK cells. However, an electron microscopic study of infected L cells showed neither extracellular virions nor budding virus particles on the cell surface, in contrast to MDCK cells which produced numerous virus particles. PAGE analysis of the plasma membrane isolated from the cells demonstrated no significant difference in the composition of viral polypeptides between permissive 1‐5C‐4 and nonpermissive L cells. It was noted that the abortiveness of influenza B virus infection in L cells may be due to a defect in host cell function involved in the initiation of virus budding.
Nature of Adult T‐Cell Leukemia‐Associated Membrane Antigen on the Surface of Adult T‐Cell Leukemia Virus‐Carrying Cells as Revealed by Membrane Immunofluorescence
Authors: Chosa Tom , Yamamoto Naoki , Koyanagi Yoshio , Kohno Michiaki , Nagata Kinya , Hinuma Yorio
Published: November 2013

Adult T‐cell leukemia‐associated membrane antigen (ATLMA) expressed on the surface of living ATL virus (ATLV)‐carrying cells was investigated by an indirect membrane immunofluorescence method using natural antibodies to ATLV in human sera. All the ATLV‐positive cell lines tested that had cytoplasmic ATL‐associated antigen (ATLA) detectable in acetone‐fixed cell smears were also positive for ATLMA, but ATLMA was not detected in any ATLV‐negative cell lines. The frequencies of ATLA‐ and ATLMA‐bearing cells in seven cell lines tested were roughly parallel. The frequency of expression of both ATLMA and ATLA in cultures of MT‐1 cells increased in the presence of 5‐iodo‐2′‐deoxyuridine. All human sera having ATLA antibody had ATLMA antibody and the titers of the two were similar in most of the sera. The anti‐ATLMA titers of human sera determined by using an ATLV‐bound non‐ATL T‐cell line as antigen were also similar to the anti‐ATLA titers. Absorption of anti‐ATLMA‐positive sera with living MT‐2 cells, in which almost 100% of the cells express ATLA and ATLMA, caused parallel decreases in the anti‐ATLA and anti‐ATLMA titers. Analysis of the 125I‐labeled surface of MT‐2 cells by immunoprecipitation with anti‐ATLMA‐positive human serum followed by gel electrophoresis revealed that p19, p24, p28, and p46 polypeptides were specifically precipitated. These data suggest that ATLMA on the cell surface is not distinguishable from ATLA in the cytoplasm.
Characteristics of Chromosomal DNA Isolated from Escherichia coli Spheroplasts
Authors: Tada Yoshifumi , Yamaguchi Junji
Published: November 2013

The chromosomal DNA of Escherichia coli spheroplasts induced by penicillin G was studied biochemically and electron microscopically. Although the spheroplasts were unable to divide, they continued to synthesize chromosomal DNA for several hours even in the presence of penicillin G. Some differences were observed between the chromosomal DNA of the parent cells and that of the spheroplasts in sucrose gradient centrifugation and electron microscopy; two types of chromosomal DNA, a slower sedimenting form and a faster sedimenting form, were released from the gently lysed parent cells. The former was membrane‐free folded chromosome and the latter was membrane‐associated chromosome. In contrast, the chromosome from the spheroplast showed a single intermediate value of sedimentation coefficient between those of the chromosomal DNA from the parent cell. Cytochrome spreading for electron microscopy showed that the spheroplast chromosomal DNA formed an aggregated mass consisting of several chromosome‐molecules of the parent cell.
Community Outbreak of Yersinia pseudotuberculosis
Authors: Inoue Masanao , Nakashima Hiroshi , Ueba Osamu , Ishida Tatsuo , Date Hiroko , Kobashi Shizuko , Takagi Kanji , Nishu Tatsuo , Tsubokura Misao
Published: November 2013

An outbreak of Yersinia pseudotuberculosis in Kurashiki, Japan is described. This is the first conclusive report of a community outbreak of this microorganism. A total of 535 pupils, five teachers, and one food attendant contracted the organism. Causative organisms were detected in 19 out of 30 patients. All isolated strains belonged to serotype VA. Out of 653 sera of the pupils, 488 showed elevated agglutinin titers ranging from 1: 80 to 1: 1,280 or more within a period of 3 months.
Hepatic Drug‐Metabolizing Enzyme System and Endotoxin Tolerance: Structural Requirement of LPS in Induction of an Early Tolerance
Authors: Egawa Kiyoshi , Yoshida Michiko , Sakaino Rie , Kasai Nobuhiko
Published: November 2013

The alteration of hepatic drug‐metabolizing enzyme activities in mice given Salmonella endotoxin by single or multiple intraperitoneal injections was investigated. An essentially the same biphasic, early and late phase, endotoxin tolerance was observed in the animals receiving a single injection of endotoxin or repetitive daily injections. The results of reciprocal cross tolerance tests using lipopolysaccharide and free lipid A preparations derived from Salmonella minnesota, Salmonella typhimurium, E. coli, Pseudomonas aeruginosa, and Chromobacterium violaceum suggested that lipid A moiety plays an important role in the induction of early endotoxin tolerance to endotoxin response.
Electron Microscopic Studies on Intracellular Multiplication of Rickettsia tsutsugamushi in L Cells
Authors: Urakami Hiroshi , Tsuruhara Takashi , Tamura Akira
Published: November 2013

The 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

To 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

Both 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

A 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

Biosynthetic 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

Experiments 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

A 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

To 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

A 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

Mycobacterium 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

The 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

The 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

The 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

Unfractionated 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

In 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

The 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

Twenty 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

The 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.
IgE Antibody Responses against Japanese Cedar Pollen in the Mouse
Authors: Tamura Shin‐Ichi , Kobayashi Toshiaki , Kikuta Kiyoshi , Nakagawa Masaro , Sakagughi Masahiro , Inouye Sakae
Published: November 2013

IgE antibody responses against Japanese cedar pollen in the mouse were investigated to develop a mouse model of human allergy for combinations of factors including pollen administration routes, elicitation antigens and inbred mouse strains. Daily short term inhalation of native pollen or intratracheal administration of pollen suspended in saline induced IgE antibody responses in DBA/2, BDF1 and Balb/c mice, but failed to induce any detectable responses in C57BL/6 and C57BL/10 mice. Intraperitoneal injection of pollen suspension also induced IgE antibody responses in DBA/2, BDF1 and Balb/c mice but not in C57BL/6 mice. IgE antibody responses against pollen described above were detected by passive cutaneous anaphylaxis (PCA) reactions using crude extract of pollen as an elicitation antigen. On the other hand, IgE antibodies specific for antigen Sugi basic protein (AgSBP), which is a major allergen of pollen in humans (Yasueda, H., Yui, Y., Shimizu, T., and Shida, T., 1983. Isolation and partial characterization of the major allergen from Japanese cedar (Cryptomeria japonica) pollen. J. Allergy Clin. Immunol. 71: 77–86), were also detected by PCA reactions using AgSBP in the sera from mice which received secondary or the tertiary stimulation by pollen. These results suggest that IgE antibody responses against Japanese cedar pollen in the mouse can be induced by airway sensitization and that the responses are genetically controlled by H‐2‐linked immune response genes. The results also suggest that not only IgE antibody responses specific for components other than AgSBP but also responses specific for AgSBP can be induced in the mouse by repeating appropriate sensitization by pollen.
Protective Effect of Acidic Mannan Fraction of Bakers' Yeast on Experimental Candidiasis in Mice
Authors: Okawa Yoshio , Suzuki Ko , Kobayashi Makiko , Asagi Masako , Sakai Kazuhiko , Suzuki Shigeo , Suzuki Masuko
Published: November 2013

An acidic fraction of Bakers' yeast mannan, WAM025, showed a significant protective effect against Candida albicans infection in mice, but a neutral fraction of the same Bakers' yeast mannan, WNM, did not exhibit this effect. Moreover, pretreatment with WAM025 resulted in a marked reduction of proliferation of C. albicans cells in the organs of the infected mice. We investigated the stimulative effect of these mannan fractions on the function of mouse peritoneal phagocytes, and found that mice administered WAM025 showed a greater increase in the number of peritoneal exudate cells, macrophages and polymorphonuclear leucocytes (PMN), than the mice treated with WNM, especially in the proportion of PMN. Peritoneal phagocytes, PMN and macrophages obtained from WAM025‐treated mice showed marked candidacidal activity. Of the phagocytes, PMN were responsible for the larger part of the candidacidal activity. The myeloperoxidase activities of PMN and macrophages in WAM025‐treated PEC were greater than in untreated macrophages. The myeloperoxidase activity of WAM025‐treated PMN was significantly greater than that of WAM025‐treated macrophages. This activity paralleled the active oxygen‐releasing activity of the phagocytes. On the other hand, the phagocytic activity of phagocytes from mice administered WNM or WAM025 for C. albicans cells was identical to that of untreated phagocytes. WAM025 seems to cause enhance elimination of the pathogen from mice, by increasing the number and candidacidal activity of phagocytic cells.
Increased Sensitivity to Phagocytosis of Staphylococcus E‐46 after Growth in Artificial Media
Authors: Yamamoto Mitsuyo , Iwata Kazuo
Published: November 2013

Staphylococcus E‐46, which exhibits high virulence in mice mainly due to resistance to phagocytosis, gradually lost its virulence during growth in artificial media [Heart‐Infusion (HI) slants]. Staphylococcus E‐46 is coagulase‐negative and DNase‐positive, and has a capsule‐like structure, and the less‐virulent derivative (LVD) strain seemed not to be changed in this respect. The bacteria which had been exposed to HI slants for more than one year (=LVD strain) became more sensitive to phagocytosis by mouse peritoneal macrophages than bacteria which were kept in a lyophilized state. The chemiluminescent response of macrophages to the LVD strain was remarkably higher than that to the original Staphylococcus E‐46. When the LVD or the sera for opsonization were heated, the chemiluminescence to the LVD was as low as that to the original Staphylococcus E‐46. Polyacrylamide gel electrophoresis of sonication‐released membrane protein from the LVD strain showed a unique band with a molecular weight of about 40,000. The factor concerned with the virulence of Staphylococcus E‐46 is discussed based on these results.
Characterization of Purified Shiga Toxin from Shigella dysenteriae 1
Authors: Yutsudo Takashi , Honda Takeshi , Miwatani Toshio , Takeda Yoshifumi
Published: November 2013

Shiga toxin was purified from the culture supernatant of Shigella dysenteriae 1 by ammonium sulfate fractionation, DEAE‐cellulose column chromatography and repeated chromatofocusing column chromatography. About 1.6 mg of purified Shiga toxin was obtained from 15 liters of culture with a yield of about 27%. The molecular weight of purified Shiga toxin was estimated to be 62,000. The toxin consisted of A and B subunits with molecular weights of about 30,000 and 5,000–6,000, respectively. The isoelectric point of purified Shiga toxin was 7.0. Purified Shiga toxin showed the following biological activities: lethal toxicity to mice when injected intraperitoneally with an LD50 of 28 ng per mouse; cytotoxicity to Vero cells, killing about 50% of the cells at 1 pg and all of the cells at 10 pg; and fluid accumulation in rabbit ileal loops at a concentration of more than 1 μg.
Strain Differences of Interferon–Generating Capacity and Resistance in Toxoplasma–Infected Mice
Authors: Shirahata Toshikazu , Mori Akiko , Ishikawa Hiroshi , Goto Hitoshi
Published: November 2013

To explore a possible correlation between susceptibility to Toxoplasma and interferon (IFN)‐generating capacity in mice, we compared the levels of serum IFN induced by stimulation with Toxoplasma lysate antigen (TLA) in different strains of Toxoplasma‐infected and uninfected mice. Injection of TLA into five strains of mice with chronic Toxoplasma infection resulted in the release of considerable amounts of IFN into the circulation. Most of these IFN activities were acid labile and not neutralized by sheep antiserum against mouse IFN‐α/β, indicating that IFN‐γ was the dominant form produced in this system. In contrast, the majority of IFN induced in uninfected mice was characterized as IFN‐α/β by their acid stability and antigenicity. The response of IFN production in Toxoplasma‐infected and uninfected mice varied quantitatively depending on the mouse strains examined. C57BL/6 mice were found to be the best producers of both IFN‐α/β and IFN‐γ, while BALB/c mice were consistently poor producers of both IFN populations. A/J, DBA/2, and C3H/He mice could be roughly classified as intermediate producers of both IFN populations. C57BL/6 and C3H/He mice showed a significant prolongation of mean survival time following primary or secondary infection with Toxoplasma compared to that of BALB/c mice. However, there was no direct correlation between the susceptibility to Toxoplasma and the levels of serum IFN.
Epidemiological Studies on the Background of the Endemic Occurrence of Tsutsugamushi Disease in Toyama Prefecture
Authors: Ishikura Mitsuhiro , Watanabe Mamoru , Matsuura Kumiko , Nakayama Takashi , Morita Osayuki , Uetake Hisao , Sakurada Norio
Published: November 2013

With a view to clarifying the actual state of inapparent infection of tsutsugamushi disease, inhabitants of endemic and nonendemic areas were screened for anti‐Rickettsia tsutsugamushi antibody (anti‐Rt antibody) by the indirect immunofluorescence test. The anti‐Rt antibody‐positive rate in the inhabitants of the endemic area (about 50%) was statistically significantly higher than that in the nonendemic area (14.7%). The antibody titer in the inhabitants of the endemic area was 10–160, and the number of inhabitants showing a high antibody titer was 2–4 times larger than that of the nonendemic area. A total of 257 volunteers in the endemic area were analyzed for the changes in anti‐Rt antibody titer over 1.5–2 years on an individual basis. An increase in the antibody titer was found in 20 inhabitants. There was no difference in the anti‐Rt antibody‐positive rate between male and female in either the endemic or the nonendemic area. The positive rate was also compared as to the distribution by 10 years of age. In the endemic area, there were no significant differences in the positive rate between any pair of 10‐year age groups from 30s to 60s, whereas in the nonendemic area, the positive rate in the teen‐age group was significantly lower than those in the age groups of 20 years or older. In Yamada district, the numbers of serum samples obtained from each age group were about the same, and the distribution of the positive rates showed a normal distribution. The nurse students having their homes in Toyama Prefecture were plotted on the map as for their anti‐Rt antibody and geographical distribution. The results showed that many of them having homes in the endemic area were positive for the antibody, while some antibody‐positives were scattered all over Toyama Prefecture.
The Role of Interleukin 1 and 2 in Generation of Acquired Resistance against Mouse Typhoid Infection Afforded by Dialyzable Factor from Salmonella typhimurium
Authors: Kita Eiji , Emoto Masashi , Nishi Kazuyuki , Katsui Noriaki , Kashiba Shuzo
Published: November 2013

Dialyzable factor (DF) prepared from a ribosomal fraction of Salmonella typhimurium was tested for its ability to induce interleukin 1 (IL 1) and 2 (IL 2) production, in relation to acquired resistance, after an intraperitoneal injection of DF. IL 1 production in vitro by peritoneal macrophages of DF‐treated mice reached the maximum 4 days after injection, at the time when the nonspecific local resistance via macrophages directly activated with DF became apparent (Kita et al, Microbiol. Immunol. 28: 807, 1984). Concanavalin A‐induced IL 2 production by splenocytes of DF‐treated mice reached the maximal level between days 6 and 8, and it could be enhanced even on day 14. Antigen‐induced blastogenic responses of splenocytes from DF‐treated mice reached the maximal level 14 days after treatment. Although DF did not show the mitogenic activity to normal splenocytes, T cells of DF‐treated mice could respond to S. typhimurium. On the contrary, T cells of normal mice could respond to heat‐killed cells of S. typhimurium when they were cultured with macrophages which had been directly stimulated in vitro with DF. Furthermore, T cells from DF‐treated mice could respond to antigens of different species of bacteria, and especially to Listeria monocytogenes. These results suggest that T cells of DF‐treated mice, being at the intermediate stage of activation via monokines including IL 1 which is produced by macrophages stimulated with DF, are able to proliferate immediately after the administration of challenging organisms as a second signal, and also that the specificity of the response may be defined by the challenging organisms.
A Plasmid of Group Q Which Confers Resistance to Trimethoprim and Sulfonamides
Authors: Hedges R. W.
Published: November 2013

A 5.2‐Mdal plasmid, determining resistance to trimethoprim and sulfonamides, is a member of incompatibility group Q.
Electron Microprobe X‐Ray Analysis of Polyphosphate Granules in Plesiomonas shigelloides
Authors: Ogawa Junko , Amano Yasuji
Published: November 2013

Electron‐dense inclusion bodies were found in most Plesiomonas shigelloides cells, regardless of the incubation time. At the 4‐hr incubation period, the size of inclusion bodies was distributed in the range of 50 to 150 nm in diameter, and at the logarithmic phase of growth it increased up to a size visible by light microscope. By an electron microprobe X‐ray analysis, phosphorus, potassium, and magnesium were detected in the inclusion bodies which confirms the assumption of Pastian and Bromel (Appl. Environ. Microbiol. 47: 216 (1984)) that the inclusion bodies have a very similar elemental composition to the polyphosphate granules of C. diphtheriae.