bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chemical Properties of Lipopolysaccharide‐Like Substance (LLS) Extracted from Leptospira interrogans Serovar canicola Strain Moulton
Authors: Shimizu Tadayori , Matsusaka Eiichiro , Nagakura Naoki , Takayanagi Kazuhisa , Masuzawa Toshiyuki , Iwamoto Yoshihisa , Morita Tamotsu , Mifuchi Ichiji , Yanagihara Yasutake 
Published: November 2013

AbstractThe aqueous layer was isolated from Leptospira interrogans serovar canicola strain Moulton by the hot phenol‐water method. After ultracentrifugation, the precipitate was designated as lipopolysaccharide‐like substance (LLS) fraction and the chemical composition was compared with that of bacterial LPS. The LLS fraction consists of 35.2% carbohydrate, 3.8% amino sugar, 36.4% lipid, 15.2% protein, and 0.3% phosphorus. Neutral sugars were detected as rhamnose, arabinose, xylose, 4‐O‐methylmannose, mannose, galactose, and a small amount of erythrose, fucose and glucose by gas‐liquid chromatography (GLC), but 2‐keto‐3‐deoxyoctonic acid was not detected in the LLS by thiobarbituric acid test and high voltage paper electrophoresis. Fatty acids detected by GLC were decanoic acid (C10: 0), dodecanoic acid (C12: 0), dodecenoic acid (C12: 1), tridecenoic acid (C13: 1), tetradecanoic acid (C14: 0), hexadecanoic acid (C16: 0), hexadecenoic acid (C16: 1), and octadecenoic acid (C18: 1). With SDS‐polyacrylamide gel electrophoresis, bacterial LPS showed many orderly bands, while the banding pattern of the leptospiral LLS was very simple. These findings demonstrate that the physicochemical properties and chemical composition of LLS fraction from Leptospira are different from those of LPS extracted from gram‐negative bacteria such as Enterobacteriaceae, and suggesting that Leptospira has no typical LPS.
Biological Activities of Lipopolysaccharide‐Like Substance (LLS) Extracted from Leptospira interrogans Serovar canicola Strain Moulton
Authors: Shimizu Tadayori , Matsusaka Eiichiro , Takayanagi Kazuhisa , Masuzawa Toshiyuki , Iwamoto Yoshihisa , Morita Tamotsu , Mifuchi Ichiji , Yanagihara Yasutake 
Published: November 2013

AbstractThe biological activities of lipopolysaccharide‐like substance (LLS) extracted from Leptospira interrogans serovar canicola strain Moulton by the hot phenol‐water method were studied in mice. The addition of 12.5 μg/ml or more of LLS fraction increased the incorporation of [3H] thymidine into in vitro cultured spleen cells of C57BL/6 mice, while the activity of the LLS fraction was about 20 times weaker than that of Salmonella typhimurium lipopolysaccharide (LPS). Pretreatment of murine spleen cells with rabbit anti‐mouse thymocyte antiserum did not diminish the mitogenic activity of leptospiral LLS, and the LLS could not increase the incorporation of [3H] thymidine into thymocytes, suggesting that LLS acts on a B‐lymphocyte population of lymphocytes. When sheep erythrocytes and LLS fraction were injected intraperitoneally into BALB/c mice, LLS exhibited an enhancing effect on antibody response in vivo. However, lethal toxicity of the LLS fraction was about 500 times lower than that of LPS in C57BL/6 mice loaded with galactosamine. No antitumor activity of leptospiral LLS (250–1,000 μg/mouse) against the ascites form of Ehrlich carcinoma in ddY mice was observed. The biological activities of the LLS fraction from the organism were weaker than those of gram‐negative bacterial LPS, suggesting that Leptospira possesses no typical LPS.
Identification of Re Lipopolysaccharide‐Binding Protein on Murine Erythrocyte Membrane
Authors: Kirikae Teruo , Inada Katsuya , Hirata Michimasa , Yoshida Masao , Kondo Seiichi , Hisatsune Kazuhisa 
Published: November 2013

AbstractOur recent studies have suggested that bacterial lipopolysaccharide (LPS) attaches to Pronase‐sensitive proteins on the murine erythrocyte membrane. In the present study, in order to identify the LPS‐binding protein on the murine erythrocyte membrane, a unique method to detect LPS‐binding protein on a nitrocellulose membrane was developed. Murine erythrocyte membrane proteins were separated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, then transferred electrophoretically onto a nitrocellulose membrane. The membrane was incubated with LPS of Salmonella minnesota R595 (Re LPS) in phosphate‐buffered saline (PBS), after the remaining sites were blocked with gelatin in PBS. We were able to obtain a non‐background stain by adding the nonionic detergent octylglucoside at the low concentration of 0.1% to the Re LPS solution. The Re LPS bound to the protein on the nitrocellulose membrane was exposed to affinity purified anti‐Re LPS antibodies (IgG) and then to alkaline phosphatase‐conjugated anti‐IgG. The alkaline phosphatase was detected on the membrane by an enzymatic reaction. This method demonstrated that Re LPS was bound to an erythrocyte protein of 96 kDa. Treatment of erythrocytes with Pronase led to disappearance of the Re LPS‐binding protein on the erythrocyte membrane. There was no difference between LPS‐responder and LPS‐nonresponder murine erythrocyte membranes in amount and molecular weight of the Re LPS‐binding protein.
“Transformation” of Human Endothelial Cells by SV40 Virions
Authors: Ide Hideyuki , Minamishima Yoichi , Eizuru Yoshito , Okada Masanobu , Sakihama Kuniharu , Katsuki Taketo 
Published: November 2013

AbstractHuman endothelial cells derived from the umbilical vein were transformed with SV40 virions. A cell line subcultured for over 60 serial passages was characterized in comparison with its untransformed counterpart which was culturable for less than five passages.The SV40‐transformed human endothelial cells, designated SV‐HUVEC, were positive not only for tumor (T) antigen specific to the SV40‐transformed cell, but also for two markers of endothelial cells, Factor VIII‐related antigen and a receptor for Ulex europaeus agglutinin I. By transformation the growth potential of the human endothelial cells was increased and their serum requirement was decreased. The SV40‐transformed endothelial cells were, however, unable to form colonies in soft agar or to form tumors in athymic nude mice, although a small nodule was produced at the site of inoculation. Subcultivation of these cells up to the 62nd passage eventually resulted in crisis and loss of further cell division. Thus, the human endothelial cells were transformed by SV40 while retaining certain normal functions but without showing tumorigenicity.
Homologous Human Macrophage Hybridomas That Produce a Novel Cytotoxic Factor in Their Culture Supernatants
Authors: Woo Hee Jong , Shimoda Olina , Imai Yasuyuki , Osawa Toshiaki 
Published: November 2013

AbstractHomologous human macrophage hybridoma cell lines were obtained by somatic cell fusion between peripheral blood monocyte‐derived macrophages and a subclone of the myelomonocytic cell line, U937‐F9. The hybridoma cell lines grown in vitro for more than a year were confirmed by manifestations of phagocytosis, adherence, nonspecific esterase, acid phosphatase, chromosome numbers and other cell surface antigens. Cell surface antigens on hybridomas were detected by flow cytometry analysis with monoclonal antibodies. With interclonal differences, a typical phenotype of hybridoma cells was CDw14+, OKM5+, Mac‐1+ (equivalent to OKM1 and Mol), OKT9+, HLA‐DR− and CD20+. After stimulation with lipopolysaccharide and calcium ionophore A23187, culture supernatants of clones c18A and c29A showed cytotoxic activity against human melanoma A375 Met‐Mix and other cell lines which were resistant to the tumor necrosis factor, lymphotoxin and interleukin 1. This cytotoxic factor was found to be distinct from the tumor necrosis factor, lymphotoxin and interleukin 1 using the anti‐tumor necrosis factor, anti‐lymphotoxin and anti‐interleukin 1 antisera.
Identification of B‐L Antigens on Reticular Epithelial Cells of the Bursa of Fabricius
Authors: Hoshi Sumio , Nunoya Tetsuo , Ueda Susumu 
Published: November 2013

AbstractWe have established two monoclonal antibodies against B‐L antigens (chicken Ia‐like antigens). The specificity of the antibodies for B‐L antigens was determined by two criteria, the cellular expression and the molecular structure of antigens with which they reacted. They reacted with antigens expressed on bursacytes, Con A‐blast thymocytes, macrophages, and MDCC MSB1, but not with thymocytes and erythrocytes. In molecular basis, they recognized 64,000 dalton glycoprotein consisting of two polypeptides, 35,000 and 32,000 dalton, which bound non‐covalently. To investigate the distribution of B‐L antigens on non‐lymphoid cells of the bursa of Fabricius, which were thought to play important roles in the differentiation of B cells, anti‐B‐L antigen and anti‐chicken immunoglobulin (Ig) monoclonal antibodies were used. B‐L antigen‐positive cells were detected in both cortical and medullary areas, whereas Ig‐positive lymphoid cells were confined to the medullary areas of normal chicken bursal follicles. In the bursal follicles of cyclophosphamide (CY)‐treated chickens, lymphoid cells were depleted but epithelial cells remained intact. And B‐L antigen‐positive but Ig‐negative cells were easily detected in the medullary areas of almost all follicles. These cells were identified to be reticular epithelial cells (REp cells) from the result of their keratin expression.
The Structure of Marek's Disease Virus DNA: Amplification of Repeat Sequence in IRs and TRs
Authors: Hayashi Masanobu , Jessip John , Fukuchi Kunihiko , Smith Mary , Tanaka Akiko , Nonoyama Meihan 
Published: November 2013

AbstractThe structure of Marek's disease virus (MDV) DNA was investigated by using Southern blot hybridization analysis. A heterogeneous region was observed in the inverted repeats region, IRs and TRs, as well as in the TRL and IRL. The results of DNA sequencing of the heterogeneous region showed that the heterogeneity of IRs and TRs was due to amplification of a 178‐bp repeat sequence. Amplification of IRs and TRs was found in viral DNA from both pathogenic and nonpathogenic strains. The structure of DNA from the latent MDV genome present in established lymphoblastic cells was also determined. Amplification of the 132‐bp repeat sequence in IRL and TRL was not found in latent MDV DNA of established lymphoblastic cells, whereas amplification of the 178‐bp repeat sequence in IRs and TRs was found in the same DNA.
Tolerance Induction of Alloreactivity by Portal Venous Inoculation with Allogeneic Cells Followed by the Injection of Cyclophosphamide
Authors: Kokudo Shuhei , Sato Soichiro , Qian Jia‐hua , Wada Kentaroh , Ikegami Ryoichi , Hamaoka Toshiyuki , Fujiwara Hiromi 
Published: November 2013

AbstractBALB/c mice receiving allogeneic C3H/He or C57BL/6 spleen cells via portal venous (p.v.) route or a single administration of cyclophosphamide (Cy) were capable of rejecting the respective allogeneic C3H/He‐ or C57BL/6‐derived tumor cells. In contrast, the combined treatment of p.v. inoculation with allogeneic lymphocytes and Cy administration abrogated the capability of rejecting allogeneic tumor cells. Such abrogation of alloreactivity was alloantigen‐specific and associated with the suppression of potentials to generate delayed‐type hypersensitivity (DTH) and cytotoxic T lymphocyte (CTL) responses to alloantigens. This was further substantiated by the inhibition of molecular mechanisms underlying anti‐allo‐DTH and ‐CTL responses. Thus, the above combined treatment led to the decreased production of lymphokines such as macrophage‐activating factor (MAF) and interleukin 2 (IL2) following the stimulation with the relevant alloantigens. These results demonstrate that p.v. inoculation of allogeneic cells followed by a single administration of Cy results in the effective elimination of alloreactivity as verified by the suppression of cellular and molecular mechanisms of alloreactive responses.
Some Applications of the Surface‐Spreading Technique to Virological Studies
Authors: Yazaki Kazumori , Sano Toshihiko , Ikegami Masato , Urushibara Toshiyuki 
Published: November 2013

AbstractBy application of the surface‐spreading technique, virus particles in infected cells and viremia serum, and precipitates in agar plates of the double immunodiffusion technique of Ouchterlony could easily and clearly be visualized without any purification process.
Two‐Dimensional Flow Cytometric Analysis of T Cell Subsets Maintained in IL‐2 Medium after Autologous Mixed Leukocyte Reaction Activation
Authors: Goto Makoto , Miyamoto Terumasa , Nishioka Kusuki 
Published: November 2013

AbstractHuman T cells are stimulated with an autologous mixed leukocyte reaction (AMLR) and can be propagated in interleukin‐2. Staining of the cultured cells with the combination of two monoclonal antibodies was evaluated by two‐dimensional flow cytometry at weekly intervals. AMLR activation resulted in an initial preservation of the CD4+ (helper/inducer T) subset predominance over the CD8+ (suppressor/cytotoxic T) cells, noted on normal circulating blood lymphocytes. However, during culture in interleukin‐2, there was a progressive increase in the percentages of CD8+ Leu 15− cytotoxic T, CD4+ Leu 8− helper T, and CD3+ HLA‐DR+ activated T cells, and a concomitant decrease in those of CD4+ Leu 8+ suppressor inducer T and CD8+ Leu 15+ suppressor T cells if the responder sheep red blood cell (SRBC)‐rosetting T cells were made up by tris ammonium chloride, but not by hypotonic shock treatment to lyse SRBC. The significant difference between hypotonic shock‐T cells and ammonium chloride‐T cells in the phenotypic changes of T cell subsets after long‐term culture in an interleukin‐2 medium may suggest a regulatory role of the ammonium chloride‐sensitive T cells in the AMLR.
Relationship between Mycobacterial Species and Their Carotenoid Pigments
Authors: Ichiyama Satoshi , Shimokata Kaoru , Tsukamura Michio 
Published: November 2013

AbstractA study of the relationship between mycobacterial species and their carotenoid pigments was carried out. According to the carotenoid pigments contained, the mycobacterial species tested were divided into four groups: the first group ofMycobacterium kansasiiandM. marinum, which formed principally onlyβ‐carotene; the second group ofM. gordonae, M. scrofulaceum, M. szulgai, M. xenopi, M. flavescens, M. phlei, M. rhodesiae, M. neoaurum, andM. aichiense, which formedβ‐carotene and a zeaxanthin‐like substance; the third group ofM. aurumandM. obuense, which formedβ‐carotene and an eschscholtzxanthin‐like substance; and the fourth group ofM. chubuenseandM. tokaiense, which formedβ‐carotene and zeaxanthin‐ and eschscholtzxanthin‐like substances. The common carotenoid pigment throughout the genusMycobacteriumwasβ‐carotene and the hypophasic carotenoids differed according to the species.
Formation of a Hexagonal Lattice Structure by an R‐Form Lipopolysaccharide of Klebsiella: Effect of Various Divalent Cations on the Lattice Formation
Authors: Kato Nobuo , Ohta Michio , Kido Nobuo , Ito Hideo , Naito Setsuko 
Published: November 2013

AbstractThe R‐form lipopolysaccharide from Klebsiella pneumoniae strain LEN‐111 (O3‐: Kl‐), from which cationic material had been removed by electrodialysis, was previously shown to form a hexagonal lattice structure with the lattice constant of 14 to 15 nm when suspended in 50 mm tris(hydroxymethyl)aminomethane buffer at pH 8.5 containing 10 mm Mg2+. Under this experimental condition, effects of other divalent metal cations on the hexagonal assembly of the electrodialyzed LPS were compared with that of Mg2+. The Zn2+, Hg2+, Cu2+, and Ni2+ could produce essentially the same hexagonal lattice structure with the lattice constant of 14.5 to 15.0 nm as that formed with Mg2+. The Cd2+, Co2+, and Fe2+ produced the hexagonal lattice structure with the lattice constant of 15.5 to 16.0 nm, and Ba2+, Sr2+, and Ca2+ produced that with the lattice constant of 18 to 19 nm. In addition, the hexagonal lattice structures formed with the latter three cations were less orderly than those formed with the other cations. When the higher concentrations of Ba2+, Sr2+, and Ca2+ were used, the lattice constants were not shortened. The length of lattice constants of the hexagonal lattice structures formed with the divalent cations did not relate to the quantity of the cations bound to the LPS. Among the divalent cations tested, Hg2+ was bound to the LPS in the smallest amount (its atomic ratio to P, 0.07), and Zn2+ and Fe2+ were bound in very large amounts (their atomic ratios to P, 2.94 and 8.28, respectively).
T Cell‐Mediated Cytotoxicity against Dengue‐Infected Target Cells
Authors: Pang Tikki , Devi Shamala , Blanden Robert V. , Lam Sai Kit 
Published: November 2013

AbstractA cytotoxic T lymphocyte (CTL) response to dengue virus‐infected target cells is described. Effector cells were generated in an in vitro secondary culture and appeared to be T cells possessing both the Lyt 1.1 and Lyt 2.1 surface antigens. A stronger CTL response was noted with the H‐2k haplotype compared to H‐2d, and H‐2 compatibility was required between CTL and target cells. CTL generated showed some cross‐reactivity with target cells infected with Japanese encephalitis virus (JEV), another flavivirus, but not with target cells infected with an alphavirus, Sindbis. The significance and importance of these findings are discussed.
Purification and Some Properties of Cytotoxin Produced by Clostridium difficile
Authors: Katoh Takahiro , Honda Takeshi , Miwatani Toshio 
Published: November 2013

AbstractThe cytotoxin produced by Clostridium difficile was highly purified by using ammonium sulfate fractionation and successive column chromatographies of DEAE‐Sephadex A‐25, hydroxyapatite, Bio‐Gel A‐0.5m, Phenyl‐Sepharose CL‐4B, and Mono Q. The purified cytotoxin gave a single band on conventional and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis in the presence of dithiothreitol. Its molecular weight was estimated to be 260,000 and 50,000 by gel filtration and sodium dodecyl sulfate‐polyacrylamide slab gel electrophoresis in the presence of dithiothreitol, respectively. Thus it was supposed that the toxin consists of 5 subunits having molecular weight of approximately 50,000. It had an isoelectric point of 6.6. The toxin was heat‐labile (60 C for 10 min) and inactivated by treatment with trypsin and pronase, or at pH below 4 or over 10. The minimum cytotoxic dose of the cytotoxin against Chinese hamster ovary cells was 3 ng. It was also demonstrated that the toxin is antigenically different from enterotoxin of C. difficile.
Activities and Properties of Putrescine‐Biosynthetic Enzymes in Vibrio parahaemolyticus
Authors: Yamamoto Shigeo , Nakao Hiroshi , Yamasaki Keiko , Takashina Kimiko , Suemoto Yasuo , Shinoda Sumio 
Published: November 2013

AbstractThe biosynthetic pathways for putrescine (Put) in Vibrio parahaemolyticus were delineated by measuring activities of the enzymes which would be involved in its biosynthesis. Experiments with labeled arginine and ornithine revealed that both of these amino acids were converted into Put by intact cells. The activities of three enzymes, arginine decarboxylase (ADC), ornithine decarboxylase (ODC), and agmatine ureohydrolase (AUH), were detected in cell extracts. ADC and ODC of V. parahaemolyticus were similar in the following properties to the corresponding enzymes of Escherichia coli: 1) both decarboxylases showed a pH optimum at 8.25 and required pyridoxal phosphate and dithiothreitol for full activity; 2) while ODC was considerably activated by GTP, ADC was only slightly; 3) both decarboxylases were inhibited by polyamines; 4) ADC was inhibited by difluoromethylarginine, a potent inhibitor of bacterial ADC. However, in contrast to the corresponding enzymes of E. coli, the V. parahaemolyticus ADC showed no requirement for Mg2+, and the AUH was active over a wide pH range of 8.5–9.5 with a maximum at pH 9.0. Furthermore, in all 6 strains tested, the activity of ADC was obviously high compared with that of ODC, and AUH was present with a relatively high activity. Cultivation of these strains at a suboptimal NaCl concentration (0.5%) resulted in a pronounced increase in both ADC and AUH activities. These observations suggest that the important pathway for Put biosynthesis in V. parahaemolyticus is the decarboxylation of arginine by ADC and the subsequent hydrolysis of its product, agmatine, by AUH.
Effect of C‐Reactive Protein on Peritoneal Macrophages
Authors: Miyagawa Nobuyuki , Okamoto Yasuyuki , Miyagawa Sachiko 
Published: November 2013

AbstractThe effect of human C‐reactive protein (CRP) isolated and purified from pooled patients' sera on macrophage function, especially on macrophage migration, was studied. Peritoneal exudate cells (PEC) from guinea pigs were used for macrophage migration inhibition (MMI) test of capillary method. Migration of either PEC or adherent purified macrophages exposed to CRP were inhibited dose‐dependently. These findings indicate that CRP inhibits macrophage migration directly, not via activation of lymphocytes contained in PEC. As control, we examined the effect of normal human serum, anti C‐polysaccharide antibodies isolated from patients' sera, and free endotoxin at the dose contaminated in CRP preparation on macrophage migration and found that none of them were effective. The effect of CRP on MMI of sensitized PEC exposed to antigen was also studied. Large amounts of CRP inhibited MMI induced by antigen, indicating the possibility that CRP may act on macrophages competitively with migration inhibitory factor (MIF) and may modulate MMI. CRP possesses MIF‐like activity and may play a functional role at the site of tissue injury by causing the accumulation of macrophages.
Effect of C‐Reactive Protein on Peritoneal Macrophages
Authors: Miyagawa Nobuyuki , Okamoto Yasuyuki , Nakano Hiroshi 
Published: November 2013

AbstractThe effect of human C‐reactive protein (CRP) on macrophage function was studied in an assay of superoxide anion (O2−) production. Peritoneal exudate macrophages (PEM) of guinea pigs exposed in vitro to various doses of CRP for 72 hr resulted in the development of O2− production dose‐dependently, measured by increases in superoxide dismutase‐inhibitable nitro blue tetrazolium reduction. The O2−‐producing activity of PEM cultured without CRP, used as a control, decreased markedly in proportion to incubation time. The O2− production by PEM exposed to CRP for 18 hr when control PEM were still high in O2− production, was decreased by larger doses of CRP, while PEM cultured without CRP for 72 hr, when O2− production by control PEM was very low, followed by incubation with CRP for another 18 hr, produced O2− CRP‐dose‐dependently as in the case of that observed after 72‐hr incubation with CRP. These results indicate that CRP is capable of activating macrophages and acts on macrophage function as a modulator. CRP possesses migration inhibitory factor (MIF)‐like activity (as reported in the preceding paper) and also macrophage‐activating factor (MAF)‐like activity, indicating that CRP may play a functional role at the site of inflammation and tissue damage by accumulating and activating macrophages.