bluuebunny/crossref_metadata_2025_split huggingface dataset:
High‐Molecular‐Weight Hyaluronic Acids Inhibit Chemotaxis and Phagocytosis but Not Lysosomal Enzyme Release Induced by Receptor‐Mediated Stimulations in Guinea Pig Phagocytes
Authors: Tamoto Koichi , Nochi Hiromi , Tada Masahito , Shimada Sachiyo , Mori Yoki , Kataoka Syuichi , Suzuki Yoshihiro , Nakamura Tohoru 
Published: November 2013

AbstractThe effects of high‐molecular‐weight (HMW) hyaluronic acids (HAs) of 1.9 × 106 Da, 8 × 105 Da and 3 × 105 Da on the receptor‐mediated functions of guinea pig peritoneal phagocytes were studied. HMW‐HAs of 1.9 × 106 Da (HA190) and 8 × 105 Da (HA80) effectively inhibited the chemotactic activity of polymorphonuclear leukocytes (PMNs) for formyl‐Met‐Leu‐Phe (fMLP). The degree of inhibition was dose‐dependent and the concentrations of HA190 and HA80 required for 50% inhibition were 0.5–1.5 mg/ml and 1.5–2.5 mg/ml, respectively. HMW‐HA of 3 × 105 Da (HA30) hardly affected the chemotaxis within a concentration range of 0.5–5.0 mg/ml. The phagocytic activities of PMNs and macrophages (Møs) for serum‐opsonized zymosan (SOZ) and polystyrene latex particles were also inhibited by these HAs in a dose‐ and molecular‐weight‐dependent manner and HA190 was again the most inhibitory. By contrast, the release of lysosomal enzyme from Møs stimulated with SOZ was not significantly affected by HMW‐HAs at any concentration used. Furthermore, the binding of [3H]fMLP with PMNs and the rosette formation of Møs with SOZ were not influenced by the presence of HMW‐HAs. These findings suggested that the binding of HMW‐HAs to the HA receptors on PMNs and Møs might produce certain intracellular signals which would be responsible for the suppression of the chemotaxis and the phagocytosis but not for the release of lysosomal enzyme. For the generation of such signals, higher‐molecular‐weight HMW‐HAs would be more effective than lower one.
Effect of Escherichia coli Lipopolysaccharide on Bacteroides fragilis Abscess Formation and Mortality in Mice
Authors: Tanaka Yasunori , Jotwani Ravi , Watanabe Kunitomo , Tanaka Kaori , Kato Naoki , Ueno Kazue 
Published: November 2013

AbstractTo study the mechanism of synergism between Bacteroides fragilis and Escherichia coli, the effect of sublethal dose of E. coli lipopolysaccharide (LPS) (25μg/mouse) was checked on B. fragilis abscess formation. LPS was administered prior or after inoculum injection. No significant difference in the abscess size was observed at necropsy on day 6. However, all the groups receiving LPS showed higher incidence of recovery of additional intestinal bacteria (23.5–45.5%) from the abscess pus. When LPS was given 4 hr prior to inoculum administration, 83–100% mortality was observed. Detailed investigation showed autoclaved cecal contents alone could also cause similar mortality. Studies with stimulation of endogenous cytokines by E. coli LPS demonstrated induction of all of them within 3 hr in the blood stream with TNF‐α demonstrating peak at 1 hr, IL‐1α and IL‐6 at 4 hr and IFN‐γ between 6–9 hr with moderately high levels at 4 hr. This E. coli LPS‐triggered cytokine cascade possibly gets further stimulated by injection of autoclaved cecal contents containing high concentration of endotoxins (1.6 × 105 EU/ml) contributed by dead bacteria and lead to the mortality of animals.
Purification and Characterization of an Acid Phosphatase from Mycoplasma fermentans
Authors: Noda Mamoru , Shibata Ken‐ichiro , Sawa Yoshihiko , Shimokoube Hirokata , Watanabe Tsuguo 
Published: November 2013

AbstractAn acid phosphatase associated with the cell membranes of Mycoplasma fermentans was released from the membranes with Triton X‐100, then purified by ion‐exchange chromatography on DEAE‐Sephacel and CM‐Sepharose, followed by affinity chromatography on Con A‐Sepharose. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis of the purified enzyme revealed a single band with a molecular mass of 31.2 kilodaltons. The enzyme activity toward p‐nitrophenyl phosphate was enhanced remarkably by Cu2+, Co2+ and Mg2+, but the activity was not inhibited by EDTA. The enzyme dephosphorylated O‐phospho‐l‐tyrosine as well as p‐nitrophenyl phosphate, but not O‐phospho‐l‐threonine, O‐phospho‐l‐serine, glucose‐1‐phosphate, phosphoryl choline and adenosine triphosphate. The level of the O‐phospho‐l‐tyrosine phosphatase activity was the highest in Mycoplasma faucium and the second highest in Mycoplasma fermentans of all tested human mycoplasmas.
Effect of BV‐araU and Acyclovir on Varicella‐Zoster Virus Replication with Various Length and Timing of Drug Exposure
Authors: Machida Haruhiko , Nishitani Makiko , Ashida Noriyuki 
Published: November 2013

AbstractWe studied antiviral effects of 1‐β‐d‐arabinofuranosyl‐5‐[(E)‐2‐bromovinyl]uracil (BV‐araU) and acyclovir against varicella‐zoster virus (VZV) multiplication varying the length or timing of drug exposure. First, residual anti‐VZV effect of drugs, exposed to cells for various periods followed by incubation in drug‐free medium, was determined by the plaque inhibition assay. None of the drugs showed activity when removed within 24 hr of incubation. Weakened efficacy of BV‐araU was seen in 2 days of treatment. When it was removed after 3 or 4 days, the ED50 was as low as that for cultures in which the drug was not removed. Still, plaque inhibition was not complete even at high concentrations. Acyclovir inhibited plaque formation only by 50% or less in 2 days of treatment. It gave a much higher ED50 in 3 days of treatment than that observed without drug removal. In the experiments, in which BV‐araU was added to VZV‐infected cells 1 day after infection, BV‐araU immediately suppressed increase in the number of infective centers at a concentration of 0.001 μg/ml, and reduced it at concentrations of 0.01 μg/ml or higher. The reduction of infective centers was seen with a dose‐dependent manner when added 2 or 3 days after infection. BV‐araU stimulated the decrease in the number of infective centers when added 4 days after infection. This inhibitory effect of acyclovir was very weak. Microscopic observations supported the above results. BV‐araU was still much superior to acyclovir in the anti‐VZV effect when the length and timing of drug exposure were varied.
Protective Capacity of L‐Form Salmonella typhimurium against Murine Typhoid in C3H/HeJ Mice
Authors: Nishikawa Fumiko , Kita Eiji , Yamada Hideki , Nakano Akira , Kashiba Shuzo 
Published: November 2013

AbstractL forms of Salmonella typhimurium LT2 conferred strong protection to a lethal challenge with its parental bacterium on innately hypersusceptible C3H/HeJ mice, and its minimal protective dose was approximately 150 L‐forming units. Although L‐form S. typhimurium was avirulent for C3H/HeJ mice, it multiplied slowly in both the liver and spleen with the maximal growth 2–3 weeks after immunization and thereafter it persisted in the liver until 24 weeks. Protective immunity began to work between 4 and 6 weeks after immunization, and it remained active as long as the L forms colonized the liver (until 24 weeks after immunization). Vaccination with the L form induced a population of T cells responding to L‐form whole‐cell lysate (WCL), while delayed‐type hypersensitivity (DTH) to the extract of S. typhimurium was induced after the establishment of solid immunity. Moreover, neither T‐cell responses nor DTH to heat‐killed S. typhimurium was generated. In addition, antibody responses were elicited to WCL but not to heat‐killed S. typhimurium. These results indicate that protection conferred by the L forms is attributable to the persistent colonization of the L forms rather than the presence of DTH, and also that Salmonella cytoplasmic antigens are involved in induction of immunological responses by vaccination with the L forms.
Cytoplasmic Membrane‐Associated Protein (CAP) Isolated from Streptococcus pyogenes: As a New Bacterial Superantigen
Authors: Sato Hideki , Itoh Tetsuro , Rikiishi Hidemi , Kumagai Katsuo 
Published: November 2013

AbstractA protein isolated from the cytoplasmic membranes of Streptococcus pyogenes (cytoplasmic membrane‐associated protein, CAP) stimulated human T cells in vitro to induce their mitogenic response. This CAP‐induced T cell proliferation required the presence of nylon‐adherent accessory cells (AC) of either autologous or allogeneic origin in the reaction mixtures. In addition, the reaction was inhibited by monoclonal antibodies (mAbs) against major histocompatibility complex (MHC) class II molecules, HLA‐DR and ‐DQ, but not ‐DP. Human lymphoid cell lines positive for HLA‐DR but not those lacking it were also effective as AC for the reaction. A binding test using fluorescein‐labeled protein revealed that CAP bound to the adherent monocytes and HLA‐DR+ but not to ‐DR– lymphoid cell lines. The proliferative response of T cells to CAP was, however, not inhibited by the addition of the lysosomotrophic agent NH4Cl to the reaction mixtures. These results suggest that the presentation of CAP by AC to human T cells is mediated through binding of the protein to the MHC class II molecules but without being processed in the AC. The proliferative response of T cells was also found to be inhibited by addition of anti‐CD2, ‐CD3 or ‐T cell receptor (TcR) mAbs. A major population responding to CAP was CD3+4+8– T cells. CAP also appears to stimulate T cells bearing Vβ8 sequences much more selectively than T cells bearing other Vβs. These results indicate that this streptococcal membrane protein, CAP, may be a new protein belonging to a group of bacterial superantigens.
Greater Amount of HCV‐RNA in Tears Compared to Blood
Authors: Feucht Heinz‐hubert , Polywka Susanne , Zöllner Bernhard , Laufs Rainer 
Published: November 2013

AbstractPlasma, tear fluid and swabs from eye, nose and pharynx of 33 patients were examined for presence of hepatitis C virus (HCV) RNA by polymerase chain reaction (PCR). All samples from plasma, tear fluid and eyeswabs were found to show a positive reaction in HCV‐RNA PCR. Remarkably, we regularly found greater amounts of amplification products in tear fluid and eyeswabs compared to plasma using the same conditions for sample preparation.
Monoclonal Antibodies to Spirosin of Yersinia enterocolitica and Analysis of the Localization of Spirosome by Use of Them
Authors: Yamato Masayuki , Takahashi Yuko , Tomotake Hiroyuki , Ota Fusao , Hirota Katsuhiko , Yamaguchi Kazuo 
Published: November 2013

AbstractTwo hybridomas producing monoclonal antibodies (MAbs) were prepared by fusing myeloma cells (Sp2/0‐Ag14) with mouse spleen cells immunized with purified spirosin from Yersinia enterocolitica SYT‐11–72 (YE72). The antibodies produced by them were designated MAbs‐S5 and S27. They were IgG2a and IgG1, respectively, both with χ light chains. MAbs‐S5 and S27 reacted specifically with spirosin from YE72. On Western blotting after limited proteolysis with Staphylococcus aureus V8 protease, YE72 spirosin revealed peptide fragments of 35 and 37 kDa reacting markedly with MAb‐S5, which suggested the presence of an antigenic determinant on these fragments. By cellular fractionation of YE72 and subsequent EIA and Western blot analysis, spirosome was shown to be present in the cytoplasm of YE72.
Unique Properties of a Cytotoxic CD4+CD8+ Intraepithelial T‐Cell Line Established from the Mouse Intestinal Epithelium
Authors: Sasahara Takeshi , Tamauchi Hidekazu , Ikewaki Nobunao , Kubota Koichi 
Published: November 2013

AbstractGrowth factor‐dependent gut intraepithelial lymphocyte (IEL) cell lines were established from a long‐term in vitro culture of BALB/c IEL with syngeneic irradiated spleen cells in the presence of concanavalin A‐stimulated spleen supernatant fluids. The cell lines were preferentially consisted of very limited thymoindependent subsets of IEL; i.e., Thy‐1+ CD5–TCRαβ+ CD4+CD8 α+β– (double‐positive; DP) IEL and Thy‐1+ CD5– TCRαβ+ CD4–CD8α+β– (CD8 single‐positive; CD8 SP) IEL. The CD8 SP IEL cell line had cytotoxic activities and was triggered to proliferate by T‐cell receptor (TCR)‐directed stimuli. The DP IEL cell line expressed high levels of the CD3‐TCRαβ, exhibited cytotoxic activity in redirected lysis assays, and had perforin in the cytoplasm, indicating the functional maturity of this cell line. However, the DP IEL cell line did not proliferate in response to TCRαβ‐directed stimuli, which indicated that TCRαβ‐mediated signalling was able to initiate cytotoxic function but not to induce proliferation of the DP IEL cell line. Although both cell lines were shown to have functional competence, they expressed J11d antigen which marks immaturity in thymocyte differentiation pathways. These results indicate that the established thymoindependent DP and CD8 SP IEL cell lines have unique properties distinct from DP thymocytes and CD8 SP peripheral T cells. Together with a recent report on freshly isolated DP IEL (10), the unique properties of the DP IEL cell line seems to support the notion that DP IEL may undergo a unique maturation process in the gut microenvironment.
Characterization of Effector Cells against B16 Melanoma in Mice Inoculated with Allogeneic Spleen Cells
Authors: Nishi Yasuaki , Hosokawa Tomohide , Aoike Akira , Han Di , Takehara Hiroshi , Kawai Keiichi , Kamahora Takeshi 
Published: November 2013

AbstractInbred C57BL/6 (B6) mice which had received an inoculation of allogeneic spleen cells showed remarkable antitumor activity against syngeneic tumor challenge with B16 melanoma cells 3 days after the allogeneic cell inoculation. This antitumor activity was not specific to the inoculated alloantigen, since the challenging B16 cells are syngeneic to B6 mice and since it was induced by BALB/c spleen cells as well as C3H/He spleen cells. The antitumor activity was sensitive to an in vivo treatment with anti‐asialo GM1 (AGM1) antiserum or anti‐Thy.1 monoclonal antibody (mAb) just before the tumor challenge and was resistant to an in vivo treatment with anti‐CD8 (Ly. 2) mAb. These results suggest that AGM1+Thy.1+CD8– activated natural killer (NK) cells were generated by alloantigen inoculation and took an important part in the antitumor effect of the alloantigen inoculation.
Two G3 Feline Rotavirus Strains Lacking Cross‐Neutralization Reactions Represent Distinct Subtypes of Serotype G3
Authors: Mochizuki Masami , Nakagomi Osamu 
Published: November 2013

AbstractTwo feline rotavirus strains, FRV‐1 and FRV64, that have been shown to lack cross‐neutralization reactions despite the sharing of serotype G3 were examined by plaque‐reduction neutralization assays in relation to other G3 strains originating from cats, dogs, humans and monkeys. While FRV‐1 and human G3 strains constituted one subtype (G3A), FRV64, canine strains and simian strains constituted another subtype (G3B).
Molecular Cloning of the Wild‐Type and Mutant thyA Gene from Shigella flexneri Y
Authors: Nur‐E‐Kamal Mohammed Shah Alam , Mamun Abu Amar Md Al , Ahmed Zia Uddin 
Published: November 2013

AbstractThe thyA gene which codes for thymidylate synthase has been cloned and sequenced from the wild‐type Shigella flexneri Y strain SH4 and a thyA mutant TSF21 after amplifying the gene by polymerase chain reaction (PCR). The nucleotide sequence revealed 98% homology to the E. coli K‐12 thyA gene. The sequence of the wild‐type thyA gene of Shigella flexneri Y was identical with that of the thyA mutant except that the residue T at position 345 was replaced by residue A in the thyA mutant. This change would cause a predicted amino acid substitution of leucine at position 44 in the polypeptide product of the wild type by glutamine in the mutant. Thus, Leu44 may be critical in enzymatic activity of the thyA gene product thymidylate synthase.
Herpes Simplex Virus‐Induced Expression of 70 kDa Heat Shock Protein (HSP70) Requires Early Protein Synthesis But Not Viral DNA Replication
Authors: Kobayashi Kappei , Ohgitani Eriko , Tanaka Yasuyuki , Kita Masakazu , Imanishi Jiro 
Published: November 2013

AbstractThe major 70 kDa heat shock protein (HSP70), which is scarcely expressed in unstressed rodent cells, was apparently induced by infection with herpes simplex virus (HSV). Infection with HSV types 1 and 2 elevated HSP70 mRNA levels within 4 hr post‐infection. HSP70 synthesis and accumulation increased in HSV‐infected cells. Irradiation of HSV with UV‐light abolished the ability to induce HSP70 mRNA. Inhibitors of viral DNA synthesis did not affect the induction of HSP70 in infected cells. Protein synthesis within 2 hr after infection was necessary for HSP70 induction.
Amniotic Fluid Enhances Allogeneic Cytotoxic T Cell Responses, Whereas It Suppresses Mitogen‐Stimulated Lymphocyte Proliferation
Authors: Parvin Mahbuba , Isobe Ken‐ichi , Zohreh Heidari , Goto Setsuko , Nakashima Izumi , Tomoda Yutaka 
Published: November 2013

AbstractMouse amniotic fluid has been shown to suppress T lymphocyte proliferation and suggested to be important in regulating immunity during pregnancy. In allogeneic pregnancy, cytotoxic T cells in pregnant lymphocytes against paternal transplantation antigen are impaired. We examined the effect of amniotic fluid to the alloreactive CTL responses. Although the amniotic fluid suppressed Con A or LPS stimulated lymphocyte proliferation as previously reported, the amniotic fluid taken from syngeneic C57BL/6 pregnant mice or allogeneic C57BL/6 × BALB/c pregnant mice enhanced the anti‐H‐2d or anti‐H‐2k CTL responses dose‐dependently. We speculate that amniotic fluid contains not only immunosuppressive factors but also immunoenhancing factors which upregulate the allogeneic CTL responses.
Purification and Characterization of Type 1 Fimbriae of Salmonella typhi
Authors: Muscas Patrizia , Rossolini Gian Maria , Chiesurin Alessandra , Santucci Annalisa , Satta Giuseppe 
Published: November 2013

AbstractType 1 fimbriae have been purified from a Salmonella typhi strain of clinical origin. Purified fimbriae retained their ability to bind to erythrocytes in a mannose‐inhibitable fashion and, in doing so, behaved preferentially as a monovalent adhesin. SDS‐PAGE analysis of the fimbrial preparation showed the presence of a 20‐kDa major polypeptide component (fimbrillin) and of additional larger polypeptides present in smaller amounts. The amino‐terminal sequence of fimbrillin was determined and turned out to be very similar but not identical to that of type 1 fimbrillins of other Salmonella serovars. A Western blot analysis of the purified fimbrial preparation using an antiserum raised against native fimbriae suggested that fimbrial proteins did not carry any major sequential epitope and that, in native fimbriae, conformational epitopes, possibly generated between different subunits, might provide for the major immunogenic epitopes. Analysis of different S. typhi clinical isolates using the anti‐fimbrial antiserum showed an overall immunological similarity of these structures within this serovar.
Opsonic Activity of Sera and Blister Fluid from Severely Burned Patients Evaluated by a Chemiluminescence Method
Authors: Ono Yasuo , Kunii Otohiko , Suzuki Hiromasa , Ikeda Hirohito , Kobayashi Kunio , Kanegasaki Shiro 
Published: November 2013

AbstractBurn wound sepsis is the most common and severe complication in the patients with severe burn. To know the systemic and local defect in immunity of burned patients, we measured the luminol‐enhanced chemiluminescence (CL) response of normal polymorphonuclear leukocytes (PMNs) upon exposure to zymosan particles, bacteria or Candida albicans that were opsonized with any of patient's serum, blister fluid of burn wound or pooled normal serum (blood type AB). Sera from patients exhibited lower opsonic activities than those of pooled normal serum in the early postburn days. The levels of serum immunoglobulins, complement components and plasma fibronectin were found to correlate well with opsonin‐index (OI), which was determined based on the CL response data obtained during the course of infusion therapy with fresh frozen plasma. Furthermore, patient's blister fluid showed much lower opsonic activity against bacteria such as Pseudomonas aeruginosa than patient's own serum. These results indicate that blister fluid is also not effective to opsonize bacteria because of the marked depression of the levels of immunoglobulins and complement components. Destruction of the skin barrier by thermal injury and impairment of systemic or local humoral immunity may predispose these patients to burn wound sepsis.
In Vitro Antibiotic Susceptibilities of Borrelia Isolates from Erythema Migrans Lesion of Lyme Disease Patients in Japan
Authors: Masuzawa Toshiyuki , Yamada Kazuto , Kawabata Hiroki , Yanagihara Yasutake 
Published: November 2013

AbstractAntibiotic susceptibilities of twelve borrelial isolates from skin of patients with erythema migrans (EM) and ticks (Ixodes persulcatus and I. ovatus) in Japan were examined by in vitro microdilution MIC method and macrodilution MBC method. Nine EM isolates and 3 tick isolates were susceptible to amoxicillin, erythromycin, and minocycline. MICs for Japanese isolates were 0.038–0.30 μg/ml, < 0.012 μg/ml, and < 0.012–0.05 μg/ml, respectively. MBCs were as follows: 0.038–0.88 μg/ml, < 0.012–0.10 μg/ml, and <0.025–0.78 μg/ml, respectively. These antibiotics could be recommended for treatment of patients in early stage of Lyme disease in Japan.
Heat Shock Protein Produced by Helicobacter pylori
Authors: Yokota Kenji , Hirai Yoshikazu , Haque Mahmudul , Hayashi Shyunji , Isogai Hiroshi , Sugiyama Toshiro , Nagamachi Eiko , Tsukada Yutaka , Fujii Nobuhiro , Oguma Keiji 
Published: November 2013

AbstractThe cells of Helicobacter pylori were suspended in the medium containing35S‐methionine. After a heat shock of the cells at 42 C for 5, 10, and 30 min, the production of proteins was analyzed by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and autoradiography. Out of many proteins produced by the cells, only 66 kDa protein production was dramatically increased by heat treatment. The N‐terminal amino acid sequence of 66 kDa protein was quite similar to that of 62 kDa and 54 kDa proteins previously suggested as heat shock protein (HSP) of H. pylori based on the reaction with polyclonal and monoclonal antibodies against HSP 60 family proteins produced by other bacteria. Therefore, it was concluded that H. pylori produces the 66 kDa protein as its major heat shock protein which belongs to HSP 60 family.
Lidocaine Hydrochloride and Acetylsalicylate Kill Bacteria by Disrupting the Bacterial Membrane Potential in Different Ways
Authors: Ohsuka Shinji , Ohta Michio , Masuda Koji , Arakawa Yoshichika , Kaneda Toshio , Kato Nobuo 
Published: November 2013

AbstractLidocaine hydrochloride (LH), a local anesthetic, and acetylsalicylate (AcSAL), show antibacterial activity for both gram‐negative and gram‐positive bacteria. Kinetic studies indicated that antibacterial activity of LH was different from that of AcSAL. A subinhibitory concentration of LH and AcSAL enhanced the sensitivity of Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa to novobiocin and nalidixic acid. The synergistic effect of AcSAL with novobiocin and nalidixic acid was higher than that of LH. The effect of both drugs on the membrane potential of inner membrane was also studied using inverted membrane vesicles of bacteria. Both LH and AcSAL depolarized the membrane potential after the vesicles were energized with nicotinamide adenine dinucleotide. However, unlike AcSAL, pre‐treatment of vesicles with LH had no effect on the generation of membrane potential. These results suggest that depolarization of the cytoplasmic membrane, preceded by the permeabilization of the outer membrane for gram‐negative bacteria, is associated with antibacterial activity of LH and AcSAL. The difference in actions of LH and AcSAL was discussed.