bluuebunny/crossref_metadata_2025_split huggingface dataset:
Saccharomyces cerevisiae‐ and Candida albicans‐Derived Mannan Induced Production of Tumor Necrosis Factor Alpha by Human Monocytes in a CD14‐ and Toll‐Like Receptor 4‐Dependent Manner
Authors: Tada Hiroyuki , Nemoto Eiji , Shimauchi Hidetoshi , Watanabe Toshihiko , Mikami Takeshi , Matsumoto Tatsuji , Ohno Naohito , Tamura Hiroshi , Shibata Ken‐ichiro , Akashi Sachiko , Miyake Kensuke , Sugawara Shunji , Takada Haruhiko 
Published: November 2013

AbstractThe cytokine‐inducing activities of fungal polysaccharides were examined in human monocytes in culture, with special reference to CD14 and Toll‐like receptors (TLRs). Tumor necrosis factor alpha (TNF‐α) production by monocytes was markedly induced in a dose‐dependent manner upon stimulation with cell walls from Candida albicans and mannan from Saccharomyces cerevisiae and C. albicans, although relatively high concentrations (10 to 100 μg/ml) of stimulants were required for activation as compared with the reference lipopolysaccharide (LPS) (1 to 10 ng/ml). The yeast form C. albicans and its mannan and cell wall fractions exhibited higher TNF‐α production than respective preparations from the hyphal form. Only slight TNF‐α production was induced by the S. cerevisiae glucan. The TNF‐α production triggered by reference LPS and purified fungal mannans required the presence of LPS‐binding protein (LBP), and these responses were inhibited by anti‐CD14 and anti‐TLR4 antibodies, but not by anti‐TLR2 antibody. In contrast to the activity of LPS, the activity of purified S. cerevisiae mannan was not inhibited by polymyxin B. These findings suggested that the mannan‐LBP complex is recognized by CD14 on monocytes and that signaling through TLR4 leads to the production of proinflammatory cytokines in a manner similar to that induced by LPS.
Immune Responses against Allogeneic and Syngeneic Tumors in Aged C57BL/6 Mice
Authors: Win Sanda , Uenaka Akiko , Nakayama Eiichi 
Published: November 2013

AbstractAged C57BL/6 (B6) mice could reject allogeneic BALB/c RL male 1 tumor as efficiently as young B6 mice. However, in vitro analysis showed impaired generation of cytotoxic T cell response in aged B6 mice against allogeneic tumor. The reaction could be augmented by the addition of recombinant interleukin‐2 (rIL‐2). Enzyme‐linked immunospots (ELISPOT) produced by CD8+ T cells purified from spleen cells showed no reduction in aged mice. The findings suggested that the number of CD8+ T cells capable of reacting against allogeneic H‐2 antigens was similar in young and aged B6 mice. Low cytotoxic T lymphocyte (CTL) responsiveness in aged B6 mice appeared to have resulted from low responsiveness of CD4+ T cells producing IL‐2. Although CTL generation was apparently impaired, strong multiple antigenicity of allogeneic tumor evoked a rejection response in aged B6 mice. On the other hand, no rejection response was observed against syngeneic EL4 tumor in aged B6 mice even after depletion of CD4CD25+ immunoregulatory cells. Depletion of CD4+CD25+ cells caused rejection of EL4 tumor in young B6 mice. The findings suggested that aged B6 mice were incapable of inducing effector cells against weak tumor antigens. Only marginal CTL response and small number of ELISPOTs were generated in young but not aged B6 mice against EL4. Addition of rIL‐2 to the culture augmented EL4 killing and ELISPOTs in spleen cells from young and aged B6 mice.
A Novel Method Using Micropig Stratum Corneum In Vitro for the Evaluation of Anti‐Trichophyton mentagrophytes Activity
Authors: Nakashima Takuji , Nozawa Akira , Majima Toshiro 
Published: November 2013

AbstractAntifungal susceptibility testing under conditions close to clinical status is expected to provide more helpful information than that obtained by a conventional microdilution method. For this purpose, we developed a novel method to evaluate anti‐Trichophyton mentagrophytes activity of antifungal agents in vitro by using disks of micropig stratum corneum epidermis (SCE). Basal agar medium containing K2HPO4, MgSO4, CaCl2 and three kinds of antibiotics. Bifonazole (BFZ), lanoconazole (LCZ) or terbinafine (TBF) was added to the basal agar medium to give serially doubling dilutions ranging from 0.0006 to 10 μg/ml. Five‐hundred‐μl portions of the agar media thus prepared were solidified in wells of flat‐bottomed plates. SCE disks (6 mm in diameter) were placed on surfaces of the agar medium and 104 conidia of T. mentagrophytes were inoculated on each SCE disk There was very good correlation between the initial concentration of the antifungal agents added to the basal agar medium (μg/ml) and the concentration of the agents impregnated into the SCE disks (μg/g) (r2>0.99). The minimum inhibitory concentration (MIC) values of BFZ, LCZ and TBF were respectively 26‐, 10‐ and 78‐times higher than those measured by the standard microdilution method. From the correlation between the concentration of the agents in the basal medium and that in the SCE disks, the above MIC values corresponded to the concentrations in SCE disks (μg/g), 832.95 for BFZ, 1.42 for LCZ and 8.87 for TBF. This novel method of antidermatophytic susceptibility testing using SCE would be useful as an in vitro screening of proper antimycotics for topical treatment of dermatophytosis.
A Limulus Amoebocyte Lysate Activating Activity (LAL Activity) That Lacks Biological Activities of Endotoxin Found in Biological Products
Authors: Ochiai Masaki , Tamura Hiroshi , Yamamoto Akihiko , Aizawa Maki , Kataoka Michiyo , Toyoizumi Hiromi , Horiuchi Yoshinobu 
Published: November 2013

AbstractPyrogenic substances in influenza HA (IHA) vaccine have been controlled by the pyrogen test or the mouse body weight decreasing toxicity (BWD) test. We examined the possibility of replacing the animal tests with the endotoxin test. Commercial IHA vaccines were found to show considerable levels of LAL activity ranging from 0.2 to 160 EU/ml. However, a batch of the vaccine having even 100 EU/ml of LAL activity showed neither pyrogenicity in rabbits nor tumor necrosis factor alpha (TNF‐α) induction in RAW264.7 cells. The LAL activity of IHA vaccine was abolished by a monoclonal antibody that recognizes LPS‐binding epitope of LAL factor C. The activity of IHA vaccine showed different physicochemical properties from those of LAL activity of endotoxin. LAL activity of endotoxin is known to be sensitive to polymyxin B treatment and was found to be resistant to polyoxyethylene 10 cetyl ether (Brij56) treatment. On the contrary, the LAL activity of IHA vaccine was shown to be resistant to polymyxin B but sensitive to Brij56 treatment. The difference in sensitivity of the two LAL activities to polymyxin B and Brij56 might suggest the possibility of their discriminative measurements.
Genetic Background Determines Sensitivity to the Inhibitory Function of NO on T Cell Proliferation and the Amounts of NO Production Mediated through IFN‐γ
Authors: Ozaki Akemi , Fukushima Atsuki , Fukata Kazuyo , Sakamoto Shuji , Taniguchi Taketoshi , Hashiguch Masaaki , Ueno Hisayuki 
Published: November 2013

AbstractNitric oxide (NO) inhibits T cell proliferation. We demonstrate that the action of NO on T cell proliferation is different for Lewis and Brown Norway (BN) rats. Splenocytes from Lewis rats consistently showed higher proliferation against concanavalin A than splenocytes from BN rats did. In contrast, NO production was higher in BN rats than in Lewis rats. A depletion of adherent cells increased proliferation in BN rats to a level similar to that in Lewis rats. Thus NO produced by adherent splenocytes could be considered to inhibit proliferation. The addition of NG‐monomethyl‐l‐arginine, a potent inhibitor of NO production, increased proliferation in Lewis rats, but much less so in BN rats. Similar results were obtained by the addition of anti‐interferon (IFN)‐γ. It is surprising that, low doses of sodium nitroprusside, an NO donor, increased proliferation in BN rats but not in Lewis rats. To investigate the mechanism of differential NO production between the two strains, splenocytes were stimulated with IFN‐γ. The early signaling event evaluated by the phosphorylation of Stat‐1 was similar in both strains, whereas inducible NO synthase (iNOS) mRNA expression seemed more sustained in BN rats. Thus the differential production of NO might be related to the differential transcriptional regulation of iNOS. Altogether, genetic background might be involved in sensitivity to the inhibitory function of NO for T cell proliferation and NO production.
Viral Load in Epstein‐Barr Virus‐Associated Hemophagocytic Syndrome
Authors: Kimura Hiroshi , Hoshino Yo , Hara Shinya , Nishikawa Kazuo , Sako Masahiro , Hirayama Masahiro , Komada Yoshihiro , Morishima Tsuneo 
Published: November 2013

AbstractThe viral load in peripheral blood from patients with Epstein‐Barr virus (EBV)‐associated hemophagocytic syndrome was measured by real‐time quantitative PCR and compared with that in infectious mononucleosis. Patients with EBV‐associated hemophagocytic syndrome generally had larger viral burdens, although it was difficult to differentiate EBV‐associated hemophagocytic syndrome from infectious mononucleosis simply by viral load. The difference in viral load seemed to be clearer in peripheral blood mononuclear cells than in plasma.
A Structurally Variant Form of the 2B4 Antigen Is Expressed on the Cell Surface of Mouse Mast Cells
Authors: Kubota Koichi 
Published: November 2013

Abstract2B4 (CD244) is a 66‐kDa CD2 family protein expressed on natural killer (NK) cells. Mouse NK cells express two isoforms of 2B4, termed 2B4L and 2B4S, whose molecular masses are 42 kDa and 36 kDa, respectively. In this study, we biochemically characterize the 2B4 antigen that was newly found on mouse bone marrow‐derived mast cells (BMMC). Anti‐2B4 mAb immunoprecipitated glycoproteins with a molecular mass of 60 kDa from BMMC. Removal of N‐linked sugars from the antigen by N‐glycosidase F treatment yielded two protein backbones of 35 kDa and 25 kDa, indicating that BMMC express the 2B4S isoform, but not 2B4L. Nucleotide sequence analyses confirmed that BMMC transcribe 2B4S mRNA. The preferential expression of the 2B4S isoform and the detection of an additional 25‐kDa glycoprotein on BMMC indicate that differences in the structure of 2B4 antigen exist between BMMC and NK cells.
Development of an Immunofluorescence Method for the Detection of Antibodies to Ebola Virus Subtype Reston by the Use of Recombinant Nucleoprotein‐Expressing HeLa Cells
Authors: Ikegami Tetsuro , Saijo Masayuki , Niikura Masahiro , Miranda Mary E.G. , Calaor Alan B. , Hernandez Marvin , Manalo Daria L. , Kurane Ichiro , Yoshikawa Yasuhiro , Morikawa Shigeru 
Published: November 2013

AbstractAn indirect immunofluorescent assay (IFA) to detect Ebola virus subtype Reston (EBO‐R) antibodies was developed by the use of a HeLa cell line stably expressing EBO‐R nucleoprotein (NP). This IFA has a high specificity for the detection of EBO‐R IgG antibodies in both hyperimmune rabbit sera and monkey sera collected during an EBO‐R outbreak in the Philippines in 1996. Furthermore, this IFA showed a higher sensitivity for the detection of EBO‐R antibodies than did the IFA using HeLa cells expressing the NP of Ebola virus subtype Zaire. These results suggest that this new IFA is useful for seroepidemiological studies of EBO‐R infection among monkeys.
Antimicrobial Activity and Stability to Proteolysis of Small Linear Cationic Peptides with D‐Amino Acid Substitutions
Authors: Hamamoto Kumiko , Kida Yutaka , Zhang Ye , Shimizu Takashi , Kuwano Koichi 
Published: November 2013

AbstractAntimicrobial peptides contribute to innate host defense against a number of bacteria and fungal pathogens. Some of antimicrobial synthetic peptides were systemically administered in vivo; however, effective protection has so far not been obtained because the effective dose of peptides in vivo seems to be very high, often close to the toxic level against the host. Alternatively, peptides administered in vivo may be degraded by certain proteases present in serum. In this study, D‐amino acids were substituted for the L‐amino acids of antimicrobial peptides to circumvent these problems. Initially a peptide (L‐peptide) rich in five arginine residues and consisting of an 11‐amino acid peptide (residues 32–42) of human granulysin was synthesized. Subsequently, the L‐amino acids of the 11‐amino acid peptide were replaced partially (D‐peptide) or wholly (AD‐peptide) with D‐amino acids. Activity and stability to proteolysis, in particular, in the serum of antimicrobial peptides with D‐amino acid substitutions were examined. Peptides with D‐amino acid substitutions were found to lyse bacteria as efficiently as their all‐L‐amino acid parent, L‐peptide. In addition, the peptide composed of L‐amino acids was susceptible to trypsin, whereas peptides containing D‐amino acid substitutions were highly stable to trypsin treatment. Similarly, the peptide consisting of L‐amino acids alone was also susceptible to fetal calf serum (FCS), however, protease inhibitors restored the lowered antimicrobial activity of the FCS‐incubated peptide. Thus, D‐amino acid substitutions can make antimicrobial peptides resistant to proteolysis, suggesting that the antimicrobial peptides consisting of D‐amino acids are potential candidates for clinical therapeutic use.
Molecular Fingerprinting of Isoniazid‐Resistant Mycobacterium tuberculosis Isolates from Chest Diseases Hospital in Kuwait
Authors: Mokaddas Eiman , Ahmad Suhail , Abal Adnan T. 
Published: November 2013

AbstractTouchdown double‐repetitive‐element‐PCR (DRE‐PCR) was carried out for typing 38 consecutive isoniazid‐resistant Mycobacterium tuberculosis strains isolated at Chest Diseases Hospital, Kuwait, during 1998–2000. The polymorphism at codon 463 in the katG gene was also determined and correlated with genotypic relationships among the isolates. The isolates exhibited 21 distinct patterns in DRE‐PCR. Nearly half of the isolates (18 of 38) exhibited unique patterns. Majority of isolates (16 of 20) yielding multiple DNA fragments in DRE‐PCR were unique strains while most of the isolates (16 of 18) yielding a single DNA fragment in DRE‐PCR clustered together. The prevalence of L463 in the katG gene was much higher in isolates from Middle Eastern (mostly Kuwaiti) patients than is reported for this ethnic group. The data indicate the possibility of some strains of South Asian/Southeast Asian origin spreading among local populations.
Monoclonal Antibody to Shiga Toxin 1, Which Blocks Receptor Binding and Neutralizes Cytotoxicity
Authors: Nakao Hiroshi , Kataoka Chiwa , Kiyokawa Nobutaka , Fujimoto Junichiro , Yamasaki Shinji , Takeda Tae 
Published: November 2013

AbstractA monoclonal antibody, 5–5B, which neutralizes Shiga toxin 1 (Stx1) cytotoxicity of Escherichia coli, was constructed. An epitope analysis indicated that Asn55 in Stx1 B subunit was an important residue. This result and our previous results using an anti‐Stx2 monoclonal antibody indicate that the region around the cysteine residue of the disulfide bond might be important for the neutralization of Stx cytotoxicity, making it a potential vaccination candidate.
Double‐Stranded nef RNA Interferes with Human Immunodeficiency Virus Type 1 Replication
Authors: Yamamoto Takuya , Omoto Shinya , Mizuguchi Masaaki , Mizukami Hajime , Okuyama Harumi , Okada Noriko , Saksena Nitin K. , Brisibe Ebiamadon A. , Otake Kaori , Fujii Yoichi R. 
Published: November 2013

AbstractRNA interference (RNAi) has been reported to be post‐transcriptional gene silencing (PTGS) by approximately 500 nucleotide‐(nt)‐long double‐stranded (ds) RNA that specifically targets homologous sequences of messenger RNA. In this report, we describe inhibition of HIV‐1 transcription by synthetic dsRNAs constructed with mutated nef genes (nef dsRNAs) derived from long‐term non‐progressors (LTNPs) using cotransfection of the target gene‐expressing plasmid and dsRNA. The effects of nef dsRNAs were examined with luciferase (Luc) reporter which is combined with the HIV‐1 (SF2) LTR in persistently HIV‐1‐infected T cell and macrophage cell lines. At 48 hr, a defective nef dsRNA (556 nt) suppressed Luc activity more potently than did SF2 full‐length nef dsRNA (744 nt), suggesting that approximately 500 nt‐long nef dsRNA could interfere with the HIV‐1 transcription.
Human Immunodeficiency Virus Type 1 Neutralization by a Single Molecule of V3‐Targeted Antibody
Authors: Harada Shinji 
Published: November 2013

AbstractHuman immunodeficiency virus type‐1 (HIV‐1) infection generally provokes antibody responses to the viral envelope glycoprotein. Two major regions of gp120, the third variable (V3) domain and the CD4‐binding site, have been identified as neutralization targets. The precise mechanism of HIV‐1 neutralization by antibodies against the V3 domain is still unknown. It is shown that by kinetic neutralization studies, one molecule of V3‐targeted monoclonal antibody (0.5P) is enough to neutralize one virion. This antibody, which neutralized more than 99% of the virus, inhibited the binding of the virus to cells by 42%. HIV‐1 pseudotyped with G glycoprotein from vesicular stomatitis virus was also neutralized by 0.5β, suggesting that the antibody did not inhibit the viral attachment but caused some alteration in the envelope. These results indicate that the antibody plays an additional role on steric change of the envelope involved in inhibition of viral entry.
Distribution of Aeromonas hydrophila Serogroups in Different Clinical Samples and the Development of Polyclonal Antibodies for Rapid Identification of the Genus Aeromonas by Direct Agglutination
Authors: Korbsrisate Sunee , Dumnin Suwat , Chawengkirttikul Rungrawan , Gherunpong Vanchit , Eampokalap Boonchuay , Gongviseisoog Charles , Janyapoon Kritsana , Lertpocasombat Kancharee , Shimada Toshio 
Published: November 2013

AbstractWe characterized a collection of 256 Aeromonas hydrophila strains isolated from blood, discharge and stool for their serogroup designation. Of these, 2.3% were untypable and 15.2% were rough strains. Among the typable strains, about 50% comprised serogroups O:11, O:16, O:18, O:34 and O:83. To develop rapid differentiation of Aeromonas from other oxidase‐positive bacteria, antisera against Aeromonas were produced to establish a direct, genus‐specific, agglutination test. It was found that among 105 isolates of Aeromonas, 102 showed positive results with the agglutination test. The calculated sensitivity and specificity were 97.1% and 90.7%, respectively.
Antimicrobial Susceptibilities of Shiga Toxin‐Producing Escherichia coli Isolates from Pigs with Edema Disease in Japan
Authors: Uemura Ryoko , Sueyoshi Masuo , Nagayoshi Miki , Nagatomo Hiroshi 
Published: November 2013

AbstractFifty‐seven Shiga toxin‐producing Escherichia coli (STEC) strains isolated from pigs with edema disease (ED) from 1997 to 2001 in Japan were examined for antimicrobial susceptibilities. The susceptibilities were compared with those of E. coli ATCC 23546 isolated from pig with ED in the 1950's. Consequently, the isolated STECs showed high susceptibility to peptides and bicozamycin in a way similar to the reference strain. On the other hand, the STECs showed low susceptibility to beta‐lactams, tetracyclines, novobiocin, fosfomycin, trimethoprim, and old quinolones. It became clear that the susceptibilities of the isolated STECs had diminished in regard to antimicrobials.
Efficient Adeno‐Associated Virus‐Mediated Gene Expression in Human Placenta‐Derived Mesenchymal Cells
Authors: Zhang Xiaohong , Nakaoka Takashi , Nishishita Toshihide , Watanabe Nobukazu , Igura Koichi , Shinomiya Ken‐ichi , Takahashi Tsuneo A. , Yamashita Naohide 
Published: November 2013

AbstractMesenchymal cells from various sources are pluripotent and are attractive sources for cell transplantation. In this study, we analyzed recombinant adeno‐associated virus (rAAV)‐mediated gene expression in human placenta‐derived mesenchymal cells (hPDMCs), which reside in placental villi. After transduction of AV‐CAG‐EGFP, a rAAV expressing enhanced green fluorescence protein (EGFP), hPDMCs showed much higher level of EGFP expression than human umbilical vein endothelial cells or rat aortic smooth muscle cells. The number of EGFP‐positive hPDMCs infected by AV‐CAG‐EGFP alone did not increase significantly by coinfection of adenovirus, which enhanced expression level of the rAAV vector. Moreover, flow cytometric analysis showed discrete positive fraction of EGFP‐expressing hPDMCs, which is about 15–20% of the cells infected with AV‐CAG‐EGFP. Therefore, some cell population in hPDMCs might be highly susceptible to rAAV‐mediated gene transduction. In addition, stable EGFP expressions were observed in about 1% of hPDMCs infected with AV‐CAG‐EGFP at 4 weeks post‐infection. Collectively, hPDMCs have characters favorable for rAAV‐mediated gene expression.
Characterization of the F Gene of Contemporary Mumps Virus Strains Isolated in Japan
Authors: Uchida Kazue , Shinohara Michiyo , Shimada Shin‐ichi , Segawa Yukari , Kimura Kazuhiro , Hoshino Yoji 
Published: November 2013

AbstractMumps virus (MuV) strains isolated in Saitama Prefecture, Japan, from 1997 to 2001, were examined by analyzing the SH and the F gene nucleotide sequences. The results of the SH gene analysis showed that only genotype G was found in 2001 as well as in 2000, and that genotype J, which we proposed as a new genotype in a previous study, was from a different lineage than the genotype J described by Tecle et al. (J. Gen. Virol. 82, 2675–2680). We therefore, propose to rename the genotype as K to avoid confusion. Then, the F gene of genotypes G, H, and K strains were analyzed together with previously reported strains in this study. The results of phylogenetic analysis of the F gene nucleotide sequences showed that these strains formed a cluster as described by the SH gene analysis. Alignment of the F amino acid sequences showed that the F protein was well conserved among strains of different genotypes with a few amino acid differences. These results provide better information for the characterization of contemporary MuV strains in Japan.
High Molecular Weight Vimentin Complex Is Formed after Proteolytic Digestion of Vimentin by Caspase‐3: Detection by Sera of Patients with Interstitial Pneumonia
Authors: Fujita Jiro , Bandoh Shuji , Yang Yu , Wu Fei , Ohtsuki Yuji , Yoshinouchi Takeo , Ishida Toshihiko 
Published: November 2013

AbstractIn a previous study, we demonstrated anti‐vimentin antibodies in sera of patients with interstitial pneumonia. We hypothesized that antibodies in sera might detect vimentin fragments formed during the process of apoptosis. To prove this, recombinant human vimentin was digested by recombinant human caspase‐3 or caspase‐8. Then, Western blotting using several commercially available antibodies against human vimentin or patients' sera which had anti‐vimentin autoantibodies, was performed. As a result, after recombinant human vimentin was digested by caspase‐3 or caspase‐8, several vimentin fragments were formed and detected by 2 kinds of monoclonal anti‐vimentin antibodies (clone 3B4 and clone V9) as well as by polyclonal sheep anti‐human vimentin antibody. It was demonstrated that high molecular weight vimentin was formed after the digestion of vimentin by caspase‐3, which was only detected by patients' sera. The high molecular weight vimentin was not formed after digestion of vimentin by caspase‐8. Our present results show that high molecular weight vimentin was formed after the digestion of vimentin by caspase‐3. In addition, it is suggested that this high molecular weight vimentin acted as an autoantigen to form anti‐vimentin autoantibody in vivo.