bluuebunny/crossref_metadata_2025_split huggingface dataset:
Further Studies on the Hyperphosphorylated Form (p40) of the Rabies Virus Nominal Phosphoprotein (P)
Authors: Toriumi Harufusa , Eriguchi Yoshiro , Takamatsu Fumihiko , Kawai Akihiko 
Published: November 2013

AbstractWe investigated possible mechanisms involved in production of a hyperphosphorylated form (p40) of rabies virus P protein, to which two dimensional (2‐D) gel electrophoresis was applied. The P gene products produced in Escherichia coli cells could be detected as a single spot of unphosphorylated 37‐kDa form (termed as p37–0) in a 2‐D gel. The 37‐kDa proteins in the virus‐infected cells are composed of some phosphorylated forms, including a major p37–1 and more phosphorylated minor forms (e.g., p37–2, p37–3, etc.), but little p37–0 is detected (Eriguchi et al., 2002). When the E. coli‐produced P protein analogues were incubated with BHK‐21 cell lysates, heparin‐sensitive phosphorylation occurred as described previously (Takamatsu et al., 1998), giving an additional 40‐kDa spot. However, such a p40‐like derivative displayed a little more basic pI value than that of the authentic p40 produced in the infected cells; hence, the former was termed p40–0 (pI=4.78), while the latter, p40–1 (pI=4.73). In contrast, p40 produced in the P cDNA‐transfected animal cell was detected at the p40–1 position. In addition, staurosporine did not affect the p40–1 production in virus‐infected nor the P cDNA‐transfected animal cells, while the agent reduced production of hyperphosphorylated forms of p37, resulting in accumulation of p37–1, but not of p37–0. These results suggest that, although p37–0 may become a substrate for the heparin‐sensitive protein kinase (PK) in vitro, only p37–1 is a substrate for p40 production catalyzed by heparin‐sensitive PK in animal cells, and staurosporine‐sensitive PK is involved in the production of more phosphorylated forms of p37, but not in p37–1 production from p37–0.
Characterization of Mechanisms of Fluconazole Resistance in a Candida albicans Isolate from a Japanese Patient with Chronic Mucocutaneous Candidiasis
Authors: Kamai Yasuki , Maebashi Kazunori , Kudoh Michinari , Makimura Koichi , Naka Wataru , Uchida Katsuhisa , Yamaguchi Hideyo 
Published: November 2013

AbstractWe examined the mechanisms of fluconazole resistance in a fluconazole‐resistant Candida albicans isolate from a Japanese patient with chronic mucocutaneous candidiasis. It was demonstrated that the highly resistant phenotype of this strain was associated with combined mechanisms of the energy‐dependent reduced intracellular accumulation of fluconazole, presumably due to the increased expression of the ATP‐binding cassette efflux pump CDR gene(s), and the reduced affinity of the target enzyme, Erg11p, to fluconazole. In particular, the reduced affinity of Erg11p was considered to contribute largely to the fluconazole resistance in the TIMM3209 strain. Biochemical studies indicated that the Erg11p from the TIMM3209 strain showed reduced susceptibility both to fluconazole and itraconazole of cell‐free ergosterol biosynthesis, and cytochrome P‐450 also showed reduced affinity to fluconazole in the carbon monoxide‐cytochrome P‐450 complex formation assay. We identified two amino acid substitutions, Y132H and G448V, in Erg11p from the TIMM3209 strain. We found that the cytochrome P‐450 from the TIMM3209 strain decayed during incubation at 37 C without fluconazole although it is unknown whether or not the phenomenon is linked to the resistant phenotype. These mutations are thought to confer the above‐mentioned characteristics to Erg11p.
2‐Aminopurine Inhibits Lipopolysaccharide‐Induced Nitric Oxide Production by Preventing IFN‐β Production
Authors: Sugiyama Tsuyoshi , Fujita Megumi , Koide Naoki , Mori Isamu , Yoshida Tomoaki , Mori Hiroshi , Yokochi Takashi 
Published: November 2013

Abstract2‐Aminopurine (2‐AP) is widely used as a specific inhibitor for double stranded‐RNA dependent protein kinase (PKR). Here we report that 2‐AP can inhibit lipopolysaccharide (LPS)‐stimulated nitric oxide (NO) production through the prevention of interferon (IFN)‐β production. 2‐AP significantly inhibited NO production in LPS‐stimulated RAW 264 murine macrophage cells. 2‐AP also reduced the expression of IFN‐β and IFN‐inducible genes, such as IFN‐γ‐inducible protein (IP)‐10 and immune‐responsive gene (IRG)‐1, and the inducible type of NO synthase (iNOS) mRNA in response to LPS. The addition of exogenous IFN‐β restored 2‐AP‐inhibited NO production in response to LPS. On the other hand, there was only partial inhibition by 2‐AP of nuclear factor (NF)‐κB activation, IL‐6 mRNA expression and tumor necrosis factor (TNF)‐α production. These results suggested that 2‐AP inhibited LPS‐induced IFN‐β production by preventing Toll/IL‐1 receptor domain‐containing adaptor‐inducing IFN‐β (TRIF)‐dependent signaling rather than myeloid differentiation factor (MyD) 88‐dependent signaling, resulting in the inhibition of NO production.
Recombinant Cholera Toxin B Subunit Activates Dendritic Cells and Enhances Antitumor Immunity
Authors: Isomura Iwao , Yasuda Yoko , Tsujimura Kunio , Takahashi Toshitada , Tochikubo Kunio , Morita Akimichi 
Published: November 2013

AbstractActivation of dendritic cells (DC) is crucial for priming of cytotoxic T lymphocytes (CTL), which have a critical role in tumor immunity, and it is considered that adjuvants are necessary for activation of DC and for enhancement of cellular immunity. In this study, we examined an adjuvant capacity of recombinant cholera toxin B subunit (rCTB), which is non‐toxic subunit of cholera toxin, on maturation of murine splenic DC. After the in vitro incubation of DC with rCTB, the expression of MHC class II and B7–2 on DC was upregulated and the secretion of IL‐12 from DC was enhanced. In addition, larger DC with longer dendrites were observed. These data suggest that rCTB induced DC maturation. Subsequently, we examined the induction of tumor immunity by rCTB‐treated DC by employing Meth A tumor cells in mice. Pretreatment with subcutaneous injection of rCTB‐treated DC pulsed with Meth A tumor lysate inhibited the growth of the tumor cells depending on the number of DC. Moreover, intratumoral injection of rCTB‐treated DC pulsed with tumor lysate had therapeutic effect against established Meth A tumor. Immunization with DC activated by rCTB and the tumor lysate increased number of CTL precursor recognizing Meth A tumor. The antitumor immune response was significantly inhibited in CD8+ T cell‐depleted mice, although substantial antitumor effect was observed in CD4+ T cell‐depleted mice. These results indicated that rCTB acts as an adjuvant to enhance antitumor immunity through DC maturation and that CD8+ T cells play a dominant role in the tumor immunity. Being considered to be safe, rCTB may be useful as an effective adjuvant to raise immunity for a tumor in clinical application.
Autoantibodies to Peroxiredoxin I and IV in Patients with Systemic Autoimmune Diseases
Authors: Karasawa Rie , Ozaki Shoichi , Nishioka Kusuki , Kato Tomohiro 
Published: November 2013

AbstractAnti‐oxidative enzymes protect living bodies from various oxidative stresses. In the systemic autoimmune diseases, autoantibodies to oxidized molecules and to anti‐oxidative enzymes have been reported. To promote understanding of the relationships between autoimmunity and oxidative stress, we here investigate whether autoimmunity to the anti‐oxidative peroxiredoxin (Prxs) enzymes exists in patients with systemic autoimmune diseases. Specifically, we detected autoantibodies to recombinant Prx I and Prx IV respectively by ELISA and western blotting. Next, clinical parameters were compared between the anti‐Prx I or IV‐positive and ‐negative patients. We found that 33% of the 92 patients with autoimmune diseases tested possessed autoantibodies to Prx I (57% in systemic lupus erythematosus (SLE), 19% in rheumatoid arthritis (RA), 5% in Behçet disease, and 46% in primary vasculitis syndrome). In contrast, autoantibodies to Prx IV were detected in only 17% of the same patients. No significant correlation was found between occurrence of the two autoantibodies. Clinically, possession of anti‐Prx I autoantibodies correlated with lower serum levels of CH50, C3, and C4. Taken together, our data demonstrate the existence of autoantibodies to Prxs for the first time. The autoantibodies to Prx I may be involved in the pathophysiology of systemic autoimmune diseases such as SLE and vasculitis.
A Novel Diagnostic Method for Human Immunodeficiency Virus Type‐1 in Plasma by Near‐Infrared Spectroscopy
Authors: Sakudo Akikazu , Tsenkova Roumiana , Onozuka Taisuke , Morita Kazuo , Li Shuming , Warachit Jiranan , Iwabu Yukie , Li Guimei , Onodera Takashi , Ikuta Kazuyoshi 
Published: November 2013

AbstractPresently, the diagnosis of virus infections is based mainly on serological assays. Although polymerase chain reaction (PCR) and enzyme‐linked immunosorbent assay (ELISA) have been increasingly used for the diagnosis of such viral infections, the risk of transfusion‐transmitted blood‐borne viruses remains. Furthermore, PCR and ELISA are expensive and time‐consuming, and sometimes cause false‐positive or false‐negative results. Therefore, a rapid, accurate and cost‐effective diagnostic procedure is needed. We subjected plasma from individuals infected with human immunodeficiency virus type‐1 (HIV‐1), the causative agent of acquired immune deficiency syndrome (AIDS), as well as plasma from uninfected individuals as a control to near‐infrared (NIR) spectroscopy, which may provide a rapid diagnostic method for HIV‐1 infection without using any reagent. NIR spectra in the 600–1,000 nm region for plasma from pre‐serologically HIV‐1‐infected individuals and healthy donors were subjected to partial least squares (PLS) regression analysis and leave‐out cross‐validation to develop a multivariate model to estimate the concentration of HIV‐1. Simultaneously, the same plasma samples were examined for HIV‐1 p24 by ELISA. The results obtained by the NIR spectroscopy model for HIV‐1 yielded a good correlation with those obtained by the reference method (HIV‐1 p24 ELISA). These results suggest that NIR spectroscopy using plasma could provide a rapid, accurate, cost‐effective tool for large‐scale diagnosis of HIV‐1 infection.
Rapid Species Identification and Partial Strain Differentiation of Clostridium butyricum by PCR Using 16S‐23S rDNA Intergenic Spacer Regions
Authors: Nakanishi Shusuke , Kuwahara Tomomi , Nakayama Haruyuki , Tanaka Mamoru , Ohnishi Yoshinari 
Published: November 2013

AbstractSome Clostridium butyricum strains have been used as probiotics for both humans and animals. Strain‐specific identification is necessary for the manufacturing process of probiotics. The aim of this study was to determine whether there are sufficient genetic variations in 16S‐23S intergenic spacer regions (ISRs) to discriminate C. butyricum at the biovar level. We amplified ISRs from five reference strains, a probiotic strain (MIYAIRI 588) and 22 isolates, and we classified them into four groups on the basis of amplification patterns (type A through D). However, amplification of ISRs is not sufficient for discriminating strains. Moreover, we compared genetic structures of these ISRs. Sequence analysis revealed that the size variations of ISRs were generated by the insertion of tRNA genes and unique sequences into the internal portion, while the external portions were highly conserved. On the basis of the highly conserved nucleotide sequences within the ISRs, we developed a PCR primer set specific to C. butyricum. In addition, the PCR primer designed from the unique inserted sequence in type B strain was useful to differentiate probiotic strains at the biovar level.
Microarray Analysis of Host Gene‐Expression during Intracellular Nests Formation of Trypanosoma cruzi Amastigotes
Authors: Imai Kazuhide , Mimori Tatsuyuki , Kawai Makoto , Koga Hisashi 
Published: November 2013

AbstractThe intracellular pathogens utilize numerous cellular components of host cells to advance the infection as well as to enter the host cell. Analyzing the host cellular response enables us to get a better understanding of the pathogenesis, and subsequently indicate possible therapeutic targets. We therefore analyzed gene‐expression profile of NIH3T3 fibroblast cells infected by Trypanosoma, a representative intracellular pathogen similar to Leishmania, using custom‐designed cDNA microarray consisting of 1,701 mKIAA cDNAs. Focusing on intracellular nest formation of Trypanosoma cruzi amastigotes, we profiled the host gene‐expression at 8 days post‐infection and found several degrees of change in 16 mKIAA genes. Among these genes, 10 were up‐regulated and 6 were down‐regulated. Assuming that these genes had important roles in the infection's progression, we performed semi‐quantitative RT‐PCR analysis and confirmed the gene expression change of 4 genes. Furthermore, 5 genes were mapped on cadherin signaling pathway using pathway analysis software. These results indicate significance of the host cellular pathway in the proliferative stage of Trypanosoma cruzi amastigotes.
Association of Cytomegalovirus with Infantile Hepatitis
Authors: Shibata Yukiko , Kitajima Naoko , Kawada Jun‐ichi , Sugaya Naomi , Nishikawa Kazuo , Morishima Tsuneo , Kimura Hiroshi 
Published: November 2013

AbstractInfantile hepatitis is occasionally seen in apparently healthy children. In most cases, the etiology of the infection is uncertain. However, cytomegalovirus (CMV), Epstein‐Barr virus (EBV), human herpesvirus‐6 (HHV‐6), human herpesvirus‐7 (HHV‐7), human parvovirus B19, and TT virus (TTV) are considered to be associated with hepatitis in children. The objective of this study was to investigate the correlations between these viruses and infantile hepatitis. Twenty‐six children from 1 to 24 months old (median age, 7 months) who had liver dysfunction of unknown etiology were enrolled in this study. Plasma samples were examined by a real‐time PCR assay for CMV, EBV, HHV‐6, HHV‐7, parvovirus B19, and TTV DNA. The DNA of CMV was detected in the plasma of four patients (15.4%) and was detected significantly more often in the patient group than in the control group. The CMV‐infected patients were 1 to 3 months old, which was significantly younger than the remaining patients. The serological findings did not always correlate with the results of the real‐time PCR assay. The DNA of TTV was detected in four patients (15.4%), while human parvovirus B19 DNA was detected in three (11.5%). However, the detection frequencies of these viral DNAs were not significantly different from those in the control groups, and some of these patients had co‐infections. These results indicate that CMV might be one of the major pathogens responsible for infantile hepatitis; however, serological tests have limited utility for the diagnosis of CMV infection in young children.
Japanese Babesia microti Cytologically Detected in Salivary Glands of Naturally Infected Tick Ixodes ovatus
Authors: Yano Yasuhiro , Saito‐Ito Atsuko , Anchalee Dantrakool , Takada Nobuhiro 
Published: November 2013

AbstractBabesia microti protozoa were detected by light and electron microscopy in the salivary glands of field‐collected Ixodes ovatus ticks; 6 of 85 adult ticks were demonstrated to be positive for B. microti DNA by polymerase chain reaction assays. In the salivary glands of unfed ticks, B. microti existed in the sporo‐blast stage in the granular acinus cells, and developed into the sporozoite stage during feeding on the host for 2 days. The present results indicated for the first time that I. ovatus can indeed carry B. microti and is not infected mechanically with the parasites by blood‐sucking. This frequent infection of I. ovatus with B. microti demonstrates the significance of such a vector‐pathogen relationship in Japan, and strongly suggests that I. ovatus is involved in the maintenance of B. microti in the fauna of Japanese rodents.
Close Correlation of Streptococcal DNase B (sdaB) Alleles with emm Genotypes in Streptococcus pyogenes
Authors: Matsumoto Masakado , Sakae Kenji , Hashikawa Shinnosuke , Torii Keizo , Hasegawa Tadao , Horii Toshinobu , Endo Miyoko , Okuno Rumi , Murayama Shoko , Hirasawa Kyoko , Suzuki Rieko , Isobe Junko , Tanaka Daisuke , Katsukawa Chihiro , Tamaru Aki , Tomita Masaaki , Ogata Kikuyo , Ikebe Tadayoshi , Watanabe Haruo ,   The Working Group for Group A Streptococci in Japan, Ohta Michio 
Published: November 2013

AbstractDNase B is a major nuclease and a possible virulence factor in Streptococcus pyogenes. The allelic diversity of streptococcal DNase B (sdaB) gene was investigated in 83 strains with 14 emm genotypes. Of the 15 alleles identified, 11 alleles carried only synonymous nucleotide substitutions. On the other hand, 4 alleles had a non‐synonymous substitution other than synonymous substitutions, resulting in the substitution of a single amino acid. The distribution of each allele was generally emm genotype‐specific. Only sdaB7 was found in both emm2 and emm4. The promoter region was highly conserved and DNase B protein was similarly expressed in all alleles.
Detection and Quantification of Four Species of the Genus Clostridium in Infant Feces
Authors: Tonooka Toshiki , Sakata Shinji , Kitahara Maki , Hanai Michio , Ishizeki Shinobu , Takada Masaaki , Sakamoto Mitsuo , Benno Yoshimi 
Published: November 2013

AbstractTo determine the composition of Clostridium in the feces of infants approximately 30 days old, we have developed a detection and quantification method of Clostridium paraputrificum, Clostridium perfringens, Clostridium tertium, and Clostridium difficile by species‐specific primers. C. perfringens and C. difficile were detected in four fecal samples from 22 infants (18.2%), whereas C. paraputrificum was detected in three samples (16.7%). C. tertium was detected in two samples (9.1%). Moreover, the occurrences of the four species in bottle‐ and mix‐fed infants were relatively higher than in breast‐fed infants (P < 0.05). Subsequently, positive samples detected by nested PCR (polymerase chain reaction) were subjected to real‐time PCR. The results showed that the numbers of C. paraputrificum, C. perfringens, C. tertium, and C. difficile ranged from about 1 × 105 to 3 × 107 cells/g wet feces.
Analysis of the Distribution of Neuropathogenic Retroviral Antigens Following PVC211 or A8‐V Infection
Authors: Nakai Ryuhei , Takase‐Yoden Sayaka , Watanabe Rihito 
Published: November 2013

AbstractA8‐V and PVC211 are neuropathogenic strains of the Friend murine leukemia virus (Fr‐MLV) that cause spongiosis in the rat brain after infection at birth. PVC211 exhibited stronger neuropathogenicity than A8‐V, and induced more severe neurological symptoms such as hind‐leg paralysis. These symptoms correlated with the neuropathological spread and intensity, which were more severe in the spinal cord of rats infected with PVC211 than in those infected with A8‐V, without exhibiting neuropathological differences in other areas of the CNS. Interestingly, virus titers recovered from infected spinal cords were similar in PVC211 and A8‐V infected animals. However, in the spinal cord infected with PVC211, glial cells attained higher immunohistochemical expression scores for the viral surface antigen, gp70 (Env) than in the A8‐V infected spinal cord, although expression levels of viral antigens in blood vessel walls were similar in A8‐V and PVC211 infections. Furthermore, many of those glial cells which carried viral antigens were found, by double immunostaining, to be microglia. The results suggested that the spread of viral antigen positive microglia plays an important role in forming the different neuro‐pathogenicity observed in A8‐V and PVC211 infections.
Comparison of Effects of Nitric Oxide Synthase (NOS) Inhibitors on Plasma Nitrite/Nitrate Levels and Tissue NOS Activity in Septic Organs
Authors: Hayashi Yuri , Abe Masayoshi , Murai Akira , Shimizu Naomi , Okamoto Iku , Katsuragi Takeshi , Tanaka Keiichi 
Published: November 2013

AbstractAn excessive production of nitric oxide (NO) by NO synthase (NOS) is considered to contribute to circulatory disturbance, tissue damage, and refractory hypotention, which are often observed in septic disorders. It is anticipated that a selective inducible NOS (iNOS) inhibitor with excellent pharmacokinetics may be potentially effective as a novel and potent therapeutic intervention in sepsis. We examined whether or not a selective iNOS inhibitor shows iNOS selectivity at the tissue level, when administered systemically. The effects of four NOS inhibitors on plasma nitrite/nitrate (NOx) and tissue NOS levels were compared in major organs (lungs, liver, heart, kidneys, and brain) 6 hr after the injection of E. coli lipopolysaccharide (LPS) into male Wistar‐King rats. The rats treated with the three iNOS inhibitors (N‐(3‐(aminomethyl)benzyl)acetamidine (1400W), (1S, 5S, 6R, 7R)‐2‐aza‐7‐chloro‐3‐imino‐5‐methylbicyclo [4.1.0] heptane hydrochloride (ONO‐1714), and aminoguanidine) administered 1 hr after LPS injection, showed dose‐dependent decreases in plasma NOx levels and NOS activity in the lungs. The non‐selective NOS inhibitor (NG‐methyl‐L‐arginine (L‐NMMA)) had an effect only at the maximum dose. The differences in in vitro iNOS selectivity among these drugs did not correlate with iNOS selectivity at the tissue level. The relationship between plasma NOx levels and NOS activity in the lungs showed a linear relationship with or without the NOS inhibitors. In conclusion, the iNOS selectivity of these drugs does not seem to differ at the tissue level. Plasma NOx levels may be a useful indicator of lung NOS activity.
Facilitated Production of Secretory IgA against Shiga Toxin B Subunits by Intranasal Application of Antigen‐Coated Polystyrene Microspheres
Authors: Kurohane Kohta , Kobayashi Chie , Imai Yasuyuki 
Published: November 2013

AbstractWe examined the effects of microspheres as antigen carriers in mucosal immunization. Shiga toxin B subunits (Stx1B) were adsorbed on 6 μm polystyrene microspheres, which were then intranasally administered to mice together with cholera toxin (CT). Stx1B‐specific serum IgG production and secretory IgA production at local mucosal sites were enhanced by the use of microspheres. When OVA was used as a model antigen, secretory IgA production but not serum IgG production was enhanced on the use of microspheres. These results indicated that microspheres provide a useful means of potentiating the immune response against Stx1B with weak immunogenicity.
Peroral Infectivity of Toxoplasma gondii in Bile and Feces of Interferon‐γ Knockout Mice
Authors: Piao Lian‐Xun , Aosai Fumie , Mun Hye‐Seong , Yano Akihiko 
Published: November 2013

AbstractToxoplasama gondii appeared in the bile and feces of interferon‐γ knockout (GKO) but not wild type mice on days 7–8 after peroral infection with T. gondii cysts of Fukaya strain. Both tachyzoite‐specific SAG1 and bradyzoite‐specific T.g.HSP30 mRNAs were detected in the bile and feces of GKO mice. Tachyzoites converted to bradyzoites by culturing in the bile. By feeding uninfected mice with the bile and washed feces of T. gondii‐infected GKO mice, T. gondii‐specific antibody formation in the serum and cyst formation in the brain were observed. The novel migration route of T. gondii from liver to bile and feces in GKO mice was confirmed.
Influence of Additional Acylation Site(s) of Influenza B Virus Hemagglutinin on Syncytium Formation
Authors: Ujike Makoto , Nakajima Katsuhisa , Nobusawa Eri 
Published: November 2013

AbstractWe studied the effects of an increase in the hydrophobicity of the transmembrane domain (TM) and cytoplasmic tail (CT) of influenza B virus hemagglutinin (BHA) on fusion activities. For this purpose, we created mutant HAs with novel acylation site(s) in the TM and/or CT. All mutants were able to induce hemifusion and to form fusion pores as well as could wild type (wt) BHA. However, the ability of these mutants to form syncytia was impaired, indicating that the increase in the hydrophobicity of these domains (especially the CT) affected fusion pore dilation.