bluuebunny/crossref_metadata_2025_split huggingface dataset:
Carbohydrate‐Dependent Biological Activities of Glycosylated Human Interferon‐β on Human Hepatoblastoma Cells In Vitro
Authors: Utsumi Jun , Matsuo‐Ogawa Etsuko , Nagahata Takemitsu , Kasama Kyoko , Kagawa Yasuhiro , Satoh Yu‐ichiro 
Published: November 2013

AbstractTo evaluate the relationship between the sugar chain structure and biological activity, fibroblast‐derived glycosylated human interferon‐β, Chinese hamster ovary cell‐derived glycosylated recombinant human interferon‐β and Escherichia coli‐derived unglycosylated recombinant human interferon‐β were evaluated using human hepatoblastoma cells in vitro. Native fibroblast interferon‐β expressed more cell‐growth inhibitory action, 2′5′‐oligoadenylate synthetase induction, and the inhibition of hepatitis B virus DNA replication than its asialoform and two recombinant interferon‐βs. These results showed that the sugar chain structure of human interferon‐β affects its biological activity on human hepatoblastoma cells.
Characterization of an Enterotoxin Produced by Vibrio cholerae O139
Authors: Nakashima Kazutoshi , Eguchi Yukinori , Nakasone Noboru 
Published: November 2013

AbstractA cholera‐like enterotoxin was purified from Vibrio cholerae O139 strain AI‐1841 isolated from a diarrheal patient in Bangladesh. Its characteristics were compared with that of cholera toxins (CTs) of classical strain 569B and El Tor strain KT25. Al‐1841 produced as much toxin as O1 strains. The toxins were indistinguishable in terms of their migration profiles in conventional polyacrylamide gel disc electrophoresis, sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and isoelectrofocusing as well as their affinity for hydroxyapatite. The skin permeability factor activity and the fluid accumulation induced in rabbit ileal loops of the toxin of AI‐1841 were identical to those of the CTs. Three toxins equally reacted against anti‐569B CT antiserum in Western blotting, and their B subunits formed a precipitin line against any anti‐B subunit antiserum by double gel immunodiffusion. Anti‐569B CTB antibody neutralized the three toxins in their PF activities and enterotoxicities. The amino acid sequence of 1841 toxin B subunit was identical with that of KT25 CTB, corresponding to the DNA sequence of ctxB from El Tor strains of the seventh pandemic. We concluded 1841 toxin was identical to CT of the seventh pandemic El Tor vibrios.
CD3+/TCR‐αβ− Cells Are Important in Protecting Spinal Cord Tissues against Theiler's Virus Strain GD VII Infection
Authors: Kohanawa Masashi , Nakane Akio , Asano Misako , Minagawa Tomonori 
Published: November 2013

AbstractIntravenous infection with Theiler's virus strain GD VII causes acute encephalomyelitis in mice. Endogenous IFN‐γ produced in the spinal cord is important to protect the tissue in mice infected with this virus. Neither CD4+ cells nor CD8+ cells infiltrated the spinal cords of infected mice until Day 9 postinfection. However, the number of CD3+/TCR‐γδ+ cells increased in the spinal cords of mice infected with the virus. These cells resided in the spinal cords of normal mice, and produced IFN‐γ as a result of stimulation by immobilized anti‐CD3 mAb. Elimination of CD3+ cells by the administration of a specific mAb augmented viral replication and suppressed production of endogenous IFN‐γ. Depletion of TCR‐αβ+ cells and ASGM1+ cells did not affect the viral replication, and did not alter the production of IFN‐γ. Therefore, CD3+/TCR‐αβ– cells producing IFN‐γ play an important role in the protection of the spinal cord against Theiler's virus infection. These results suggest that CD3+/TCR‐αβ– cells might be identical to TCR‐γδ+ cells.
Effect of Substitution for Arginine Residues near Position 146 of the A Subunit of Escherichia coli Heat‐Labile Enterotoxin on the Holotoxin Assembly
Authors: Okamoto Keinosuke , Takatori Rie , Okamoto Kyoko 
Published: November 2013

AbstractEscherichia coli heat‐labile enterotoxin (LT) is a holotoxin which consists of one A and five B subunits. Although B subunit monomers released into periplasm can associate into pentameric structures in the absence of the A subunit, the A subunit accelerates the assembly. To express the function, A subunit constructs the proper spatial structure. However, the regions involved in the construction are unknown. To identify the regions, we substituted arginine residues near position 146 of the A subunit with glycine by oligonucleotide‐directed site‐specific mutagenesis and obtained the mutants expressing LT(R141G), LT(R143G), LT(R146G), LT(R143G, R146G), LT(R141G, R143G, R146G) and LT(R143G, R146G, R148G). We purified these mutant LTs by using an immobilized d‐galactose column and analyzed the purified mutant LTs by SDS‐PAGE to examine the amount of A subunit associated with B‐subunit oligomer. The substitution of an arginine residue at any position did not induce a significant alteration in the amount of A subunit associated with B‐subunit oligomer. However, the substitution of more than two arginine residues induced a significant decrease in the amount of A subunits associated with the B‐subunit oligomer. Subsequently, we measured the level of the intracellular B‐subunit oligomer of these mutant strains. The measurement revealed that the amount of B‐subunit oligomer in cells decreased as the number of substituted arginine residues increased. These results show that all arginine residues near position 146 are important for the construction of the functional A subunit, and thus for holotoxin formation, although each individual arginine residue is not an absolute requirement.
Comparison of the Selectivity of Anti‐Varicella‐Zoster Virus Nucleoside Analogues
Authors: Machida Haruhiko , Nishitani Makiko , Watanabe Yohko , Yoshimura Yuichi , Kano Fumitaka , Sakata Shinji 
Published: November 2013

AbstractWe compared the selectivity of six anti‐varicella‐zoster virus (VZV) drugs, which are clinically available or of which clinical efficacy for the treatment of VZV infections has been reported. Sorivudine (BV‐araU) had the most potent anti‐VZV effect in the plaque inhibition assay, followed by brivudine (BVDU) and 5‐propynyl‐arabinofuranosyluracil (Pry‐araU). All test compounds, except vidarabine (AraA), had only a very weak effect on human embryonic lung cell growth. The selectivity indexes (ID50 for cell growth/ED50 for VZV plaque inhibition) of BV‐araU, BVDU, and Pry‐araU were > 1,000,000, 20,000, and > 10,000, respectively, while those of acyclovir and penciclovir ranged from 600 to 800. AraA was much less selective than any of the other drugs tested. We measured the amount of pH] thymidine incorporated into the acid‐insoluble fraction of VZV‐infected cells to determine the ability of these drugs to selectively inhibit viral DNA synthesis. [3H]Thymidine incorporation was markedly inhibited by all anti‐VZV compounds, except BVDU. Treatment of infected cells with drugs from 32 to 38 hr after infection inhibited the DNA synthesis to the same extent as VZV plaque formation, except that AraA inhibited the DNA synthesis at a lower dose than for VZV plaque formation. DNA synthesis in non‐infected growing cells was inhibited to the same extent as cell growth. A particularly high selectivity index for the inhibition of DNA synthesis was noted for BV‐araU, which was defined as the ratio of inhibitions of DNA synthesis in VZV‐infected and non‐infected. The highest selectivity indexes were recorded for BV‐araU > Pry‐araU > acyclovir ≥ penciclovir > AraA.
Inhibition of Varicella‐Zoster Virus Glycoprotein Expression by Peripheral Blood Mononuclear Cells
Authors: Mizutani Ken‐ichi , Ito Masahiro , Kamiya Hitoshi , Sakurai Minoru 
Published: November 2013

AbstractThe effect of peripheral blood mononuclear cells (PBMC) on expression of varicella‐zoster virus (VZV) glycoproteins (Gps) was analyzed by flow cytometry. PBMC from VZV seropositive and seronegative donors and supernatant of PBMC co‐cultured with VZV‐infected human embryonic fibroblasts reduced VZV Gp expression. Neutralization of supernatant fluid with mixture of anti‐interferons (IFN)‐α, ‐β, ‐γ, and tumor necrosis factor (TNF)‐α partially reduced inhibitory activity of supernatant on VZV Gp expression. Deletion of natural killer (NK) cells and adherent cells from PBMC reduced inhibitory activity of PBMC on VZV Gp expression. These results suggest that IFN‐α, ‐β, ‐γ, TNF‐α and other soluble factors released from NK cells and monocytes by co‐cultivation with VZV‐infected fibroblasts inhibit VZV Gp expression.
Differentiation of Clostridium difficile, Clostridium bifermentans, Clostridium sordellii, and Clostridium perfringens from Diarrheal Stool by API ZYM and API LRA Oxidase Test
Authors: Fontana Carla , Jezzi Teresa , Testore Gian Piero , Dainelli Benedetto 
Published: November 2013

AbstractA simple, rapid and reliable outline for identification of clostridia isolates from human infections was developed. It consists of a combination of API ZYM and API LRA Oxidase tests. The enzymatic activities were performed with strains sub‐cultured onto carbohydrate‐free medium (Columbia blood agar). Fifty‐five strains of Clostridium difficile, C. bifermentans, C. sordellii, and C. perfringens from clinical specimens and eight reference standard strains representing different species of the same genus were analyzed. The accuracy of the new method was evaluated by comparison with the results obtained by DNA/DNA analysis.
Frequent Detection of Hepatitis C Virus Subtype 3a (HCV‐3a) Isolates in Thailand by PCR Using Subtype‐Specific Primers
Authors: Apichartpiyakul Chatchawann , Miyajima Hirofumi , Doi Hisaya , Mizokami Masashi , Homma Morio , Hotta Hak 
Published: November 2013

AbstractBy means of a polymerase chain reaction (PCR) method using subtype‐specific primers for hepatitis C virus (HCV) subtypes 1a, 1b, 2a, 2b and 3a, the prevalence of each subtype among HCV isolates in Chiang Mai, Thailand, was determined. HCV‐3a appeared to be the most common subtype in blood donors, and was also frequently found in patients with liver disease. HCV‐1b, but not HCV‐2a or −2b, was also commonly found in this area, while a considerable percentage of the total HCV isolates still remained unclassifiable by the above methods. Serotype analysis of the HCV isolates using C14‐1 and C14‐2 recombinant peptides revealed that HCV‐3a was likely to carry an antigenic determinant(s) different from those of the major types 1 (HCV‐1a and −1b) and 2 (HCV‐2a and −2b).
Identification of Phosphocholine‐Containing Glycoglycerolipids Purified from Mycoplasma fermentans‐Infected Human Helper T‐Cell Culture as Components of M. fermentans
Authors: Matsuda Kazuhiro , Harasawa Ryo , Li Jin‐liang , Kasama Takeshi , Taki Takao , Handa Shizuo , Yamamoto Naoki 
Published: November 2013

AbstractPreviously, we have reported the occurrence of novel phosphocholine‐containing glycoglycerolipids (GGPLs: GGPL‐I and GGPL‐III) in human helper T‐cell culture (MT‐4 cell line) (Matsuda et al, Glycoconjugate J. 10: 340). However, the GGPLs disappeared from the MT‐4 cells after treatment with an antimycoplasma agent. This disappearance suggested the involvement of microorganisms in the GGPL expression. In this paper, we show that the novel lipids are components of Mycoplasma fermentans itself. The supernatant fluid of the antimycoplasma agent‐untreated MT‐4 cell culture produced mycoplasma‐like colonies on PPLO agar plates, and PCR and immunological methods revealed the presence of M. fermentans. GGPLs were expressed again in the treated MT‐4 cells after infection with the isolated M. fermentans. The isolated M. fermentans had glycoglycerolipids corresponding to GGPL‐I and GGPL‐III. Thin‐layer chromatography‐mass spectrometry and immunological analyses showed that these glycoglycerolipids which were derived from the isolated M. fermentans were identical with GGPL‐I and GGPL‐III previously obtained. This is the first report that shows mycoplasma has phosphocholine‐containing glycoglycerolipids.
Typing of Staphylococcus epidermidis Colonizing in Human Nares by Pulsed‐Field Gel Electrophoresis
Authors: Hu Lan , Umeda Akiko , Amako Kazunobu 
Published: November 2013

AbstractFrom the nares of 11 healthy adults, 253 strains of coagulase negative staphylococcus were isolated and 88% of them were identified as Staphylococcus epidermidis using the API STAPH system. Chromosomal DNA fingerprinting of the isolated strains revealed that each person carried multiple types of S. epidermidis in his or her nares. The colonization of the strains was not stable; the types of the isolates changed in the first and the second examinations 5 months apart. The results contrasted with previous findings in which only one strain of S. aureus colonized persistently in the nares of healthy adults.
Heat Shock Proteins in the Human Periodontal Disease Process
Authors: Ando Tomohiro , Kato Tetsuo , Ishihara Kazuyuki , Ogiuchi Hideki , Okuda Katsuji 
Published: November 2013

AbstractThe production of HSP by periodontopathic Gram‐negative bacteria was examined by SDS‐PAGE, two dimensional gel electrophoresis, and Western blotting using monoclonal antibodies against HSPs. Strains of Actinobacillus actinomycetemcomitans, Eikenella corrodens, Fusobacterium nucleatum, Prevotella intermedia, Prevotella nigrescens, Prevotella melaninogenica, and Treponema socranskii species produced HSP which reacted with anti‐Yersinia enterocolitica HSP 60 and/or mycobacterial 65‐kDa HSP monoclonal antibodies. It was found that gingival homogenate samples from patients with adult periodontitis reacted with anti‐human HSP 60 and bovine brain HSP 70 monoclonal antibodies. Antibodies which reacted with bacterial HSP were also found in a serum sample from a periodontitis patient. The present study suggests that HSPs are implicated in the human periodontal disease process.
Seroepidemiological Survey ofBartonella (Rochalimaea) henselaein Domestic Cats in Japan
Authors: Ueno Hiroshi , Muramatsu Yasukazu , Chomel Bruno B. , Hohdatsu Tsutomu , Koyama Hiroyuki , Morita Chiharu 
Published: November 2013

AbstractA total of 199 domestic cat serum samples from 3 geographical areas (northeastern, central and southwestern) of Japan collected between 1992 to 1994 were examined for serum antibody againstBartonella henselaeusing an immunofluorescent assay. The antibody prevalence was 15.1% (30/199). A significant difference in the prevalence ofB. henselaeantibody was observed between the northeastern area (6.3%:3/48) and the central area (22.0%: 13/59) in Japan. There was no significant difference between the average age of seropositive cats (4.39 ±3.26 years) and that of seronegative cats (4.03 ±3.84 years), and also between the frequency of seropositive male cats (16.5%: 15/91) and that of seropositive female cats (11.8%:9/76). This is the first report ofB. henselaeantibodies in cats in Japan.
Correlation between the Presence of Virulence‐Associated Genes as Determined by PCR and Actual Virulence to Mice in Various Strains of Listeria Spp.
Authors: Nishibori Takeaki , Cooray Karven , Xiong Huabao , Kawamura Ikuo , Fujita Masashi , Mitsuyama Masao 
Published: November 2013

AbstractFive chromosomal genes, prfA, plcA, hlyA, mpl and plcB, are implicated in the virulence of Listeria monocytogenes and some of these genes have been used for the identification of bacteria by polymerase chain reaction (PCR). Using 6 strains of L. monocytogenes and 3 L. innocua strains, the relationship was examined between the presence of five virulence‐associated genes and actual virulence to mice in terms of 50% lethal dose (LD50), bacterial viability in the organ of infected mice and the intracellular growth in cultured macrophages. None of the five genes could be amplified by PCR in all the L. innocua strains and they were actually avirulent to mice. All L. monocytogenes strains were shown to be virulent and to have intact virulence‐associated genes except for the strain ATCC15313. This particular strain was revealed to be avirulent and defective in hlyA and plcA in PCR amplification. It was suggested that PCR detection of genes prfA, mpl, or plcB may not be sufficient to detect virulent strains of L. monocytogenes. It appeared that the ability to produce listeriolysin O (LLO), which is encoded by hlyA, was critical for the expression of virulence regardless of the amount of LLO produced.
Optimum Culture Conditions for Production of Exfoliative Toxin by Staphylococcus hyicus
Authors: Watanabe Takao , Tanabe Taishi , Sato Hisaaki , Fuse Yoko , Ueda Kengo , Nakano Katsushige , Saito Hiroshi , Maehara Nobutoshi 
Published: November 2013

AbstractOptimum culture conditions for the production of exfoliative toxin by Staphylococcus hyicus (shET) were examined. High shET activity was obtained from the culture filtrate of HI and TY broth inoculated with S. hyicus. The pH in these two media ranged from 7 to 8.5 during bacterial culture, while the lowest pH in TS and BHI broth was less than 6. shET activity in the culture filtrate from TY broth inoculated with 107 CFU of S. hyicus per ml was higher than that in TY broth inoculated with 106 and 108 CFU of bacteria per ml. When shET activity in the culture filtrate was measured under various shaking conditions, the culture filtrate shaken at 75 oscillations per min had the highest shET activity of the five shaking conditions. shET activity of the culture filtrate of TY broth to which protease inhibitor had been added was the same as that of TY broth without inhibitor. shET activity in a shaking culture in an Erlenmeyer flask was also the same as that in sac culture and that in shaking culture using a shaking (Sakaguchi) flask. shET activity in TY broth supplemented with 100 mM glucose was significantly lower than that in TY broth without glucose. Based on the above results, the optimum culture conditions for the production of shET were as follows: inoculation of 3 × 109 CFU of S. hyicus strain P‐1 into 300 ml of TY broth in a 2,000‐ml Erlenmeyer flask, and incubation at 37 C with shaking at 75 oscillations per min. Then shET activity of the culture filtrate under appropriate culture conditions was measured after various incubation periods. shET activity was detected 6 hr after inoculation, reached the maximum (253 exfoliative unit/0.1 ml) at 16 hr and decreased between 20 and 48 hr. Thus, the optimum incubation period was determined to be 16 hr. Then the optimum concentration of ammonium sulfate for isolation of shET from the culture filtrate under appropriate culture conditions was examined. The greatest shET activity was obtained from the fraction salted out with 90% saturated ammonium sulfate. Thus, the optimum concentration of ammonium sulfate for the isolation of shET was determined to be 90% saturation.
Production of Tumor Necrosis Factor‐α in Granulocytopenic Mice with Pulmonary Candidiasis and Its Modification with Granulocyte Colony‐Stimulating Factor
Authors: Futenma Mitsuhiko , Kawakami Kazuyoshi , Saito Atsushi 
Published: November 2013

AbstractIn the present study, a lethal model of pulmonary candidiasis was established using granulocytopenic mice with cyclophosphamide. These mice started to die 1 day after infection and had all died within the next 48 hr. The counts of live C. albicans in the lung gradually increased with time, while the organisms were quickly eliminated in the normal mice. From the histology and measurements on bronchoalveolar lavage fluid (BALF), polymorphonuclear cells (PMN) response was almost zero up to 24 hr, and then a weak but significant response was observed at 48 hr, while a marked accumulation of PMN was detected from as early as 6 hr in normal mice. In contrast, macrophages had accumulated in BALF by 48 hr in granulocytopenic mice, but not in normal mice. Both in serum and BALF, a considerable level of tumor necrosis factor‐α (TNF‐α) was detected from 6 hr, peaking at 24 to 48 hr, while in normal mice the quantity was under the detection limit in serum and very low in BALF. The effects of administering granulocyte colony‐stimulating factor (G‐CSF) on these parameters were next examined. G‐CSF significantly prolonged the survival time of granulocytopenic mice, promoted the clearance of organisms through increasing the counts of PMN in the lung, and strongly inhibited the production of TNF‐α both in BALF and serum. These results suggest that this cytokine does not protect them, but plays some role in their death due to candidial infection in granulocytopenic mice.
Nucleotide Sequence Analysis of Recent Epidemic Strains of Enterovirus 70
Authors: Yamazaki Kenji , Oishi Isao , Minekawa Yoshiichi 
Published: November 2013

AbstractNucleotide sequences of the genome of enterovirus 70 (EV70) isolated in Osaka in 1993 were determined, and compared with those of the past epidemic strains. Nucleotide substitution rates in 332 bp, 174 bp, and 178 bp of the genes encoding viral protein (VP)1, VP2, or VP3 were 9.0, 7.5, and 5.6% between Kumoi‐2/93 and J670/71, respectively. Likewise, the putative amino acid substitution rates were 1.8, 0, and 0%. It seems that the epidemic strains of EV70 in Japan have been evolving at a constant high nucleotide substitution rate but almost all the substitutions were synonymous.
Characterization of Nontoxic‐Nonhemagglutinin Component of the Two Types of Progenitor Toxin (M and L) Produced by Clostridium botulinum Type D CB‐16
Authors: Ohyama Tohru , Watanabe Toshihiro , Fujinaga Yukako , Inoue Kaoru , Sunagawa Hiroyuki , Fujii Nobuhiro , Inoue Katsuhiro , Oguma Keiji 
Published: November 2013

AbstractA 9.8‐kbp DNA fragment which contained a neurotoxin gene and its upstream region was cloned from Clostridium botulinum type D strain CB‐16. Nucleotide sequencing of the fragment revealed that genes encoding for hemagglutinin (HA) subcomponents and one for a nontoxic‐nonhemagglutinin (NTNH) component were located upstream of the neurotoxin gene. This strain produced two toxins of different molecular size (approximately 300 kDa and 500 kDa) which were designated as progenitor toxins (M and L toxins). The molecular size of the NTNH component of L toxin was approximately 130 kDa on SDS‐PAGE and its N‐terminal amino acid sequence was M‐D‐I‐N‐D‐D‐L‐N‐I‐N‐S‐P‐V‐D‐N‐K‐N‐V‐V‐I which agreed with that deduced from the nucleotide sequence. In contrast, the M toxin had a 115‐kDa NTNH component whose N‐terminal sequence was S‐T‐I‐P‐F‐P‐F‐G‐G‐Y‐R‐E‐T‐N‐Y‐I‐E, corresponding to the sequence from Ser141 of the deduced sequence. A 15‐kDa fragment, which was found to be associated with an M toxin preparation, possessed the same N‐terminal amino acid sequence as that of the 130‐kDa NTNH component. Furthermore, five major fragments generated by limited proteolysis with V8 protease were shown to have N‐terminal amino acid sequences identical to those deduced from the nucleotide sequence of 130‐kDa NTNH. These results indicate that the 130‐kDa NTNH of the L toxin is cleaved at a unique site, between Thr and Ser, leading to the 115‐kDa NTNH of the M toxin.