bluuebunny/crossref_metadata_2025_split huggingface dataset:
PCR‐RFLP Analysis of Cytomegalovirus Infections Associated with Bone Marrow Transplantation in Japanese Children
Authors: Kawasaki Satoshi , Oshitani Hitoshi , Suzuki Hiroshi , Arakawa Masaaki , Mizuta Katsumi , Imaizumi Masue , Tsuchiya Shigeru , Konno Tasuke 
Published: November 2013

AbstractIn order to investigate the longitudinal molecular epidemiology of cytomegalovirus (CMV) infections associated with bone marrow transplantation (BMT) in Japanese children, we analyzed 36 CMV strains from 11 cases. Three regions (DNA polymerase, glycoprotein H, and immediate‐early regions) of CMV DNA were amplified by polymerase chain reaction (PCR), and amplified products were each digested with two restriction enzymes, followed by electrophoresis. These restriction fragment length polymorphism (RFLP) analyses allowed the differentiation of 36 strains into 13 genotypes. Each patient excreted his or her own CMV with distinct genotype over the study period of up to one year. CMVs of two different genotypes were recovered during a one‐month study from one recipient, who received a peripheral blood stem cell transplantation. Although the majority of patients and donors were CMV‐seropositive before BMT, multiple CMV infections might not be common and the reactivation of latently infected CMV might be prominent in Japanese children receiving transplants.
Significance of Anaerobic Preincubation of Helicobacter pylori tor Measuring Metronidazole Susceptibility by the Etest
Authors: Chida‐Sakata Noriko , Baba Masaru , Inagawa Hiroko , Wada Akihito , Tanaka Tatsuro , Hoshihara Yoshio , Takemoto Tadayoshi 
Published: November 2013

AbstractThe Etest is widely used for measuring the susceptibility of Helicobacter pylori to metronidazole. By using 55 H. pylori isolates from 55 patients and a standard H. pylori strain, NCTC11637, we compared metronidazole susceptibility results obtained from the Etest with or without anaerobic preincubation to those obtained from the agar dilution method. Mueller Hinton agar plates supplemented with 5% horse blood were used for both methods. For the Etest, plates were incubated for 72 hr at 35 C under microaerophilic conditions after 0‐, 4‐ or 24‐hr periods of anaerobic preincubation. For the agar dilution method, the plates were incubated at the same microaerophilic conditions as those for the Etest. Without anaerobic preincubation for the Etest, 39 of the 56 (70%) H. pylori isolates were categorized as resistant to metronidazole (minimal inhibitory concentration >8 mg/liter), whereas only one of the 56 (1.8%) isolates was resistant according to the agar dilution method. The resistant and susceptible agreement rate was 32%. Four‐hour anaerobic preincubation did not alter the readings of the Etest significantly. However, when the Etest was performed with 24‐hr anaerobic preincubation, the number of isolates categorized as resistant was reduced to six (11%), improving the agreement rate to 91%. For measuring the metronidazole susceptibility of H. pylori by the Etest, 24‐hr anaerobic preincubation is necessary to agree with the results obtained by the agar dilution test.
Human Immunodeficiency Virus Type 1 Vpr Modifies Cell Proliferation via Multiple Pathways
Authors: Yamaguchi Tomoyuki , Watanabe Nobumoto , Nakauchi Hiromitsu , Koito Atsushi 
Published: November 2013

AbstractVpr, one of the accessory molecules of HIV‐1, has been demonstrated to arrest the cell cycle at the G2 phase. This Vpr‐mediated cell cycle arrest is implicated to have an important role in the viral life cycle. In the present study, we quantitate the extent of Vpr‐mediated cell cycle arrest with the use of a bicistronic vector consisting of a vpr gene and a green fluorescence protein sequence. Using this system, we examined the effect of several Vprs on cell cycle progression and growth of cells from different species quantitatively. We found that Vpr from the T‐cell line‐adapted HIV‐1SF2 strain (Vpr2) could not significantly induce G2 arrest in HeLa cells but was able to induce it in 293T cells. However, strong inhibition of cell proliferation in HeLa cells as well as in 293T cells was observed by Vpr2. This ability of Vpr2 to inhibit cell proliferation without G2 arrest was also observed when expressed in monkey cell line. Analyses of chimeric Vprs revealed that this species‐non‐specific growth inhibitory activity of Vpr was not mediated solely by the C‐terminal region of Vpr. These results indicated that the growth inhibitory activity of Vpr is independent of its G2 arresting activity. In addition, the species‐non‐specific nature of this activity suggests that Vpr has a novel mechanism to retard cell proliferation by influencing basic cellular functions.
Association of a Cellular 21‐kDa Transmembrane Protein (VAP21) with Enveloped Viruses
Authors: Yamamoto Kentaro , Tochikura Tadafumi S. , Xiao Sa , Sakurai Akira , Kawai Akihiko 
Published: November 2013

AbstractWe reported previously that the rabies virions contained a 21‐kDa cellular transmembrane protein (referred to as VAP21) as a minor component (Sagara, J. et al, Microbiol. Immunol. 41(12): 947‐955, 1997). In this study, we further examined the possible interactions of VAP21 with other enveloped viruses, including the vesicular stomatitis virus (VSV; negative‐stranded RNA virus), Sindbis virus (positive‐stranded RNA virus) and herpes simplex virus type 1 (HSV‐1; double‐stranded DNA virus). An immunoblot analysis demonstrated that all of these enveloped viruses contained VAP21 in the virion as a minor component. Immunoprecipitation studies suggested that VAP21 was associated with certain viral proteins in the cell, such as the matrix (M) protein of VSV, a capsid protein of Sindbis virus, and at least a capsid protein (VP5) of HSV‐1. The association was disrupted by treatment with 0.5% sodium dodecyl sulfate, but resistant to the treatment with 1% NP‐40 plus 1% deoxycholate. These results suggest that: 1) VAP21 is not primarily associated with the viral transmembrane glycoprotein but rather with the internal viral protein, and, 2) this association would cause the efficient incorporation of VAP21 into the virion.
Identification of Copper‐Zinc Superoxide Dismutase Gene from Enteroaggregative Escherichia coli
Authors: Kumar Rakesh , Sidhu Maninder Kaur , Ganguly Nirmal Kumar , Chakraborti Anuradha 
Published: November 2013

AbstractWe describe here the identification of sodC gene from enteroaggregative Escherichia coli (EAggEC). A 294 bp gene‐specific fragment was amplified from the organism by DNA as well as RT‐PCR using primers from bacterial sodC sequences. The metal co‐factor present in the protein was confirmed by running samples in native gels and inhibiting with 2 mM potassium cyanide. However, the nonpathogenic E. coli possesses the gene but does not express it. Thus, the presence of copper‐zinc superoxide dismutase encoded by sodC was demonstrated for the first time in EAggEC, which means it could be a novel candidate for a virulence marker.
Potentiation of Immune Response against the RESA Peptides of Plasmodium falciparum by Incorporating a Universal T‐Cell Epitope (CS. T3) and an Immunomodulator (Polytuftsin), and Delivery through Liposomes
Authors: Kumar Pawan , Biswas Sukla , Rao Donthamsetty Nageswara 
Published: November 2013

AbstractSynthetic peptides representing repeat sequences of ring‐infected erythrocyte surface antigen (RESA) of Plasmodium falciparum have shown poor immunogenicity and protection. In this study, the RESA peptides [(EENVEHDA)2 and (DDEHVEEPTVA)2] were chemically linked to a universal T‐cell determinant, CS.T3, derived from the CS protein of P. falciparum. Polytuftsin (TKPR)40, a polymer of naturally occurring immunomodulator “tuftsin,” was physically mixed with these conjugates. These preparations in alum and liposomes were immunized in four inbred strains of mice with different genetic backgrounds to study the humoral response. In the case of liposome‐entrapped preparations, a 10 μg dose of antigen showed the optimum antibody response. Mice immunized with liposome containing RESA peptide(s)‐CS.T3 conjugate along with polytuftsin showed the highest antibody levels in all the strains, whereas the RESA peptide(S) alone, adsorbed on alum or entrapped in liposomes, showed either poor or moderate antibody levels. The antibodies raised against liposome‐entrapped preparations in both high‐responder strain (SJL/J H‐2s) and low‐responder strain (FVB/J H‐2q) showed 24‐fold lower Kd values as compared to the alum adsorbed preparations, suggestive of high affinity antibodies. All the antigen preparations predominantly induced IgG2a and IgG2b isotype response, suggesting that the T‐helper response involved is of the CD4+ Th1 type. The in vitro merozoite reinvasion inhibition assay showed 5092% inhibition with sera raised against different antigen formulations. The highest percentage inhibition was observed with the RESA peptide‐CS.T3 conjugate containing polytuftsin in liposomes. Thus, the incorporation of peptide antigens inside liposomes not only reduced the antigen dose by 5‐fold but also elicited a high titre with high affinity antibodies and the inhibition of merozoites to RBC in vitro. Therefore, we conclude that the incorporation of these synthetic constructs in liposomes could be a useful strategy for the development of a subunit immunogen against malaria.
Isolation of Feline Parvovirus from Peripheral Blood Mononuclear Cells of Cats in Northern Vietnam
Authors: Miyazawa Takayuki , Ikeda Yasuhiro , Nakamura Kazuya , Naito Risako , Mochizuki Masami , Tohya Yukinobu , Vu Dat , Mikami Takeshi , Takahashi Eiji 
Published: November 2013

AbstractFeline parvovirus (FPV) was isolated rather frequently from the peripheral blood mononuclear cells (PBMCs) of cats in northern Vietnam by coculturing with MYA‐1 cells (an interleukin‐2‐dependent feline T lymphoblastoid cell line) or Crandell feline kidney (CRFK) cells (a feline renal cell line). Efficiency of virus isolation was higher in MYA‐1 cells than in CRFK cells. Interestingly, among the 17 cats from which FPV was isolated, 9 cats were positive for virus neutralizing (VN) antibody against FPV, indicating that FPV infected PBMCs and was not eliminated from PBMCs even in the presence of VN antibodies in the cats.
Differences in Genomic Macrorestriction Patterns of Arabinose‐Positive (Burkholderia thailandensis) and Arabinose‐Negative Burkholderia pseudomallei
Authors: Chaiyaroj Sansanee C. , Kotrnon Kanokorn , Koonpaew Surapong , Anantagool Narisara , White Nick J. , Sirisinha Stitaya 
Published: November 2013

AbstractWe reported previously two biochemically and antigenically distinct biotypes of Burkholderia pseudomallei. These two distinct biotypes could be distinguished by their ability to assimilate l‐arabinose. Some B. pseudomallei isolated from soil samples could utilize this substrate (Ara+), whereas the other soil isolates and all clinical isolates could not (Ara). Only the Ara− isolates were virulent in animals and reacted with monoclonal antibody directed at the surface envelope, most likely the exopolysaccharide component. In the present study, pulsed‐field gel electrophoresis was employed for karyotyping of these previously identified B. pseudomallei strains. We demonstrate here that the DNA macrorestriction pattern allows the differentiation between B. pseudomallei, which can assimilate l‐arabinose, and the proposed B. thailandensis, which cannot do so. Bacterial strains from 80 melioidosis patients and 33 soil samples were examined by genomic DNA digestion with NcoI. Two major reproducible restriction patterns were observed. All clinical (Ara) isolates and 9 Ara soil isolates exhibited macrorestriction pattern I (MPI), while 24 soil isolates (Ara+) from central and northeastern Thailand displayed macrorestriction pattern II (MPII). The study here demonstrated pulsed‐field gel electrophoresis to be a useful tool in epidemiological investigation possibly distinguishing virulent B. pseudomallei from avirulent B. thailandensis or even identifying closely related species of Burkholderia.
Phylogenetic Analysis of Saccharolytic Oral Treponemes Isolated from Human Subgingival Plaque
Authors: Sakamoto Mitsuo , Koseki Takeyoshi , Umeda Makoto , Ishikawa Isao , Benno Yoshimi , Nakase Takashi 
Published: November 2013

AbstractA total of 74 strains of oral treponemes, which were isolated from subgingival plaque samples from patients with periodontitis, were taxonomically studied on the basis of biochemical characteristics, DNA‐DNA hybridization, and 16S rRNA gene sequences. These organisms fermented carbohydrates and required rumen fluid or short‐chain volatile fatty acids for growth. The isolates were divided into seven subgroups based on their biochemical characteristics. The levels of DNA relatedness among the representative strains of each subgroup and Treponema socranskii (including three subspecies) were greater than 78%, while the levels of DNA relatedness among these strains and other Treponema species, including T. denticola and “T. vincentii”, were less than 15%. DNA‐DNA hybridization indicated that all subgroups belonged to T. socranskii. This result correlated well with the cluster on the phylogenetic trees based on 16S rRNA sequences.
Phenotypic and Genotypic Homogeneity of the Strains of Rickettsia japonica Isolated from Patients with Oriental Spotted Fever
Authors: Uchiyama Tsuneo 
Published: November 2013

AbstractNine pathogenic strains of Rickettsia japonica isolated from patients with Oriental spotted fever were compared phenotypically and genotypically. Constitution and antigenicity of the proteins demonstrated to be the same among strains. Polymerase chain reaction (PCR) amplification of the two major outer membrane protein genes (ompA and ompB) and an intracellular spotted fever group‐common antigen protein gene (rps120) produced the same sizes of products for all strains. Restriction fragment length polymorphism of the PCR products showed the same pattern among strains with each endonuclease. Thus, these strains belong to a single type, the same as the type strain YH (=ATCC VR‐1363).
Detection of Clostridium difficile Toxin A by Reversed Passive Latex Agglutination
Authors: Toma Claudia , Nakamura Shinichi , Kamiya Shigeru , Nakasone Noboru , Iwanaga Masaaki 
Published: November 2013

AbstractA reversed passive latex agglutination (RPLA) assay for detecting Clostridium difficile toxin A is presented. Purified monoclonal antibody (mAb 37B5) was used for latex sensitization. The culture supernatants of 93 strains of C. difficile were tested by RPLA assay and the results compared with those of a commercially available latex agglutination test, PCR and cytotoxin assay with Vero cells. There was agreement between RPLA, cytotoxicity and PCR assays, but 29 strains were positive in the RPLA assay while 35 were positive in the cytotoxicity test and PCR using primer pair NK3‐NK2 directed to the nonrepeating portion of the C. difficile toxin A gene. The 6 cytotoxic but RPLA‐negative strains were demonstrated to be toxin A‐negative/toxin B‐positive strains in the PCR assay by using primer pair NK11‐NK9 directed to the repeating portion of the C. difficile toxin A gene. There were no cross‐reactions with culture supernatants of the other clostridial strains except for two strains of C. sordelli that produced hemorrhagic toxin (which is immunologically related to C. difficile toxin A).
Production and Partial Characterization of Antibody to Cord Factor (Trehalose 6,6′‐Dimycolate) in Mice
Authors: Fujiwara Nagatoshi , Oka Shiro , Ide Michio , Kashima Kazutoshi , Honda Takeshi , Yano Ikuya 
Published: November 2013

AbstractAntibody production against the trehalose 6,6′‐dimycolate (TDM, cord factor) of Rhodococcus ruber, a non‐pathogenic species of the Actinomycetales group, was investigated in mice by repeated intraperitoneal injection of TDM in water‐in‐oil‐in‐water micelles without carrier protein. The antigenic TDM was isolated and purified chromatographically from the chloroform‐methanol extractable lipids of R. ruber. The hydrophobic moiety of this TDM was composed of two molecules of monoenoic or dienoic α‐mycolic acids with a carbon chain length ranging from C44 to C48 centering at C46. To detect the antibody, an enzyme‐linked immunosorbent assay (ELISA) system was employed using plastic plates coated with TDM. The antibody reacted against the TDM of R. ruber. The antibody was reactive in similar fashion against glycosyl monomycolates differing in the carbohydrate moiety, such as that of glucose mycolate (GM) and mannose mycolate (MM), obtained from R. ruber. Moreover, the antibody reacted against mycolic acid methyl ester itself when it was used as the antigen in ELISA, and trehalose did not absorb the antibody to TDM or inhibit the reaction. These results indicate that the epitope of TDM recognized by the antibody is mycolic acid, an extremely hydrophobic part of the molecule. Next, we prepared monoclonal anti‐TDM antibody (moAb) in mice myeloma cells to examine its biological activities and the role of humoral immunity in mycobacterial infection. MoAb reacted against the TDM, glycosyl mycolate, and mycolic acid methyl ester in ELISA in the same manner as our polyclonal antibody did. The administration of moAb suppressed granuloma formation in the lungs, spleen, and liver induced by TDM and inhibited the production of interleukin‐1 (IL‐1) and chemotactic factor, which is reported to precede granuloma formation.
Production of Murine Collagen‐Induced Arthritis Using Klebsiella pneumoniae O3 Lipopolysaccharide as a Potent Immunological Adjuvant
Authors: Takahashi Kazuko , Kato Yutaka , Sugiyama Tsuyoshi , Koide Naoki , Kawai Makoto , Fukada Masako , Yoshida Tomoaki , Yokochi Takashi 
Published: November 2013

AbstractCollagen‐induced arthritis (CIA) was produced in mice with non H‐2q and H‐2r haplotypes by repeated immunization of porcine type‐II collagen (CII) together with Klebsiella O3 lipopolysaccharide (KO3 LPS) as an immunological adjuvant. Histological changes that appeared in joints of repeatedly immunized mice were characterized by destruction of normal joint structure, synovial hyperplasia with proliferation of synovial cells, and infiltration of inflammatory cells. No such lesions were produced in mice receiving repeated injections of CII alone or KO3 LPS alone. Development of the humoral antibody and the delayed‐type hypersensitivity to CII was exclusively found in mice immunized with the mixture of CII and KO3 LPS. It was therefore suggested that arthritis lesions induced by repeated immunization with the mixture of CII and KO3 LPS might be caused by an autoimmune mechanism, and that the experimental model might be useful for characterization of human rheumatoid arthritis (RA).
Chitin Synthase 2 Gene Sequence of Malassezia Species
Authors: Kano Rui , Aizawa Tomohiro , Nakamura Yuka , Watanabe Shinichi , Hasegawa Atsuhiko 
Published: November 2013

AbstractNucleotide sequences of the chitin synthase 2 (CHS2) gene of seven species, Malassezia furfur, M. globosa, M. obtusa, M. pachydermatis, M. restricta, M. slooffiae and M. sympodialis, were analyzed for their phylogenetic relationship. About 620‐bp genomic DNA fragments of the CHS2 gene were amplified from these Malassezia species by polymerase chain reaction (PCR) and sequenced. The CHS2 nucleotide sequences of these Malassezia species showed more than 95% similarity between the species. A phylogenetic analysis of the nucleotide sequences of CHS2 gene fragments of seven Malassezia species revealed that the species were genetically distinct from each other.
Development of Restriction Fragment‐Length Polymorphism Method to Differentiate Five Subtypes of Feline Immunodeficiency Virus
Authors: Kurosawa Kyoko , Ikeda Yasuhiro , Miyazawa Takayuki , Izumiya Yoshihiro , Nishimura Yorihiro , Nakamura Kazuya , Sato Eiji , Mikami Takeshi , Kai Chieko , Takahashi Eiji 
Published: November 2013

AbstractFeline immunodeficiency virus (FIV) isolates have been classified into five subtypes (A to E) based on the sequences of the env variable V3 to V5 region. In this study, we sequenced a partial gag region of 4 and 3 isolates belonging to subtypes C and E, respectively. Phylogenetic analysis revealed that the branching pattern based on the region was similar to that based on the env V3 to V5 region. Here, we propose a protocol to differentiate five subtypes by polymerase chain reaction amplifying 329 bp within the region followed by restriction fragment‐length polymorphism analysis using four restriction enzymes.
Characterization of an Outer Membrane Protein Gene, pgmA, and Its Gene Product from Porphyromonas gingivalis
Authors: Hongo Hirohisa , Osano Etsuo , Ozeki Masami , Onoe Takaya , Watanabe Koji , Honda Okahito , Tani Hiroshi , Nakamura Hiroshi , Yoshimura Fuminobu 
Published: November 2013

AbstractA gene upstream from fimA, the gene encoding fimbrilin, on the chromosome of Porphyromonas gingivalis was sequenced and shown to be the gene encoding an outer membrane protein in this organism based on homology and biochemical analyses. Therefore, the gene (formerly ORF5) was designated pgmA, the P. gingivalis outer membrane protein A gene. The gene product, PgmA, was sensitive to protease, and was detected as a 60‐kDa protein from wild‐type strains with trichloroacetic acid treatment, which was carried out to destroy intrinsic proteases, and from protease‐deficient mutants without this treatment prior to electrophoresis. PgmA was indeed present in the membrane fraction. Its nature was determined to be that of outer membrane proteins in gram‐negative bacteria based on attempts at differential extraction of inner membrane proteins with detergents. No evidence has been found thus far from functional analyses that this protein is related to fimbrial morphogenesis and functions or to serum resistance of this organism.
A Clostridium perfringens hem Gene Cluster Contains a cysGB Homologue That Is Involved in Cobalamin Biosynthesis
Authors: Koyama Michio , Katayama Seiichi , Kaji Masato , Taniguchi Yuki , Matsushita Osamu , Minami Junzaburo , Morita Shushi , Okabe Akinobu 
Published: November 2013

AbstractThe hem gene cluster, which consists of hemA, cysGB, hemC, hemD, hemB, and hemL genes, and encodes enzymes involved in the biosynthetic pathway from glutamyl‐tRNA to uroporphyrinogen III, has been identified by the cloning and sequencing of two overlapping DNA fragments from Clostridium perfringens NCTC8237. The deduced amino acid sequence of the N‐terminal region of C. perfringens HemD is homologous to those reported for the C‐terminal region of Salmonella typhimurium CysG and Clostridium josui HemD. C, perfringens CysGB is a predicted 220‐residue protein which shows homology to the N‐terminal region of S. typhimurium CysG. Disruption of the cysGB gene in C. perfringens strain 13 by homologous recombination reduced cobalamin (vitamin B12) levels by a factor of 200. When grown in vitamin B12‐deficient medium, the mutant strain showed a four‐fold increase in its doubling time compared with that of the wild‐type strain, and this effect was counteracted by supplementing the medium with vitamin B12. These results suggest that C. perfringens CysGB is involved in the chelation of cobalt to precorrin II as suggested for the CysGB domain of S. typhimurium CysG, enabling the synthesis of cobalamin.
Decreased Prevalence of Orientia tsutsugamushi in Trombiculid Mites and Wild Rodents in the Primorye Region, Far East Russia
Authors: Urakami Hiroshi , Tamura Akira , Tarasevich Irina V. , Kadosaka Teruki , Shubin Felix N. 
Published: November 2013

AbstractThe isolation of Orientia tsutsugamushi was attempted from 249 rodents and approximately 14,000 trombiculid mites captured in the Primorye region, Far East Russia in 1993 and 1994, where high infection rates were recorded in both rodents and mites in the 1960s. However, no rickettsia was isolated from the samples. Low antibody titers against O. tsutsugamushi were detected in 7.1% of the rodents. These results indicate that the prevalence of O. tsutsugamushi in the Primorye region has decreased considerably in the past 30 years.