bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enhancement of the Growth of Helicobacter pylori in Brucella Broth by Hydrogen Peroxide
Authors: Murano Akiyuki , Miyake Masami , Kato Jiro , Tanzawa Hideki , Takeo Kanji , Noda Masatoshi 
Published: November 2013

AbstractWe found that a sub‐lethal concentration of hydrogen peroxide (HPOx) enhanced the growth of Helicobacter pylori in Brucella broth supplemented with 10% fetal bovine serum (BB/FBS). The enhancement was evident at 0.1 mM HPOx and reached a maximun at 3.5 mM. The growth stimulation was dependent on the basal media used; when brain heart infusion broth (BHIB) was used instead of BB, the growth was not altered regardless of the presence or absence of HPOx. Furthermore, the growth in BHIB/FBS was comparable to that in BB/FBS plus 3.5 mM HPOx. This suggested that the enhancement of growth by HPOx resulted from the derepression of the inhibitory factor existing in BB by HPOx. The inhibitory substance seemed to be bisulfite salt since the bacteria grew to a similar extent in bisulfite‐less Brucella broth (BLBB)/FBS compared to the bacterial growth in BHIB/FBS and BB/FBS plus HPOx. These results indicate that the detoxification of bisulfite in BB can be easily achieved by simply adding HPOx to the medium, which causes the oxidation of bisulfite to bisulfate, a less‐toxic compound to the bacterial growth. Since we also found that the morphology and cellular protein profile of BB/FBS‐cultured bacteria were apparently different from those cultured in BLBB/FBS, we propose that the use of BB for primary isolation and cultivation of H. pylori should be limited on certain occasions, or if necessary, BB can be used after detoxification of the bisulfite by the addition of a low concentration of HPOx.
Genetic Variation in the Capsid Region of Human Astrovirus Serotype 4 Isolated in Japan
Authors: Hachiya Masahiko , Matsui Mihoko , Sanogo Mussa , Oseto Mitsuaki , Morooka Keiichi , Ushijima Hiroshi 
Published: November 2013

AbstractThe entire capsid regions of 12 serotype‐4 astroviruses from Japan were sequenced and compared with those of other serotypes. Serotype‐4 isolates were divided into two new subgroups. The intrasubgroup nucleic acid and deduced amino acid sequences were quite homologous (more than 93%), but slightly less so between subgroups (almost 85%). However, the serotype‐4 sequences differed from those of serotypes 1,2,3,5,6,7 and 8 (less than 50%). Determining whether these differences significantly alter the epidemiology and antigenicity will require further investigation.
Intracellular Salmonella typhimurium Induce Lysis of Human Polymorphonuclear Leukocytes Which Is Not Associated with the Salmonella Virulence Plasmid
Authors: Chiu Cheng‐hsun , Ou Jonathan T. 
Published: November 2013

AbstractThe interaction between Salmonella typhimurium and human polymorphonuclear leukocytes (PMNs) was analyzed in vitro. Three S. typhimurium strains, the wild‐type strain OU5043, its isogenic virulence plasmid‐cured strain OU5048, and LT2, which represented the types that exhibited three mouse virulence levels, respectively, were used in this study. There was no correlation between the recovery of intracellular S. typhimurium from PMNs and the presence or absence of the virulence plasmid, or the strains' mouse virulence level. When the oxygen‐dependent response of PMNs upon phagocytosis of S. typhimurium was examined by checking the intracellular reduction of nitroblue tetrazolium (NBT), the fraction of PMNs that reduced NBT on phagocytosis of the three strains was around 80%, whereas it was 58% with Escherichia coli, 95% with phorbol 12‐myristate 13‐acetate and 15% with a negative control. Thus there were no significant differences among the three Salmonella strains in terms of their ability to induce the oxidative response in PMNs. Microscopic analysis of Salmonella‐infected PMNs indicated that the intracellular Salmonella induced lysis of PMNs. Both OU5043 and OU5048 exhibited a significant intracellular cytotoxic effect on PMNs after 24 hr of infection and this effect was not associated with the presence or absence of the virulence plasmid. On the other hand, lysis of PMNs was related to the intracellular survival of Salmonella, as ofloxacin, an antibiotic, appeared to be able to protect human PMNs from Salmonella‐induced cytotoxicity when this agent was added into the medium to inactivate the intracellular organism. The ability to induce lysis of PMNs by either wild‐type or plasmid‐cured strains of S. typhimurium may play a crucial role in the pathogenesis of non‐typhoid Salmonella. The contribution of pSTV to humam salmonellosis is likely to be limited. Furthermore, early institution of antibiotics with a high intracellular activity against Salmonella, such as fluoroquinolones, may be useful to prevent the dissemination of Salmonella infection.
Secretion of Hemolysin of Aeromonas sobria as Protoxin and Contribution of the Propeptide Region Removed from the Protoxin to the Proteolytic Stability of the Toxin
Authors: Nomura Tomohiko , Fujii Yoshio , Okamoto Keinosuke 
Published: November 2013

AbstractThe sequence at the amino terminus region of the hemolysin of Aeromonas sobria is homologous with that of aerolysin of A. hydrophila. However, there is no homology between the two toxins in the sequence at the carboxy terminal region. It has been shown that aerolysin is secreted into culture supernatant as a protoxin. This proaerolysin is activated by the proteolytic removal of a carboxy terminal peptide. However, the role of the carboxy terminal region, which is removed in the activation process, has not been elucidated. In this study, we showed that hemolysin is also secreted as a protoxin into culture supernatant and that prohemolysin is cleaved by the protease of A. sobria between Ser‐446 and Ala‐447, resulting in the removal of a 42 amino acid peptide. The removal of the peptide converts the prohemolysin into active hemolysin. Subsequently, we mutated the hemolysin gene to delete the last several amino acid residues and expressed the genes in Escherichia coli, in order to examine the role of the carboxy terminal region of prohemolysin. The amounts of these mutant hemolysins accumulated in the periplasmic space of E. coli were very low compared with that of the wild‐type. This observation indicated that the carboxy terminal region of prohemolysin contributes to the proteolytic stability of the toxin.
Suppression by the DNA Fragment of the motX Promoter Region on Long Flagellar Mutants of Vibrio alginolyticus
Authors: Furuno Masaaki , Nishioka Noriko , Kawagishi Ikuro , Homma Michio 
Published: November 2013

AbstractThe axial length of the polar flagellum (Pof) of Vibrio alginolyticus is about 5 μm. We previously isolated mutants that make abnormally long flagella. The swarm sizes of these mutants in a soft agar plate are smaller than that of a wild‐type strain. We cloned a DNA fragment into the pMF209 plasmid that restored the swarming ability of the long‐Pof strain VIO578. The swimming speed and flagellar length of these transformants were almost equal to the wild‐type values. The amounts of PF47 flagellin and PF60 sheath‐associated protein, which increased in the long‐Pof mutants, were retrieved to almost the wild‐type level in the transformants. The plasmid pMF209 contained only a 143 bp chromosomal fragment whose sequence is about 80% similar to that of the motX promoter region of V. parahaemolyticus. We speculate that this sequence interacts with a regulatory protein that controls Pof expression. The mutation causing the long‐Pof phenotype may be in the gene encoding this protein or in the control region of a structural gene that is regulated by this protein.
Evaluation of an Enzyme‐Linked Immunosorbent Assay Based on Binding Inhibition for Type‐Specific Quantification of Poliovirus Neutralization‐Relevant Antibodies
Authors: Hashido Madoka , Horie Hitoshi , Abe Shinobu , Doi Yutaka , Hashizume So , Agboatwalla Mubina , Isomura Shin , Nishio Osamu , Hagiwara Akio , Inouye Sakae 
Published: November 2013

AbstractTo detect neutralization‐relevant antibodies against 3 types of poliovirus (PV) without using tissue cultures and live viruses, an enzyme‐linked immunosorbent assay (ELISA) based on monoclonal antibody‐binding inhibition was evaluated using sera from 80 vaccinated Japanese children and 60 Pakistani poliomyelitis patients. Compared with the neutralization test, the sensitivity of the inhibition ELISA was 100% (111/111) for detection of anti‐PV1 antibody, 98.3% (118/120) for anti‐PV2, and 96.5% (82/85) for anti‐PV3, and the specificity was 93.1% (27/29), 100% (20/20), and 92.7% (51/55), respectively. Thus, the inhibition ELISA showed excellent potential as a seroepidemiologic tool in both vaccinated and naturally‐infected populations.
Chromosome‐Determined Zinc‐Responsible Operon czr in Staphylococcus aureus Strain 912
Authors: Kuroda Makoto , Hayashi Hideo , Ohta Toshiko 
Published: November 2013

AbstractA novel operon, czrAB (zinc‐responsible genes), was identified in the chromosome of Staphylococcus aureus. The operon consists of two genes, czrA and czrB. The czrA gene, coding for an 11.5 kDa protein, was homologous to cadC, arsR of S. aureus plasmid pI258 and smtB of Synechococcus PCC7942. The czrB, coding for a 36 kDa membrane spanning protein, was homologous to the czcD gene, cobalt, zinc and the cadmium‐resistant factor of Bacillus subtilis and Alcaligenes eutrophus. In the presence of zinc (0.1‐10 mM), the transcription of czrAB was enhanced in a concentration‐dependent manner. Other heavy metals, such as cobalt, copper, manganese and nickel showed no effect on czrAB expression. The disruptant of the czrB gene became sensitive to zinc ion (MIC, 2 mM; MBC, 10 mM), and the complementation with the plasmid recovered the resistance to zinc at the same concentration as a parental strain (MIC, 5 mM; MBC, 20 mM). The disruptant accumulated intracellular zinc up to 0.4 mg per g dry weight of the organism, while that of the parental strain was 0.25 mg per g dry weight. The findings indicated that the novel operon czrAB should play a role in the transportation of zinc across the cell membrane to maintain the proper intracellular concentration.
Anti‐HIV‐1 Activity of an lonically Modified Porous Polypropylene Membrane Determined by Filtration of a Viral Suspension
Authors: Owada Takashi , Miyashita Yuko , Motomura Tadahiro , Onishi Makoto , Yamashita Shuzo , Yamamoto Naoki 
Published: November 2013

AbstractWe describe here a unique anti‐HIV‐1 membrane, derived from a chemically modified porous polypropylene (PP) membrane, which lowers viral infectivity upon the filtration of HIV‐1 suspension. A cationic polymer, polyethyleneimine (PEI) was graft‐polymerized onto the PP filter membrane (PP‐PEI), and infectious HIV‐1HTLV‐IIIB derived from MOLT‐4/HIV‐1HTLV‐IIIB cells (HIV‐1HTLV‐IIIB(MOLT‐4)) was applied. When a viral suspension of high titer (103.93 TCID50 ml 1) was filtered, efficient reduction (>99%) of gag p24 antigen levels and infectious titer resulted. In a viral suspension of medium titer (102.37 TCID50 ml 1), a significant decrease in the p24 antigen did not occur, although the titer was markedly reduced (>95%). Electron microscopic observation suggested that PEI induced viral aggregations under high titer conditions, and under medium titer conditions, PEI deprived HIV‐1HTLV‐IIIB of its infectivity alone to avoid virus adsorption. In contrast, HIV‐1 propagated in human peripheral blood mononuclear cells (PBMC) such as HIV‐1HTLV‐IIIB(PBMC) was more efficiently trapped by PP‐PEI at lower titers as compared with HIV‐1HTLV‐IIIB(MOLT‐4) from MOLT‐4/HIV‐1HTLV‐IIIB cells. These data suggest host cell modification in the interactions between PP‐PEI and HIV‐1 strains. Since HIV‐1HTLV‐IIIB(MOLT‐4) and HIV‐1HTLV‐IIIB(PBMC) were almost electrically neutral and negative, respectively, we concluded that the divergent effect of PEI on each HIV‐1HTLV‐IIIB resulted from their different electrical characteristics.
Functional Characteristics of Human Peripheral Blood α/βTCR+, CD4‐ and CD8‐Double‐Negative (DN) T Cells
Authors: Niehues Tim , Eichelbauer Dirk , Schneider E. Marion 
Published: November 2013

AbstractThe function of human peripheral blood α/βTCR positive, CD4‐ and CD8‐ double‐negative T lymphocytes (DN cells) in vivo is not completely understood. The response of immunomagnetically isolated DN cells to PHA and anti‐CD3 was compared to the response of single‐positive (SP) CD4 and CD8 subsets. Proliferation of DN cells in response to PHA was largely independent of APC. This suggests activation requirements for DN cells that are different from SP cells. Upon activation, HLA‐DR was found to be upregulated early on DN cells, and IL‐4 and IL‐10 were detected in the supernatants of DN cells. These observations in vitro could correspond with an immunoregulatory role of human DN cells in vivo.
Changes in Prevalence of Herpes Simplex Virus Type 1 and 2 Antibodies from 1973 to 1993 in the Rural Districts of Japan
Authors: Hashido Madoka , Kawana Takashi , Matsunaga Yasuko , Inouye Sakae 
Published: November 2013

AbstractUsing the gG‐capture ELISA, changes in the seroprevalence of HSV‐1 and HSV‐2 from 1973 to 1993 were studied for 614 sera collected from general adults living in rural Japan. The HSV‐1 seroprevalence for men and women decreased from 75.3 and 80.6% in 1973 to 54.4 and 59.6%, respectively, in 1993. The HSV‐2 seroprevalence also decreased from 10.2 and 9.9% in 1973 to 1.8 and 1.2%, respectively, in 1993. Although the decrease in HSV‐2 prevalence seemed to be correlated with the general decrease of sexually transmitted diseases in Japan since the 1950s, these findings should not be interpreted as typical, as HSV‐2 infections are particularly known to distribute unevenly among populations, according to sexual activity and cohorts.
Growth of Starved Escherichia coli O157 Cells in Selective and Non‐Selective Media
Authors: Sata Shin , Osawa Ro , Asai Yoshio , Yamai Shiro 
Published: November 2013

AbstractEscherichia coli O157 strains starved in sterile deionized water (SDW) and filter‐sterilized natural river water (SRW) were investigated with specific reference to their culturability in selective and non‐selective media. Growth of the strains starved in both SDW and SRW were markedly suppressed with time in selective liquid media such as modified trypticase soy broth supplemented with novobiocin (mTSB+n) and modified E. coli broth supplemented with novobiocin (mEC+n). This suppression was more pronounced when incubated at 42 C than at 37 C, especially with mEC+n. By contrast, such growth suppression was seldom observed when cultured at 37 C in non‐selective liquid media such as trypticase soy broth (TSB) and buffered peptone water. In mEC+n at 42 C, the non‐starved cells from overnight cultures with an initial density of less than 103 colony‐forming units (CFU)/ml grew to the density of over 107 CFU/ml after 24 hr incubation, whereas those starved for 6 weeks in SRW were only to maintain their initial density or died off after 24 hr incubation under the same culturing conditions. These results indicated that the isolation of starved cells of E. coli O157 from water samples would be most difficult with selective enrichment or direct plating on the selective plate media. It is thus highly recommended that a “resuscitation” of the cells with non‐selective enrichment should be performed as a routine practice for maximum recovery of E. coli O157 from water systems.
Genotypic Characterization of Human and Environmental Isolates of Salmonella choleraesuis Subspecies choleraesuis Serovar Infantis by Pulsed‐Field Gel Electrophoresis
Authors: Murakami Koichi , Horikawa Kazumi , Otsuki Koichi 
Published: November 2013

AbstractTo determine the extent of genetic diversity of Salmonella choleraesuis subspecies choleraesuis serovar Infantis and whether environmental isolates were similar or identical to human isolates, a total of 110 isolates from humans, broiler samples, egg production facilities, riverwater, sewage, and chicken meat were analyzed epidemiologically by pulsed‐field gel electrophoresis. While the isolates showed 35 distinct pulsed‐field profiles, none had the genotype of the human isolates. One pulsed‐field profile was shared by 43 (39%) of the 110 isolates. These results indicate that relatively fewer clonal lines of S. serovar Infantis had spread widely while multiple clonal lines, including the strain involved in the outbreak, exist in Western Japan.
Proposal of Sphingomonas suberifaciens (van Bruggen, Jochimsen and Brown 1990) Comb. Nov., Sphingomonas natatoria (Sly 1985) Comb. Nov., Sphingomonas ursincola (Yurkov et al. 1997) Comb. Nov., and Emendation of the Genus Sphingomonas
Authors: Yabuuchi Eiko , Kosako Yoshimasa , Naka Takashi , Suzuki Shinnichiro , Yano Ikuya 
Published: November 2013

AbstractBased on the results of a phylogenetic analysis of 16S rRNA and the presence of sphingoglycolipid in cellular lipids of the type strains, transfer of “Rhizomonas” suberifaciens, Blastomonas natatoria and Erythromonas ursincola to the genus Sphingomonas as Sphingomonas suberifaciens (van Bruggen et al 1990) comb, nov., Sphingomonas natatoria (Sly 1985) comb, nov., and Sphingomonas ursincola (Yurkov et al 1997) comb. nov. are herein proposed together with the emendation of genus Sphingomonas, The type strain of S. suberifaciens is van Bruggen Ca1=ATCC 49382=NCPPB 3629=IFO 15211=JCM 8521, that of S. natatoria is ATCC 35951=DSM 3183=NCIMB 12085=JCM10396, and that of S. ursincola is DSM 9006=KR‐99.
Detection and Nucleotide Sequence Analysis of Human Caliciviruses (HuCVs) from Samples in Non‐Bacterial Gastroenteritis Outbreaks in Hokkaido, Japan
Authors: Ohyama Tohru , Yoshizumi Shima , Sawada Harumi , Uchiyama Yasuhiro , Katoh Yoshinobu , Hamaoka Naohiro , Utagawa Etsuko 
Published: November 2013

AbstractSamples of feces and vomit collected from patients during 13 non‐bacterial gastroenteritis outbreaks which occurred in Hokkaido between 1995 and 1998 were examined by electron microscopy (EM) and reverse‐transcription polymerase chain reaction (RT‐PCR) for evidence of infection with human caliciviruses (HuCVs). In 6 food‐borne outbreaks, oysters were the probable source of infection, while the origin of HuCVs was not found out for the other 7 outbreaks. One‐hundred‐eleven of 214 stool, vomit and oyster specimens examined gave positive results by RT‐PCR, while HuCVs were detected by EM in 36 of 121 stool specimens examined. We determined the nucleotide sequences of 470‐bp or 373‐bp PCR products amplified from the RNA polymerase region of the HuCV genomes with primer sets MR3/4 and Yuri22F/R, respectively. The sequences of different strains revealed great heterogenicity, with a range of 60 to 100% homology among strains. In a few cases, a mixed genotype was found in the same patient or same outbreak by means of nested PCR and cloning of PCR products into an appropriate vector. Of the 19 different strains found, 4 strains could be classified as Norwalk virus (genogroup 1) and the other 15 strains as Snow Mountain agent (genogroup 2) based on genotyping with homology analysis. Furthermore, the strains belonging to genogroup 2 could be classified into 4 subgroups with more than 93% homology in amino acids among strains in the subgroup.
Borna Disease Virus Infection in Two Family Clusters of Patients with Chronic Fatigue Syndrome
Authors: Nakaya Takaaki , Takahashi Hirokazu , Nakamura Yurie , Kuratsune Hirohiko , Kitani Teruo , Machii Takashi , Yamanishi Koichi , Ikuta Kazuyoshi 
Published: November 2013

AbstractA high rate of Borna disease virus (BDV) infection has been demonstrated in patients with chronic fatigue syndrome (CFS). Herein, we focused on BDV infection in two family clusters of patients with CFS: a father, mother, two sons and one daughter (family #1); and a father, mother, two daughters and one son (family #2). All members, except for the elder son in family #1 and the father and son in family #2, were diagnosed with CFS. The results supported that all the family members with CFS were infected with BDV, as evidenced by the presence of antibodies to viral p40, p24 and/or gp18 and BDV p24 RNA in peripheral blood mononuclear cells. The healthy members, except for the father of family #2 who was positive for antibody to p24, were all negative by both assays. Follow‐up studies in family #1 continued to reveal BDV antibodies and BDV RNA, except in the mother, who lost the RNA upon slight recovery from the disease.
Salt‐Sensitive Growth of Staphylococcus aureus: Stimulation of Salt‐Induced Autolysis by Multiple Environmental Factors
Authors: Ochiai Toshiro 
Published: November 2013

AbstractThe growth of Staphylococcus aureus 209P became extremely sensitive to a high NaCl concentration following lowered temperature, reduced air‐supply, and decreased Ca2+ concentration in the medium. Cells in high‐NaCl and low‐Ca2 concentration media either autolyzed or transformed into protoplast‐like forms during growth when grown standing below 37 C. The abnormal growth, however, was invariably avoided by preliminary supplementation with polyanetholesulfonate (autolysin inhibitor) in the growth media. These results suggested that the autolytic activity of this organism was precisely controlled by multiple environmental factors such as ionic strength, temperature, air supply, and the concentration of Ca2+.
Characterization of the Coxiella burnetii sucB Gene Encoding an Immunogenic Dihydrolipoamide Succinyltransferase
Authors: Van Nguyen Sa , To Ho , Yamaguchi Tsuyoshi , Fukushi Hideto , Hirai Katsuya 
Published: November 2013

AbstractThe Coxiella burnetii sucB gene encoding the dihydrolipoamide succinyltransferase (E2o) enzyme was cloned by immunological screening of a λ EMBL3 genomic library prepared from strain Nine Mile DNA and sequenced. The homology of the cloned gene product to the counterpart in Escherichia coli was 54.3%, but the homology of the N‐terminal region was only 42%. The gene was expressed in E. coli as an independent unit from its own promoter, producing an immunoreactive protein of about 50 kDa on SDS‐PAGE which reacted with antisera from laboratory animals and sera from human patients with acute Q fever. The study results suggest that the C. burnetii E2o enzyme may serve as a potential target antigen for diagnostic assays for Q fever.
Survey on Antibody against Parapoxvirus among Cattle in Japan
Authors: Sentsui Hiroshi , Inoshima Yasuo , Minami Akihiro , Yamamoto Yasunori , Murakami Kenji , Shimizu Shinya 
Published: November 2013

AbstractA seroepidemiological survey was performed on antibody against parapoxvirus among cattle in Japan using the agar gel immunodiffusion test and enzyme‐linked immunosorbent assay. A total 1,819 sera were collected from cattle in various parts of Japan for the survey. The positive rates were in the range of 40 to 98%, and the reactors increased gradually in number with advancing age. These results indicate that parapoxvirus infection is already prevalent among cattle in Japan. It remains to be elucidated, however, whether the antibodies detected have been produced by infection of the same virus or other viruses that belong to the genus parapoxvirus.
Isolation of Orientia tsutsugamushi from Leptotrombidium fuji and Its Characterization
Authors: Tamura Akira , Makisaka Yukie , Kadosaka Teruki , Enatsu Teruyuki , Okubo Koji , Koyama Shoichiro , Yu Qiang , Urakami Hiroshi 
Published: November 2013

AbstractIn our attempts to isolate Orientia tsutsugamushi from trombiculid mites, a strain was successfully isolated from Leptotrombidium fuji collected in Aichi Prefecture, Japan. This is the first case of isolation of O. tsutsugamushi from L. fuji. A phylogenetic analysis based on the base‐sequence homology of the 56‐kDa type‐specific antigen‐gene indicated that the strain is a new type which is not closely related to any strains analyzed previously. Three strains isolated from Leptotrombidium pallidum harvested at the same area were identified as being closely related to the JP‐2 type (subtype‐2 of Karp type in Japan) by phylogenetic analysis.