bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of Vibrio cholerae O139 Bengal Isolated from Different Geographical Areas Using Macrorestriction DNA Analysis
Authors: Kurazono Hisao , Yamasaki Shinji , Ratchtrachenchai Orn‐anong , Nair G. Balakrish , Takeda Yoshifumi 
Published: November 2013

AbstractVibrio cholerae O139 isolated from different countries, as well as from different locations within a country, were examined using macrorestriction DNA analysis to determine the clonality of the O139 strains. NotI digests of genomic DNA of representative strains from Nepal, India, Bangladesh, China, Thailand, and Malaysia revealed very similar but not identical patterns. Examinations of the banding patterns generated by pulsed‐field gel electrophoresis of strains isolated within countries revealed complete homogeneity. These results further reiterate the spread of an identical clone of V. cholerae O139 although it appears that genetic polymorphism among the O139 strains is becoming apparent.
Cryptic Appearance of a New Clone of Vibrio cholerae Serogroup O1 Biotype El Tor in Calcutta, India
Authors: Yamasaki Shinji , Nair G. Balakrish , Bhattacharya S.K. , Yamamoto Shingo , Kurazono Hisao , Takeda Yoshifumi 
Published: November 2013

AbstractIn this study, pulsed‐field gel electrophoresis (PFGE) was applied to determine if the Vibrio cholerae O1 strains which reappeared after being temporarily displaced in Calcutta by the O139 serogroup were different from those isolated before the advent of the O139 serogroup. NotI digestion generated a total of 11 different patterns among the 24 strains of V. cholerae randomly selected to represent different time frames. Among the V. cholerae O1 strains isolated after July 1993, 4 PFGE banding patterns designated as H through K were observed with pattern H dominating. Pattern H was distinctly different from all other patterns encountered in this study including patterns A, B and C of V. cholerae O1 El Tor, which dominated before November 1992, and pattern F, which was the dominant V. cholerae O139 pattern. Further, pattern H was also different from the NotI banding patterns of the representative strains of the 4 toxigenic clonal groups of V. cholerae O1 El Tor currently prevailing in different parts of the world. NotI fragments of the new clone of V. cholerae O1 did not hybridize with an O139 specific DNA probe, indicating that there was no O139 genetic material in the new clone of V. cholerae O1. Hybridization data with an O1‐specific DNA probe again differentiated between the clones of V. cholerae O1 existing before the genesis of the O139 serogroup and the O1 strains currently prevalent.
Location and Ultrastructure of Borrelia japonica in Naturally Infected Ixodes ovatus and Small Mammals
Authors: Yano Yasuhiro , Takada Nobuhiro , Ishiguro Fubito 
Published: November 2013

AbstractThe internal organs of Ixodes ovatus and the ears of wild rodents (Apodemus speciosus, Eothenomys smithii) and an insectivore (Crocidura dsinezumi) were cultured to isolate borreliae; positive samples were examined for the distribution and dissemination of spirochetes in the host tissues using electron microscopy. Seven isolates were derived from the unfed ticks and the three species of mammals. These isolates were identified as Borrelia japonica judging from the outer surface protein profile using sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and reactivity to a B. japonica‐specific monoclonal antibody. Borreliae were found only in the midgut lumen of the tick in close contact with the microvilli on the midgut epithelium; on the other hand, borreliae found in the ears of the mammals existed freely in the collagenous intercellular substances of connective tissues or in close contact with fibrocytes. The ultrastructural disparities between the borreliae in ticks and mammals appeared to correspond to differences in motility. Interestingly, the borrelia which invaded through the perineurium appeared to contact the basement membrane of a Schwann cell that enclosed several nonmyelinated nerve fibers. This may offer important information regarding the involvement of the nervous system in Lyme disease.
Optical Configuration Analysis of Hydroxy Fatty Acids in Bacterial Lipids by Chiral Column High‐Performance Liquid Chromatography
Authors: Nakagawa Yoji , Kishida Kenichi , Kodani Yoshifumi , Matsuyama Tohey 
Published: November 2013

AbstractThrough the adoption of a chiral stationary phase in high‐performance liquid chromatography and a simple derivatization method for hydroxy fatty acids, it became easy to separate and identify the optical isomers of 2‐ and 3‐hydroxy fatty acids composing several kinds of microbial lipids. The 2‐ and 3‐hydroxy fatty acids were converted with dinitrophenyl isocyanate to their 3, 5‐dinitrophenyl urethane derivatives (DU‐derivatives), which were analyzable by HPLC using a chiral column. By varying the composition of an eluent, separation of the DU‐derivatives of hydroxy fatty acids differing in optical configuration, chain length and position of hydroxyl group was achieved. The general elution orders of these DU‐derivatives were determined with authentic 2‐ and 3‐hydroxy fatty acids. Small amounts (~300 μg) of ornithine‐containing lipids isolated from the Serratia marcescens strains were examined by this method to identify 3‐hydroxy fatty acids of the lipids as D isomers.
Alterations in the Expression of ELAM‐1, ICAM‐1 and VCAM‐1 after In Vitro Infection of Endothelial Cells with a Clinical Isolate of Human Cytomegalovirus
Authors: Shahgasempour Shapour , Woodroffe Salwa B. , Garnett Helen M. 
Published: November 2013

AbstractHuman cytomegalovirus (HCMV) infection of endothelial cells resulted in increased adhesion of the cells to peripheral blood leukocytes. It was demonstrated by flow cytometry that increased adhesiveness parallels the increased expression of cell surface adhesion molecules (ELAM‐1, ICAM‐1, VCAM‐1). The increased adhesion of PMN and T‐lymphocytes was due to upregulation in the expression of ELAM‐1 and ICAM‐1. The upregulation of VCAM‐1 resulted in the increased adhesiveness of monocytes and T‐lymphocytes to HCMV‐infected HUVEC. The increased adhesiveness to leukocytes was caused by HCMV replication since endothelial cells exposed to HCMV‐free supernatants and UV‐inactivated HCMV did not show any increase in adhesiveness to any of the leukocytes tested.
Analysis of the Structural Gene Encoding a Hemolysin in Vibrio mimicus
Authors: Rahman Md. Monzur , Miyoshi Shih‐Ichi , Tomochika Ken‐Ichi , Wakae Hitoshi , Shinoda Sumio 
Published: November 2013

AbstractAn environmental isolate of V. mimicus, strain E‐33, has been reported to produce and secrete a hemolysin of 63 kDa. The hemolysin is enterotoxic in test animals. The nucleotide sequence of the structural gene of the hemolysin was determined. We found a 2,232 bp open reading frame, which codes a peptide of 744 amino acids, with a calculated molecular weight of 83,903 Da. The sequence for the structural gene was closely related to the V. cholerae el tor hlyA gene, coding an exocellular hemolysin. The amino terminal amino‐acid sequence of the 63 kDa hemolysin, purified from V. mimicus, was determined by the Edman degradation method and found to be NH2‐S‐V‐S‐A‐N‐N‐V‐T‐N‐N‐N‐E‐T. This sequence is identical from S‐152 to T‐164 predicted from the nucleotide sequence. So, it seems that the mature hemolysin in V. mimicus is processed upon deleting the first 151 amino acids, and the molecular mass is 65,972 Da. Analyzing the deduced amino‐acid sequence, we also found a potential signal sequence of 24 amino acids at the amino terminal. Our results suggest that, like V. cholerae hemolysin, two‐step processing also exists in V. mimicus hemolysin.
Domain‐Specific Rearrangement between the Two Arg‐Gingipain‐Encoding Genes in Porphyromonas gingivalis: Possible Involvement of Nonreciprocal Recombination
Authors: Nakayama Koji 
Published: November 2013

AbstractPorphyromonas gingivalis has two functional genes (rgpA and rgpB) encoding an arginine‐specific cysteine proteinase (Arg‐gingipain, RGP). Several RGP‐encoding genes have been cloned and sequenced from various P. gingivalis strains, but all of the genes seem to be essentially equivalent to rgpA. In this study, we cloned and sequenced the second rgp gene (rgpB). A comparison of the rgpB gene and the rgp1 gene, one of the rgpA‐equivalent genes, revealed that their gene structures were very similar to each other, except that the rgpB gene did not possess most of the hemagglutinin domain present in the C‐terminal region of the rgpl gene, and provided strong evidence for homologous recombination between the proteinase domain regions of the two rgp genes. The presence of nonreciprocal recombination in P. gingivalis was experimentally proven using suicide/integration plasmid systems. The results provide one of the hypothetical scenarios of the generation of the two rgp genes; that is, they have been generated through the duplication of an ancestor rgp gene, insertion of the hemagglutinin domain region into one copy of the two resulting rgp genes (or deletion of the region from one rgp) and homologous recombination between the proteinase domain regions of the two rgp genes.
Supernatant from Cultured Intestinal Cells Inhibits the Growth of Gram‐Negative Bacteria
Authors: Kops Sandra Kraeuter , Chao David , West A. Brian 
Published: November 2013

AbstractBacterial growth chambers of Transwell units bearing intestinal epithelial monolayers (C2BBe) was consistently observed to be stagnant during the course of transmigration studies with Salmonella typhi. This limitation could not be explained by varying the bacterial load in the inoculum. Conditioned media produced by cultured C2BBe cells would not support bacterial growth. Growth support of the media was restored by heating to 95 C for 10 min. C2BBe conditioned media had bacteriostatic activity for a large variety of gram‐negative, enteropathogenic bacteria but had no effect on gram‐positive bacteria.
Infection of an HIV‐1/SIVmac Chimeric Virus Having HIV‐1 env to Macaque Monkeys via the Vaginal Cavity
Authors: Ido Eiji , Okada Mineyuki , Enose Yoshimi , Kuwata Takeo , Igarashi Tatsuhiko , Sata Tetsutaro , Terao Keiji , Chen Jiangli , Hayami Masanori 
Published: November 2013

AbstractWe previously reported that an HIV‐1/SIVmac chimeric virus (designated as NM‐3rN) having HIV‐1 env efficiently infected macaque monkeys by intravenous inoculation. In this study, this chimeric virus was atraumatically inoculated into the vaginal cavity of two rhesus and one cynomolgus monkeys. Although antibody response and detection of proviral genome by PCR were observed in both rhesus monkeys, virus recovery was only once from PBMC in one of them. In the cynomolgus monkey, no virus was recovered and proviral DNA detection was rare. Thus, vaginal inoculation with NM‐3rN resulted in poor systemic infection, implying the presence of selective pressure while passing through mucosal membranes.
Presence of IgM Antibodies Which Sensitize HIV‐1‐Infected Cells to Cytolysis by Homologous Complement in Long‐Term Survivors of HIV Infection
Authors: Okada Noriko , Wu Xiaoshan , Okada Hidechika 
Published: November 2013

AbstractAlthough human cells are resistant to homologous human complement due to the presence of species‐specific membrane inhibitors, a naturally occurring IgM antibody which recognizes an asialo‐oligosaccharide can sensitize HIV‐1‐infected cells for complement‐mediated cytolysis. Therefore, we investigated whether long‐term survivors of HIV‐1 infection harbor such antibodies in their sera. Thirty of 31 sera from HIV‐1 seropositive hemophilia patients who have survived HIV‐1 infection 10 years or more showed appreciable cytolytic activity, while only 2 sera of 10 seropositive patients presumed to have been infected with HIV‐1 (due to sexual contact) more recently showed cytolytic activity. On the other hand, only 7 out of 43 sera from seronegative hemophilia patients showed cytolytic activity. Immunofluorescence staining for IgM on HIV‐L‐infected cells essentially correlated with the cytolytic capacity of the sera. Therefore, naturally occurring IgM antibodies and/or generated IgM antibodies reactive with the HIV‐L‐infected cells in patients might have been responsible for long‐term survival due to complement‐mediated immune cytolysis which may, in conjunction with cytotoxic T lymphocytes, synergistically suppress the infected cells in vivo. Therefore, the transfusion of such IgM antibodies could be effective for the treatment of HIV‐L‐infected individuals.
Cytokines Released by Human Peripheral Blood Mononuclear Cells Inhibit the Production of Early and Late Cytomegalovirus Proteins
Authors: Torigoe Sadayoshi , Campbell Donald E. , Starr Stuart E. 
Published: November 2013

AbstractCytomegalovirus‐infected human fibroblasts are susceptible to lysis by natural killer cells and cytotoxic T cells. The purpose of this study was to determine whether non‐lytic mechanisms might also contribute to the control of cytomegalovirus infection. The appearance of cytomegalovirus proteins in infected fibroblasts was determined by flow cytometry. Infected fibroblasts incubated with peripheral blood mononuclear cells for 3 days expressed less early and late proteins than fibroblasts incubated without peripheral blood mononuclear cells. Supernatants generated by the cocultivation of peripheral blood mononuclear cells with cytomegalovirus‐infected fibroblasts inhibited the production of cytomegalovirus early and late proteins. The soluble factors in supernatants which contributed to the inhibitory effect were identified as interferons α, β and γ, and tumor necrosis factors α and β. The ability of supernatants to inhibit the production of cytomegalovirus early protein was mimicked by combinations of corresponding recombinant cytokines. The inhibition of cytomegalovirus protein production by cytokines produced by peripheral blood mononuclear cells may contribute to early containment of cytomegalovirus infection.
Herpes Simplex Virus‐Specific IgM, IgA and IgG Subclass Antibody Responses in Primary and Nonprimary Genital Herpes Patients
Authors: Hashido Madoka , Kawana Takashi 
Published: November 2013

AbstractTo clarify the humoral immunity in herpes simplex virus (HSV) infection, HSV‐specific IgM, IgA and IgG subclass antibody responses were studied in patients with genital herpes: 17 primary, 13 recurrent and 6 nonprimary first episode. A total of 181 serum samples serially collected from the patients, 5 per patient until 213 days after the onset of disease (on average), were analyzed by an enzyme‐linked immunosorbent assay. IgGl, IgG3 and IgA were detected in all patients with primary and nonprimary infections, whereas IgG4 was detected in 74% of only those with nonprimary infections and IgG2 was detected in none. IgM was detected in 100% of the patients with primary infections, but also in 68% of those with nonprimary infections. IgA showed a peak similar to that of IgM in patients with primary infections. No significant difference was observed in the detection rate or pattern of antibody responses between the recurrent and nonprimary first episode infections, nor between the HSV‐1 and HSV‐2 infections. These findings may be useful to improve the diagnostic potential of HSV serology.
Isolation of a New Orientia tsutsugamushi Serotype
Authors: Seong Seung‐yong , Park Sae‐gwang , Kim Hang‐rae , Han Tae‐hee , Kang Jae‐seung , Choi Myung‐sik , Kim Ik‐sang , Chang Woo‐hyun 
Published: November 2013

AbstractOrientia tsutsugamushi, the etiological agent of scrub typhus, is an antigenically diverse organism and many serologically distinct strains have been identified. The 56 kDa protein of O. tsutsugamushi, a major protein in the outer membrane, has been thought to be responsible for this antigenic variability. A strain of O. tsutsugamushi isolated in Korea cross‐reacted with both Gilliam strain‐specific and Karp strain‐specific monoclonal antibodies. When its 56 kDa protein gene was cloned and analyzed, its sequence showed variation especially between 1,200 and 1,250 bp, showing that this isolate is a new O. tsutsugamushi strain.
Genetic Identification of Members of the Genus Corynebacterium at Genus and Species Levels with 16S rDNA‐Targeted Probes
Authors: Hou Xiao‐gang , Kawamura Yoshiaki , Sultana Ferdousi , Hirose Kenji , Miyake Masaki , Otsuka Yoshihito , Misawa Shigeki , Oguri Toyoko , Yamamoto Hiroyuki , Ezaki Takayuki 
Published: November 2013

Abstract16S rRNA gene‐targeted probes were designed for the identification of corynebacteria at the genus and species levels. The genus‐specific probe hybridized all clinically important members of the genus Corynebacterium and could distinguish them from other coryneform bacteria and phylogenetically related high G + C% gram‐positive bacteria, including Actinomyces, Rhodococcus, Gordona, Nocardia, Streptomyces, Brevibacterium and Mycobacterium. The species‐specific probes for C. jeikeium and C. diphtheriae could differentiate these two species from other members of this genus. The probes were used to select corynebacteria among gram‐positive clinical isolates which had been tentatively identified as corynebacteria by biochemical tests. We screened 59 strains with the genus‐specific probe; 51 strains hybridized to the genus‐specific probe, 8 did not. Of the 51 strains that hybridized to the genus‐specific probe, 1 hybridized to the C. diphtheriae species probe and 13 hybridized to the C. jeikeium species probe. The 8 strains that did not hybridize to the genus probe were further characterized by analyzing cell wall diaminopimelic acid and partial 16S rRNA sequencing. The results indicated that these strains were distributed in the genera Arthrobacter and Brevibacterium.
Amplification of a Full‐Length Borna Disease Virus (BDV) cDNA from Total RNA of Cells Persistently Infected with BDV
Authors: Shoya Yuko , Kobayashi Takeshi , Koda Toshiaki , Lai Patrick K. , Tanaka Hidetoshi , Koyama Tsukasa , Ikuta Kazuyoshi , Kakinuma Mitsuaki , Kishi Masahiko 
Published: November 2013

AbstractWe have developed a novel reverse transcriptase‐polymerase chain reaction (RT‐PCR) to amplify the full‐length 8.9 kilobase (kbp) cDNA of the Borna disease virus (BDV) RNA genome from the total cellular RNA of MDCK cells persistently infected with BDV (MDCK/BDV). Antigenomic BDV cDNA was reverse transcribed using a 53‐mer oligonucleotide primer, corresponding to the 5′‐terminus of a putative 3′‐leader sequence of the BDV RNA genome, for 2 hr at 42 C followed by 30 min at 55 C. PCR was performed in the presence of this 53‐mer antigenomic primer and a 25‐mer primer, corresponding to the 3′‐terminus of the BDV antigenomic cDNA, by use of an rTth DNA polymerase with proof‐reading activity. The amplified full‐length BDV cDNA was detected in as little as 20 ng of total cellular RNA of MDCK/BDV. This RT‐PCR method should be a useful technique to study the molecular quasispecies of BDV.
Cellular Immune Responses to Cell Wall Peptidoglycan Associated Protein Antigens in Tuberculosis Patients and Healthy Subjects
Authors: Dhiman Neelam , Verma Indu , Khuller Gopal Krishan 
Published: November 2013

AbstractWe have isolated cell wall peptidoglycan associated proteins (CW‐Pr) of Mycobacterium tuberculosis H37Ra by chemical treatment with trifluoromethanesulfonic acid:anisole (2:1), which further resolved into 71, 60 and 45 kDa proteins on SDS‐PAGE. A study was carried out to investigate the immunoreactivity of these proteins with blood samples from 4 categories, including 15 tuberculous patients (TB), 5 tuberculous patients on ATT (TBT), 10 PPD non‐reactive healthy controls (HPPD−) and 11 PPD reactive healthy controls (HPPD+). Comparing the proliferative responses to cell wall protein antigens, it was observed that the 71 kDa protein gave maximum stimulation with PBMCs from the TB and HPPD+ groups. The adherent PBMCs from the TB group also demonstrated enhanced phagocytosis, particularly in the presence of 71 and 45 kDa proteins, and the phagocytic index was significantly higher (P < 0.05) than the TBT group. However, PBMCs from of the groups recognized the 60 kDa cell wall antigen. Our results suggest that the 71 kDa protein from the cell wall of M. tuberculosis is highly immunogenic.
Sequence Analysis of the Influenza Virus Strain A/Shearwater/Australia/1/72 (H6N5)
Authors: Ward Alister C. , Harley Vincent R. 
Published: November 2013

AbstractAnalysis of the NS and M genes of the archetype H6N5 influenza virus strain A/shearwater/Australia/1/72 shows it to be a typical example of the avian host reservoir, containing Old World/Eurasian internal proteins with divergent surface proteins, which is a potential source of new pandemic strains.
Effects of Chronic Isoproterenol Treatment or Submandibular and Sublingual Ablation on Microflora of Mouse Tooth Surfaces
Authors: Maeda Nobuko , Okamoto Masaaki , Nakagawa Yoichi , Fujita Hiroshi , Humphreys‐Beher Michael G. 
Published: November 2013

AbstractBALB/cA mice were examined for the effects of chronic isoproterenol treatment or submandibular‐sublingual gland ablation on the natural patterns of oral bacterial colonization on tooth surfaces. Indigenous microflora on the tooth surfaces of BALB/cA mice was relatively simple. The predominant bacterial groups were Enterobacteriaceae (45.9%), enterococci (29.4%) and staphylococci (15.7%). Isoproterenol, which resulted in the induced synthesis of proline‐rich proteins, caused a decrease in the total cultivable bacteria on the tooth surfaces. The proportion of Enterobacteriaceae in the isoproterenol‐treated mice decreased, although the proportion of other bacterial groups increased. Salivary gland ablation, which caused the loss of mucins in saliva, showed essentially the same number of total bacteria as the control. Salivary gland ablation resulted in a decrease in the proportion of Enterobacteriaceae, while the proportion of Gram‐positive rods and staphylococci increased.
Heat Shock Cognate Protein 71‐Associated Peptides Function as an Epitope for Toxoplasma gondii‐Specific CD4+ CTL
Authors: Yang Tian‐hui , Aosai Fumie , Norose Kazumi , Mun Hye‐seong , Yano Akihiko 
Published: November 2013

AbstractHLA‐DR‐restricted CD4+ cytotoxic T‐lymphocyte (CTL) lines specific for Toxoplasma gondii (T. gondii)‐infected melanoma cells have been established from peripheral blood lymphocytes (PBLs) of a patient with chronic toxoplasmosis. The role of heat shock cognate protein (HSC) 71 in antigen (Ag) processing and presentation of T. gondii‐infected melanoma cells to these CD4+ CTL lines was investigated. A human melanoma cell line (P36) pulsed with T. gondii‐infected P36 cell‐derived HSC71 was lysed by a T. gondii‐specific CD4+ CTL line (Tx‐HSC‐1). The Tx‐HSC‐1 also killed T. gondii‐infected P36 cells. The lytic activity of Tx‐HSC‐1 against P36 cells pulsed with T. gondii‐infected P36 cell‐derived HSC71 was inhibited by monoclonal antibodies (mAbs) against HSC71. Anti‐human leukocyte antigen (HLA)‐DR mAb also partially blocked the lytic activity, whereas anti‐HLA‐A,B,C mAb did not block the lytic activity. In addition, a flow cytometric analysis with these specific mAbs against HSC71 showed HSC71 to be expressed on the cell surface of T. gondii‐infected P36 cells as well as uninfected P36 cells. These data indicate that HSC71 molecules are expressed on human melanoma cell line P36, and that HSC71 may play a potential role in Ag presentation and processing of T. gondii‐infected P36 cells to CD4+ CTL.