bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cloning and Sequence Analysis of Another Shiga‐Like Toxin IIe Variant Gene (slt‐IIera) from an Escherichia coli R107 Strain Isolated from Rabbit
Authors: Kim Sang‐hyun , Cha In‐ho , Kim Kyoung‐sook , Kim Yong‐hwan , Lee Young‐choon 
Published: November 2013

AbstractAn Escherichia coli R107 strain (O26 serotype) producing a Shiga‐like toxin IIe variant (SLT‐IIera) was isolated from the mesenteric lymph node of a freshly dead rabbit carcass. The entire structural gene for this SLT‐IIera was cloned from chromosomal DNA by PCR using primers based on previously published slt‐IIe sequences. Nucleotide sequence analysis indicated that the slt‐llera gene was very similar to slt‐IIe (formerly called slt‐IIy) from E. coli strains S1191 and 412; five and one nucleotide changes were detected in A and B subunits, respectively, which resulted in changes in amino acid sequences of the corresponding subunits by three and one residues. Recombinant SLT‐IIera and SLT‐IIe produced using an E. coli host‐vector system showed similar cytotoxicity, suggesting that the variations in the structural gene of SLT‐IIera have no significant effect on cytotoxic level.
Direct Detection by PCR of Escherichia coli O157 and Enteropathogens in Patients with Bloody Diarrhea
Authors: Takeshi Kouichi , Ikeda Tetsuya , Kubo Akiko , Fujinaga Yukako , Makino Souichi , Oguma Keiji , Isogai Emiko , Yoshida Shin‐ichi , Sunagawa Hiroyuki , Ohyama Tohru , Kimura Hiroo 
Published: November 2013

AbstractDirect detection of Escherichia coli O157 and foodborne pathogens associated with bloody diarrhea were achieved using polymerase chain reaction (PCR) after the preparation of DNA from stool specimens using the microspin technique. PCR was compared with cultivation and toxin production tests with respect to the efficiency of detection of each pathogen; E. coli O157, Vibrio parahaemolyticus, Salmonella serovar Enteritidis and Campylobacter jejuni. Detection of some or all of the above pathogens in clinical stool specimens was achieved using PCR. The minimum number of cells required for the detection of the above pathogens by PCR was 101 CFUs/0.5 g of stool sample. PCR was completed within 6 hr. The above pathogens were also detected in cultivation and toxin production tests. Partial purification of the template DNA using the microspin technique was essential for the elimination of PCR inhibitors from the DNA samples. This PCR method is an accurate, easy‐to‐read screening method for the detection of Shiga‐like toxin producing E. coli O157 and enteropathogens associated with bloody diarrhea in stool specimens.
Prevalence of Herpes Simplex Virus Type 1‐ and 2‐ Specific Antibodies among the Acute, Recurrent, and Provoked Types of Female Genital Herpes
Authors: Hashido Madoka , Lee Francis K. , Nahmias Andre J. , Kawana Takashi 
Published: November 2013

AbstractSixty‐eight sera from the acute, recurrent, and provoked types of female genital herpes were compared for the seroprevalence of herpes simplex virus (HSV) types 1 and 2 by immunodot assay using HSV glycoprotein G. In the HSV‐1‐isolated patients, no HSV‐2 antibodies were detected, whereas in the HSV‐2‐isolated patients, HSV‐1 seroprevalence was 9% for the acute type, 89% for the provoked type (P< 0.005), and 55% for the recurrent type (P<0.05). The natural history of female genital herpes and the possible protective role of pre‐existing antibodies in preventing the acquisition or clinical manifestation of a subsequent HSV infection are discussed.
Sho‐Saiko‐To, a Traditional Kampo Medicine, Enhances the Anti‐HIV‐1 Activity of Lamivudine (3TC) In Vitro
Authors: Piras Giovanna , Makino Masahiko , Baba Masanori 
Published: November 2013

AbstractSho‐saiko‐to (SST), a traditional Kampo medicine, has been examined for its inhibitory effect on human immunodeficiency virus type 1 (HIV‐1) replication in peripheral blood mononuclear cells (PBMCs). SST alone moderately inhibited HIV‐1 replication at a concentration of 25 μg/ml. When SST was combined with zidovudine (AZT), lamivudine (3TC) or AZT plus 3TC, SST enhanced the anti‐HIV‐1 activity of 3TC. In contrast, SST slightly enhanced the anti‐HIV‐1 activity of AZT plus 3TC but did not enhance the activity of AZT alone. These results suggest that the combination of SST and 3TC has potential as a chemother‐apeutic modality of HIV‐1 infection.
Immune Response to Neutralizing Epitope on Human Cytomegalovirus Glycoprotein B in Japanese: Correlation of Serologic Response with HLA‐Type
Authors: Wada Kaoru , Mizuno Shinichi , Ohta Hirotoshi , Nishiyama Yukihiro 
Published: November 2013

AbstractAntigenic domain 1 (AD‐1), located between amino acids 608 and 625 of human cytomegalovirus (CMV) gB protein, is the major domain recognized by neutralizing antibodies. Amino acids 552 to 630 are essential for the binding of neutralizing antibodies. We developed an enzyme‐linked immunosorbent assay (ELISA) to detect antibodies against a fusion protein containing amino acid residues 549 to 644 of the gB polypeptide and maltose binding protein (MBP). Of 180 seropositive samples, 106 (58.9%) showed positive immuno‐reactivity against the fusion protein. None of the seronegative samples reacted with the fusion protein. Among 57 seropositive individuals typed for HLA, subjects with HLA‐DR9 had a higher positive rate against the fusion protein (13/14=92.9%) than those without HLA‐DR9 (25/43=58.1%). In addition, subjects with HLA‐DR15 had a lower positive rate against the fusion protein (7/16=43.3%) than those without HLA‐DR15 (31/41 = 75.6%). Mean OD values of HLA‐DR15‐positive individuals were significantly lower than those of HLA‐DR15‐negative individuals. Thus, among CMV‐infected individuals, HLA‐DR9 may be associated with responders for neutralizing antibodies and HLA‐DR15 may be associated with non/low‐responders.
Molecular Epidemiology of Vibrio cholerae O1 Isolated from Sporadic Cholera Cases in Okinawa, Japan
Authors: Iwanaga Masaaki , Honma Yasuko , Enami Mika 
Published: November 2013

AbstractIn July 1994, 6 cholera cases due to Vibrio cholerae O1 El Tor Ogawa sporadically appeared in Okinawa. All 6 patients had no history of traveling abroad. In the period of this cholera outbreak, a strain of V. cholerae O1 El Tor Ogawa was detected from an imported fish at the Naha port quarantine station. The isolates were characterized to clarify whether or not, they belonged to a common clone. Phenotypes were identical except that one strain revealed cured Celebes and the others were original Celebes in kappa phage typing. The restriction fragment patterns of DNA of the isolates hybridized with an enzyme‐labeled oligonucleotide probe for cholera toxin gene (ctx) were identical. Randomly amplified polymorphic DNA of the isolates were identical when a primer was used, but 2 patterns were seen when another primer was used. Pulsed‐field gel electrophoresis of the chromosomal DNA digested with NotI restriction enzyme showed 3 patterns. The DNA fragment pattern of the strain isolated from the imported fish was different from the clinical isolates. These results suggested that there was no epidemiological relation among the strains of V. cholerae O1 isolated during this period.
Evaluation of Enzyme‐Labeled Oligonucleotide Probes to Identify Enterohaemorrhagic Escherichia coli
Authors: Yoh Myonsun , Takagi Kazuhiro , Eda Junji , Ohtomo Masato , Takarada Yutaka , Shibata Shuji , Honda Takeshi 
Published: November 2013

AbstractAlkaline phosphatase‐conjugated oligonucleotide probes were developed to detect the gene coding for Vero toxin 1 (VT1) and Vero toxin 2 (VT2). Using these probes, 3 hr was enough to detect VT genes when suspicious colonies of enterohaemorrhagic Escherichia coli (EHEC) were obtained on an agar plate. The results of a hybridization test with 144 isolates of EHEC O157 and one isolate of Shigella dysenteriae Type 1 agreed exactly with the immunological detection, reversed passive latex agglutination (RPLA) test, of VTs in their culture supernatants. The sensitivity levels of these probes for the detection of VT genes were 100%. The specificity of these probes were also tested with a total of 1,002 strains of Escherichia coli other than EHEC and 8 strains of Shigella sp. other than Shigella dysenteriae Type 1; the results showed 100% specificity.
A Thymic Hormone Protects Mice from Enteropathy during Acute Graft‐versus‐Host Disease
Authors: Inagaki‐Ohara Kyoko , Sakai Tetsu , Koya Goyo , Awaya Akira , Yoshikai Yasunobu 
Published: November 2013

AbstractWe have previously reported that a nonapeptide thymic hormone, facteur thymique serique (FTS), is involved in the differentiation and activation of intestinal intraepithelial lymphocytes (i‐IEL) in mice. In this study, we examined the effect of FTS treatment on enteropathy in a murine model for acute graft‐vs.‐host disease (GVHD) induced by injection of parental C57BL/6 splenocytes into unirradiated (C57BL/6XDBA/2) F1 hybrids. FTS treatment significantly protected mice from developing acute GVHD as assessed by mortality rate, splenomegaly and enteropathy. The infiltration of donor‐derived TCRαβ i‐IEL bearing CD8αβ was significantly inhibited in the small intestine of FTS‐treated mice, and the frequencies of apoptosis of crypt cells in the intestinal mucosa were decreased in these mice during acute GVHD. These results suggest that FTS treatment contributes to protection against enteropathy of acute GVHD. Thus, FTS may provide a useful approach to control acute GVHD after blood transfusion or bone marrow transplantation.
Molecular Typing of Staphylococcus epidermidis and Other CNS with Repetitive Element Sequence‐Based PCR
Authors: Hu Xiaohong , Totake Kyoko 
Published: November 2013

AbstractThe chromosomal distribution of the repetitive DNA sequence found in Mycoplasma pneumoniae (REP‐MP2) provides an ideal target for detecting DNA fragment patterns specific to individual Staphylococcus epidermidis and S. haemolyticus strains. A REP‐MP2 sequence‐based PCR (rep‐PCR) was developed and applied to CNS isolates. We identified a 450 bp genomic DNA fragment which was common and specific to S. epidermidis isolates and not found in other CNS. In addition, S. epidermidis isolates showed several bands that could be grouped into 14 different fragment patterns. Similarly, S. haemolyticus isolates were classified into 10 groups. Significant correlations between the typing patterns of S. epidermidis and resistance to oxacillin (P<0.05), gentamicin (P<0.01), erythromycin (P<0.02), and sulfamethoxazole‐trimethoprim (P<0.001) were found. The rep‐PCR method is a rapid and reproducible discriminatory means for molecular typing of S. epidermidis and other CNS.
Production of TNFα and IL‐6 by Activated Whole Blood from HIV‐1 Infected Patients Detected by a One‐Stage Procedure: Relationship with the Phenotype of HIV‐1 Isolates
Authors: Benyoucef Samira , Hober Didier , Shen Lu , Ajana Faïza , De Groote Donat , Bocket‐Mouton Laurence , Gérard Yann , Lion Georges , Vilain Virginie , Wattré Pierre 
Published: November 2013

AbstractDiluted whole blood (WB) culturing may be the most appropriate milieu in which to study cytokine production in vitro. We tested TNFα and IL‐6 production using small volumes of WB (25 μl) from HIV‐1 positive patients with a one‐step procedure that combines WB stimulation with LPS, PHA and cytokine measurement. We studied 49 patients without secondary infection or at distance of secondary infection staged according to the 1993 classification of the CDC and 12 healthy seronegative subjects. Heparinized blood from 5 control subjects had been collected sequentially during a period of 5 months. The individual variations of TNFα and IL‐6 production were limited for all these individuals. In 1 out of 20 CDC group A patients, 6 out of 17 CDC group B patients and 3 out of 12 CDC group C patients, we obtained higher values of TNFα than the mean + 2 S.D. of the control group. In 3 out of 20 CDC group A patients, 1 out of 17 CDC group B patients without AIDS and 5 out of 12 CDC group C patients, the TNFα values were lower than the mean −2 S.D. of the control group. Low IL‐6 values were obtained in 1 out of 20 CDC group A patients and 1 out of 17 CDC group B patients and 3 out of 12 CDC group C patients. There was no correlation between TNFα production in vitro and plasma level of TNFα. We found no correlation between the levels of cytokines and monocyte count or between the levels of cytokines and CD4 T‐cell count in peripheral blood. Our data point out a disarray in TNFα and IL‐6 production by WB from HIV‐1 infected patients. The relationship between the disarray of cytokine production and cytopathogenicity of HIV‐1 isolates in the P4 cell line was investigated in this study. We found a correlation between the high level of TNFα produced by WB and the phenotype of HIV‐1 isolates isolated from patients. The one‐stage procedure used in this work is of potential value to investigate the activation status of cells for monitoring HIV‐1 positive individuals and predicting HIV‐1 phenotype.
Bioactive Fragment of Human IL‐1β [163‐171] Modulates the Immune Response to Synthetic Peptides of HIV
Authors: Gokulan Kuppan , Rao D. Nageswara 
Published: November 2013

AbstractThe activation of T helper cells specific for viral antigens is critical for antibody production and the generation of cytotoxic T cells during retroviral infection. In this study, we examined the effect of linking HIV peptides with a bioactive fragment of human interleukin‐1β (IL‐1β) (163–171) on the induction of immune response to the peptides. A panel of highly purified synthetic peptides representing defined regions of gp41, Gag and gp120 were used as antigens. Mouse spleen cells primed with the peptide conjugates produced greater proliferation on in vitro stimulation than spleen cells primed with peptide alone. In addition, antibody production as assessed by ELISA was observed after immunization with conjugated peptides but not with peptide alone, indicating B‐cell activation. We also found that a high level of IgG2a antibody production correlated with a high level of IFN‐γ production. These findings favor the notion that IL‐1β plays an important role in immune responses. These observations support the formulation and design of synthetic vaccines against HIV using synthetic HIV peptides conjugated with immunomodulators. Such an approach may provide an effective vaccination against other infectious agents.
Bactericidal Activity of a Fermented Hot‐Water Extract from Stevia rebaudiana Bertoni towards Enterohemorrhagic Escherichia coli O157:H7 and Other Food‐Borne Pathogenic Bacteria
Authors: Tomita Toshio , Sato Naohiko , Arai Tomio , Shiraishi Hiroyuki , Sato Minoru , Takeuchi Masaaki , Kamio Yoshiyuki 
Published: November 2013

AbstractA fermented aqueous extract from Stevia rebaudiana Bertoni showed strong bactericidal activity towards a wide range of food‐borne pathogenic bacteria including enterohemorrhagic Escherichia coli O157:H7. The colony‐forming ability of the food‐borne pathogenic bacteria tested so far was reduced to < 10−7 when exposed to ⩾40% (v/v) solutions of the fermented extract at 37 C for 2 hr. Secretion of verocytotoxin 1 and 2 by enterohemorrhagic E. coli was also diminished by fermented extract at a concentration of ⩾10% (v/v). In contrast, the fermented extract did not significantly kill Bifidobacteria or Lactobacilli. The active principle(s) of the fermented Stevia extract were bactericidal under acidic conditions.
Adhesive Property of Toxin‐Coregulated Pilus of Vibrio cholerae O1
Authors: Tamamoto Tohru , Nakashima Kazutoshi , Nakasone Noboru , Honma Yasuko , Higa Naomi , Yamashiro Tetsu 
Published: November 2013

AbstractThe adhesive property of toxin‐coregulated pilus (TCP) to the human intestine (jejunum), and whether or not TCP mediates the adhesion of Vibrio cholerae 395 organisms to the intestinal epithelium were investigated using visually proving methods. The purified TCP did not agglutinate human erythrocytes nor adhere to the surface of human intestinal epithelium. V. cholerae 395 adhered to the epithelium, but the adhesion was not inhibited by blocking the pili with the Fab fraction of anti‐TCP IgG. The organisms adhered to the intestine treated with purified TCP in advance, as well as to the intact intestine. These findings suggest that TCP is not involved in the adhesion of these organisms to the intestinal epithelium.
Detection of Coxiella burnetii from Dust in a Barn Housing Dairy Cattle
Authors: Yanase Tsuyoshi , Muramatsu Yasukazu , Inouye Iwao , Okabayashi Tamaki , Ueno Hiroshi , Morita Chiharu 
Published: November 2013

AbstractWe attempted to detect Coxiella burnetii in dust samples collected from a barn housing dairy cattle by the polymerase chain reaction (PCR) method. Ten dust samples (five from ventilation fans and five from crossbeams) were collected from two areas in a barn on a farm near Sapporo, Hokkaido. C. burnetii was detected in 5 of the 10 dust samples. It was believed that aerial contamination by C. burnetii occurred in the barn.
Tinctorial Properties of Spherical Bodies in Broth Cultures of Nocardia asteroides GUH‐2
Authors: Kohbata Shunro 
Published: November 2013

AbstractIn yeast extract‐supplemented brain heart infusion (BHI) broth cultures of Nocardia asteroides GUH‐2, many spherical bodies (SBs) were frequently seen nearby filamentous cells. They showed no Gram‐positivity when Gram stain was applied. When acridine orange stain was applied, many of them showed different green fluorescence from bright orange fluorescence of the filamentous nocardiae under ultraviolet light. Their acid‐fastness appeared to depend on the presence of paraffin. Using the polymerase chain reaction (PCR) method, 16S rRNA genes were detected in SB‐containing broth cultures inoculated with culture filtrates from broth cultures of the strain and identical to that of N. asteroides. These results suggest that SBs are cell wall‐defective (CWD) forms which result from the spontaneous mutation of N. asteroides GUH‐2.
The Use of Heterologous Promoters for Adeno‐Associated Virus (AAV) Protein Expression in AAV Vector Production
Authors: Ogasawara Yoji , Urabe Masashi , Ozawa Keiya 
Published: November 2013

AbstractAlthough adeno‐associated virus (AAV) vectors are potentially useful gene transfer vehicles for gene therapy, the vector production system is currently at the developmental stage. We constructed AAV helper plasmids (Rep and Cap expression plasmids) by replacing a native AAV promoter, p5, with various heterologous promoters to examine whether the efficiency of AAV vector production was influenced by modulating the AAV protein expression pattern. The helper plasmids containing heterologous promoters (EF, CMV, SV40, B19p6, and CAG promoters, respectively) expressed Rep78/68 more efficiently than a conventional helper plasmid (pIM45), but the expression of Rep52/40 and Cap decreased, resulting in a significant reduction in AAV vector production. Furthermore, the efficiency of vector production never fully recovered even if the Cap proteins were supplied by an additional expression plasmid. A large amount of Rep78/68 and/or a reduced level of Rep52/40 may have deleterious effects on AAV vector production. The present findings will aid in the development of a more efficient AAV vector production system.
Antigenic and Functional Analyses of Glycoprotein of Rabies Virus Using Monoclonal Antibodies
Authors: Luo Ting Rong , Minamoto Nobuyuki , Hishida Miyuki , Yamamoto Keiko , Fujise Toru , Hiraga Shinya , Ito Naoto , Sugiyama Makoto , Kinjo Toshio 
Published: November 2013

AbstractThirty‐five monoclonal antibodies (MAbs) against glycoprotein (G protein) of the RC‐HL strain of the rabies virus have been established. Using these MAbs, two antigenic sites (I and II) were delineated on the G protein of the RC‐HL strain in a competitive binding assay. Of these, 34 MAbs recognized the epitopes on site IL Site II was further categorized into 10 subsites according to their patterns in a competitive binding assay. Each site II‐specific MAb showed 5 to 23 nonreciprocal competitions. The reactivities of 35 MAbs to rabies and rabies‐related viruses in an indirect immunofluorescent antibody test showed that six MAbs in group A binded to rabies and rabies‐related viruses and eight MAbs in group E reacted only with rabies viruses, considering that the former represent the genus‐specific of Lyssavirus and the latter are rabies virus‐specific. From biological assays, 28 of the 35 MAbs showed neutralization activity, 31 showed hemagglutination inhibition (HI) activity, and 18 showed immunolysis (IL) activity. The MAbs recognizing neutralization epitopes fell into at least three groups: those exhibiting both HI and IL activity, those showing only HI activity, and those showing neither HI nor IL activity. All IL epitopes overlap with HA epitopes. Five of the nine MAbs which reacted with the antigen treated by sodium dodecyl sulfate in ELISA were not reduced, or reduced only slightly, in the titer. None of the MAbs reacted with 2‐mercaptoethanol‐treated antigen. Only one MAb that recognized site I reacted with the denatured G protein in a Western blotting assay, indicating that its epitope is linear. These results suggest that almost all of the epitopes on the G protein of the rabies virus are conformation‐dependent and the G protein forms a complicated antigenic structure.
Protective Effect of Administration of Skim Milk on Exogenous and Endogenous Infection in Mice
Authors: Kobayashi Toshiya , Ohmori Toshrhiro , Yanai Minoru , Takeshita Yasuyoshi , Mitsuyama Masao 
Published: November 2013

AbstractIn order to minimize the denaturation of proteins in milk, normal cow's milk was pasteurized at 61 C for 20 min. The protective effects of the thus prepared skim milk (low‐heat skim milk) on exogenous and endogenous infection were examined as compared with conventional skim milk which was pasteurized at 121 C for 2 sec. The antibody titers to Listeria monocytogenes and Escherichia coli of low‐heat skim milk were almost equal to that of raw milk, while no antibody was detected in the conventional skim milk. When mice were given low‐heat skim milk or conventional skim milk, the incidence of the translocation of orally inoculated Listeria monocytogenes to the spleen was lower in the low‐heat skim milk group than that in the conventional skim milk group. The life span of 7 Gy X‐ray irradiated mice given low‐heat skim milk was significantly prolonged in comparison to that of mice given conventional skim milk. However, there were no differences in the number of bacteria in the feces or IgA production by Peyer's patch cells between the two groups. These results suggest that antibodies in low‐heat skim milk, which still have reactivity to exogenous or indigenous bacteria, may contribute to the protective effects against bacterial infection.
Discrimination among the Clinical Isolates of Candida albicans by Amplification of the Repetitive Sequences, alts
Authors: Doi Matsuko , Chibana Hiroji , Nakagawa Yoshiyuki , Tanaka Kenji 
Published: November 2013

AbstractA primer pair, PB and BSH, which amplified alts, a portion of Candida albicans‐specific repetitive sequence, RPS, gave stable and reproducible fingerprint patterns of the strains by polymerase chain reaction (PCR). We applied this method to clinical isolates of C. albicans for strain discrimination. Using PCR fingerprint patterns, we could analyze the relatedness of C. albicans strains including those isolated from children with leukemia and their bedside parents. The results indicated that PCR analysis targeting an alt region gives rise to the same conclusion as the previous study obtained by SmaI RFLP analysis.