bluuebunny/crossref_metadata_2025_split huggingface dataset:
Design of Species‐Specific Primers to Identify 13 Species of Clostridium Harbored in Human Intestinal Tracts
Authors: Kikuchi Eisaku , Miyamoto Yukiko , Narushima Seiko , Itoh Kikuji 
Published: November 2013

AbstractThe genus Clostridium is dominant in human intestinal tracts and plays an important role in human health. We designed species‐specific primers to identify 13 species of Clostridium (C. perfringens, C. paraputrificum, C. bifermentans, C. difficile, C. clostridiiforme, C. nexile, C. sphenoides, C. indolis, C. ramosum, C. cocleatum, C. butyricum, C. sordellii, and C. innocuum) easily and rapidly. The PCR annealing temperature was set at a uniform 60 C for application to all strains at the same time. To confirm the specificities of these primers, 85 intestinal bacteria in total, including type strains, reference strains, and isolates were used. Ten primers (including those for C. perfringens to C. cocleatum) indicated high specificities. Although there were some cross‐reactions with the other three primers, the target species were distinguishable from other bacteria by the different sizes of PCR products.
emm Typing of Group A Streptococcus Clinical Isolates: Identification of Dominant Types for Throat and Skin Isolates
Authors: Tanaka Daisuke , Gyobu Yotaku , Kodama Hirohide , Isobe Junko , Hosorogi Shiho , Hiramoto Yuko , Karasawa Tadahiro , Nakamura Shinichi 
Published: November 2013

AbstractT and emm types were determined for group A streptococci isolated from patients with various infections during 1990–1999 in Toyama Prefecture, Japan. Out of 906 isolates, 872 isolates were divided into 20 T serotypes, and 34 isoltes were T nontypeable (TNT). T12, T1, and T4 were dominant among 699 throat isolates; on the other hand, T11, T28, TB3264, and TNT were dominant among 80 skin isolates. The emm types of 190 isolates were determined following specific PCR amplification and sequencing of the products. Twenty T serotypes were divided into 34 T type/emm type combinations. Thirty‐four TNT isolates were divided into 14 emm types, in which emm58 was the most common (38%). Among 82 throat isolates randomly selected, predominant T types T12, T1, and T4 isolates were of the respective same numbers in emm type. T11/emm89, T28/emm28, TB3264/emm 13w, and TNT/emm58 were predominant among 80 skin isolates. emm‐type distribution observed in the present study was that usually reported in the western world. To our knowledge, 3 T/emm is a novel combination. These results show that emm typing allows the characterization of group A streptococci from various sources.
Antibacterial Effect of Kampo Herbal Formulation Hochu‐Ekki‐To (Bu‐Zhong‐Yi‐Qi‐Tang) on Helicobacter pylori Infection in Mice
Authors: Yan Xiaoqun , Kita Masakazu , Minami Masato , Yamamoto Toshiro , Kuriyama Hiroko , Ohno Tomoyuki , Iwakura Yoichiro , Imanishi Jiro 
Published: November 2013

AbstractBecause Helicobacter pylori (H. pylori) infection is a major cause of gastroduodenal diseases in humans, the eradication of H. pylori using antibiotics is very effective for the treatment of gastroduodenal diseases. However, it has recently been reported that resistance to these antibiotics is developing. In the present study, the antibacterial effect of a Kampo (traditional Japanese medicine) herbal formulation, Hochu‐ekki‐to (HET; Formula repletionis animalis et supletionis medii), against H. pylori was examined in vitro and in vivo. HET inhibited the growth of antibiotic‐resistant strains of H. pylori as well as antibiotic‐sensitive strains at a dose of 2.5 mg/ml in vitro. When 1,000 mg/kg of HET was administered orally to C57BL/6 mice for 7 days before or after inoculation with H. pylori, H. pylori in the stomach was significantly reduced in the HET‐pre‐treatment group compared with the control group. Furthermore, HET in combination with antibiotics completely eradicated the bacteria in mice. The expression of interferon (IFN)‐γ was induced in the gastric mucosa of the mice pre‐treated with HET. There were no significant differences between the colonization of H. pylori in the control and HET treatment groups in IFN‐γ gene‐deficient mice. These results suggest that the antibacterial effect of HET may be partly due to IFN‐γ induction, and that HET may be clinically useful for treatment of H. pylori infection.
Comparison of Four Microbial Enzymes in Clostridia and Bacteroides Isolated from Human Feces
Authors: Nakamura Joe , Kubota Yoshihiko , Miyaoka Masaaki , Saitoh Toshihiko , Mizuno Fumio , Benno Yoshimi 
Published: November 2013

AbstractThe activities of four microbial enzymes (azoreductase, nitroreductase, β‐glucuronidase, and β‐glucosidase) in major anaerobic members of human fecal microflora were quantified and the influence of the host factors on expression of these microbial enzyme activities was also investigated. Clostridium paraputrificum and C. clostridiiforme showed much higher activities than other fecal anaerobes tested. Nitroreductase activity in C. paraputrificum isolated from fecal specimens of patients with colon cancer was significantly (P<0.05) higher than that in the clostridia isolated from healthy subjects and the subjects given high beef diets. However, the activities of some microorganisms tested showed marked differences in each strain.
Effects of Recombinant Cholera Toxin B Subunit on IL‐1β Production by Macrophages In Vitro
Authors: Maeyama Jun‐ichi , Isaka Masanori , Yasuda Yoko , Matano Keiko , Taniguchi Tooru , Morokuma Kazunori , Ohkuma Kunio , Tochikubo Kunio , Goto Norihisa 
Published: November 2013

AbstractRecombinant cholera toxin B subunit (rCTB) is a safe and potent mucosal adjuvant. As a clue to the mechanism of the adjuvant effect of rCTB, the profile of cytokines secreted in vitro by the mouse peritoneal macrophage (MΦ) treated with rCTB was examined. IL‐1β secretion, intracellular production, and expression of its mRNA of LPS‐stimulated MΦ was greatly enhanced by treatment with rCTB. IL‐1β production in response to other microbial stimulators, such as Pansorbin, Sansorbin, insoluble peptidoglycan, and Taxol, was also potentiated by rCTB. MΦ pretreated with rCTB before 24 hr could maintain the ability to produce a high level of IL‐1β, suggesting that this ability may be involved in the adjuvant activity of rCTB on MΦ stimulation. The possibility of close association between rCTB and signal transduction of a Toll‐like receptor family in MΦ is discussed.
Purification, Cloning and Characterization of Variant LukE‐LukD with Strong Leukocidal Activity of Staphylococcal Bi‐Component Leukotoxin Family
Authors: Morinaga Naoko , Kaihou Yuriko , Noda Masatoshi 
Published: November 2013

AbstractStaphylococcus aureus produces bi‐component leukotoxins composed of non‐associated soluble proteins, S and F. Neither S nor F component alone is cytotoxic, but components together are active. These include Panton‐Valentine leukocidin (PVL), γ‐hemolysin, LukE‐LukD and others. Purification of leukotoxin from Staphylococcus aureus V8 strain (ATCC 27733) which does not have PVL genes, identified an F component with 100% identical to that of PVL in the first twenty‐five N‐terminal amino acids. Molecular cloning of this toxin obtained 2,595 nucleotides sequences containing two novel open reading frames for S and F. Deduced amino acid sequences of the S and F were respectively 91 and 94% identical to those of LukE and LukD. These were named variant of LukE‐LukD (LukEv‐LukDv). The activity of the recombinant LukEv‐LukDv to rabbit leukocytes was similar to that of recombinant PVL. LukEv‐LukDv was hemolytic to rabbit red blood cells although the activity was only 8% of γ‐hemolysin, but PVL was not. These activities were quite different from the LukE‐LukD which was reported no hemolytic and poorly cytotoxic to leukocytes compared to PVL. The lukEv‐lukDv was found in 87% of clinical isolates of Staphylococcus aureus but lukE‐lukD was not detected. These data demonstrate the existence of variant LukE‐LukD in V8 strain (ATCC 27733).
Epstein‐Barr Virus (EBV) Nuclear Antigen Leader Protein (EBNA‐LP) Forms Complexes with a Cellular Anti‐Apoptosis Protein Bcl‐2 or Its EBV Counterpart BHRF1 through HS1‐Associated Protein X‐1
Authors: Matsuda Go , Nakajima Kaori , Kawaguchi Yasushi , Yamanashi Yuji , Hirai Kanji 
Published: November 2013

AbstractEpstein‐Barr virus (EBV) nuclear antigen leader protein (EBNA‐LP) plays a critical role in EBV‐induced transformation. An earlier report (Y. Kawaguchi et al., J. Virol. 74: 10104–10111, 2000) showed that EBNA‐LP interacts with a cellular protein HS1‐associated protein X‐1 (HAX‐1). The predicted amino acid sequence of HAX‐1 exhibits similarity to that of another cellular protein Nip3 which has been shown to interact with cellular and viral anti‐apoptotic proteins such as Bcl‐2 and BHRF1, an EBV homolog of Bcl‐2. Here we investigated whether HAX‐1, like Nip3, interacts with Bcl‐2 proteins and report the following. (i) A purified chimeric protein consisting of gluthathione S‐transferase (GST) fused to BHRF1 (GST‐BHRF1) or Bcl‐2 (GST‐Bcl‐2) specifically pulled down HAX‐1 transiently expressed in COS‐7 cells. (ii) GST‐BHRF1 or GST‐Bcl‐2 was not able to pull down EBNA‐LP transiently expressed in COS‐7 cells, whereas each of the GST fusion proteins formed complexes with EBNA‐LP in the presence of HAX‐1. These results indicated that EBNA‐LP interacts with the viral and cellular Bcl‐2 proteins through HAX‐1, suggesting that EBNA‐LP possesses a potential function in the regulation of apoptosis in EBV‐infected cells.
Adhesion and Colonization of Enterohemorrhagic Escherichia coli O157:H7 in Cecum of Mice
Authors: Nagano Keiji , Taguchi Kazuki , Hara Takumi , Yokoyama Shin‐ichiro , Kawada Kenji , Mori Hiroshi 
Published: November 2013

AbstractInfectious diseases due to enterohemorrhagic Escherichia coli (EHEC) are characterized by diarrhea, hemorrhagic colitis and hemolytic uremic syndrome. The adherence of EHEC on intestinal epithelial cells is a first step for developing these diseases. In the present study, we examined whether EHEC O157: H7 adhere to intestinal epithelial cells of mice and cause F‐actin accumulation in the epithelial cells following the intragastric inoculation of the pathogen. Fecal shedding of the EHEC O157:H7 strain was observed in ICR mice up to 3 weeks. Fecal shedding periods of the type III secretion system‐related gene (espA and sepL) deletion mutants were clearly shorter than that of the wild‐type EHEC O157:H7 strain. The EHEC O157:H7 colonies were found on the epithelial surfaces of the ceca in association with F‐actin accumulation beneath the attached bacteria.
Terminal Restriction Fragment Length Polymorphism Analysis for Human Fecal Microbiota and Its Application for Analysis of Complex Bifidobacterial Communities
Authors: Sakamoto Mitsuo , Hayashi Hidenori , Benno Yoshimi 
Published: November 2013

AbstractTerminal restriction fragment length polymorphism (T‐RFLP) analysis was used to characterize and compare human fecal microbiota among individuals. T‐RFLP patterns of fecal 16S ribosomal DNA (rDNA) PCR products from three adults revealed host‐specific bacterial communities and were in good agreement with those reported in our previous study. In addition, we applied T‐RFLP analysis for the analysis of complex bifidobacterial communities in human fecal samples. The developed method based on Bifidobacterium genus‐specific PCR and T‐RFLP could identify more than one bifidobacterial species. T‐RFLP patterns of Bifidobacterium genus‐specific PCR products from the fecal samples were host‐specific as well as those of fecal 16S rDNA PCR products. These results were confirmed by PCR‐denaturing gradient gel electrophoresis (DGGE) with primers specific for the genus Bifidobacterium and Bifidobacterium species‐ and group‐specific PCR. Our study demonstrates that T‐RFLP analysis is useful for assessment of the diversity of the human fecal microbiota and rapid comparison of the community structure among individuals, and that the applied method is useful for rapid and sensitive analysis of bifidobacterial community.
The Suppressive Effect of Bifidobacteria on Bacteroides vulgatus, a Putative Pathogenic Microbe in Inflammatory Bowel Disease
Authors: Shiba Tadahiko , Aiba Yuji , Ishikawa Hiroki , Ushiyama Atsushi , Takagi Atsushi , Mine Tetsuya , Koga Yasuhiro 
Published: November 2013

AbstractBacteroides, a predominant commensal bacteria in the gut, are thought to be responsible for the development of inflammatory bowel disease (IBD). In the present study, we examined whether or not bifidobacteria suppress B. vulgatus, a representative pathogenic Bacteroides species, in both the coculture system and the gnotobiotic murine model. As a result, Bifidobacterium infantis 1222 highly inhibited the growth of B. vulgatus in the coculture and also significantly suppressed the systemic antibody response raised by B. vulgatus colonizing the gut in gnotobiotic mice. Colonization of the mice by B. vulgatus increased the number of Peyer's patch (PP) cells bearing PNA (peanut agglutinin)+/anti‐κ+ phenotype, which represents plasma cell‐like B cells. Moreover, treatment of those B. vulgatus‐implanted mice with B. infantis 1222 abrogated such increase in the number of PNA+/anti‐κ+ cells. These results thus suggested that B. infantis 1222 protected the gut epithelial layer including the PP from being invaded by Bacteroides, thereby suppressing the systemic antibody response raised by Bacteroides.
Proinflammatory Cytokines in Bovine Colostrum Potentiate the Mitogenic Response of Peripheral Blood Mononuclear Cells from Newborn Calves through IL‐2 and CD25 Expression
Authors: Yamanaka Hitoki , Hagiwara Katsuro , Kirisawa Rikio , Iwai Hiroshi 
Published: November 2013

AbstractBovine colostrum contains high concentrations of cytokines, and colostral cytokines are considered to be an important factor in stimulation of maturation of the immune system in newborns. In this study, 5 proinflammatory cytokines (IL‐1β, IL‐6, TNF‐α, IFN‐γ and IL‐1 receptor antagonist, IL‐1ra) present in colostrum were tested for their potential to enhance mitogenic response and to elicit expression of IL‐2 mRNA and CD25 in peripheral blood mononuclear cells (PBMC) from newborn calves before being fed colostrum. PBMC were pretreated with each recombinant bovine cytokine for 2 hr before stimulation with concanavalin A (ConA). Pretreatment of PBMC from newborn calves with IL‐1β, TNF‐α or IFN‐γ significantly enhanced the ConA response, whereas IL‐1ra inhibited the response. The degree of enhancement or inhibition of mitogenic response by these cytokines was more pronounced in PBMC from newborn calves than in those from adult cows. Although IL‐2 mRNA expression in ConA‐stimulated PBMC from newborn calves was weaker than that in those from adult cows of ConA‐stimulated controls, the expression levels became comparable after pretreatment with IL‐1β, TNF‐α or IFN‐γ. The CD25 expression in PBMC from newborn calves was also enhanced by pretreatment with IL‐1β, TNF‐α and IFN‐γ. These results suggest that pretreatment of neonatal PBMC with IL‐1β, TNF‐α or IFN‐γ promotes mitogenic response to ConA through up‐regulating the production of IL‐2 and the expression of the mature IL‐2 receptor.
Structure and Biological Activities of β‐Glucans from Yeast and Mycelial Forms of Candida albicans
Authors: Miura Noriko N. , Adachi Yoshiyuki , Yadomae Toshiro , Tamura Hiroshi , Tanaka Shigenori , Ohno Naohito 
Published: November 2013

AbstractWe have achieved the extraction of cell wall β‐glucan from the mycelial form of Candida albicans (C. albicans) IFO 0579 (M‐CSBG) by using acetic acid, sodium hypochlorite (NaClO), and dimethylsulfoxide (DMSO) treatments. The yield of M‐CSBG was significantly lower (7.5% from dried mycelial cells) than that of the yeast form from C. albicans IFO 1385 (Y‐CSBG, 25.9% from dried yeast cells). The properties of M‐CSBG were similar to those of Y‐CSBG in terms of nuclear magnetic resonance (NMR) spectra and limulus reactivity. Molecular weight (Mw) of M‐CSBG was slightly higher than that of Y‐CSBG. Both Y‐CSBG and M‐CSBG induced the production of comparable amounts of macrophage inflammatory protein‐2 (MIP‐2), a chemotactic factor, from mouse peritoneal exudate cells (PEC) in vitro. These findings suggest that the structure and properties of CSBG from yeast and mycelial cells are similar to each other.
Modulation of Procarboxypeptidase R (ProCPR) Activation by Complementary Peptides to Thrombomodulin
Authors: Shimomura Yasuyo , Kawamura Takeshi , Komura Hidefumi , Campbell William , Okada Noriko , Okada Hidechika 
Published: November 2013

AbstractWe designed complementary peptides (C‐peptides) using a novel computer program (MIMETIC), which generates a series of peptides designed to interact with a target peptide sequence. Carboxypeptidase R (CPR) is an unstable basic carboxypeptidase found in fresh serum in addition to carboxypeptidase N (CPN) which is stable. CPR is generated from its precursor form (proCPR) by trypsinlike enzymes, and its activation is mediated by thrombin generated in the coagulation cascade. The efficiency of activation is enhanced approximately 1,200‐fold when thrombin (T) is bound to thrombomodulin (TM). We attempted to generate C‐peptides which recognize the T‐binding site within TM assuming that some of these might interfere with the generation of T and TM complexes (T‐TM). Among three peptides designed, two inhibited the enhancement in activation of proCPR by T in the presence of TM. One of the peptides at 16 μM reduced the activation of proCPR to the level obtained by T alone.
Identification of the Spotted Fever Group Rickettsiae Detected from Haemaphysalis longicornis in Korea
Authors: Lee Jung‐Hee , Park Hyo‐Soon , Jung Kwang‐Don , Jang Won‐Jong , Koh Seong‐Eun , Kang Shin‐Seok , Lee In‐Yong , Lee Won‐Ja , Kim Bum‐Joon , Kook Yoon‐Hoh , Park Kyung‐Hee , Lee Seung‐Hyun 
Published: November 2013

AbstractSeven Haemaphysalis ticks were found positive in PCR assay of gltA gene to detect the spotted fever group (SFG) rickettsiae DNA from 100 ticks. The nucleotide sequence of 16S rRNA gene was determined from 5 ticks and compared to those of other Rickettsia strains. The nucleotide sequence from 4 ticks showed high homologies (99.7 to 100%) with that of R. japonica YH, and that from 1 tick (tick no. 48) was identical with that of R. rickettsii R, suggesting that SFG rickettsiae exists in Korea. This is the first documentation of SFG rickettsiae in Korea.
Chemistry and Biology of Angiitis Inducer, Candida albicans Water‐Soluble Mannoprotein‐β‐Glucan Complex (CAWS)
Authors: Ohno Naohito 
Published: November 2013

AbstractDeep mycoses have been clearly demonstrated to release β‐glucans into the blood. Structure of the β‐glucan was, at least in part, suggested to be a mannoprotein β‐glucan complex (CAWS) as assessed by biochemical and immunochemical analyses of the extracellular macromolecular fraction of Candida albicans. Half clearance time of i.v. administered CAWS was about 30 min in mice. In addition to the reactivity with limulus G‐test, CAWS was found to exhibit various biological activities, such as cytokine synthesis by leukocyte, platelet aggregation, lethal toxicity, enhancement of side effect of indomethacin, induction of coronary arteritis in mice, and so on. In this review, the chemical properties and biological activities of CAWS are discussed.
Molecular Analysis of Fecal Microbiota in Elderly Individuals Using 16S rDNA Library and T‐RFLP
Authors: Hayashi Hidenori , Sakamoto Mitsuo , Kitahara Maki , Benno Yoshimi 
Published: November 2013

AbstractFecal microbiota in six elderly individuals were characterized by the 16S rDNA libraries and terminal restriction fragment length polymorphism (T‐RFLP) analysis. Random clones of 16S rRNA gene sequences were isolated after PCR amplification with universal primer sets from total genomic DNA extracted from feces of three elderly individuals. These clones were partially sequenced (about 500 bp). T‐RFLP analysis was performed using 16S rDNA amplified from six subjects. The lengths of the terminal restriction fragment (T‐RF) were analyzed after digestion by HhaI and MspI. Among 240 clones obtained, approximately 46% belonged to 27 known species. About 54% of the other clones were 56 novel “phylotypes” (at least 98% homology of clone sequence). These libraries included 83 species or phylotypes. In addition, about 13% (30 phylotypes) of these phylotypes were newly discovered in these libraries. A large number of species that are not yet known exist in the feces of elderly individuals. 16S rDNA libraries and T‐RFLP analysis revealed that the majority of bacteria were Bacteroides and relatives, Clostridium rRNA cluster IV, IX, Clostridium rRNA subcluster XIVa, and “Gammaproteobacteria”. The proportion of Clostridium rRNA subcluster XIVa was lower than in healthy adults. In addition, although Ruminococcus obeum and its closely related phylotypes were detected in high frequency in healthy young subjects, hardly any were detected in our elderly individuals. “Gammaproteobacteria” were detected at high frequency.
Improved Recovery of Rabies Virus from Cloned cDNA Using a Vaccinia Virus‐Free Reverse Genetics System
Authors: Ito Naoto , Takayama‐Ito Mutsuyo , Yamada Kentaro , Hosokawa Junji , Sugiyama Makoto , Minamoto Nobuyuki 
Published: November 2013

AbstractTo improve efficiency of recovery of rabies virus from cloned cDNA, we established a BHK cell clone that stably expresses T7 RNA polymerase, which we named BHK/T7–9. We also constructed new helper plasmids for expression of nucleoprotein and RNA polymerase of the RC‐HL strain using the pTM1 plasmid vector, which makes the T7 RNA polymerase‐transcripts from the plasmid cap‐independent for translation. After co‐transfection of these helper plasmids and the previously constructed full‐length genome plasmid of the RC‐HL strain to BHK/T7–9 cells, recombinant rabies virus was efficiently recovered from the cloned cDNA.
Monoclonal Antibody #3‐9‐16 Recognizes One of the Two Isoforms of Rabies Virus Matrix Protein That Exposes Its N‐Terminus on the Virion Surface
Authors: Ameyama Satoshi , Toriumi Harufusa , Takahashi Takuo , Shimura Yuji , Nakahara Tomomi , Honda Yoshikazu , Mifune Kumato , Uchiyama Tsuneo , Kawai Akihiko 
Published: November 2013

AbstractWe investigated behaviors of the rabies virus matrix (M) protein using a monoclonal antibody (mAb), #3‐9‐16, that recognized a linear epitope located at the N‐terminus of the protein. Based on the reactivity with this mAb, M proteins could be divided into at least two isoforms; an ordinary major form (Mα) whose 3‐9‐16 epitope is hidden, and an N‐terminal‐exposed epitope‐positive form (Mβ). The Mβ protein accounted for about 25–30% of the total M proteins in the virion, while its content in the cell ranged from 10 to 15% of total M protein. Fluorescent antibody (FA) staining showed that the Mβ antigen distributed in the Golgi area where the colocalized viral glycoprotein antigen was also detected. Mβ antigen was shown to be exposed on the surface of infected cells by both immunoprecipitation and FA staining with the mAb, whereby the cells might have become sensitive to the mAb‐dependent complement‐mediated cytolysis. Similarly, the Mβ antigen was shown to be exposed on the virion surface, and the infectivity of the virus was destroyed by the mAb in the presence of a complement. Together with these results, we think that the M protein molecule takes either of two conformations, one (Mβ) of which exposes the 3‐9‐16 epitope located in the N‐terminal region of the M protein, that are also exposed on the surface of the virion and infected cells, whereby it might play a certain important role(s) in the virus replication process differently from the other form (Mα), probably through its intimate association with the Golgi area and/or the cell membrane.