bluuebunny/crossref_metadata_2025_split huggingface dataset:
PCR Detection of Capnocytophaga Species in Dental Plaque Samples from Children Aged 2 to 12 Years
Authors: Hayashi Fumiko , Okada Mitsugi , Zhong Xiaobo , Miura Kazuo 
Published: November 2013

AbstractThe purpose of this study was to detect the presence of Capnocytophaga sputigena, C. ochracea, and C. gingivalis in plaque samples from the toothbrushes of 122 children, using a polymerase chain reaction (PCR) method. The subjects were 25, 85, and 12 children with healthy gingiva, gingivitis, and periodontitis, respectively, ranging in age from 2–12 years old. Plaque samples were collected from all erupted tooth sites using a sterile toothbrush. The mean amount of DNA recovered from the samples was approximately 19.3 μg, which was deemed sufficient for performing a PCR‐based survey. C. sputigena prevalence in healthy, gingivitis, and periodontitis subjects was 48.0%, 36.5% and 25.0%, respectively, that for C. ochracea was 100%, 89.4%, and 50.0%, respectively, and that for C. gingivalis was 96.0%, 84.7%, and 75.0%, respectively. The lowest age of positive subjects was approximately 2 years. Our results showed that C. sputigena was moderately prevalent, whereas C. ochracea and C. gingivalis were commonly detected in the oral cavities of the tested children, suggesting that all of these species become established in the early years.
Relatedness between the Coagulase Gene 3′‐End Region and Coagulase Serotypes among Staphylococcus aureus Strains
Authors: Kanemitsu Keiji , Yamamoto Hiroyuki , Takemura Hiromu , Kaku Mitsuo , Shimada Jingoro 
Published: November 2013

AbstractThe 3′‐end region of the coagulase gene from 22 strains of Staphylococcus aureus including 10 standard serotype strains was sequenced, and five subgroups with 48 tandem repeating units were distinguished among the tested strains. Phylogenetic analysis of the 3′‐end region of the coagulase gene indicated that strains belonging to the same serotype were clustered in the same branch. A phylogenetic tree of the deduced amino acid sequences revealed that the C‐terminal region might not be responsible for the epitope of the coagulase protein.
Intraperitoneal Immune Cell Responses to Eubacterium saphenum in Mice
Authors: Salam Mohammad Abdus , Sato Michiko , Hoshino Etsuro 
Published: November 2013

AbstractOral asaccharolytic Eubacterium saphenum, which are newly isolated Gram‐positive rods and one of the predominant microorganisms in human periodontal pockets, were injected intraperitoneally in mice to elucidate their pathogenicity in periodontal diseases. Infiltrating immune cells in the peritoneal exudate were quantitated and intracellular T cell (CD4+/CD8+/γδ+) production of cytokines IL‐4 and IFN‐γ which are related to cellular and humoral immunity, respectively, was determined. Neutrophils appeared first in peritoneal exudates, followed by macrophages and lymphocytes, after the injection of either E. saphenum or Porphyromonas gingivalis. Intracellular IL‐4+ and IFN‐γ+ γδ T cells were detected in the exudates after the injection of E. saphenum (4.6±0.8% and 10.1±1.4%, respectively) and P. gingivalis (5.3±1.6% and 10.1±2.1%, respectively). The intracellular production of IL‐4/IFN‐γ in CD4+/CD8+ T cells was rather low indicating that the main response was from γδ T cells which initiated the immune reactions in mouse peritoneal cavities after injection of E. saphenum or P. gingivalis. Serum IgG and IgM levels were elevated in animals injected with E. saphenum and similarly with P. gingivalis. The present study showed that with slight differences, similar modes of cell response and cytokine and Ig production were observed after intraperitoneal injection of both E. saphenum and P. gingivalis, indicating that E. saphenum may play just as important a role in periodontal diseases as P. gingivalis.
A Comparison of Complete Genome Sequences of the Attenuated RC‐HL Strain of Rabies Virus Used for Production of Animal Vaccine in Japan, and the Parental Nishigahara Strain
Authors: Ito Naoto , Kakemizu Misako , Ito Kumi A. , Yamamoto Atsuko , Yoshida Yoshio , Sugiyama Makoto , Minamoto Nobuyuki 
Published: November 2013

AbstractIn order to establish the molecular basis of the pathogenicity of the attenuated RC‐HL strain of rabies virus used for the production of animal vaccine in Japan, the complete genome sequence of this strain was determined and compared with that of the parental Nishigahara strain which is virulent for adult mice. The viral genome of both strains was composed of 11,926 nucleotides. The nucleotide sequences of the two genomes showed a high homology of 98.9%. The homology of the G gene was lower than those of N, P, M and L genes at both nucleotide and deduced amino acid levels, and the percentage of radical amino acid substitutions on the G protein was the highest among the five proteins. These findings raise the possibility that the structure of the G protein is the most variable among the five proteins of the two strains. Furthermore, we found two clusters of amino acid substitutions on the G and L proteins. The relevance of these clusters to the difference in the pathogenicity between the two strains is discussed.
Hemadsorption Expressed by Cloned H Genes from Subacute Sclerosing Panencephalitis (SSPE) Viruses and Their Possible Progenitor Measles Viruses Isolated in Osaka, Japan
Authors: Furukawa Kyoko , Ayata Minoru , Kimura Masatsugu , Seto Toshiyuki , Matsunaga Isamu , Murata Ryosuke , Yamano Tsunekazu , Ogura Hisashi 
Published: November 2013

AbstractMost subacute sclerosing panencephalitis (SSPE) viruses, including our Osaka‐1, −2, and −3 strains isolated in Osaka, have shown negative hemadsorption (HAD) by African green monkey red blood cells. This property has been thought to be characteristic of SSPE virus as compared to the positive reaction of the standard Edmonston strain of measles virus (MV). However, this assumption has become quite obscure because MV mutates frequently at the genetic level during its multiplication and also because recent field strains isolated by lymphoblastoid cell lines have shown negative HAD. To investigate the above issue, the nucleotide sequences of the hemagglutinin (H) genes from SSPE virus Osaka‐1, −2, or −3 strains were compared to those of various MV field strains isolated in Osaka by Vero cells. The H gene sequences of three SSPE strains were relatively conserved without such biased hypermutation as had been observed in the matrix (M) gene of three SSPE strains. However, this analysis of the H gene sequence of the SSPE viruses enabled us to deduce possible progenitor MVs, which are in agreement with the deduction from the M gene analysis we reported previously. The HAD of Vero cells transfected with the cloned H cDNAs from the SSPE strains and their progenitors suggested that negative HAD of the SSPE viruses has been maintained as one of original properties of the progenitor MVs rather than having been acquired as an altered one during long‐term persistent infection in the brains of patients with SSPE.
Cytokine Responses to Recombinant Cholera Toxin B Subunit Produced by Bacillus brevis as a Mucosal Adjuvant
Authors: Maeyama Jun‐ichi , Isaka Masanori , Yasuda Yoko , Matano Keiko , Kozuka Satoshi , Taniguchi Tooru , Ohkuma Kunio , Tochikubo Kunio , Goto Norihisa 
Published: November 2013

AbstractWe attempted to clarify the mechanism of the mucosal adjuvanticity of recombinant cholera toxin B subunit (rCTB), which is inherently uncontaminated with the holotoxin produced by Bacillus brevis and has a powerful mucosal adjuvant activity, on cytokine responses compared with that of cholera toxin (CT). rCTB had no ability to stimulate cyclic AMP formation in mouse peritoneal macrophages (Mφ). Cytokine production by non‐immunized Mφ cultured with rCTB or CT and by the spleen cells of mice co‐immunized intranasally with ovalbumin (OVA) and rCTB or CT was examined. rCTB alone did not induce interleukin (IL)‐1α/β or IL‐6 production by Mφ, but combination of rCTB with lipopolysaccharide (LPS) enhanced both IL‐1α/β production. Conversely, CT plus LPS suppressed IL‐1α/β production more than LPS alone. Both rCTB and CT suppressed IL‐12 secretion induced by interferon γ (IFN γ) plus LPS. IL‐2, IL‐4, IL‐5, and IL‐10 were secreted by mouse spleen cells restimulated with OVA after intranasal co‐administration of OVA together with rCTB, and in response to CT, the same cytokines were secreted. The different effect of rCTB on Mφ from that of CT may mean a difference between the mechanisms of rCTB and CT during the early stage of an immune response.
Identification and Characterization of the sodA Genes Encoding Manganese Superoxide Dismutases in Vibrio parahaemolyticus, Vibrio mimicus, and Vibrio vulnificus
Authors: Kimoto Ryoko , Funahashi Tatsuya , Yamamoto Noriko , Miyoshi Shin‐ichi , Narimatsu Shizuo , Yamamoto Shigeo 
Published: November 2013

AbstractSequencing of Fur titration assay‐positive clones obtained from genomic DNA libraries of Vibrio parahaemolyticus, V. mimicus and V. vulnificus revealed open reading frames encoding proteins of 202, 205 and 202 amino acid residues, respectively. Each open reading frame was preceded by a predicted Fur box which overlaps a likely promoter with similarity to the −10 and −35 consensus sequence of Escherichia coli. The deduced amino acid sequences shared considerable homology with bacterial Mn‐containing superoxide dismutases (MnSODs). Consistent with this, these Vibrio strains produced proteins with SOD activity resistant to inhibition by H2O2 and KCN only when grown under iron‐limiting conditions. Primer extension analysis of the total RNA from these vibrios revealed iron‐repressible expression of the genes. Furthermore, when grown under iron‐limiting conditions, E. coli carrying a plasmid with each cloned gene over‐expressed protein with the same electrophoretic mobility and insensitivity of SOD activity to H2O2 and KCN. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis followed by N‐terminal amino acid sequencing revealed that proteins (MnSODs) having N‐terminal amino acid sequences consistent with those deduced from the corresponding genes were present in cell lysates of the vibrios grown under these iron‐limited conditions. These results demonstrate that the genes cloned in this study are sodA homologs encoding MnSODs, whose expression is regulated by the iron status of the growth medium. PCR using a primer set based on the V. parahaemolyticus sodA sequence revealed the presence of homologous genes in certain other Vibrio species.
Plasminogen Activator Production in a Rat Model of Pneumocystis carinii Pneumonia
Authors: Angelici Elena , Contini Carlo , Spezzano Massimiliano , Romani Roberto , Carfagna Paolo , Serra Pietro , Canipari Rita 
Published: November 2013

AbstractSeveral studies have indicated that the serine protease urokinase‐plasminogen‐activator (uPA) is an important factor in host defense against pulmonary pathogens. To gain a better insight into the role of uPA in Pneumocystis carinii (P. carinii) pneumonia (PCP), we evaluated PA production in alveolar macrophages (AMs) obtained from rats with steroid‐induced PCP. Treatment with cortisone acetate favored PCP in 91% of rats. In the bronchoalveolar lavage (BAL) samples of immunosuppressed rats both with and without PCP, we observed a decrease in uPA activity as well as a decrease in cell number. Urokinase‐PA production by AMs was reduced in rats treated with cortisone alone. However, an increase in cell‐associated uPA was observed in rats with PCP. This increase appears to be produced in response to P. carinii infection. In fact, when AMs obtained from untreated healthy or immunosuppressed uninfected rats were challenged with P. carinii, a significant increase in PA activity in cell lysates was observed, though a lower response was obtained in cortisone‐treated animals. Our results suggest that healthy AMs respond to the presence of P. carinii with an increase in uPA production and that this response in immunodepressed rat‐AMs is partially impaired.
Enterocolitis Caused by Methicillin‐Resistant Staphylococcus aureus: Molecular Characterization of Respiratory and Digestive Tract Isolates
Authors: Watanabe Hiroshi , Masaki Hironori , Asoh Norichika , Watanabe Kiwao , Oishi Kazunori , Kobayashi Shinobu , Sato Akiyoshi , Nagatake Tsuyoshi 
Published: November 2013

AbstractWe investigated the mechanism of outbreak of enterocolitis caused by methicillin‐resistant Staphylococcus aureus (MRSA). Five epidemiological markers [coagulase type, enterotoxin type, toxic shock syndrome toxin‐1 (TSST‐1) production, β‐lactamase production and pulsed‐field gel electrophoresis (PFGE)] of 45 strains of MRSA isolated simultaneously from the respiratory tract (nasal cavity and/or pharynx and/or sputum) and stool (plus one sample of gastric juice) in 13 patients (8 males and 5 females, mean age, 77.1 years) were compared retrospectively. Forty‐four of the 45 isolates of MRSA were positive for enterotoxin C and TSST‐1 production, and the remaining isolate was positive for enterotoxin A and negative for TSST‐1 production. All isolates were coagulase type II, and 27 showed β‐lactamase production. The patterns of coagulase type, enterotoxin type, TSST‐1 and β‐lactamase production of MRSA isolated from the respiratory tract were similar to those of MRSA isolated from the intestine in 12 of 13 patients. Molecular typing by PFGE demonstrated that the pattern of respiratory tract isolates was identical to those of stool isolates in 9 (69.2%), similar in 3 (23.1%), and different in 1 (7.7%). The data suggested that enterocolitis might be caused by the MRSA colonized in the respiratory tract and incorporated into the digestive tracts. Therefore, we propose that early eradication of MRSA in the respiratory tract is important for protection of patients against the development of enterocolitis, particularly in susceptible patients, e.g., immunocompromised or pre‐operated patients with digestive diseases, especially malignant disease.
DNA Substrates Influence the Recombination Efficiency Mediated by FLP Recombinase Expressed in Mammalian Cells
Authors: Nakano Masakazu , Ishimura Masakazu , Chiba Joe , Kanegae Yumi , Saito Izumu 
Published: November 2013

AbstractThe FLP recombinase derived from Saccharomyces cerevisiae mediates precise site‐specific recombination between a pair of FLP recognition targets (FRTs). Like the Cre/loxP system derived from bacteriophage P1, the FLP/FRT system has recently been applied to gene regulation systems using an FLP‐expressing recombinant adenovirus (rAd) (Nakano et al, Nucleic Acids Res. 29: e40, 2001). In an attempt to improve the FLP/FRT system by altering its DNA substrates, we compared the recombination efficiency among different substrates by a quantitative in vitro assay using FLP expressed in mammalian cells. Unexpectedly, we found that one linearized DNA substrate showed 4‐ to > 20‐fold lower recombination efficiency than other substrates, which phenomenon has not been observed in the Cre/loxP system. The quantitative in vitro assay using truncated DNA substrates suggested that the recombination efficiency seemed to be influenced not only by the linearized position of the substrate, but also by the length between a pair of FRTs. Such substrate preference of FLP expressed in mammalian cells should probably be noted when designing versatile applications of the FLP/FRT system as a gene regulation system in mammalian systems. Fortunately, however, we demonstrated that no substrate preference was observed when using a particular substrate (pCAFNF5) and the preference was reduced when using a certain pair of mutant FRTs (f72), which will also be a promising tool for simultaneous gene regulation in combination with wild‐type FRT.
Bacterial Arylsulfate Sulfotransferase as a Reporter System
Authors: Yun Hee‐Jeong , Kwon Ae‐Ran , Choi Eung‐Chil 
Published: November 2013

AbstractTo investigate whether the arylsulfate sulfotransferase (ASST) is suitable as a reporter system for monitoring gene expression, a reporter vector carrying the fragments of the astA coding region without the promoter region was constructed and designated as pSY815. As a test of the ASST reporter system's suitability, the regulatory regions of ermC and lacZ were inserted upstream of the coding region of the reporter gene to generate pSY815‐EC and pSY815‐LZ, respectively. In the absence of the inserted regulatory regions, the plasmids displayed very low background activities in Bacillus subtilis and Escherichia coli. The ASST activity under the control of the ermC regulatory region was increased 4.4‐fold in B. subtilis when induced by 0.1 μgml–1 of erythromycin. These results were consistent with a lacZ reporter gene assay of the ermC regulatory region. Furthermore, we confirmed that the lacZ promoter in E. coli was strongly induced to a 17.9‐fold increase by 0.05 mM of isopropyl‐β‐D‐thiogalactopyranoside (IPTG) in this reporter system. These results indicate that the ASST is a suitable reporter system. The lack of endogenous activity, the simple detection of enzyme activity in the living cell, the commercially available non‐toxic substrates, and the high sensitivity make ASST a useful genetic reporter system for monitoring gene expression and understanding gene regulation.
Menadione‐Catalyzed Luminol Chemiluminescent Assay for the Viability of Escherichia coli ATCC 25922
Authors: Yamashoji Shiro , Takeda Mitsue 
Published: November 2013

AbstractEscherichia coli ATCC 25922 produced O2− in the presence of menadione, and O2−‐dependent luminol chemiluminescence intensity was proportional to colony‐forming unit (CFU) in the exponential phase. CFU was determined by using a 96‐well plate at a range of 3 × 103 to 8 × 107 CFU/well (0.1 ml) after a 10‐min incubation with menadione, followed by chemiluminescent assay for 5 s. After a 4‐hr incubation of E. coli (105 CFU/0.1 ml) with menadione and an antimicrobial agent inhibiting the synthesis of peptidoglycan, protein, and DNA, the inhibitory concentration (IC) of the antimicrobial agent determined by menadione‐catalyzed luminol chemiluminescent assay was in good agreement with minimal inhibitory concentration (MIC) of the NCCLS (National Committee for Clinical Laboratory Standard) method requiring 18 hr. Menadione‐catalyzed luminol chemiluminescent assay is expected to be useful for the rapid determination of cell viability under the conditions of various cell growths and stresses.
Role of Interleukin (IL)‐1 Type 1 Receptor in Mycobacterial Infection
Authors: Sugawara Isamu , Yamada Hiroyuki , Hua Shucheng , Mizuno Satoru 
Published: November 2013

AbstractIt is important to gain a better understanding of IL‐1‐mediated signaling events in mycobacterial infection. In order to clarify the role of IL‐1 receptor type 1 (IL‐1 R1) in IL‐1 R1, knockout (KO) mice were infected with either Mycobacterium tuberculosis H37Rv or Kurono strain by the respiratory route, and their ability to control mycobacterial growth, pulmonary granuloma formation, and cytokine mRNA expression was investigated. IL‐1 R1 KO mice developed significantly larger (P < 0.01) granulomatous lesions with neutrophil infiltration in their lungs than wild‐type mice did after infection with the M. tuberculosis Kurono strain. The number of mycobacterial colonies in lungs and spleen increased from five weeks post‐infection. Interferon‐γ production by spleen cells was low in IL‐1 R1 KO mice. It is concluded that the IL‐1 R1 is essential for IL‐1‐mediated signaling events in mycobacterial infection.
Staphylococcus aureus Requires Increased Level of Ca2+ or Mn2+ to Grow Normally in a High‐NaCl/Low‐Mg2+ Medium
Authors: Ochiai Toshiro 
Published: November 2013

AbstractMg2+‐availability in Staphylococcus aureus cells decreased significantly with increasing NaCl concentration in growth media. Cells grew in a NaCl‐free, Chelex resin‐treated complex medium only if the medium was supplemented with 50 μM MgCl2, while, growth was limited when the medium was further supplemented with 1.0 M NaCl. Cells grown in such a high‐NaCl/low‐Mg2+ medium exhibited the morphologic abnormality of larger than normal cells. Both sufficient growth and normal cell morphology were restored by increasing Mg2+ concentration in a high‐NaCl medium, or by supplementation with either CaCl2 or MnSO4 in a high‐NaCl/low‐Mg2+ medium. Supplementing with BaCl2, SrCl2 or FeSO4, however, had no effect. These results indicate that Ca2+ and Mn2+ might play some essential role in the growth of Staphylococcus aureus in a high‐NaCl/low‐Mg2+ environment.
Delayed Onset of Systemic Bacterial Dissemination and Subsequent Death in Mice Injected Intramuscularly with Streptococcus pyogenes Strains
Authors: Saito Mitsumasa , Kajiwara Hideko , Ishikawa Takahiko , Lida Ken‐ichiro , Endoh Miyoko , Hara Toshiro , Yoshida Shin‐ichi 
Published: November 2013

AbstractStreptococcus pyogenes causes severe invasive diseases in humans, including necrotizing fasciitis, sepsis, and streptococcal toxic shock syndrome (STSS). We found that mice infected intramuscularly (i.m.) with S. pyogenes strains developed bacteremia and subsequent sudden death after at least 10 days of a convalescent period. Mostly, it occurred more than 21 days after muscle infection. We provisionally designate this phenomenon as “delayed death.” Just after muscle infection, all the mice lost weight and activity, but recovered completely within 3 days. They had kept good activity and a fine coat of fur till one or two days before their death. Some of the dead mice were found to have soft‐tissue necrosis. There was no correlation between the virulence leading to the delayed death and the severity of diseases from which strains were isolated. It was also found that the production of neither streptococcal pyrogenic exotoxin (SPE) A nor B correlated to the virulence leading to delayed death. The bacteria obtained from the organs of the mice with delayed death expressed capsule. We suggest that the mice with delayed onset of systemic bacterial dissemination and subsequent death after muscle infection with S. pyogenes are the animal models of STSS, because the pathophysiology is extremely similar to that of human STSS.
Ultrastructural Features of Borrelia garinii in Naturally Infected Voles, Clethrionomys rufocanus, with Special Reference to the Relationship with Peripheral Nerves
Authors: Yano Yasuhiro , Ishiguro Fubito , Nakata Keisuke , Takada Nobuhiro 
Published: November 2013

AbstractThe ultrastructural features of Borrelia garinii found in the ear tissues of the vole, Clethrionomys rufocanus, are described. The spirochetes were observed in the interstitium of connective tissue or in contact with fibroblasts and were occasionally situated in the endoneurium of the peripheral nerves. The spirochetes did not injure or enter into the fibroblasts or Schwann cells. The cytotoxicity and migration of the spirochetes are also discussed.