bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sequence Analysis of the Gene Encoding a Spotted Fever Group‐Specific Intracytoplasmic Protein PS120 of Rickettsia japonica
Authors: Uchiyama Tsuneo 
Published: November 2013

AbstractThe 3,438‐nucleotide (nt) sequence containing a 3,054‐nt open reading frame of the gene (rps120) encoding an antigenic, intracytoplasmic, spotted fever group‐specific and heat‐stable 120‐kilodalton protein (PS120) of Rickettsia japonica was determined. The nt and deduced 1,018 amino‐acid (aa) sequences were compared to those of R. conorii since only those of this species had been determined among SFG rickettsiae. The homologies of these sequences between R. japonica and R. conorii were considerably high at 97 and 95%, respectively. These high homologies were comparable to those of β‐peptides encoded by the ompB genes among SFG rickettsiae. It was also found that the genome of R. prowazekii contained a nt sequence with 68% homology to that of the rps120 gene of R. japonica.
Complete Nucleotide Sequence and the Genome Organization of Tobacco Leaf Curl Geminivirus from Japan
Authors: Shimizu Sachiko , Ikegami Masato 
Published: November 2013

AbstractThe genomic DNA of tobacco leaf curl geminivirus (TLCV) from tomato plants with leaf curl disease in Japan has been sequenced. The single circular DNA molecule comprises 2,761 nucleotides. TLCV DNA contains six open reading frames (ORFs) capable of encoding proteins with a molecular weight greater than 10 K. In total nucleotide sequence comparisons with other geminiviruses, TLCV was most closely related to tomato leaf curl virus from Taiwan (TwToLCV) (76% identity), tomato leaf curl virus from Bangalore (ToLCV‐Ba) (74%) and agerantum yellow vein virus (AYVV) (74%), all possessing a monopartite genome. The significant but relatively low sequence similarity in the genomic DNA between TLCV and other geminiviruses suggests it is a distinct geminivirus in genus Begomovirus.
Genetic Diversity and the Absence of Regional Differences of Borrelia garinii as Demonstrated by ospA and ospB Gene Sequence Analysis
Authors: Yabuki Mihe , Nakao Minoru , Fukunaga Masahito 
Published: November 2013

AbstractUnfed adult Ixodes persulcatus ticks were collected from four locations of Nagano and Hokkaido in Japan. Infected Borrelia garinii were investigated by PCR‐RFLP of the ospA and ospB gene sequences. The primer set amplified an approximately 1.6‐kb DNA fragment (0.7‐kb in some strains), and BsrI, BstYI, or NlaIII digestion of the product resulted in six distinctively different PCR‐RFLP groups and two independent borrelial strains. The representatives in each PCR‐RFLP group and individuals from the borrelial strains were sequenced, and their deduced amino acid sequences were aligned. A neighbor‐joining phylogenetic analysis showed that the B. garinii OspA or OspB sequences were each divided into three major clusters including isolates from both the Nagano and Hokkaido locations. There was no local difference in OspA/B sequences between Nagano and Hokkaido. The osp gene of Borrelia burgdorferi sensu lato is highly heterogeneous, and this was also confirmed by our sequence analysis. Some strains of the different PCR‐RFLP groups had closely related OspA sequences, while the OspB sequences of these strains were quite different. These findings suggested intraspecies gene exchange and recombination events between the two genes in B. garinii.
Complement Receptor Type 3 Plays an Important Role in Development of Protective Immunity to Primary and Secondary Corynebacterium pseudotuberculosis Infection in Mice
Authors: Lan Dinh Thi Bich , Makino Sou‐Ichi , Shirahata Toshikazu , Yamada Manabu , Nakane Akio 
Published: November 2013

AbstractThe present study was performed to investigate the role of CR3, the type 3 complement receptor, in host defense against primary and secondary Corynebacterium (C.) pseudotuberculosis infection in mice. Treatment of mice with 5C6, an anti‐CR3 monoclonal antibody (mAb), resulted in unrestricted multiplication of bacteria in the organs and dramatically increased mortalities of the infected mice. Histological examinations showed the inflammation, degeneration and necrosis of organs and revealed that the infection‐enhancing effect of 5C6 mAb was associated with the failure of mice to focus mononuclear phagocytes at sites of bacterial multiplication. These results suggest that CR3 plays an important role in host defense against primary as well as secondary C. pseudotuberculosis infection in mice.
Simple Preparation of Parapoxvirus Genome DNA for Endonuclease Analysis
Authors: Inoshima Yasuo , Morooka Akira , Murakami Kenji , Sentsui Hiroshi 
Published: November 2013

AbstractWe compared five methods for improved extraction of very‐large parapoxvirus DNA from infected cells: (i) alkaline‐lysis procedure followed by phenol extraction; (ii) modified Hirt procedure, which was a neutral lysis procedure followed by phenol extraction; (iii) Hirt procedure; (iv) method used for extraction of vaccinia virus DNA; and (v) standard procedure using virus purification with an ultracentrifuge and protease‐sodium dodecyl sulfate‐phenol treatment. The alkaline‐lysis procedure was more rapid, inexpensive and simpler than the other methods. Moreover, with this method it is not necessary to prepare any special facilities, reagents and kits. Although the extracted DNA was still crude, we could reproducibly prepare viral DNA from 2 × 106 infected cells in less than 2 hr and it could be readily digested by restriction endonuclease. This method will aid rapid genetic classification of parapoxvirus.
CPR‐Total (TAFI and Activated TAFI) Levels in Plasma/Serum of Hemophiliacs
Authors: Guo Xiaoyan , Okada Noriko , Okada Hidechika 
Published: November 2013

AbstractArginine carboxypeptidase (CPR) is a single‐chain plasma protein generated during coagulation from a precursor (proCPR). proCPR is the same molecule as thrombin activable fibrinolysis inhibitor (TAFI), which retards fibrin clot lysis in vitro and most likely modulates fibrinolysis in vivo. In this study, the amount of CPR‐total, which includes proCPR (TAFI) and CPR (activated TAFI), in hemophiliac patients was evaluated using a newly developed enzyme linked immunosorbent assay (ELISA). The amount of CPR‐total in plasma or serum of most of the hemophiliac patients was in the range of healthy individuals. There was no significant difference in hemophiliac patients with or without HIV‐1 infection. However, two out of the 74 hemophiliac patients showed a significantly high level. The upregulation of CPR‐total might contribute to compensate for inefficient coagulation in some hemophiliac individuals.
Molecular Characterization and Functional Analysis of a secA Gene Homolog in Actinobacillus actinomycetemcomitans
Authors: Novak Karen F. , Nonnemacher Michael R. , Pourhamidi Jaleh 
Published: November 2013

AbstractResults of Southern blot analyses and polymerase chain reaction revealed that the Gram‐negative pathogen, Actinobacillus actinomycetemcomitans, harbored DNA homologous to the secA gene of Escherichia coli. In E. coli, the secA gene product is essential for translocation of proteins across the inner membrane via the Sec system. This A. actinomycetemcomitans secA homolog was cloned and its nucleotide sequence determined. Amino acid sequence analysis of the cloned gene revealed significant homology to the SecA proteins of Haemophilus influenzae, E. coli, Caulobacter crescentus and Bacillus subtilis. Although the cloned gene did not complement a temperature sensitive mutation in the E. coli secA gene, strains harboring the cloned gene did produce a protein that cross‐reacted with anti‐SecA antibody. In addition, the cloned gene did restore sensitivity to sodium azide in an E. coli azideR mutant. These data support the hypothesis that A. actinomycetemcomitans may use a system similar to the Sec system of E. coli to transport proteins across the cytoplasmic membrane, but suggest that the A. actinomycetemcomitans gene product may require genera‐specific Sec proteins to complement some Sec mutations in E. coli.
Analysis of Vibrio mimicus Clinical Strains by Arbitrarily Primed Polymerase Chain Reaction
Authors: Bi Kewei , Shi Lei , Maehara Yoko , Miyoshi Shin‐Ichi , Tomochika Ken‐Ichi , Shinoda Sumio 
Published: November 2013

AbstractA total of 51 Vibrio mimicus clinical strains from different geographic locations were examined by arbitrarily primed polymerase chain reaction (AP‐PCR). The primer VMH‐3 divided them into 28 groups, although 18 groups consisted of a single strain at present. All groups had a common 1.0‐kb amplification fragment. Most of the groups consisted of strains from same region, although two exceptional groups showed a few amplification fragments including strains from different regions. AP‐PCR groups were not consistently associated with serogroups. AP‐PCR is thought to be a valuable and easy method for the epidemiological study of V. mimicus.
Phylogenetic Analysis of the Small Hydrophobic (SH) Gene of Mumps Virus in Korea: Identification of a New Genotype
Authors: Kim Sang Hyun , Song Ki‐joon , Shin Young Kyoo , Kim Jee Hee , Choi Soon Mi , Park Kwang Sook , Baek Luck Ju , Lee Yong Ju , Song Jin‐won 
Published: November 2013

AbstractViral RNAs extracted from fifteen mumps virus isolated from throat swab, saliva, blood, urine or CSF during mumps epidemics between 1997–1998 in Korea were amplified by reverse transcriptase‐polymerase chain reaction (RT‐PCR) and compared by nucleotide sequencing of the small hydrophobic (SH) gene. The deduced amino acid sequences of the SH gene were aligned with the published sequences of mumps virus isolated in different geographic areas. A comparison of the SH gene of mumps viruses in Korea indicated 96.2–100% and 91.2–100% similarity at the nucleotide and amino acid levels, respectively. Phylogenetic analysis, using the neighbor‐joining method, showed that Korean mumps virus strains formed a genetically distinct monophyletic group from previously reported genotypes based on the 315‐bp length nucleotide and 57 deduced amino acid sequences of the SH gene, and possibly be designated as a new genotype (I).
Disintegration of Mg2+‐Induced Hexagonal Assembly of an R‐Form Lipopolysaccharide from Klebsiella pneumoniae by Treatment with CaCl2
Authors: Ito Hideo , Naito Setsuko , Kato Nobuo 
Published: November 2013

AbstractR‐form lipopolysaccharide (LPS) from Klebsiella pneumoniae strain LEN‐111 (O3—: K1—), which was precipitated by the addition of 2 volumes of ethanol containing 10 mm MgCl2 for the purification process, ultrastructurally exhibited membrane pieces consisting of an ordered hexagonal lattice structure with a lattice constant of 14 to 15 nm. When the R‐form LPS was suspended in 50 mm tris (hydroxymethyl) aminomethane buffer (at pH 8.5) containing 1 mm or higher concentrations of CaCl2 and kept at 4 C for 10 hr, the ordered hexagonal lattice structure of the R‐form LPS was disintegrated and changed to an irregular rough, mesh‐like structure. By treatment with CaCl2, the content of Mg in the LPS was markedly decreased, and conversely, the content of Ca was increased to a level depending upon the concentration of CaCl2. Results indicate that the addition of CaCl2 to suspensions of the Mg‐bound R‐form LPS result in a tighter binding of Ca2 to the R‐form LPS and the release of Mg2 from the R‐form LPS, and as a consequence, destroys the Mg2+‐induced ordered hexagonal lattice structure of the R‐form LPS.
Chemical and Serological Properties of Lipopolysaccharides from Vibrio parahaemolyticus O‐Untypeable Strains Isolated from Patients
Authors: Hashii Noritaka , Kondo Seiichi , Iguchi Takehiro , Nishibuchi Mitsuaki , Hisatsune Kazuhito 
Published: November 2013

AbstractChemical and serological studies have been carried out on the O‐antigenic lipopolysaccharides (LPS) of six strains, U‐6443, W‐90144, X‐3972, AD‐7999, 90A‐6611 and KX‐V212, of Vibrio parahaemolyticus isolated from patients. The O‐serotypes of these strains have not been identified because they were not agglutinated by any diagnostic antisera against known O‐serotype strains. A compositional sugar analysis of their LPS revealed that out of the six O‐untypeable (OUT) strains, U‐6443, W‐90144 and AD‐7999 strains belonged to chemotype II (chemotype of O2), 90A‐6611 and KX‐V212 strains to chemotype III (chemotype of O3, O5, O11 and O13) and X‐3972 strain to chemotype IV (chemotype of O4). A structural analysis of LPS isolated from KX‐V212 revealed that the inner core region of the LPS consisted of only one mole of 2‐keto‐3‐deoxy‐d‐manno‐octonic acid, which carried a phosphate group at position C4 and the outer core at position C5. In passive hemolysis tests performed by using LPS as the antigen to sensitize sheep red blood cells (SRBC), and diagnostic antisera (O1 to O11) or anti‐whole‐cell rabbit antisera raised against O12, O13 and the six OUT strains, strong cross‐reactivity was observed among LPS derived from the strains belonging to chemotype II (U‐6443, W‐90144, AD‐7999 and O2). Strong cross‐reactivity was also observed between X‐3972 (chemotype IV) and O4 LPS. In contrast, LPS from two of the strains belonging to chemotype III (90A‐6611 and KX‐V212) did not react with any of the antisera raised against known O‐serotypes. Cross‐absorption tests showed that the O‐antigens of U‐6443, W‐90144 and AD‐7999 were identical to that of O2, and the O‐antigen of X‐3972 to that of O4. On the other hand, after the absorption of antisera raised against 90A‐6611 and KX‐V212 with O2 cells, the hemolytic activities against SRBC sensitized with homologous LPS were still retained at a high titer, whereas the hemolytic activities against SRBC sensitized with LPS from other O‐serotype strains were completely eliminated. A cross‐absorption test revealed that the O‐antigens of these two strains were identical to each other. Thus, it was demonstrated that the O‐serotype of OUT strains 90A‐6611 and KX‐V212 was not involved in the known O‐serotypes; rather it represented a novel serotype which has not hitherto been reported.
Acyclovir Resistance in Herpes Simplex Virus Isolates from Keratitis Cases: An Analysis from a Developing Country
Authors: Pramod Naranatt Padmanabhan , Thyagarajan Sadras Panchatcharam , Mohan Keta Venkata Krishna , Anandakannan Kanagasabai 
Published: November 2013

AbstractSeven herpes simplex type‐1 (HSV‐1) isolates from herpes simplex keratitis (HSK) cases clinically resistant to acyclovir (ACV) were analyzed for the mechanism of ACV resistance in them. The purpose of the study was to focus the attention of ophthalmologists on the frequency of occurrence of ACV resistance in HSK and to characterize such a phenomenon. We employed in‐vitro plaque reduction assay, thymidine kinase assay, polymerase chain reaction, single‐strand confirmation polymorphism analysis and sequencing to detect any mutation(s) in thymidine kinase gene in this analytical study. Four of the seven HSV‐1 isolates proved ACV resistant by plaque reduction assay and three of them showed reduced thymidine kinase activity. Altered mobility pattern indicative of mutation within 335 base pair PCR product bracketing the suggested homopolymer mutational hotspot (7 Guanosine) was detected in 2 of these 3 isolates. DNA sequencing showed a deletion at nucleotide position 336 from the tk gene transcription start in both the isolates. This mutation has generated the first TGA stop codon 27 nucleotides downstream in the tk open reading frame. Our study also suggests the need of clinical/molecular surveillance of ACV resistance in HSV types in a given geographic location for better management of HSV infections.
Cloning and Sequencing of the Gene Encoding the Candidate HtrA of Rickettsia typhi
Authors: Kim Jong‐hyun , Hahn Myong‐joon 
Published: November 2013

AbstractWe screened a phage library of Rickettsia typhi with a polyclonal antiserum to clone genes which encode immunogenic proteins of R. typhi. Among several clones obtained, one clone codes for a 466‐amino‐acid protein similar to the heat‐shock protein, HtrA. The deduced rickettsial HtrA contains a putative signal peptide sequence at the N‐terminus, a serine protease‐like domain, and two PDZ domains. The recombinant protein of rickettsial HtrA reacted with sera from patients with murine typhus and tsutsugamushi disease. We suggest that this gene and its recombinant protein would be valuable for the immunologic diagnosis of rickettsial diseases.
Protective Immune Responses Induced by a Non‐Pathogenic Simian/Human Immunodeficiency Virus (SHIV) against a Challenge of a Pathogenic SHIV in Monkeys
Authors: Yoshino Naoto , Ami Yasushi , Someya Kenji , Ando Shuji , Shinohara Katsuaki , Tashiro Fumio , Lu Yichen , Honda Mitsuo 
Published: November 2013

AbstractA simian/human immunodeficiency virus (SHIV)‐NM3n containing the human nef, but not the monkey nef, and vpr genes of SIV was inoculated into two cynomolgus monkeys, resulting in systemic infection with a minimum level of transient virus load. In order to study the nature of immune responses associated with the prevention of a pathogenic SHIV, the SHIV‐NM3n‐inoculated monkeys and three naive monkeys were intravenously challenged with a pathogenic SHIV containing the envelope gene of HIV‐1 89.6. After the heterologous virus challenge, all of the SHIV‐NM3n‐inoculated animals completely avoided the loss of CD4+ T lymphocytes in PBMC as well as lymphoid tissues compared to pathogenic SHIV‐injected control animals. The inhibition of CD4+ cell depletion was associated with maintaining the proliferative response of helper T‐cells against SIV p27 in the previously nonpathogenic virus‐inoculated animals following the pathogenic virus challenge. Furthermore, the decline of CD28+ cells, the increase in CD95+ cells, and the enhancement of in vitro apoptosis in PBMC were inhibited in the non‐pathogenic virus‐inoculated animals. These results suggest that nonpathogenic SHIV‐NM3n infection induces the protection of monkeys from heterologous pathogenic viruses that may be associated with blocking the change in immune responses and the cell loss induced by a pathogenic virus.
Isolation and Characterization of a Major Antigenic Component of Malassezia globosa to IgE Antibodies in Sera of Patients with Atopic Dermatitis
Authors: Koyama Tomoki , Kanbe Toshio , Ishiguro Ayako , Kikuchi Akihiko , Tomita Yasushi 
Published: November 2013

AbstractThree major components of Malassezia globosa were isolated from 2‐ME extracts of this fungus by ion‐exchange column chromatography and are referred to as Malg46a, Malg46b and Malg67, respectively. IgE antibodies to these components in the sera of patients with AD were detected by immunoblots. In Western blot, IgE antibodies to Malg46b were most frequently detected in the sera of AD patients. Dot blot with the Malg46b‐containing fraction immunologically reacted with 69% of the sera of the patients, and with 83% of the sera of the patients who were positive for IgE antibodies to the 2‐ME extract of M. globosa in the Western blot. The intensities generated for each dot correlated well with the total intensities generated for the 2‐ME extract of M. globosa in the Western blot (r=0.763). In the lectin blot, Con A reacted with both Malg46a and Malg46b but not with Malg67. The polyclonal antibody to Malg46b reacted strongly only with the 2‐ME extract of M. globosa and reacted slightly with M. restricta. In conclusion, a glycoprotein, Malg46b of M. globosa, is dominantly expressed in this fungus and is a possible major antigen for IgE antibodies in patients with AD.
Effect of the Sialyl Lewis X (SLe) Moiety on Splenic Accumulation of SLex‐Carboxymethylpullulan Conjugate
Authors: Horie Kazutoshi , Sakagami Masahiro , Kuramochi Kentaro , Ito Teruomi , Hamana Hiroshi 
Published: November 2013

AbstractSialyl Lewis X (SLex), an E‐selectin ligand, was conjugated with carboxymethylpullulan (CMPul) and the disposition characteristics of this conjugate after intravenous administration were investigated using mice with ear edema. The concentration of 3H‐labeled SLex‐CMPul in the spleen was significantly high. When CMPul was modified with a saccharide unable to bind to E‐selectin, this splenic accumulation was not observed. The uptake of radiolabeled SLex‐CMPul by the spleen was completely inhibited by a 100‐fold molar of cold SLe‐CMPul but not by a sialyl N‐acetyllactosamine‐CMPul conjugate (SLN‐CMPul). Microautoradiography analyses revealed that SLex‐CMPul accumulated in the marginal zone of the spleen.
Dithiothreitol Enhances Listeria monocytogenes Mediated Cell Cytotoxicity
Authors: Westbrook David G. , Bhunia Arun K. 
Published: November 2013

AbstractThe hybridoma Ped‐2E9 based cytotoxicity assay was developed to distinguish virulent from avirulent Listeria species in 6 hr. The cytotoxicity effect on Ped‐2E9 was reported to be primarily due the cytolytic action of listeriolysin O (LLO), produced by L. monocytogenes. In this study, the effect of a reducing agent, dithiothreitol (DTT, 0–2 mm) that is known to activate LLO was investigated to make the Ped‐2E9 based cytotoxicity assay an even more sensitive and rapid. Also, we examined the effect of fetal bovine serum (FBS, 0–50%), a common ingredient of tissue culture media on cytotoxicity. A DTT concentration of 0.5 mm gave an optimum cytotoxicity effect, which could be measured by both alkaline phosphatase (AP) and lactate dehydrogenase (LDH) assays in just 1.5–2 hr. FBS, at levels between 10 to 50%, significantly inhibited Listeria‐mediated cytotoxicity. Concentrated culture filtrates from L. monocytogenes or LLO producing recombinant L. innocua (prfA+hlyA+) strain also caused cytotoxicity effects, which were observed by scanning electron microscopy or a cytotoxicity assay in 2–3 hr. Interestingly, addition of DTT to culture filtrates produced 100% cell cytotoxicity in just 15 min. This indicated that LLO activity, which is responsible for Ped‐2E9 cytotoxicity, was augmented several folds with the addition of a reducing agent. Examination of Listeria isolates belonging to different serogroups from clinical sources or naturally contaminated meat products with DTT gave cytotoxicity results in 2 hr, which were comparable to the 5‐hr assay analyzed concurrently without DTT. These results indicated that DTT, which activated the LLO, could be used in the cytotoxicity assay to enhance Listeria‐mediated Ped‐2E9 cell cytotoxicity. This knowledge will greatly assist us to develop a user‐friendly rapid assay to screen cytopathogenic properties of Listeria species.
Blockade ofSalmonella enteritidisPassage across the Basolateral Barriers of Human Intestinal Epithelial Cells by Specific Antibody
Authors: Sugita‐Konishi Yoshiko , Ogawa Michinaga , Arai Soichi , Kumagai Susumu , Igimi Shizunobu , Shimizu Makoto 
Published: November 2013

AbstractAntibodies specific toSalmonella enteritidis(S.E.) were obtained from immunized egg yolk, and their protective effects against S.E. were studied by using monolayer‐cultured human intestinal epithelial cells, Caco‐2 and T84. TheSalmonellaadherence and entry to the cells were partially inhibited by the antibodies. The antibodies inhibited the decrease in transepithelial electrical resistance (TEER) of the intestinal epithelial monolayers and IL‐8 secretion of the cells induced by S.E. invasion. Also, the antibodies blocked the penetration of bacteria through the cell layer although they did not inhibit the growth of bacteria in the cells. Confocal microscopic photographs revealed the bacteria in the infected monolayer cells were bound to antibodies. These results indicate that anti‐S.E. antibodies may protect the cells from destruction induced by S.E. invasion in intestinal epithelial cells in addition to the partial inhibition of adhesion and invasion of S.E. at the cell surface. Passive antibodies against invasive bacteria would be useful to prevent the migration of S.E. to blood not only at the cell surface but also inside of intestinal epithelial cells.