bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quantitative SNP‐Detection Method for Estimating HIV‐1 Replicative Fitness: Application to Protease Inhibitor‐Resistant Viruses
Authors: Ibe Shiro , Fujisaki Saeko , Fujisaki Seiichiro , Morishita Takayuki , Kaneda Tsuguhiro 
Published: November 2013

AbstractWe have improved the methods for the standard competitive growth assay of human immunodeficiency virus type 1 (HIV‐1). The cloning step for the mixed viral population and subsequent genotype analysis for arbitrary numbers of clones were excluded from procedures. Instead, a single nucleotide polymorphism (SNP)‐detection step was devised for the determination of viral populations. The quantitative SNP‐detection method can rapidly estimate the proportion of wild‐type and mutant populations with high reproducibility. Consequently, this method allows manipulation of many samples within a short period. Using this new competitive growth assay, replicative fitness of drug‐resistant HIV‐1 containing an M46I amino acid mutation in the protease was assessed in the presence or absence of indinavir. Without indinavir, replicative fitness of wild‐type HIV‐1 surpassed that of M46I‐mutated HIV‐1, and the fraction of mutated virus was reduced to about 10% at passage #9. In contrast, the fraction of M46I‐mutated virus increased to >90% at passage #5 in the presence of 26.4 nM indinavir. Almost identical results were obtained for L90M‐mutated HIV‐1 with or without saquinavir. HIV‐1 can survive under indinavir pressure by acquiring M46I mutation, as with acquisition of the L90M mutation under saquinavir pressure. However, these mutations damage viral replicative fitness under natural conditions without any drugs. Subtle differences between wild‐type and mutant viruses are thus easily detected using the improved method.
Proteolytic Activation of Vibrio mimicus (Vm) Major Outer Membrane Protein Haemagglutinin (HA) with Vm‐HA/Protease: Implication for Understanding Bacterial Adherence
Authors: Alam Munirul , Miyoshi Shin‐ichi , Ahmed Kabir Uddin , Hasan Nur A. , Tomochika Ken‐ichi , Shinoda Sumio 
Published: November 2013

AbstractVibrio mimicus (Vm) haemagglutinins (HAs), such as an extracellular HA/protease (Vm‐HA/protease) and a major outer membrane protein‐HA (Vm‐OMPHA), have been recognized as the putative adherence factors for the bacterium. However, the mechanism by which HAs coordinate the adherence function of the bacterium remains as yet unknown. We report herein the positive interaction between Vm‐HA/protease and Vm‐OMPHA resulting in significant enhancement of the haemagglutinating ability. In this interaction, no cleaved polypeptide was detected; however, limited proteolysis of Vm‐OMPHA was confirmed by SDS‐PAGE. The proteolytic activation of the native cell‐associated Vm‐OMPHA by limited proteolysis was also demonstrated in several V. mimicus strains. Proteolytic activation of OMPHA was also achieved with various proteases from bacterial and eukaryotic sources. These findings may indicate a novel coordination of V. mimicus HAs in the adherence of the bacterium.
The ABCs (Antibody, B Cells, and Carbohydrate Epitopes) of Cholera Immunity: Considerations for an Improved Vaccine
Authors: Provenzano Daniele , Kováč Pavol , Wade William F. 
Published: November 2013

AbstractCholera, a diarrheal disease, is known for explosive epidemics that can quickly kill thousands. Endemic cholera is a seasonal torment that also has a significant mortality. Not all nations with extensive rural communities can achieve the required infrastructure or behavioral changes to prevent epidemic or endemic cholera. For some communities, a single‐dose cholera vaccine that protects those at risk is the most efficacious means to reduce morbidity and mortality. It is clear that our understanding of what a protective cholera immune response is has not progressed at the rate our understanding of the pathogenesis and molecular biology of cholera infection has. This review addressesV. choleraelipopolysaccharide (LPS)‐based immunogens because LPS is the only immunogen proven to induce protective antibody in humans. We discuss the role of anti‐LPS antibodies in protection from cholera, the importance and the potential role of B cell subsets in protection that is based on their anatomical location and the intrinsic antigen‐receptor specificity of various subsets is introduced.
CrossLinking of the CD69 Molecule Enhances S100A9 Production in Activated Neutrophils
Authors: Shimada Sonoko , Nakamura Manabu , Tanaka Yasuhiko , Tsutsumi Kouichiro , Katano Masayoshi , Masuko Kayo , Yudoh Kazuo , Koizuka Izumi , Kato Tomohiro 
Published: November 2013

AbstractExpression of CD69 on neutrophils and generation of anti‐CD69 autoantibodies in patients with rheumatoid arthritis (RA) have been reported. Thus natural ligands for CD69 not yet identified and/or the anti‐CD69 autoantibodies possibly affect neutrophils by evoking CD69 signaling, which may further affect joint‐composing cells in RA. However, the effect of the CD69 signaling in neutrophils remains largely unclear. To elucidate the issue, we tried to identify proteins affected by the crosslinking of CD69 on neutrophils using a proteomic approach. Specifically, CD69 on granulocyte‐macrophage colony stimulating factor (GM‐CSF)‐activated neutrophils was crosslinked by anti‐CD69 monoclonal antibodies, and then intracellular proteins were detected using 2‐dimensional electrophoresis and further identified by mass spectrometry and subsequent protein database searching. As a result, we successfully identified multiple proteins that increased their production by the CD69 signaling. Among the proteins, we focused on one of the up‐regulated proteins, S100A9 calcium binding protein (S100A9), and investigated proteome changes brought by a recombinant S100A9 in a human synovial sarcoma cell line (SW982), a human chondrosarcoma cell line (OUMS‐27), and a human T leukemia cell line (Jurkat). This revealed that the recombinant S100A9 altered proteomes of SW982 and OUMS‐27, and to a lesser extent, that of the Jurkat cells. Further, S100A9 induced IL‐1β production from neutrophils and the SW982 cells. These data suggest that unidentified natural ligands for CD69 and/or the anti‐CD69 autoantibodies possibly affect joint‐composing cell types through the increased production of S100A9 in neutrophils, providing a new insight into functions of CD69 on neutrophils in RA.
Direct Application of AvaII PCR Restriction Fragment Length Polymorphism Analysis (AvaII PRA) Targeting 644 bp Heat Shock Protein 65 (hsp65) Gene to Sputum Samples
Authors: Mun Ho‐Suk , Kim Hyun‐Ju , Oh Eun‐Ju , Kim Hong , Park Young‐Gil , Bai Gil‐Han , Do Junghwan , Cha Chang‐Yong , Kook Yoon‐Hoh , Kim Bum‐Joon 
Published: November 2013

AbstractTo evaluate the usefulness of the AvaII PRA method targeting 644‐bp hsp65 gene for the direct detection of pathogenic mycobacteria from clinical specimens, we applied this method to 40 sputum samples and compared the results to those obtained by IS6110 PCR. Although this method showed a sensitivity slightly lower than IS6110 PCR (97.5% vs. 100%), it detected infections of M. avium complex (MAC) in two patients, which was not possible by IS6110 PCR. We conclude that AvaII PRA is a highly effective method for directly detecting pathogenic mycobacteria in primary clinical specimens.
Diversity in the Oligomeric Channel Structure of the Multidrug Efflux Pumps in Pseudomonas aeruginosa
Authors: Yoshihara Eisaku , Eda Shima 
Published: November 2013

AbstractMexAB‐OprM, the multidrug efflux pump of Pseudomonas aeruginosa, contributes to the high resistance of this organism to a wide variety of antibiotics. To investigate the structure and function of OprM, the outer membrane channel of MexAB‐OprM, we examined the oligomeric states of OprM and its homologues OprJ and OprN. These proteins were treated with crosslinking reagent after their reconstitution into liposome membranes. The crosslinked products indicated that OprM and OprN formed trimers, while OprJ unexpectedly appeared to form a tetramer. In order to test whether differences in oligomeric structure might be intimately related to channel function, we examined the channel‐forming activity of these proteins by liposome swelling assay. However, no significant differences in channel characteristics were detected among OprM, OprJ, and OprN. We proposed the probable explanation for the diversity in the oligomeric structure of the channel proteins.
Mycobacterial Mammalian Cell Entry Protein 1A (Mce1A)‐Mediated Adherence Enhances the Chemokine Production by A549 Alveolar Epithelial Cells
Authors: Kohwiwattanagun Juthaporn , Kawamura Ikuo , Fujimura Takao , Mitsuyama Masao 
Published: November 2013

AbstractMycobacterial mammalian cell entry protein 1A (Mce1A) is involved in the uptake of bacteria in non‐phagocytic cells and also possibly in granuloma formation. However, it has not been clarified whether the interaction between mycobacterial Mce1A and epithelial cell induces chemokine and cytokine production which is required for granuloma formation. To this end, we infected A549 alveolar epithelial cells in vitro with E. coli expressing Mce1A on the cell surface and examined the resultant chemokine/cytokine production. Mce1A promoted bacterial adherence and internalization of E. coli into A549 cells, and these recombinant bacteria induced high levels of MCP‐1 and IL‐8 production, compared to E. coli harboring the plasmid vector alone. Chemokine production was enhanced by the internalization of recombinant E. coli expressing Mce1A because cytochalasin D treatment partially inhibited MCP‐1 and IL‐8 production. However, Mce1A‐coated latex beads did not induce the chemokine production. These results suggest that although Mce1A does not induce production of chemokines, it may promote chemokine induction by augmenting the interaction between bacteria and epithelial cells.
Genotyping and Quantitation of Noroviruses in Oysters from Two Distinct Sea Areas in Japan
Authors: Nishida Tomoko , Nishio Osamu , Kato Masahiko , Chuma Takehisa , Kato Hirotomo , Iwata Hiroyuki , Kimura Hirokazu 
Published: November 2013

AbstractNorovirus (NV) is a causative agent of acute gastroenteritis in humans, and shellfishes including oysters act as major vehicles of the virus. To investigate the genetic characteristics of NVs, we collected 1,512 oysters for raw consumption between October 2002 and March 2005 from two distinct areas (area A: the Sanriku Sea area; area B: the Setouchi Sea area). We detected the capsid gene and subjected it to phylogenetic analysis. By further quantification of the copy number of the genome by using real‐time PCR, the NV capcid gene was detected in approximately 5% of the oysters, and they showed wide diversity. Two percent of the oysters from area B showed relatively large number of NVs, i.e., over 100 copies of capsid gene/oyster, whereas this was not observed in area A. Most of the detected NVs from oysters and humans were genetically related when the capsid region was compared. These results suggested that NVs obtained from humans and those obtained from oysters showed a potential relationship to each other and that some populations of Japanese oysters accumulated a relatively large number of NVs.
Infiltration of H‐2d‐Specific Cytotoxic Macrophage with Unique Morphology into Rejection Site of Allografted Meth A (H‐2d) Tumor Cells in C57BL/6 (H‐2b) Mice
Authors: Nomi Hayahito , Tashiro‐Yamaji Junko , Miura‐Takeda Sayako , Shimizu Tetsunosuke , Azuma Haruhito , Ueda Haruhiko , Katsuoka Yoji , Kubota Takahiro , Yoshida Ryotaro 
Published: November 2013

AbstractIt is assumed that CD8+ cytotoxic T lymphocytes (CTLs) mediate direct lysis of allografts and that their growth, differentiation, and activation are dependent upon cytokine production by CD4+ helper T lymphocytes. In the present study, the effector cells responsible for the rejection of i.p. allografted, CTL‐resistant Meth A tumor cells from C57BL/6 mice were characterized. The cytotoxic activity was associated exclusively with peritoneal exudate cells and not with the cells in lymphoid organs or blood. On day 8, when the cytotoxic activity reached a peak, 3 types of cells (i.e., lymphocytes, granulocytes, and macrophages) infiltrated into the rejection site; and allograft‐induced macrophages (AIM) were cytotoxic against the allograft Bacterially‐elicited macrophages also exhibited cytotoxic activity (≈1/2 of that of AIM) against Meth A cells, whereas the cytotoxic activity of AIM against these cells but not that of bacterially‐elicited macrophages was completely inhibited by the addition of donor (H‐2d)‐type lymphoblasts, suggesting H‐2d‐specific cytotoxicity of AIM against Meth A cells. In contrast, resident macrophages were inactive toward Meth A cells. Morphologically, the three‐dimensional appearance of AIM showed them to be unique large elongated cells having radiating peripheral filopodia and long cord‐like extensions arising from their cytoplasmic surfaces. The ultrastructural examination of AIM revealed free ribosomes in their cytoplasm, which was often deformed by numerous large digestive vacuoles. These results indicate that AIM are the H‐2d‐specific effector cells for allografted Meth A cells and are a more fully activated macrophage with unique morphological features.
Molecular Characterization of Equine Herpesvirus 1 (EHV‐1) Isolated from Cattle Indicating No Specific Mutations Associated with the Interspecies Transmission
Authors: Pagamjav Ochir , Yamada Souichi , Ibrahim El‐Sayed M. , Crandell Robert A. , Matsumura Tomio , Yamaguchi Tsuyoshi , Fukushi Hideto 
Published: November 2013

AbstractInterspecies trasmission of equine herpesvirus 1 (EHV‐1) from horse to cattle was shown by Crandell et al. (1988). Specific mutations related to the transmission were studied by comparison of five EHV‐1 isolates in cattle (BH1247, 3M20‐3, G118, G1753, and 9BSV4) using polymerase chain reaction and restriction fragment length polymorphism analysis with added sequencing. G118 and 3M20–3 were the genome type EHV‐1 P, while G1753 was the genome type EHV‐1 B. BH1247 and 9BSV4 might be other genome types. We could not identify specific mutations related to the interspecies transmission.
Gene Expression Profiling of Polymorphonuclear Leukocytes Treated with the Culture Filtrate of Aspergillus fumigatus and Gliotoxin
Authors: Higurashi Hiromi , Arai Makoto , Watanabe Akira , Igari Hidetoshi , Seki Naohiko , Kamei Katsuhiko , Kuriyama Takayuki 
Published: November 2013

AbstractPathogens of the Aspergillus species are frequently seen in deep‐seated mycoses. We previously demonstrated that the culture filtrate of Aspergillus fumigatus (CF) has immunosuppressive effects on polymorphonuclear leukocytes (PMNs), which act as the main phagocytes to hyphae of Aspergillus fumigatus (A. fumigatus). But little is known about the gene expression profiles involved in it. Therefore we investigated the changes in gene expression in human PMNs treated with CF or gliotoxin at two time points, using microarray analysis. CF and gliotoxin changed the expression of 548 and 381 genes, respectively. Only 51 genes showed the same expression patterns with the two stimulants, and CF‐induced changes in gene expression occurred comparatively earlier than those induced by gliotoxin. Among 31 genes encoding apoptosis, which were up‐ or down‐regulated in this assay, only 3 genes were similarly changed by both kinds of stimulation. Apoptosis was detected and quantified using two apoptosis assays. CF and gliotoxin changed the expessions of only 3 out of 19 regulated genes related to inflammatory mediators and receptors similarly. The up‐regulation of the gene encoding annexin 1 (ANXA1), which is known to be involved in extravasation and apoptosis of neutrophils, may play a role in the immunosuppressive effect of A. fumigatus. The difference in expression changes between CF and gliotoxin is presumed to be caused by the interaction among the components of CF and therefore the interaction is an area of interest for further investigation.
Simultaneous Detection and Genogroup‐Screening Test for Norovirus Genogroups I and II from Fecal Specimens in Single Tube by Reverse Transcription‐Loop‐Mediated Isothermal Amplification Assay
Authors: Fukuda Shinji , Sasaki Yukie , Kuwayama Masaru , Miyazaki Kazuo 
Published: November 2013

AbstractWe developed a reverse transcription‐loop‐mediated isothermal amplification (RT‐LAMP) assay for the rapid detection of noroviruses (NoVs) in a genogroup‐specific manner in a previous study. In this study, to detect NoVs more easily and simply, we have developed an RT‐LAMP assay for the simultaneous detection of NoV genogroup I (GI) and II (GII) genomes in a single tube. The genogrouping was achieved by using fluorescence‐labeled primers, and the green and red colors for GI and GII, respectively, sometimes with yellow color for GI and GII mixture, were indicated on the agarose gel under UV light at 312 nm.
A Murine Model of Esophageal Candidiasis with Local Characteristic Symptoms
Authors: Ishibashi Hiroko , Hisajima Tatsuya , Hu Weimin , Yamaguchi Hideyo , Nishiyama Yayoi , Abe Shigeru 
Published: November 2013

AbstractA simple method to establish a murine esophageal candidiasis model that displayed characteristic symptoms of the condition was developed using the sedative agent, chlorpromazine. Mice were immunosuppressed with prednisolone and were given tetracycline hydrochloride. One day later, the mice received chlorpromazine to keep them in a sedated state for about 3 hr. Under the sedated condition, they were infected with 4× 107 viable cells of Candida albicans by intra‐esophageal injection with a round‐head needle on syringe. From day 3 to day 6 post inoculation, 105‐106 colony forming units of C. albicans were recovered from the esophageal tube of each mouse and whitish, curd‐like patches were observed on most of the inner surface of the tube. Histological examination showed that C. albicans in esophageal lesions grew mainly in mycelial form. In this experimental model, intragastric administration of an itraconazole oral solution (20 mg/kg/day) was clearly effective. This model would provide a useful tool to investigate the pathogenesis of C. albicans esophageal infection and the efficacy of various antifungal agents microbiologically and symptomatically.
Serotypes, Antimicrobial Susceptibility, and gyr A Gene Mutation of Campylobacter jejuni Isolates from Humans and Chickens in Thailand
Authors: Boonmar Sumalee , Morita Yukio , Fujita Masahiro , Sangsuk Leelaowadee , Suthivarakom Karun , Padungtod Pawin , Maruyama Soichi , Kabeya Hidenori , Kato Masahiko , Kozawa Kunihisa , Yamamoto Shigeki , Kimura Hirokazu 
Published: November 2013

AbstractIn Thailand, 51% (36/70) Campylobacter jejuni isolates from humans and 68% (47/69) isolates from poultry were classified into 10 Penner serotypes (serotype B, C, R, E, G, A, K, D, I, and L) and 9 serotypes (serotype A, C, I, K, B, E, S, D, and L), respectively. The rate of antimicrobial drug resistance to nalidixic acid, ciprofloxacin, ampicillin, tetracycline, and erythromycin shown by human isolates were 96%, 96%, 29%, 57%, and 14%, while that shown by poultry isolates were 77%, 77%, 22%, 26%, and 17%, respectively. All quinolone‐resistant strains contained a mutation in the gyrA gene (T86→I86), suggesting that the strains were already widespread in Thailand.
Cryptococcus arboriformis Sp. Nov., a Novel Anamorphic Basidiomycetous Yeast Species Isolated from a Patient's Urine
Authors: Sugita Takashi , Takashima Masako , Sano Ayako , Nishimura Kazuko , Kinebuchi Takahiro , Yamaguchi Satoshi , Osanai Hiroaki 
Published: November 2013

AbstractA basidiomycetous yeast strain isolated from the urine of a 73‐year‐old Japanese patient with chronic renal failure was revealed to be a novel species by sequencing the D1/D2 26S rDNA and ITS regions of the rRNA gene. The name Cryptococcus arboriformis sp. nov. is proposed for the isolate, with IFM 54862T (=CBS 10441T =JCM 14201T) as the type strain. A phylogenetic analysis positioned Cryptococcus arboriformis in the Trichosporonales lineage and showed that it is closely related to C. haglerorum.
Duration of Neutralizing Antibody Titer after Japanese Encephalitis Vaccination
Authors: Abe Motoharu , Okada Kenji , Hayashida Kenshi , Matsuo Fujio , Shiosaki Kouichi , Miyazaki Chiaki , Ueda Kohji , Kino Yoichiro 
Published: November 2013

AbstractIn paired serum samples collected from 17 children, we measured neutralizing antibody (NTAb) titers after the second series of routine Japanese encephalitis (JE) vaccination in Japan to estimate the duration of NTAb titer when children did not receive the third series of routine vaccination by applying a random coefficient model. We also measured NTAb titers in adult serum samples to confirm the duration of NTAb titer estimated in the analysis of pediatric serum samples. In the absence of the third series of routine vaccination, 18% (3/17), 47% (8/17), 82% (14/17) and 100% (17/17) of children were estimated to become NTAb negative at 5, 10, 15, and 20 years after the second series of routine vaccination, respectively. Of 38 adults, 39.5% (15/38) became NTAb negative; the percentage was somewhat lower than that of antibody‐negative children. The results suggested that JE vaccination schedule should be reevaluated in the future.
Real‐Time PCR Assay to Detect DNA in Sera for the Diagnosis of Deep‐Seated Trichosporonosis
Authors: Mekha Nanthawan , Sugita Takashi , Ikeda Reiko , Nishikawa Akemi , Poonwan Natteewan 
Published: November 2013

AbstractDeep‐seated trichosporonosis due to Trichosporon asahii is life‐threatening and has high mortality. A real‐time PCR assay to detect T. asahii DNA in sera for diagnosis of this fungal infection was developed. The assay showed a higher sensitivity than polysaccharide antigen detection method. Our new real‐time PCR assay may be used for diagnosing deep‐seated trichosporonosis due to T. asahii.
Characterization of Prion Susceptibility in Neuro2a Mouse Neuroblastoma Cell Subclones
Authors: Uryu Masahide , Karino Ayako , Kamihara Yukiko , Horiuchi Motohiro 
Published: November 2013

AbstractIn this study, we established Neuro2a (N2a) neuroblastoma subclones and characterized their susceptibility to prion infection. The N2a cells were treated with brain homogenates from mice infected with mouse prion strain Chandler. Of 31 N2a subclones, 19 were susceptible to prion as those cells became positive for abnormal isoform of prion protein (PrPSc) for up to 9 serial passages, and the remaining 12 subclones were classified as unsusceptible. The susceptible N2a subclones expressed cellular prion protein (PrPC) at levels similar to the parental N2a cells. In contrast, there was a variation in PrPC expression in unsusceptible N2a subclones. For example, subclone N2a‐1 expressed PrPC at the same level as the parental N2a cells and prion‐susceptible subclones, whereas subclone N2a‐24 expressed much lower levels of PrP mRNA and PrPC than the parental N2a cells. There was no difference in the binding of PrPSc to prion‐susceptible and unsusceptible N2a subclones regardless of their PrPC expression level, suggesting that the binding of PrPSc to cells is not a major determinant for prion susceptibility. Stable expression of PrPC did not confer susceptibility to prion in unsusceptible subclones. Furthermore, the existence of prion‐unsusceptible N2a subclones that expressed PrPC at levels similar to prion‐susceptible subclones, indicated that a host factor(s) other than PrPC and/or specific cellular microenvironments are required for the propagation of prion in N2a cells. The prion‐susceptible and ‐unsusceptible N2a subclones established in this study should be useful for identifying the host factor(s) involved in the prion propagation.