bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genetic and Immunological Analysis of a Novel Variant of Shiga Toxin 1 from Bovine Escherichia coli Strains and Development of Bead‐ELISA to Detect the Variant Toxin
Authors: Ohmura‐Hoshino Mari , Ho Soon‐Tae , Kurazono Hisao , Igarashi Kazuei , Yamasaki Shinji , Takeda Yoshifumi 
Published: November 2013

AbstractA novel variant of Shiga toxin 1 (Stx1) was identified from bovine Escherichia coli strains. The stx1 variant genes designated as stx1v51 and stx1v52 were cloned and sequenced. The two variant genes differed each other by 2 bp, but the deduced amino acid sequences of the two Stx1 variant toxins were the same and had 94% and 92% homology to that of prototype A and B subunits of Stx1, respectively. The variant toxin designated as Stx1v52 was purified to homogeneity. Although inhibition of protein synthesis in vitro by purified Stx1v52 was almost equal to that of purified Stx1, Vero cell cytotoxicity and mouse lethality of Stx1v52 were several folds lower than those of prototype Stx1. In Ouchterlony double gel diffusion test, the precipitin line between Stx1v52 and Stx1 formed a spur against anti‐Stx1 serum but was fused against anti‐Stx1v52 serum. Stx1v52 and Stx1v52‐specific‐bead‐ELISA was developed, and both Stx1 and Stx1v52 could be detected with high sensitivity using Stx1v52 conjugate. However, Stx1v52 but not Stx1 could be detected with Stx1v52‐specific bead‐ELISA.
Electron Microscopic Observations of Orientia tsutsugamushi in Salivary Gland Cells of Naturally Infected Leptotrombidium pallidum Larvae during Feeding
Authors: Kadosaka Teruki , Kimura Eisaku 
Published: November 2013

AbstractWe performed a detailed electron microscopic observation on the escaping process of Orientia tsutsugamushi from the salivary gland cells of naturally infected trombiculid larvae into the acinar lumen of the gland during feeding on mice. In unfed larvae, many O. tsutsugamushi were intermingled with secretory granules in the cytoplasm of the salivary gland cell. O. tsutsugamushi was neither found in the acinar lumen nor observed escaping from the apical surface of the gland cell. In contrast, in the larvae fed on mice, many O. tsutsugamushi were observable in the acinar lumen. They were enveloped with the host glandular cell membrane. In salivary gland cells, secretory granules changed the distribution and accumulated in the apical region. In such cells, the majority of O. tsutsugamushi were found at the base of the cell. Some O. tsutsugamushi were pushing the glandular cell membrane outward in various degrees, showing different stages of escape. These findings suggest that larval feeding induced O. tsutsugamushi escape from salivary gland cells, that the escape was by budding, during which O. tsutsugamushi were enveloped in the host cell membrane, and that O. tsutsugamushi would be injected into the mouse skin as a mixture with mite saliva. The study also revealed the presence of many small vesicles that had the same cell wall structure as O. tsutsugamushi in the cytoplasm of the salivary gland cell. Most of them seemed to be products from degenerated Orientia.
A Novel Type of Two‐Component Regulatory System Affecting Gingipains in Porphyromonas gingivalis
Authors: Hasegawa Yoshiaki , Nishiyama So‐ichiro , Nishikawa Kiyoshi , Kadowaki Tomoko , Yamamoto Kenji , Noguchi Toshihide , Yoshimura Fuminobu 
Published: November 2013

AbstractWe surveyed the Porphyromonas gingivalis W83 genome database for homologues of FimS, the first two‐component sensor histidine kinase, which could possibly control virulence factors. Including fimS, we found six putative sensor kinase genes in the genome. The gene encoding one of the homologues was cloned from a P. gingivalis plasmid library, sequenced, and analyzed using its mutants. Two gene‐disruption mutants were created in strain ATCC 33277 by introducing a drug cassette into the gene. The mutants formed nonpigmented colonies, indicating that they might be defective in proteinase production, a characteristic of this organism. Proteinase activities, measured as arginine‐ and lysine‐specific (Rgp and Kgp gingipains, respectively) activities, of the mutants were almost half those of the parent strain. Unlike the parent and wild‐type strains, most of the gingipain activities were detected in the culture supernatant, not in cells, of the mutants. Abnormal production of gingipains was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analyses. These results strongly suggest that this newly‐discovered two‐component sensor kinase is involved in maturation and proper localization of gingipains to the outer membrane through an unknown mechanism. The gene encoding the sensor histidine kinase was designated gppX, which represents regulation (X) of gingipains and black pigmentation in P. gingivalis.
Quantities of Interleukin‐12p40 in Mature CD8α Negative Dendritic Cells Correlate with Strength of TCR Signal and Determine Th Cell Development
Authors: Fujimoto Noriki , Ishida Hideaki , Nakamura Ichiro , Ogasawara Kazumasa , Itoh Yasushi 
Published: November 2013

AbstractStrength of T cell antigen receptor (TCR) signaling drives the development of Th1 and Th2 subsets from naive T helper precursors. The quantity of interleukin‐12 (IL‐12) from antigen presenting cells (APC) is also profoundly involved in Th development. TCR signal strength and IL‐12 production from dendritic cells (DCs) are linked by CD40 ligand (CD40L) expression on activated T cells. CD40L on the activated T cells interacts with CD40 on DC, resulting in induction of IL‐12 from DCs. However, the subsets of DC in spleen that produce the IL‐12 have not been clearly identified. Purification of DC subsets itself may provide maturation signals to immature DCs. Thus, we used non‐purified mouse spleen cells to analyze IL‐12 producing cells, near to steady states, during the interaction of naive T cells either with or without agonist. Mature CD86highCD8α−DCs in spleen mainly produced IL‐12p40 after stimulation of high dose agonist. The ratio of CD40L positive T cells and IL‐12p40 secreting CD86highDCs correlated with the concentration of agonist and Th1 development. However, anti‐IL‐12 did not completely inhibit the Th1 development. Altogether, strength of TCR signaling directs Th cell development by regulating CD40L expression on T cells which determines production of IL‐12p40 from CD86highCD8α−DC via CD40.
Need to Differentiate Lethal Toxin‐Producing Strains of Burkholderia gladioli, Which Cause Severe Food Poisoning: Description of B. gladioli Pathovar cocovenenans and an Emended Description of B. gladioli
Authors: Jiao Zhenquan , Kawamura Yoshiaki , Mishima Noriko , Yang Ruifu , Li Na , Liu Xiumei , Ezaki Takayuki 
Published: November 2013

AbstractBurkholderia cocovenenans produces a lethal toxin (Bongkrekic acid) that leads to high fatality in food poisoning cases. However, B. cocovenenans was combined in Burkholderia gladioli in 1999. B. gladioli was originally described as a phytopathogenic bacteria that sometimes causes pneumonia in humans and that acts as an opportunistic pathogen. We thought that it was clinically dangerous to describe these two species without considering their pathogenicities. From our data of 16S rRNA sequence analysis, DNA‐DNA hybridization, and fatty acid analysis, we could confirm that B. cocovenenans and B. gladioli should be categorized as a single species. However the species really weaved lethal toxin‐producing strains with non‐lethal strains. To emphasize that B. gladioli contains two different pathogens, we describe a new pathovar, B. gladioli pathovar cocovenenans, for the lethal toxin‐producing strains. We provide characteristics that differentiate this lethal toxin‐producing pathovar from other phytopathogenic pathovars within B. gladioli, together with an emended description of B. gladioli.
A Multi‐Prefectural Outbreak of Shigella sonnei Infections Associated with Eating Oysters in Japan
Authors: Terajima Jun , Tamura Kazumichi , Hirose Kenji , Izumiya Hidemasa , Miyahara Michiko , Konuma Hirotaka , Watanabe Haruo 
Published: November 2013

AbstractAmong roughly one thousand incidents of shigellosis annually in Japan, approximately 70% of the cases are estimated to be associated with overseas travel. However, at the end of 2001, reports of domestically acquired Shigella sonnei infections suddenly increased. We report here the first multi‐prefectural outbreak of Shigella sonnei infections linked to the consumption of imported oysters in Japan at the end of 2001. Isolates of S. sonnei from patients epidemiologically linked to eating contaminated oysters and from the imported oysters themselves showed an indistinguishable pulsed‐field gel electrophoresis pattern and drug resistance pattern.
Contribution of a Single Hydrogen Bond between βHis81 of MHC Class II I‐Ek and the Bound Peptide to the pH‐Dependent Thermal Stability
Authors: Saito Keigo , Oda Masayuki , Sarai Akinori , Azuma Takachika , Kozono Haruo 
Published: November 2013

AbstractTo determine the energetic contribution of the hydrogen bond between βHis81 of the major histocompatibility complex class II (MHC II) molecule, I‐Ek, and the bound hemoglobin peptide (Hb), we analyzed the thermal stability of the hydrogen bond‐disrupted mutant, I‐Ek‐Hb βH81Y, in which the βHis81 residue was replaced with Tyr, by differential scanning calorimetry. The thermal stability of the I‐Ek‐Hb βH81Y mutant was lower than that of the I‐Ek‐Hb wild‐type, mainly due to the decreased enthalpy change. The difference in the denaturation temperature of the I‐Ek‐Hb βH81Y mutant as compared with that of the I‐Ek‐Hb wild‐type at pH 5.5 was only slightly smaller than that at pH 7.4, in agreement with the increased stability at an acidic pH, a unique characteristic of MHC II. Thus, the hydrogen bond contributed by βHis81 is critical for peptide binding, and is independent of pH, which can alter the hydrophilicity of the His residue.
Toll‐Like Receptor 3: A Link between Toll‐Like Receptor, Interferon and Viruses
Authors: Matsumoto Misako , Funami Kenji , Oshiumi Hiroyuki , Seya Tsukasa 
Published: November 2013

AbstractProduction of type I interferon (IFN‐α/β) by virus‐infected cells is the central event in their antiviral immune responses. In mammalian cells, IFN‐α/β gene transcription is induced through distinct signaling pathways by viral infection or by treatment with double‐stranded (ds) RNA, which is an intermediate of virus replication. Toll‐like receptor 3 (TLR3) was found to recognize dsRNA and transmit signals to activate NF‐κB and the IFN‐β promoter. Recent identification of the TLR3‐adaptor protein and its downstream signaling molecules, which are involved in IFN‐α/β production, revealed a novel IFN‐inducing pathway for an anti‐viral immune response. Here, we summarize the current knowledge of TLR3‐mediated immune responses.
Nocardia testaceus Sp. Nov. and Nocardia senatus Sp. Nov., Isolated from Patients in Japan
Authors: Kageyama Akiko , Yazawa Katsukiyo , Nishimura Kazuko , Mikami Yuzuru 
Published: November 2013

AbstractTwo actinomycete strains isolated from sputum between 1999 and 2001 in Japan were provisionally assigned to the genus Nocardia based on morphological criteria. These isolates were further studied in order to determine their specific taxonomic status. Detailed chemotaxonomic characterization and 16S rDNA gene sequence analysis of these isolates also confirmed that they belong to the genus Nocardia. The 16S rDNA sequence data of the two strains showed that they are most similar to that of Nocardia carnea and Nocardia flavorosea. However, DNA‐DNA relatedness data showed that the two strains could be distinguished from N. carnea and N. flavorosea and therefore represented two new species within the genus Nocardia. The designation of the two isolated strains are Nocardia testaceus for IFM 0937T (=JCM 12235T, DSM 44765T) and Nocardia senatus for IFM 10088T (=JCM 12236T, DSM 44766T).
Expression of Equine Interleukin‐18 by Baculovirus Expression System and Its Biologic Activity
Authors: Wu Donglai , Murakami Kenji , Liu Nihong , Konishi Misako , Muneta Yoshihiro , Inumaru Shigeki , Kokuho Takehiro , Sentsui Hiroshi 
Published: November 2013

AbstractThe equine interleukin‐18 (IL‐18) cDNA that contains the coding sequence was cloned and a recombinant baculovirus, named AcEIL‐18, was constructed. The recombinant protein of the equine IL‐18 was expressed by AcEIL‐18 and its expression was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) and Western blotting. Insect cells infected with AcEIL‐18 secreted a precursor IL‐18 with 24 kilo dalton (kDa) into the culture supernatant. Western blot analysis showed that mature equine IL‐18 about 18 kDa was also confirmed without co‐expression of caspase‐1. Culture supernatant from AcEIL‐18 infected cells showed a synergistic effect with recombinant human lnterleukin‐12 for induction of interferon‐γ gene expression in equine peripheral mononuclear cells, indicating that the recombinant equine IL‐18 expressed in this study also has biological activity without any treatment.
Development and Evaluation of a Rapid Flow‐Through Immuno Filtration Test Using Recombinant Filarial Antigen for Diagnosis of Brugian and Bancroftian Filariasis
Authors: Baskar Lakshmi Kanthan Vijaya , Srikanth Tokuluva Ramalingam , Suba Salahudeen , Mody Himansu Chirag , Desai Pradeep Kumar , Kaliraj Perumal 
Published: November 2013

AbstractThere is an imperative need to develop a rapid antibody test that can be used for diagnosis of clinical cases in travelers and expatriates, primary surveillance in areas of unknown endemicity, detection of early infection in childhood and for monitoring chemotherapeutic programs. A rapid‐format, simple and qualitative flow through immuno filtration test has been developed for the identification of total IgG antibodies to recombinant filarial antigen WbSXP‐1. This test system employs colloidal gold‐protein A reagent as the antibody capture reagent. The sensitivity and specificity of the test was evaluated in a total of 1,230 serum samples. The sensitivity of the test was found to be 90.8% with brugian (n=70) and 91.4% with bancroftian (n = 140) microfilaraemic subjects. The test showed minimum reactivity (4/10) with Loa loa microfilaria (MF) positive sera and no reactivity (0/20) with Onchocerca MF positive sera. This rapid diagnosis is found to be non‐reactive with individuals having other parasitic diseases including schistosomiasis (n = 10), soil‐transmitted helminthiases (n=34) and protozoan infections (n=33) indicating the potential of this test as a prospective method of diagnosis for both brugian and bancroftian lymphatic filariasis. Stability kinetics was studied at different temperatures and different time periods. The rapid flow‐through immuno filtration test is advantageous since it can be stored at room temperature, is user friendly and is particularly applicable in the field as an initial screening method, for epidemiological monitoring of filarial infections in bancroftian and brugian endemic regions of the world.
Analysis of Major Virulence Factors in Porphyromonas gingivalis under Various Culture Temperatures Using Specific Antibodies
Authors: Murakami Yukitaka , Masuda Takashi , Imai Masashi , Iwami Jun , Nakamura Hiroshi , Noguchi Toshihide , Yoshimura Fuminobu 
Published: November 2013

AbstractPorphyromonas gingivalis is implicated in the occurrence of adult periodontitis. We have previously identified major outer membrane proteins from P. gingivalis, which include representative virulence factors such as gingipains, a 75 kDa major protein, RagA, RagB, and putative porin. Fimbriae, another important virulence factor, exist on the cell surface. In this study, we identified major supernatant proteins. They were fimbrilin, the 75 kDa major protein, gingipains and their adhesin domains. Microscopic examination showed that supernatant proteins formed vesicle‐like and fimbrial structures. To learn more about the character of this bacterium, we examined effects of growth temperature on localization and expression of these virulence factors. In general, localization of major virulence factors did not change at the various growth temperatures used. Most of the 75 kDa major protein, RagA, RagB, and putative porin were found in the envelope fraction, not in cell‐free culture supernatant. Gingipains were found in both the envelope fraction and supernatant. More than 80% of fimbriae were associated with cells, less than 20% migrated to the supernatant. Most fimbriae existed in the whole cell lysate, although there was a small amount in the envelope fraction. When the growth temperature was increased, expression of fimbriae, gingipains, the 75 kDa major protein, RagA, and RagB decreased. However, temperature had almost no effect on expression of putative porin. The tendency for expression of major virulence factors to decrease at higher temperatures may enable P. gingivalis to survive under hostile conditions.
The Participation of Nitric Oxide in Peritoneal Exudate Cell Cytotoxicity of Mice by Fusobacterium nucleatum
Authors: Kato Chihomi , Mikami Masato , Saito Kazuko 
Published: November 2013

AbstractPreviously we reported that mice infected recurrently with live Fusobacterium nucleatum (Fn) synthesize a significant amount of NO between 12 hr and 24 hr after Fn injection. Fn is a gram‐negative rod periodontal pathogen. NO could not be induced by heat‐killed Fn or in untreated mice. This NO, derived from the iNOS after infection of live Fn, was not involved in the Fn reduction because Fn clearance occurs within 6 hr. We investigated in this study whether this NO was involved in cytotoxicity in peritoneal exudate cells (PEC) in vivo. The mice were divided into two groups: those treated with live Fn (immune) and those left untreated (normal). PEC number, NO production, detection of apoptosis or death cells, and lactate dehydrogenase (LDH) release activity after injection of live Fn were compared in these groups. In the immune group, the increase of the total cell numbers caused by an increase in neutrophils, a significant NO production only after injection of live Fn at 24 hr and identification of iNOS positive macrophages were confirmed. The apoptotic rate was very low and did not increase at 24 hr in vivo. Therefore, apoptosis was seldom relevant to the NO. In the immune group, LDH activity was remarkable high at 24 hr, and dead cells and macrophages phagocytizing cell fragments increased at the same time. Pretreatment of L‐NMMA, an inhibitor of iNOS, suppressed LDH activity and cell death. Therefore, the NO derived from the iNOS is involved in the cytotoxicity. These results suggest that NO may contribute to the inflammatory response during Fn infection in periodontitis.
Acetate‐Mediated Production of Orotic Acid by ura3 Mutants of Candida albicans
Authors: Nezu Takashi , Shimokawa Osamu 
Published: November 2013

AbstractCandida albicans ura3 mutants were found to produce large amounts of orotic acid when the growth medium was supplemented with sodium acetate. Experiments with 13C‐labeled acetate showed that the acetate served as a precursor of orotic acid. This system of acetate‐mediated production of orotic acid is similar to other documented microbial producers in yield but unique for its acetate requirement.
Genotype Analysis of Malassezia restricta as the Major Cutaneous Flora in Patients with Atopic Dermatitis and Healthy Subjects
Authors: Sugita Takashi , Tajima Mami , Amaya Misato , Tsuboi Ryoji , Nishikawa Akemi 
Published: November 2013

AbstractLipophilic yeasts of the genus Malassezia colonize the skin surface of humans and are an exacerbating factor in atopic dermatitis (AD). Two species, M. restricta and M. globosa are major cutaneous microflora in both AD patients and healthy subjects. We compared the DNA sequences of the intergenic spacer (IGS) region, located between the 26S and 5S rRNA genes of M. restricta colonizing the skin surfaces of 13 AD patients and 12 healthy subjects, and of three CBS stock strains as references. The IGS 1 sequences were divided into two major groups, corresponding to AD patients and healthy subjects. These findings suggest that a specific genotype of M. restricta plays a significant role in AD, although M. restricta commonly colonizes both AD patients and healthy subjects.
Variation of Toxigenic Vibrio cholerae O1 in the Aquatic Environment of Bangladesh and Its Correlation with the Clinical Strains
Authors: Islam Mohammad Sirajul , Talukder Kaisar Ali , Khan Nurul Huda , Mahmud Zahid Hayat , Rahman Mohammed Ziaur , Nair Gopinath Balakrish , Siddique Abul K.M. , Yunus Mohammad , Sack David Allen , Sack Richard Bradley , Huq Anwar , Colwell Rita R. 
Published: November 2013

AbstractThe diversity of toxigenic V. cholerae O1 in the aquatic environment of Bangladesh is not known. A total of 18 environmental and 18 clinical strains of toxigenic V. cholerae O1 were isolated simultaneously from four different geographical areas and tested for variation by the pulsed‐field gel electrophoresis method. Environmental strains showed diversified profiles and one of the profiles was common to some environmental strains and most clinical strains. It appears that one clone has an advantage over others to cause disease. These findings suggest that the study of the molecular ecology of V. cholerae O1 in relation to its environmental reservoir is important in identifying virulent strains that cause disease.
Rapid Identification of Borrelia burgdorferi Sensu Lato by PCR‐Restriction Fragment Length Polymorphism Analysis of groEL Gene
Authors: Park Hyo‐Soon , Lee Jung‐Hee , Jeong Eun‐Ju , Park Tae‐Kyu , Jang Won‐Jong , Park Kyung‐Hee , Kim Bum‐Joon , Kook Yoon‐Hoh , Lee Seung‐Hyun 
Published: November 2013

AbstractThirty‐one reference strains and 23 Korean isolates of the genus Borrelia were identified through the PCR‐RFLP analysis using the groEL gene. This will be useful for the rapid differentiation of B. burgdorferi sensu lato and complements one of the 5S–23S intergenic spacers.
Nocardia aobensis Sp. Nov., Isolated from Patients in Japan
Authors: Kageyama Akiko , Suzuki Shin‐ichi , Yazawa Katsukiyo , Nishimura Kazuko , Kroppenstedt Reiner M. , Mikami Yuzuru 
Published: November 2013

AbstractFive clinical isolates, strains IFM 0137, 0372T, 0496, 0556, and 0952, were provisionally assigned to the genus Nocardia based on morphological criteria. Nearly complete 16S rDNA sequences were determined for these strains. These data showed that they are most similar to that of Nocardia africana, Nocardia cerradoensis and Nocardia veterana. However, DNA‐DNA relatedness data showed that the five strains were of a single species and were distinguishable from N. africana, N. cerradoensis and N. veterana. Therefore, these strains represent a new species within the genus Nocardia. The designation of these five strains is Nocardia aobensis sp. nov. The type strain is IFM 0372T (=NBRC 100429T=JCM 12352T=DSM 44805T).