bluuebunny/crossref_metadata_2025_split huggingface dataset:
The trpF Nucleotide Sequence and Its Promoter Analysis in Pseudomonas aeruginosa
Authors: Murata Takahiro 
Published: November 2013

AbstractThe nucleotide sequence of the gene for the phosphoribosyl anthranilate isomerase (PRAI) (trpF) in Pseudomonas aeruginosa was determined. The gene encoded a polypeptide of 211 amino acid residues and its initiation codon was UUG. Northern hybridization analysis showed only one transcript for trpF (about 700 nucleotides). Primer extension analysis revealed three transcription start sites which were at positions 52, 55 and 57 bases upstream of the initiation codon. A highly GC‐rich 9‐base‐pair inverted repeat with 5‐base‐pair spacing was found upstream of a putative Shine‐Dalgarno sequence. This inverted repeat could form a 23‐nucleotide stem‐loop structure in the transcript. Deletion of the inverted repeat slightly decreased the promoter activity of trpF. The presence of tryptophan had no effect on transcription of trpF.
The Prevalence of Listeria monocytogenes in the Environment of Dairy Farms
Authors: Ueno Hiroshi , Yokota Keiichi , Arai Takanori , Muramatsu Yasukazu , Taniyama Hiroyuki , Iida Takashi , Morita Chiharu 
Published: November 2013

AbstractThe prevalence of Listeria monocytogenes in the environment of dairy farms was surveyed from December 1993 to June 1994 in one city of Hokkaido. L. monocytogenes was isolated from 3 out of 5 farms investigated. Serovar 4b organism was isolated from the brain stem of a cow from one farm which was clinically diagnosed as having listeriosis. The same serovar of L. monocytogenes was also isolated from the rectal contents of a healthy cow, straw on the floor, straw in the barn, and silage scattered around the silo from the same farm. At another farm, with no reported cases of bovine listeriosis, serovar 1/2 organism was isolated from the same types of samples as the above mentioned farm except from straw on the floor. The difference in the isolation rates of the organism from straw on the floor between the two farms (22%: 5/23 vs 0%: 0/24) is considered to be caused by the different feeding methods of silage between the two farms.
Kinetics of Anti‐Mannan Antibodies Useful in Confirming Invasive Candidiasis in Immunocompromised Patients
Authors: van Deventer A. Johanna M. , Goessens Wil H.F. , van Zeijl Jan H. , Mouton Jan W. , Michel Marc F. , Verbrugh Henri A. 
Published: November 2013

AbstractIn studying the anti‐mannan antibodies longitudinally in serial serum samples of three immunocompromised patients, it was observed that anti‐mannan antibodies started to increase shortly after the moment that cultures of deep‐tissue sites became positive with Candida albicans. The mean anti‐mannan antibody titers determined in a group of 36 immunocompromised patients with invasive candidiasis increased within two weeks after the probable onset of invasive candidiasis. In contrast, anti‐mannan antibody levels in serial serum samples of 14 immunocompromised patients who were only colonized with C. albicans remained stable or decreased over time. The HA test measuring the anti‐mannan antibodies was 64% sensitive and 89% specific in determining invasive candidiasis. In contrast, antibodies specific for candidal cytoplasmic antigens or enolase alone were of little value in confirming invasive candidiasis in these immunocompromised patients.
Threonine‐74 Is a Key Site for the Activity of Clostridium perfringens Alpha‐Toxin
Authors: Nagahama Masahiro , Sakurai Jun 
Published: November 2013

AbstractA mutant toxin (MT) that abolished almost 99% of the hemolytic activity of alpha‐toxin was isolated by random polymerase chain reaction (PCR) mutagenesis of the gene for Clostridium perfringens alpha‐toxin. In the mutant toxin, the amino acids at Tyr (Y)‐62, Thr (T)‐74 and Ile (I)‐345 were substituted with His, Ile and Met, respectively. Replacement of T‐74 with Ile by site‐directed mutagenesis resulted in the loss of hemolytic, phospholipase C and sphingomyelinase activities by 1/250‐fold of that of the wild‐type. The replacement of Y‐62 with Ile or I‐345 with Met alone did not affect the activities of the toxin. T74I mutant bound to sheep erythrocyte membranes and specifically bound [65Zn]2+ in Tris‐buffered saline, in the same manner as the wild‐type, and contained 2 mol of zinc ions per mol of protein. These results suggest that the T‐74 residue plays a key role in these biological activities of C. perfringens alpha‐toxin.
Detection of the Disease Severity‐Related Molecular Differences among New Thai Dengue‐2 Isolates in 1993, Based on Their Structural Proteins and Major Non‐Structural Protein NS1 Sequences
Authors: Thant Kyaw‐Zin‐ , Morita Kouichi , Igarashi Akira 
Published: November 2013

AbstractWe determined the nucleotide sequences of the whole structural protein gene of four new dengue‐2 viruses by the primer extension dideoxy chain termination method, using multiple cDNA clones for six overlapping gene regions. The nucleotide sequences of the major non‐structural protein NS1 gene of these viruses were also determined by direct sequencing of the reverse‐transcription polymerase chain reaction products. These viruses were isolated from dengue patients with different clinical severities in Nakhon Phanom, Northeastern Thailand in 1993. The results were compared with the sequences of prototype New Guinea C strain and other reference strains. All four viruses revealed highest homology to New Guinea C strain. The homology between each of the four strains and New Guinea C strain varies from 95.09% to 95.29% in its nucleotide sequences, and from 97.24% to 97.78% in its amino acid sequences covering all structural proteins and NS1 protein. The PreM region shows the highest divergence (6.59% to 7.32%) in its nucleotide sequence, whereas C protein is most highly conserved (only 1.75% to 2.63% divergence). Our data showed that there are certain molecular differences in the genomic structure of these four new isolates, which indicate the possibility that these changes are related with the virulence of the virus strains.
Fgr Expression Restricted to Subpopulation of Monocyte/Macrophage Lineage in Resting Conditions Is Induced in Various Hematopoietic Cells after Activation or Transformation
Authors: Hatakeyama Shigetsugu , Iwabuchi Kazuya , Ato Manabu , Iwabuchi Chikako , Kajino Kiichi , Takami Kimitaka , Katoh Masahito , Ogasawara Kazumasa , Good Robert A. , Onoé Kazunori 
Published: November 2013

AbstractThe c‐fgr gene product (Fgr) is a member of the src‐family of protein tyrosine kinases. We have established a monoclonal antibody (2H2) which recognizes the unique N‐terminal domain of the murine Fgr. In the present study, using immunohistochemical analysis and immune complex kinase assay with the 2H2, we investigated expression of Fgr in various cell populations and tissues in a murine system. In resting conditions, Fgr expression was confined to subsets of a monocyte/macrophage lineage. Thus, Fgr+ cells were detected in paracortical areas and medullas of lymph nodes, but seen only in marginal zones of the spleen and the medulla of the thymus. No Fgr+ macrophage was detected in other tissues, Peyer's patches, brain, heart, lung, liver, pancreas, kidney and peritoneal cavity. However, immune complex kinase assay revealed that, upon stimulation, T and B cells as well as peritoneal macrophages expressed significant levels of Fgr molecules. Transformed cell lines of lymphoid origin, EL‐4 and LK35.2, which are T and B lineage lymphomas, respectively, also expressed Fgr molecules. Thus, various cells of hematopoietic origin appeared to possess a potentiality to express Fgr following activation or transformation. The present findings may help elucidate the functional significance of Fgr in immunologically committed cells in either activated or non‐activated conditions.
Immunochemical Characteristics and Localization on Cells of Protective Antigen (PAg) Prepared from Leptospira interrogans Serovar lai
Authors: Masuzawa Toshiyuki , Nakamura Ryuta , Beppu Yoshihito , Yanagihara Yasutake 
Published: November 2013

AbstractImmuno‐electron microscopic methods revealed that the protective antigen (PAg) of Leptospira interrogans serovar lai exists on the outer envelope sheathing the leptospiral cell body. PAg lost its protective activity after treatment by hydrolysis with 2 M formic acid at 100 C for 2 hr, or oxidation with periodate at 4 C for 40 hr. The antigenic oligosaccharide fraction was further purified from the hydrolyzed PAg by immunoaffinity column coupled with protective monoclonal antibody, LW2, and by gel filtration of HPLC. The antigenic oligosaccharide fraction contained two unknown sugars and 4‐O‐methylmannose (molar ratio 3:5:1). These findings suggested that these sugars are components of an antigenic determinant contributing to the protective immunity against serovar lai infection.
Isolation and Characterization of Teichoic Acid‐Like Substance as an Adhesin of Staphylococcus aureus to HeLa Cells
Authors: Matsuura Takashi , Miyake Yoichiro , Nakashima Seiji , Komatsuzawa Hitoshi , Akagawa Yasumasa , Suginaka Hidekazu 
Published: November 2013

AbstractA cell wall component that bound to HeLa cells (HeLa cell‐binding CWC) was isolated from a clinical isolate of Staphylococcus aureus. The HeLa cell‐binding CWC was resistant to heat (100 C, 1 hr) and proteases, did not stain with Coomassie Brilliant Blue R‐250 on SDS‐PAGE but stained as a broad band with antiserum against the strain on Western blots. These data suggest that the HeLa cell‐binding CWC is not a protein, and may be teichoic acid. Purified teichoic acid bound to HeLa cells, whereas fractions without teichoic acid did not. In Western blots, HeLa cell‐binding CWC appeared as a broad band of less than 35 kDa, similar to that of purified teichoic acid. These data suggest that the HeLa cell‐binding CWC obtained in this study is teichoic acid. Teichoic acid inhibited S. aureus adherence to HeLa cells and bound to the cells time and dose dependently, in a saturable and reversible manner, and therefore appears to be an adhesin of S. aureus to HeLa cells.
Analysis of the Phospholipase C Gene of Clostridium perfringens KZ1340 Isolated from Antarctic Soil
Authors: Kameyama Kohtaro , Matsushita Osamu , Katayama Seiichi , Minami Junzaburo , Maeda Masazumi , Nakamura Shinichi , Okabe Akinobu 
Published: November 2013

AbstractClostridium perfringens KZ1340 isolated from Antarctic soil was first classified as Clostridium plagarum and later as a lecithinase‐negative variant of C. perfringens. Although the strain produced no detectable lecithinase (phospholipase C, PLC) activity in the culture supernatant, it was shown by Southern blot hybridization to possess a PLC‐encoding gene (plc). To determine the cause of the PLC deficiency, we cloned and sequenced the plc gene from KZ1340. The deduced amino acid sequence consists of 398 amino acid residues, coinciding with those of the plc genes previously determined. Tyrosine was substituted for histidine at amino acid position 148, which is thought to bind a zinc ion essential for PLC activity. Northern blot analysis revealed that KZ1340 expressed the plc gene at an extremely low level. Furthermore, the plc gene cloned from C. perfringens strain 13 into a plasmid was expressed weakly in KZ1340, compared to that in strain 13. This indicates that the former strain represses plc gene expression in trans. When a phylogenetic tree of plc genes was constructed, the KZ1340 plc gene formed a monophyletic branch along with those of various other C. perfringens strains, supporting the classification of the strain as a variant of C. perfringens.
Evidence That Two Distinct Populations of Rabbit Anti‐Idiotypic Antibodies Are Induced by Three Monoclonal Antibodies Specific for Bluetongue Virus Core Protein VP7
Authors: Lin Min , Zhou En‐Min 
Published: November 2013

AbstractThree groups of anti‐idiotypic antibodies (anti‐Id or Ab2), designated RAb2‐A, RAb2‐B1, and RAb2‐B2, were isolated from rabbit antiserum raised against three monoclonal antibodies (mAbs) (M1875, M1877, and M1886) specific for the bluetongue virus core protein, VP7. RAb2‐A was specific for the idiotype of M1875. RAb2‐B1 and RAb2‐B2, isolated through the M1877 and M1886 affinity columns, respectively, were directed against the common idiotype that is shared by M1877 and M1886 and therefore classified in the same population (RAb2‐B). Further characterization revealed that the two anti‐Id populations, RAb2‐A and RAb2‐B, were significantly different. RAb2‐A was an Ab2β type of anti‐Id since (i) its reaction with M1875 was inhibited by the antigen; (ii) it inhibited the M1875‐VP7 interaction; and (iii) it elicited anti‐VP7 antibody response in Balb/c mice. In contrast, RAb2‐B may represent an Ab2α type of anti‐Id since its reactions with M1877 or M1886 were not inhibited by the antigen, even though it inhibited mAbs from binding to the antigen. These results indicated that RAb2‐A and RAb2‐B represent two distinct populations of anti‐Ids to anti‐VP7 mAbs with similar epitope specificity.
Characterization of Glycoprotein H and L of Human Herpesvirus 7
Authors: Mukai Tetsu , Hata Atsuko , Isegawa Yuji , Yamanishi Koichi 
Published: November 2013

AbstractThe genes encoding the glycoproteins H (gH) and L (gL) of human herpesvirus 7 (HHV‐7) have been identified. The gH open reading frame (ORF) was 2,070 base pairs in length and encoded a predicted 690 amino‐acid protein. The gH contained characteristics of a transmembrane glycoprotein including 10 consensus N‐linked glycosylation sites, 12 cysteine residues, a potential amino‐terminal signal sequence and a predicted transmembrane segment located near the carboxyl terminus. The gL ORF was 738 base pairs in length and encoded a predicted 246 amino‐acid protein. Four possible N‐glycosylation sites and 6 cysteine residues existed within gL. The predicted amino‐acid sequences of the HHV‐7 gH and human herpesvirus 6 variant A (HHV‐6A) gH gene products exhibited 23.6% identity to each other, and those of the gL gene products had 26.0% identity. Upon in vitro translation of the gL gene, the addition of microsomal membranes resulted in two modified products with molecular weights of 32 kDa and 35 kDa from the unmodified initial translation product of 26 kDa. An amino‐terminal portion of gH and the full length of gL were expressed as glutathione S‐transferase fusion proteins, and these proteins were used to raise immune sera in mice. Lysates of cells infected with HHV‐7 were subjected to immunoprecipitation analysis. Approximate molecular weights of 33, 37, 80 and 90 kDa polypeptides were immunoprecipitated with antibodies against the gH protein. Antibodies against the gL protein polypeptides with the same molecular weights were also precipitated, and were observed with the antibodies against the gH protein. These results suggest that HHV‐7 gH and gL may form a heterodimeric complex with each other in HHV‐7 infected cells, as has been reported for other herpesviruses.
Suppressed Induction of 2‐5AS in Cells with HTLV‐I Is Not Associated with the Fluctuation of IFN‐Receptor, IFN‐Regulatory Factors (IRF‐1 and IRF‐2) or STAT‐1α
Authors: Yokosawa Noriko , Indoh Tomokazu , Kubota Toru , Yashiki Teruo , Fujii Nobuhiro 
Published: November 2013

AbstractSome viruses seem to be capable of suppressing interferon (IFN)‐induced 2′, 5′‐oligoadenylate synthetase (2‐5AS) induction. Cells infected with human T‐lymphotropic virus type‐I (HTLV‐I) show different natures for the constitutive production of IFN‐γ or sensitivity to IFN. Poor induction of 2‐5AS was found in IFN‐γ producer cells carrying HTLV‐I (MT‐1, MT‐2 and SMT‐1). On the other hand, in non‐ or low‐producing cell lines of IFN‐γ such as HUT102 and OKM‐2, significant activity of 2‐5AS was induced by treatment with IFN‐α. A satisfactory level of IFN receptor was detectable in SMT‐1 cells in spite of the poor induction of 2‐5AS. There were no differences in either the interferon regulatory factor‐2 (IRF‐2) mRNA transcript or the level of STAT‐1α between SMT‐1 and HUT102 cells. However, the transcription of IRF‐1 mRNA was slightly reduced in SMT‐1 cells as compared with that of HUT102 cells.
Adjuvanticity of an Ornithine‐Containing Lipid of Flavobacterium meningosepticum as a Candidate Vaccine Adjuvant
Authors: Kato Hiroshi , Goto Norihisa 
Published: November 2013

AbstractOrnithine‐containing lipids (OrnL) extracted from Flavobacterium meningosepticum have been reported to have various biological activities such as B‐cell mitogenicity and macrophage activation to generate interleukin‐1 and prostaglandin E2. We, using ovalbumin (OVA) as an antigen, evaluated the adjuvant activity of OrnL as an immunological adjuvant in BALB/c mice. OrnL showed the function of forming liposome‐like vesicles retaining biological activities when prepared as either small unilamellar or dehydration‐rehydration vesicles. Although OrnL was not shown to have enough entrapping efficacy for use as a vaccine adjuvant, phosphatidylglycerol (PG) and cholesterol (CHOL) added to stabilize the vesicle membrane increased the entrapping efficacy to the same extent as that of conventional liposomes. Furthermore, the stabilized OrnL vesicles tolerated centrifugation to remove non‐entrapped antigens. Completely antigen‐entrapped OrnL vesicles including PG and CHOL induced a significantly greater enhancement of IgG antibody production than did aluminum hydroxide gel in BALB/c mice from week 6. These results indicate that OrnL can be utilized as an immunological adjuvant for vaccines.
Studies of Polymorphic DNA Fingerprinting and Lipid Pattern of Mycobacterium tuberculosis Patient Isolates in Japan
Authors: Chaicumpar Kunyaluk , Fujiwara Nagatoshi , Nishimura Osamu , Hotta Hisako , Pan Jiong Wei , Takahashi Mitsuyoshi , Abe Chiyoji , Yano Ikuya 
Published: November 2013

AbstractStrain differentiation by DNA restriction fragment length polymorphism (RFLP) has been used mainly for the epidemiological purpose of Mycobacterium tuberculosis infection. In this study, we tried to connect the molecular and phenotypic characteristics of M. tuberculosis patient isolates by comparing the DNA fingerprints obtained by RFLP using IS6110 and lipid patterns using two‐dimensional thin‐layer chromatography (2‐D TLC) with silica gel, since M. tuberculosis has a lipid‐rich cell envelope which contributes to the virulence and immunomodulatory properties. We found that 66 isolates of M. tuberculosis from tuberculosis patients showed that the occurrence of IS6110 varied from 1 to 24 copies. The IS6110 patterns were highly variable among isolates. Fifty different RFLP patterns were observed, and 12 RFLP patterns were shared by two or more strains. By computerized analysis of the RFLP patterns of M. tuberculosis patient isolates, we found that 95% of the isolates fell into seven clusters, from A to G, with at least two isolates in each (> 30% similarity). Among the cellular lipids, the phospholipid composition did not differ by strain, whereas the glycolipid pattern differed markedly. Especially, the relative concentration of cord factor and sulfolipid, both of which were known as virulent factors, varied by strain. The fingerprints of some strains showed an association between the DNA and glycolipid patterns, even though some of the same DNA fingerprint strains showed differences in lipid patterns. Among the patient isolates, M. tuberculosis strain 249 possessed a specific glycolipid with 2‐O‐methyl‐L‐rhamnose and L‐rhamnose, which is rarely found in other strains. This glycolipid showed serological activity against the sera of tuberculosis patients, even if the reactivity was not as strong as trehalose dimycolate. It also showed the inhibition of phagosome‐lysosome fusion in macrophages, suggesting involvement with virulence. These results suggest that RFLP analysis using IS6110 is useful for clustering the human isolates of M. tuberculosis, however, for further strain differentiation on virulence, a lipid analysis provides more information.
Father‐to‐Mother‐to‐Infant Transmission of HIV‐1: Clonally Transmitted Isolate of Infant Mutates More Rapidly than That of the Mother and Rapidly Loses Reactivity with Neutralizing Antibody
Authors: Okamoto Yukari , Shiosaki Koichi , Eda Yasuyuki , Tokiyoshi Sachio , Yamaguchi Yumi , Gojobori Takashi , Hachimori Takashi , Yamazaki Shudo , Honda Mitsuo 
Published: November 2013

AbstractThe sequences of the V3 loop and surrounding regions of human immunodeficiency virus type‐1 from a father‐to‐mother‐to‐infant trimmer were studied and the horizontal and vertical transmissions compared. The father's virus was variable for reactivity with neutralizing antibody and sequences of the V3 loop central core sequence. In contrast, the mother's viral sequences were much less diverse and reacted with a virus neutralizing antibody. The infant's viral sequences were also less diverse than those of the father, and N‐glycosylation sites were conserved. By phylogenetic analysis, the major clone, of which V3‐peptide reacted with the neutralizing antibody, was found to be transmitted from the mother to her infant; however, the mutated minor clones did not bind to the antibody. These findings suggest that both horizontal and vertical virus transmission were selective, and that the clonally transmitted virus in infants mutates more rapidly than viruses in the mother, to whom the virus was horizontally transmitted.
Two Distinct Populations of Primary Cytotoxic Cells Infiltrating into Allografted Tumor Rejection Sites: Infiltration of Macrophages Cytotoxic against Allografted Tumor Precedes That of Multiple Sets of Cytotoxic T Lymphocytes with Distinct Specificity to Alloantigens
Authors: Yoshida Ryotaro , Matsuura Akihiro , Einaga Kuniko , Ushio Yumiko , Yamamoto Naoki , Yoneda Yukio 
Published: November 2013

AbstractIt has been reported that the rejection of tumor allografts is mainly mediated by cytotoxic T lymphocytes (CTLs). Here, we characterized the cytotoxic effector cells of C57BL/6 (B6; H‐2b) mice infiltrating into the rejection site of the i.p. allografted Meth A fibrosarcoma (or P815 mastocytoma) cells of H‐2d origin. Two types of cytotoxic cells (i.e., CD8+ CTLs and macrophages (Mφs)) were identified by flow cytometric fractionation of the infiltrates or by specific in vitro elimination of cells either with antibody (Ab)‐coated beads or with an Ab‐plus complement. Of particular interest, these effector cells showed distinct and unique target specificities. First, the CTLs were inactive against transplanted tumor (e.g., Meth A) cells, whereas they were cytotoxic against donor‐related concanavalin A (Con A) blasts as well as CTLL‐2 (H‐2b) cells transfected with a class I gene of H‐2d origin. A cold target competition assay suggested that the CTLs were composed of multiple sets of T cells, each of which specifically recognized different allo‐antigens. Second, the Mφs lysed the allografted tumor cells but were inert toward the Con A blasts and the CTLL‐2 transfectants. Unexpectedly, the infiltration of Mφs preceded the infiltration of CTLs by several days during the course of rejection. These results indicate that two distinct populations of unique cytotoxic cells (i.e., CTLs and Mφs) are induced in the allografted tumor rejection site, and that the infiltration of cytotoxic Mφs responsible for rejection precedes that of the CTLs cytotoxic against cells expressing donor‐related allo‐antigens.
Seasonal Variations in the Presence of Antibodies against Coxiella burnetii in Dairy Cattle in Hokkaido, Japan
Authors: Yanase Tsuyoshi , Muramatsu Yasukazu , Ueno Hiroshi , Morita Chiharu 
Published: November 2013

AbstractThe prevalence and seasonal variations of infection by Coxiella burnetii in cattle were investigated seroepidemiologically on a farm in Hokkaido, Japan, by an immunofluorescent antibody test. A total of 364 serum samples from 28 cows were collected from August 1993 to October 1995 in two barns on the farm. It was found that the number of antibody‐positive cows and their antibody titers were significantly elevated in winter and decreased in summer. In addition, antibodies were detectable in seroconverted cows for about five months.