bluuebunny/crossref_metadata_2025_split huggingface dataset:
Seroepidemiology of Spotted Fever Group and Typhus Group Rickettsioses in Humans, South Korea
Authors: Jang Won‐Jong , Choi Yeon‐Joo , Kim Jong‐Hyun , Jung Kwang‐Don , Ryu Ji‐Sun , Lee Seung‐Hyun , Yoo Cheon‐Kwon , Paik Hyung‐Suk , Choi Myung‐Sik , Park Kyung‐Hee , Kim Ik‐Sang 
Published: November 2013

AbstractThe prevalence of spotted fever group (SFG) and typhus group (TG) rickettsioses was investigated in 3,362 sera by immunofluorescence assay. The serum samples were obtained from patients with acute febrile episodes in South Korea from December 1992 to November 1993. The number of polyvalent positive sera against SFG rickettsial agents at the level of 1:40 dilution was 269 (8%) in Rickettsia sibirica, 482 (14.34%) in R. conorii, and 546 (16.24%) in R. akari. Many of the positive sera contained immunoglobulin (Ig) M antibodies rather than IgG antibodies. These results strongly suggest that SFG rickettsioses are prevalent in Korea. For TG rickettsial agents, the number of positive sera was 1,096 (32.60%) in R. typhi and 951 (28.29%) in R. prowazekii. Only a few epidemic typhus positive sera contained IgM antibodies. The result suggests that recent and/or primary infections of epidemic typhus were very rare in Korea during the said period. Among seven patients who had high titers (1:5,120) of IgG antibody to R. prowazekii, six were over 50 years old. The result suggests that Brill‐Zinsser disease was prevalent in Korea.
Quantitative Analysis of Cereulide, an Emetic Toxin of Bacillus cereus, by Using Rat Liver Mitochondria
Authors: Kawamura‐Sato Kumiko , Hirama Yumi , Agata Norio , Ito Hideo , Torii Kenzo , Takeno Akira , Hasegawa Tadao , Shimomura Yoshiharu , Ohta Michio 
Published: November 2013

AbstractAn emetic toxin cereulide, produced by Bacillus cereus, causes emetic food poisonings, but a method for quantitative measurement of cereulide has not been well established. A current detection method is a bioassay method using the HEp‐2 cell vacuolation test, but it was unable to measure an accurate concentration. We established a quantitative assay for cereulide based on its mitochondrial respiratory uncoupling activity. The oxygen consumption in a reaction medium containing rat liver mitochondria was rapid in the presence of cereulide. Thus uncoupling effect of cereulide on mitochondrial respiration was similar to those of uncouplers 2,4‐dinitrophenol (DNP), carbonylcyanide m‐chlorophenylhydrazone (CCCP), and valinomycin. This method gave constant results over a wide range of cereulide concentrations, ranging from 0.05 to 100 μg/ml. The minimum cereulide concentration to detect uncoupled oxygen consumption was 50 ng/ml and increased dose‐dependently to the maximum level. Semi‐log relationship between the oxygen consumption rate and the cereulide concentraiton enables this method to quantify cereulide. The results of this method were highly reproducible as compared with the HEp‐2 cell vacuolation test and were in good agreement with those of the HEp‐2 cell vacuolation test. The enterotoxin of B. cereus or Staphylococcus aureus did not show any effect on the oxygen consumption, indicating this method is specific for the identification of cereulide as a causative agent of emetic food poisonings.
A Comparison of Mycolic Acid Analysis for Nontuberculous Mycobacteria Identification by Thin‐Layer Chromatography and Molecular Methods
Authors: Leite Clarisse Queico Fujimura , Rocha Adalgiza da Silva , de Andrade Leite Sergio Roberto , Ferreira Rosa Maria Carvalho , Suffys Philip N. , de Souza Fonseca Leila , Saad Maria Helena Féres 
Published: November 2013

AbstractThe development of fast, inexpensive, and reliable tests to identify nontuberculous mycobacteria (NTM) is needed. Studies have indicated that the conventional identification procedures, including biochemical assays, are imprecise. This study evaluated a proposed alternative identification method in which 83 NTM isolates, previously identified by conventional biochemical testing and in‐house M. avium IS1245‐PCR amplification, were submitted to the following tests: thin‐layer chromatography (TLC) of mycolic acids and PCR‐restriction enzyme analysis of hsp65 (PRA). High‐performance liquid chromatography (HPLC) analysis of mycolic acids and Southern blot analysis for M. avium IS1245 were performed on the strains that evidenced discrepancies on either of the above tests. Sixty‐eight out of 83 (82%) isolates were concordantly identified by the presence of IS1245 and PRA and by TLC mycolic acid analysis. Discrepant results were found between the phenotypic and molecular tests in 12/83 (14.4%) isolates. Most of these strains were isolated from non‐sterile body sites and were most probably colonizing in the host tissue. While TLC patterns suggested the presence of polymycobacterial infection in 3/83 (3.6%) cultures, this was the case in only one HPLC‐tested culture and in none of those tested by PRA. The results of this study indicated that, as a phenotypic identification procedure, TLC mycolic acid determination could be considered a relatively simple and cost‐effective method for routine screening of NTM isolates in mycobacteriology laboratory practice with a potential for use in developing countries. Further positive evidence was that this method demonstrated general agreement on MAC and M. simiae identification, including in the mixed cultures that predominated in the isolates of the disseminated infections in the AIDS patients under study. In view of the fact that the same treatment regimen is recommended for infections caused by these two species, TLC mycolic acid analysis may be a useful identification tool wherever molecular methods are unaffordable.
Characterization of Effector Cells with Anti‐Candida Activity Obtained from Murine Bone Marrow Cells Cultured in the Presence of rhG‐CSF: Comparison between Normal and CY‐Treated Mice
Authors: Morofuji Shinichiro , Abe Shigeru , Okinaga Kota 
Published: November 2013

AbstractBone marrow cells (BMC) obtained from normal and cyclophosphamide (CY)‐treated mice were cultured in the presence of recombinant human granulocyte‐colony stimulating factor (rhG‐CSF) and their effector cell activities inhibiting growth of C. albicans were examined. When BMC from CY‐treated mice were preincubated with 0.05 ng/ml of rhG‐CSF, effector cells with enhanced anti‐C. albicans activity were recovered in the adherent cell population, whereas anti‐C. albicans activity of BMC from normal mice was found in the non‐adherent cell population. During culture without the presence of rhG‐CSF, non‐adherent BMC, seemingly granulocytes, from normal mice showed apoptotic change, but addition of rhG‐CSF clearly inhibited this change. On the other hand, when BMC from CY‐treated mice were cultured with rhG‐CSF, adherent cells as the main effector had the appearance of monocytes. These differences between the effectors with anti‐C. albicans activity obtained from normal and CY‐treated mice are discussed.
Analysis of Polymorphism of 18S rRNA Gene in Wuchereria bancrofti Microfilariae
Authors: Bhandari Yashodhar , Dabir Pankaj , Nandhakumar Krithika , Dayananda Kannayakanahalli Maheshwarappa , Shouche Yogesh Shripad , Reddy Maryada Venkata Rami 
Published: November 2013

AbstractThe polymorphism of the 18S rRNA gene in Wuchereria bancrofti microfilariae (mf) collected from three different zones in India was analyzed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). The RFLPs of the amplified products obtained after digestion with restriction enzymes SspI, MspI and HhaI showed no difference in the banding patterns among the mf isolates from different endemic zones. Further the sequencing of PCR products did not show any difference in the nucleotide sequence either. The phylogenetic analysis of the sequences of W. bancrofti mf isolates from different endemic zones has shown branching with the earlier reported sequences of W. bancrofti and its close relative Brugia malayi.
Lactobacillus acidophilus Expressing Recombinant K99 Adhesive Fimbriae Has an Inhibitory Effect on Adhesion of Enterotoxigenic Escherichia coli
Authors: Chu Hyuk , Kang Seungha , Ha Seckho , Cho Kwangkeun , Park Sung‐moo , Han Kyung‐ho , Kang Sang Kee , Lee HongGu , Han Seung Hyun , Yun Cheol H. , Choi Yunjaie 
Published: November 2013

AbstractThe most common enteric colibacillosis in neonatal and newborns is caused by enterotoxigenic Escherichia coli (ETEC). Colonization of ETEC in the small intestine is associated with adhesions using fimbriae, which is known as a specific adhesion factor and provides highly specific means for anchoring and prerequisite for an infectious agent. In the present study we have engineered Lactobacillus acidophilus to produce recombinant K99 fimbriae, which is used for the colonization to the intestine of pigs. The expression of K99 fimbrial protein was confirmed using SDS‐PAGE, immunoblot and agglutination analyses. To evaluate a function of the K99 fimbrial protein, inhibition and competition tests were performed on pre‐screened intestinal brush border from pigs. The tests showed that recombinant L. acidophilus, not control L. acidophilus, had a significant inhibitory effect to and competition against K99+ E. coli in a dose dependent manner. In conclusion, we demonstrated that recombinant K99 fimbriae producing L. acidophilus was able to prevent E. coli binding to intestinal brush border.
Effect of Orally Administered Non‐Viable Lactobacillus Cells on Murine Humoral Immune Responses
Authors: He Fang , Morita Hirotsugu , Kubota Akira , Ouwehand Arthur C. , Hosoda Masataka , Hiramatsu Masaru , Kurisaki Jun‐ichi 
Published: November 2013

AbstractBALB/c mice were immunized intraperitoneally with the food antigen ovalbumin (OVA) while they were fed with Lactobacillus GG heated killed cell preparation. The oral administration of Lactobacillus GG did not appear to modify the antigen‐augmented serum IgE in the tested mice but significantly augmented serum OVA specific IgG in the tested mice fed with a diet containing 0.1% Lactobacillus GG as the non‐viable cell preparation (P<0.05). The fecal OVA specific IgA of the tested mice fed with non‐viable Lactobacillus GG cells was also significantly elevated (P<0.05) compared to those from OVA immunized mice. The spleen cells of mice fed with non‐viable Lactobacillus GG cells secreted more IL‐6 (P<0.01). These results suggest that the non‐viable Lactobacillus GG can augment the systemic and mucosal immune responses in a host animal favoring secretory IgA but not IgE in an adjuvant‐like manner.
Gene Cloning and Properties of the RND‐Type Multidrug Efflux Pumps MexPQ‐OpmE and MexMN‐OprM from Pseudomonas aeruginosa
Authors: Mima Takehiko , Sekiya Hiroshi , Mizushima Tohru , Kuroda Teruo , Tsuchiya Tomofusa 
Published: November 2013

AbstractWe cloned two operons for putative RND‐type multidrug efflux pumps from Pseudomonas aeruginosa by a PCR method. We designated the genes in one operon mexPQ(‐opmE) and in another operon mexMN. Introduction of the mexPQ‐opmE into drug hypersensitive cells resulted in elevated MICs of macrolides, fluoroquinolones and some other drugs. Introduction of the mexMN into the hypersensitive cells possessing oprM, but not into cells not possessing oprM, resulted in elevated MICs of chloramphenicol and thiamphenicol. Thus, we conclude that MexPQ‐OpmE and MexMN‐OprM are functional multidrug efflux pumps when expressed in P. aeruginosa.
Mycobacterial RNase E‐Associated Proteins
Authors: Kovacs Lorand , Csanadi Agnes , Megyeri Klara , Kaberdin Vladimir R. , Miczak Andras 
Published: November 2013

AbstractRNase E and its complex with other proteins (‘degradosome’) play an important role in RNA processing and decay in Escherichia coli and in many other bacteria. To identify the proteins which can potentially interact with this enzyme in mycobacteria, Mycobacterium tuberculosis H37Rv RNase E was cloned and expressed as a 6HisFLAG‐tagged fusion protein. Analysis of the mycobacterial RNase E overexpressed and purified from M. bovis BCG revealed the presence of GroEL and two other copurified proteins, products of the Mb1721 (inorganic polyphosphate/ATP‐NAD kinase) and Mb0825c (acetyltransferase) genes. Identical copies of these two genes can be found in M. tuberculosis H37Rv.
Inhibitory Effect of RNAi on Japanese Encephalitis Virus Replication In Vitro and In Vivo
Authors: Murakami Manabu , Ota Takahide , Nukuzuma Soichi , Takegami Tsutomu 
Published: November 2013

AbstractFlaviviruses include many insect‐mediated small viruses and still cause serious problems in the world. In humans, JEV can cause acute meningioencephalomyelitis, resulting in fatality rates of 5 to 40%. RNA‐interference (RNAi) as an antiviral mechanism was originally discovered in plants and then found in the specific suppression of gene expression of other organisms such as Caenorhabditis elegans, Drosophila and vertebrates. As JEV is an RNA virus, RNAi could be a reasonable approach for therapeutic purposes to use against Japanese encephalitis. In this study, we examined the effect of RNAi on JEV replication. Viral reproduction in Vero cells was decreased to 7.2% and 39.0% of control by the transfection of small interference RNAs, JCR and JN3R at 250 NM, respectively. Under the transfection of 5 μg/ml pJRi which produces stem‐loop RNAi, viral reproduction was decreased to about 10% of control. Western blot analysis indicated that RNAi inhibited the translation level. We used pJRi in the animal experiment. After the inoculation of viruses at 5× 103 PFU, pJRi at 1.0 and 5.0 μg/g was injected into mice i.p. JEV‐infected control mice (n=5) died within 15 days. pJRi (1.0 or 5.0 μg/g)‐medicated mice survived 40 or 80% at 15 days. The data clearly indicate that pJRi has highly potent inhibitory activity against JEV replication in vivo. The results in vivo and in vitro provide evidence that JEV replication was efficiently inhibited by RNAi and RNAis could be used as an antiviral drug against JEV infection.
Susceptibility Loci to Coronary Arteritis in Animal Model of Kawasaki Disease Induced with Candida albicans‐Derived Substances
Authors: Oharaseki Toshiaki , Kameoka Yosuke , Kura Fumiaki , Persad Amanda S. , Suzuki Kazuo , Naoe Shiro 
Published: November 2013

AbstractWe have established an animal model of coronary arteritis which is histopathologically similar to that observed in cases of Kawasaki disease (KD), is a well‐known childhood vasculitis syndrome. Coronary arteritis in this mouse model has been induced by intraperitoneal injection of Candida albicans‐derived substances (CADS). Arteritis varied by mouse strain with the highest incidence by 71.1% (27/38) found in C3H/HeN mice, but absent in CBA/JN mice (0%, 0/27), suggesting association of genomic background to develop the disease. The present study aims to elucidate the susceptibility loci associated with coronary arteritis by using this animal model. The association of the onset of arteritis with polymorphic microsatellite markers between the two strains was examined using one hundred and fifteen of N1 backcross progeny [(CBA×C3H)F1×C3H]. Based on our analysis, arteritis‐susceptibility loci with suggestive linkage were mapped on D1Mit171 and D1Mit245 (map position 20.2 cM) on chromosome 1 (P=0.0019). These loci include several kinds of inflammatory cytokine receptors, such as interleukin 1 receptor and tumor necrosis factor receptor. We also found the cytokine response against CADS, levels of inflammatory cytokines interleukin‐1β, tumor necrosis factor‐α, and interleukin‐6 in sera increased within 24 hr after CADS injection. Our results may indicate based on genomics that ligand‐receptor interaction between these inflammatory cytokines and the receptors of these cytokines may affect the onset of arteritis.
Genotyping of Norovirus Strains Detected in Outbreaks between April 2002 and March 2003 in Osaka City, Japan
Authors: Seto Yoshiyuki , Iritani Nobuhiro , Kubo Hideyuki , Kaida Atsushi , Murakami Tsukasa , Haruki Kosuke , Nishio Osamu , Ayata Minoru , Ogura Hisashi 
Published: November 2013

AbstractNoroviruses (NVs) are the major cause of food‐ and waterborne nonbacterial gastroenteritis in Japan. Between April 2002 and March 2003, a total of 111 fecal specimens from 40 outbreaks of acute nonbacterial gastroenteritis in Osaka City, Japan were subject to NV detection. Seventy‐two samples (64.9%) from 31 outbreaks (77.5%) were NV positive by a real time reverse transcription (RT)‐PCR assay. To further determine the genotype of individual NV strains, we sequenced the capsid N‐terminal/shell (N/S) domain of some representative strains from each outbreak. The 51 NV strains detected in this study were segregated into 15 genotypes (6 in genogroup I and 9 in genogroup II), and GII/5 genotype NV was a dominant outbreak genotype.
Duplex Shuttle PCR for Differential Diagnosis of Budgerigar Fledgling Disease and Psittacine Beak and Feather Disease
Authors: Ogawa Hirohito , Yamaguchi Tsuyoshi , Fukushi Hideto 
Published: November 2013

AbstractTwo common viral diseases in psittacine birds including budgerigar fledgling disease (BFD), generally called avian polyomavirus (APV) infection, and psittacine beak and feather disease (PBFD) have similar clinical manifestations characterized by feather disorders. A duplex shuttle PCR was developed for detection of APV and PBFD virus (PBFDV). Two pairs of oligonucleotide primers were designed to amplify a 298‐bp fragment of the t/T antigen region of APV genome and a 495‐bp fragment of the capsid protein region encoded by open reading frame (ORF) C1 of PBFDV genome, respectively. In the present study, APV and PBFDV were detected simultaneously in one tube by duplex shuttle PCR using these two pairs of primers. The detection limits were 2 viral copies of APV and 3 viral copies of PBFDV. In the clinical application, we detected 16 APV‐positive, 15 PBFDV‐positive, and 3 mixed infected samples in 39 samples examined. Sequences of the amplified products were read. The t/T antigen region was conserved in the APV‐positive samples as expected. ORF C1 of PBFDV genome showed diversity. Phylogenic analysis indicated that PBFDV ORF C1 consisted of 6 clusters which were related to subfamilies of psittacine birds. Our duplex shuttle PCR could be a useful method for differential diagnosis and molecular epidemiology of BFD and PBFD.
Quantitative Microarray‐Based DNA‐DNA Hybridization Assay for Measuring Genetic Distances among Bacterial Species and Its Application to the Identification of Family Enterobacteriaceae
Authors: Amano Makoto , Ohkusu Kiyofumi , Kusaba Koji , Ikeda Hironori , Nagasawa Zenzo , Aoki Yosuke , Kawamura Yoshiaki , Kobatake Shinzo , Tanaka Takumi , Matsuura Shuuji , Ezaki Takayuki 
Published: November 2013

AbstractQuantitative DNA‐DNA hybridization to measure the genetic distances among bacterial species is indispensable for taxonomical determination. In the current studies, we developed a method to determine bacterial DNA relatedness on a glass microarray. Reference DNAs representing a total 93 species of Enterobacteriaceae were arrayed on a glass microplate, and signal intensities were measured after 2 hr of hybridization with Cy3‐labeled bacterial DNAs. All immobilized DNAs from members of the family Enterobacteriaceae were identified by this method except for DNAs from Yersinia pseudotuberculosis and Y. pestis. These results suggest that quantitative microarray hybridization could be an alternative to conventional DNA‐DNA hybridization for measuring chromosome relatedness among bacterial species.
Nonstructural Proteins 4A and 4B of Hepatitis C Virus Transactivate the Interleukin 8 Promoter
Authors: Kadoya Hiroyasu , Nagano‐Fujii Motoko , Deng Lin , Nakazono Naoki , Hotta Hak 
Published: November 2013

AbstractInterleukin 8 (IL‐8) is induced in many cell types by various stimuli including virus infection. It was reported that nonstructural protein 5A (NS5A) of hepatitis C virus (HCV) was involved in induction of IL‐8 expression at both mRNA and protein levels in cultured human cells. In this study, we aimed to determine whether or not another HCV protein(s) transactivates the IL‐8 gene expression, by means of an IL‐8 promoter‐driven luciferase reporter assay and measurement of endogenous IL‐8 mRNA and secreted IL‐8 protein levels. We observed that NS4B, and NS4A to a lesser extent, significantly transactivated the IL‐8 promoter, which resulted in enhanced production of IL‐8 protein. Also, the IL‐8 expression was augmented in Huh‐7 cells harboring an HCV subgenomic RNA replicon, compared with the control cells. Deletion mutational analysis of the IL‐8 promoter revealed the possible involvement of the transcription factor AP‐1 in both NS4A‐ and NS4B‐mediated IL‐8 gene activation. In addition, the IL‐8 gene activation by NS4B, but not that by NS4A, was likely to involve NF‐κB and/or NFIL‐6. The degree of the transactivation by NS4B and NS4A varied with different human cell lines, with HeLa cells showing the strongest activation followed by Huh‐7 cells, and with HepG2 cells exhibiting a marginal level of activation. Taken together, our present results suggest the possibility that NS4B and NS4A play an important role in inducing the IL‐8 gene expression under certain cellular conditions, which might be one of the strategies to establish persistent HCV infection.
The Time‐Dependent Serial Gene Response to Zeocin Treatment Involves Caspase‐Dependent Apoptosis in HeLa Cells
Authors: Hwang Jooyeon , Kim Young‐Youl , Huh Sungjin , Shim Junghee , Park Chan , Kimm Kuchan , Choi Dong Kug , Park Tae‐Kyu , Kim Soonhag 
Published: November 2013

AbstractZeocin, a member of the bleomycin/phleomycin family of antibiotics, is known to bind DNA and to induce apoptosis in cervical cancer cells, but the mechanism underlying this apoptotic response is poorly understood. The present study was undertaken to elucidate time‐dependent serial transcript patterns in the HeLa cervical carcinoma cell line, following treatment with Zeocin. The HeLa cell proliferation rate was found to gradually decrease following Zeocin exposure, in a time‐ and dose‐dependent manner. RNA transcript level measurements, for time‐dependent serial gene expression profiling, were determined at 0, 6, 12, 18 and 24 hr using a 0.5 k apoptosis functional microarray chip. Further statistical analysis, using a significance test at a 95% confidence level, for transcripts with a greater than 2‐fold change on the array chips, identified 49 up‐regulated and 57 down‐regulated genes. Our gene expression profile data indicate that Zeocin treatment induces an initial release of cytochrome c, the down‐regulation of Bcl‐XL, ENDOG, DAXX and MDM2, and the up‐regulation of CASP and BID. This suggests that a p53‐independent mitochondrial caspase cascade pathway is primarily involved in Zeocin‐induced apoptosis. Such caspase‐dependent cytotoxic activity also implies that this cell death pathway occurs via the caspase 8 and BID genes. However, disruption of either FAS or TNFR1 signaling did not interfere with the Zeocin induced apoptotic response in our experimental system. We hypothesize that Zeocin could be active against cervical cancer cell resistance to conventional chemotherapy and postulate that Zeocin is a novel candidate for the development of new chemotherapeutic treatments of gynecological cancers.
Effect of Cytokine‐Encoding Plasmid Delivery on Immune Response to Japanese Encephalitis Virus DNA Vaccine in Mice
Authors: Bharati Kaushik , Appaiahgari Mohan Babu , Vrati Sudhanshu 
Published: November 2013

AbstractWe have previously shown that immunization of mice with plasmid pMEa synthesizing Japanese encephalitis virus (JEV) envelope protein induced anti‐JEV humoral and cellular immune responses. We now show that intra‐muscular co‐administration of mice with pMEa and pGM‐CSF, encoding murine granulocyte‐macrophage colony‐stimulating factor or pIL‐2, encoding murine interleukin‐2 given 4 days after pMEa, augmented anti‐JEV antibody titers. This did not enhance the level of protection in immunized mice against JEV. However, intra‐dermal co‐administration of pMEa and pGM‐CSF in mice using the gene gun, enhanced anti‐JEV antibody titers resulting in an increased level of protection in mice against lethal JEV challenge.