• Title/Summary/Keyword: Resistant Genes

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Establishment and characterization of bortezomib-resistant U266 cell line: Constitutive activation of NF-κB-mediated cell signals and/or alterations of ubiquitylation-related genes reduce bortezomib-induced apoptosis

  • Park, Juwon;Bae, Eun-Kyung;Lee, Chansu;Choi, Jee-Hye;Jung, Woo June;Ahn, Kwang-Sung;Yoon, Sung-Soo
    • BMB Reports
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    • v.47 no.5
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    • pp.274-279
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    • 2014
  • Bortezomib has been known as the most promising anti-cancer drug for multiple myeloma (MM). However, recent studies reported that not all MM patients respond to bortezomib. To overcome such a stumbling-block, studies are needed to clarify the mechanisms of bortezomib resistance. In this study, we established a bortezomib-resistant cell line (U266/velR), and explored its biological characteristics. The U266/velR showed reduced sensitivity to bortezomib, and also showed cross-resistance to the chemically unrelated drug thalidomide. U266/velR cells had a higher proportion of CD138 negative subpopulation, known as stem-like feature, compared to parental U266 cells. U266/velR showed relatively less inhibitory effect of prosurvival NF-${\kappa}B$ signaling by bortezomib. Further analysis of RNA microarray identified genes related to ubiquitination that were differentially regulated in U266/velR. Moreover, the expression level of CD52 in U266 cells was associated with bortezomib response. Our findings provide the basis for developing therapeutic strategies in bortezomib-resistant relapsed and refractory MM patients.

Characterization of tet(M) and tet(G) Genes among Tetracycline-resistant Aeromonas spp. Isolated from Imported Ornamental Fishes (수입산 관상어로부터 분리된 tetracycline 내성 Aeromonas spp.에 tet(M) 및 tet(G) 유전자의 특성 분석)

  • Park, Shin-Hoo;Jun, Lyu-Jin;Cho, Ki-Taek;Jin, Ji-Woong;Jeong, Hyun-Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.45 no.3
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    • pp.238-245
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    • 2012
  • In this study, the molecular structures of tet(M) and tet(G) carried by tetracycline (Tc) resistant bacteria in intestinal microflora from the imported ornamental fish were characterized and compared with each other depend on the imported countries. Of the total isolates, approximately 8.9% of the Ent-lac+(lactose fermentative bacteria on coliform media) Tc resistant isolates in fish from three different countries, Singapore, Taiwan and Brazil, were appeared to contain tet(M). Three representative isolates of different countries, Aeromonas spp. JSM-1 (Singapore), JTM-1 (Taiwan) and JBM-1 (Brazil), were isolated and analyzed the molecular structures of tet(M) gene. Interestingly, partial sequence of tet(M) genes (1099 bp) in JBM-1 (Brazil) showed 99.5% homology with the tet(M) found in the Vibrio spp. RV16 isolate, obtained from marine fish in Korea and known to carry Tn1545 parent type of tet(M). In contrast, tet(M) gene in JSM-1 and JTM-1 showed mosaic structure of Tn1545 and Tn916, and 100% homology with each other. It may suggest the presence of various characteristics in terms of tet(M) gene structure. The determined sequence of the tet(G) from Aeromonas spp. JSG-1 and JBG-1 isolated from Singapore and Indonesia ornamental fish respectively showed similar nucleotide sequence homology but revealed a few nucleotide changes in comparison with the sequence of the prototype tet(G) gene (S52437 in GenBank).

Persistence of Multidrug-Resistant Acinetobacter baumannii Isolates Harboring blaOXA-23 and bap for 5 Years

  • Sung, Ji Youn;Koo, Sun Hoe;Kim, Semi;Kwon, Gye Cheol
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1481-1489
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    • 2016
  • The emergence and dissemination of carbapenemase-producing Acinetobacter baumannii isolates have been reported worldwide, and A. baumannii isolates harboring blaOXA-23 are often resistant to various antimicrobial agents. Antimicrobial resistance can be particularly strong for biofilm-forming A. baumannii isolates. We investigated the genetic basis for carbapenem resistance and biofilm-forming ability of multidrug-resistant (MDR) clinical isolates. Ninety-two MDR A. baumannii isolates were collected from one university hospital located in the Chungcheong area of Korea over a 5-year period. Multiplex PCR and DNA sequencing were performed to characterize carbapenemase and bap genes. Clonal characteristics were analyzed using REP-PCR. In addition, imaging and quantification of biofilms were performed using a crystal violet assay. All 92 MDR A. baumannii isolates involved in our study contained the blaOXA-23 and bap genes. The average absorbance of biomass in Bap-producing strains was much greater than that in non-Bap-producing strains. In our study, only three REP-PCR types were found, and the isolates showing type A or type B were found more than 60 times among unique patients during the 5 years of surveillance. These results suggest that the isolates have persisted and colonized for 5 years, and biofilm formation ability has been responsible for their persistence and colonization.

Effect of antibiotic resistant factors in effluent of wastewater treatment plant on stream (하수처리장 방류수에 존재하는 항생제 내성인자가 하천에 미치는 영향)

  • Jang, Yejin;Yoo, Yong-Jae;Sul, Woo Jun;Cha, Chang-Jun;Rhee, Ok-Jae;Chae, Jong-Chan
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.316-319
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    • 2017
  • The antibiotic resistant genes (ARG) and mobile genetic elements (MGE) were investigated with the effluent of waste-water treatment plant (WWTP), and river waters of upstream and downstream in order to elucidate the effect of effluent on antibiotic resistance in a natural river. Total numbers of 134~183 of ARG and MGE were detected and the abundance of ARG and MGE was 0.063~0.422 copies per one of 16S rRNA gene in three water samples. Effluent sample contained the highest amount of the total number and abundance of ARG and MGE whereas total viable cells were observed in the lowest amount among the three samples. This indicated that the genes were originated from cells died during the wastewater treatment process. In addition, the co-relationship of abundance between ARG and MGE suggested that acquired resistance was a prevalent mechanism among the antibiotic-resistant bacteria existing in WWTP.

Expression of Cell Proliferation-Related PCNA and E2F Genes in Drosophila Gut and Inhibitory Effect of Nitric Oxide

  • Choi, Na-Hyun;Kim, Young-Shin;Hwang, Mi-Sun;Nam, Hyuck-Jin;Kim, Nam-Deuk;Chung, Hae-Young;Yoo, Mi-Ae
    • Animal cells and systems
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    • v.5 no.1
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    • pp.59-64
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    • 2001
  • To understand the late gut development and differentiation, identification and characterization of target genes of homeotic genes involved in gut development are required. We have previously reported that homeodomain proteins can regulate expression of the cell proliferation-related genes. We investigated here the expression of the Drosophila proliferating cell nuclear antigen(PCNA) and E2F(dE2F) genes in larval and adult guts using transgenic flies bearing lacz reporter genes. Both PCNA and dE2F genes were expressed strongly in whole regions of the larval and adult guts including the esophagus, proventriculus, midgut and hindgut, showing higher expression in foregut and hindgut imaginal rings of larva. Nitric Oxide(NO) has been known to be involved in cell proliferation and tumor growth and also to have an antiproliferative activity. Therefore, we also investigated effects of NO on the expression of PCNA and dE2F genes in gut through analyses of lacz reporter expression level in the SNP (NO donor)-treated larval guts. Expressions of both PCNA and dE2F were greatly declined by SNP. The inhibitory effect of NO was shown in whole regions of the gut, especially in hindgut, while the internal region of proventriculus, esophagus, foregut imaginal ring and hindgut imaginal ring was resistant. Our results suggest that this inhibitory effect may be related with the antiproliferative activity of NO.

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Identification of p54nrb and the 14-3-3 Protein HS1 as TNF-α-Inducible Genes Related to Cell Cycle Control and Apoptosis in Human Arterial Endothelial Cells

  • Stier, Sebastian;Totzke, Gudrun;Grunewald, Elisabeth;Neuhaus, Thomas;Fronhoffs, Stefan;Schoneborn, Silke;Vetter, Hans;Ko, Yon
    • BMB Reports
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    • v.38 no.4
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    • pp.447-456
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    • 2005
  • TNF-$\alpha$ plays a pivotal role in inflammation processes which are mainly regulated by endothelial cells. While TNF-$\alpha$ induces apoptosis of several cell types like tumor cells, endothelial cells are resistant to TNFa mediated cell death. The cytotoxic effects of TNF-$\alpha$ on most cells are only evident if RNA or protein synthesis is inhibited, suggesting that de novo RNA or protein synthesis protect cells from TNF-$\alpha$ cytotoxicity, presumably by NF-${\kappa}B$ mediated induction of protective genes. However, the cytoprotective genes involved in NF-${\kappa}B$ dependent endothelial cell survival have not been sufficiently identified. In the present study, the suppression subtractive hybridization (SSH) method was employed to identify rarely transcribed TNF-$\alpha$ inducible genes in human arterial endothelial cells related to cell survival and cell cycle. The TNF-$\alpha$-induced expression of the RNA binding protein $p54^{nrb}$ and the 14-3-3 protein HS1 as shown here for the first time may contribute to the TNF-$\alpha$ mediated cell protection of endothelial cells. These genes have been shown to play pivotal roles in cell survival and cell cycle control in different experimental settings. The concerted expression of these genes together with other genes related to cell protection and cell cycle like DnaJ, $p21^{cip1}$ and the ubiquitin activating enzyme E1 demonstrates the identification of new genes in the context of TNF-$\alpha$ induced gene expression patterns mediating the prosurvival effect of TNF-$\alpha$ in endothelial cells.

Analysis of miRNA expression in the trachea of Ri chicken infected with the highly pathogenic avian influenza H5N1 virus

  • Suyeon Kang;Thi Hao Vu;Jubi Heo;Chaeeun Kim;Hyun S. Lillehoj;Yeong Ho Hong
    • Journal of Veterinary Science
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    • v.24 no.5
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    • pp.73.1-73.16
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    • 2023
  • Background: Highly pathogenic avian influenza virus (HPAIV) is considered a global threat to both human health and the poultry industry. MicroRNAs (miRNA) can modulate the immune system by affecting gene expression patterns in HPAIV-infected chickens. Objectives: To gain further insights into the role of miRNAs in immune responses against H5N1 infection, as well as the development of strategies for breeding disease-resistant chickens, we characterized miRNA expression patterns in tracheal tissues from H5N1-infected Ri chickens. Methods: miRNAs expression was analyzed from two H5N1-infected Ri chicken lines using small RNA sequencing. The target genes of differentially expressed (DE) miRNAs were predicted using miRDB. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis were then conducted. Furthermore, using quantitative real-time polymerase chain reaction, we validated the expression levels of DE miRNAs (miR-22-3p, miR-146b-3p, miR27b-3p, miR-128-3p, miR-2188-5p, miR-451, miR-205a, miR-203a, miR-21-3p, and miR-200a3p) from all comparisons and their immune-related target genes. Results: A total of 53 miRNAs were significantly expressed in the infection samples of the resistant compared to the susceptible line. Network analyses between the DE miRNAs and target genes revealed that DE miRNAs may regulate the expression of target genes involved in the transforming growth factor-beta, mitogen-activated protein kinase, and Toll-like receptor signaling pathways, all of which are related to influenza A virus progression. Conclusions: Collectively, our results provided novel insights into the miRNA expression patterns of tracheal tissues from H5N1-infected Ri chickens. More importantly, our findings offer insights into the relationship between miRNA and immune-related target genes and the role of miRNA in HPAIV infections in chickens.

luxS and smcR Quorum-Sensing System of Vibrio vulnificus as an Important Factor for In Vivo Survival

  • SHIN NA-RI;BAEK CHANG-HO;LEE DEOG-YONG;CHO YOUNG-WOOK;PARK DAE-KYUN;LEE KO-EUN;KIM KUN-SOO;YOO HAN-SANG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1197-1206
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    • 2005
  • Vibrio vulnificus is an opportunistic pathogen that causes a septicemia and expresses numerous virulence factors, in which luxS and smcR are genes encoding for components responsible for quorum-sensing regulation. In the present study, null mutants were constructed with lesions in each or both of these two genes from the V. vulnificus Vv$\Delta$Z strain, which is a lacZ$^{-}$ and chloramphenicol/streptomycin-resistant derivative of the wild-type ATCC29307 strain, and their phenotypes related to virulence were compared with those of the parental cells. $LD_{50}$ and histopathological findings of luxS-, smcR-, or luxS- smcR- deficient mutant were not different from those of the parent strain, a lacZ-deficient streptomycin-resistant strain in mice. However, time of death in mice was delayed, and numbers of bacteria survived in bloodstream after intraperitoneal injection in mice were decreased by mutation, especially luxS and smcR double mutant (VvSR$\Delta$ZSR). These phenomena were supported by increased serum sensitivity and delayed bacterial proliferation in both murine blood and iron-restricted medium. These results suggest that the luxS and luxR homologous genes in V. vulnificus could playa role in bacterial survival in host by enhancing proliferation and adjusting to changed environment.

Species Transferability of Klebsiella pneumoniae Carbapenemase-2 Isolated from a High-Risk Clone of Escherichia coli ST410

  • Lee, Miyoung;Choi, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.30 no.7
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    • pp.974-981
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    • 2020
  • Sequence type 410 (ST410) of Escherichia coli is an extraintestinal pathogen associated with multi drug resistance. In this study, we aimed to investigate the horizontal propagation pathway of a high-risk clone of E. coli ST410 that produces Klebsiella pneumoniae carbapenemase (KPC). blaKPC-encoding E. coli and K. pneumoniae isolates were evaluated, and complete sequencing and comparative analysis of blaKPC-encoding plasmids from E. coli and K. pneumoniae, antimicrobial susceptibility tests, polymerase chain reaction, multilocus sequence typing, and conjugal transfer of plasmids were performed. Whole-genome sequencing was performed for plasmids mediating KPC-2 production in E. coli and K. pneumoniae clinical isolates. Strains E. coli CPEc171209 and K. pneumoniae CPKp171210 were identified as ST410 and ST307, respectively. CPEc171209 harbored five plasmids belonging to serotype O8:H21, which is in the antimicrobial-resistant clade C4/H24. The CPKp171210 isolate harbored three plasmids. Both strains harbored various additional antimicrobial resistance genes. The IncX3 plasmid pECBHS_9_5 harbored blaKPC-2 within a truncated Tn4401a transposon, which also contains blaSHV-182 with duplicated conjugative elements. This plasmid displayed 100% identity with the IncX3 plasmid pKPBHS_10_3 from the K. pneumoniae CPKp171210 ST307 strain. The genes responsible for the conjugal transfer of the IncX3 plasmid included tra/trb clusters and pil genes coding the type IV pilus. ST410 can be transmitted between patients, posing an elevated risk in clinical settings. The emergence of a KPC-producing E. coli strain (ST410) is concerning because the blaKPC-2-bearing plasmids may carry treatment resistance across species barriers. Transgenic translocation occurs among carbapenem-resistant bacteria, which may spread rapidly via horizontal migration.

Antimicrobial Resistance and Minimum Inhibitory Concentrations of Vibrio parahaemolyticus Strains Isolated from Gomso Bay, Korea (곰소만 해역 해수에서 분리한 장염비브리오(Vibrio parahaemolyticus)의 항균제 내성 및 최소발육억제농도의 구명)

  • Kim, Tae-Ok;Um, In-Seon;Kim, Hee-Dai;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.5
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    • pp.582-588
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    • 2016
  • Seventy-nine Vibrio parahaemolyticus isolates from surface seawater from Gomso Bay, west coast of Korea, were analyzed for the presence of virulence genes and their susceptibility to 30 different antimicrobials. All 79 isolates were examined for the presence of two virulence genes (tdh or trh) using polymerase chain reaction (PCR); however, no isolates possessed either the tdh or trh gene. According to a disk diffusion susceptibility test, all of the strains studied were resistant to oxacillin, penicillin, and vancomycin, followed by ticarcillin (97.5%), ampicillin (96.2%), clindamycin (86.1%), erythromycin (10.1%), streptomycin (7.6%), cefoxitin (6.3%), amikacin (2.5%), and cephalothin (2.5%). However, all of the strains were susceptible to 19 other antimicrobials including cefepime, cefotaxime, chloramphenicol, gentamycin, nalidixic acid, sulfamethoxazole/trimethoprim, and trimethoprim. All 79 isolates (100%) were resistant to four or more classes of antimicrobials, and two strains exhibited resistance to eight antimicrobial agents. The average minimum inhibitory concentrations (MICs) for V. parahaemolyticus for ampicillin, penicillin, ticarcillin, and vacomycin were 946.5, 1,305.9, 1,032.3, and 45.0 µg/mL, respectively.