• Title/Summary/Keyword: multiple-resistance gene

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Antibacterial Effect of Various Fermentation Products and Identification of Differentially Expressed Genes of E.coli (다양한 발효액의 항균효과와 대장균의 유전적 변화에 미치는 영향)

  • Heo, Jihye
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.2
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    • pp.119-124
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    • 2022
  • Pseudomonas aeruginosa (P. aeruginosa), Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) are typical opportunistic pathogens. Moreover, these bacteria are known to possess multidrug-resistant (MDR) properties. This study investigates the antimicrobial activity of six fermented products, which have varying efficacies against P. aeruginosa, E. coli, and S. aureus. To identify novel candidate genes, differential expression analysis was performed using an annealing control primer. In the disk diffusion method, Fig vinegar (FV) and Diospyros kaki Thunb vinegar (DTV) showed the greatest increase in inhibition compared to other fermented products, whereas fermented Korean traditional nature herb (FKTNH) had no antibacterial effect. This study identified down-regulation of Escherichia coli O157:H7 ompW gene for outer membrane protein W, whereas gene for synthetic construct Lao1 gene for L-amino acid oxidase were up-regulated in E. coli treated with 5% FV. Consuming fermented vinegar helps prevent bacterial infections. Especially, FV and DTV are potentially useful alternative natural products for multidrug resistance. Furthermore, both are expected to be used as effective natural antimicrobial agents, such as disinfectants.

Hypoxia Induced Multidrug Resistance of Laryngeal Cancer Cells via Hypoxia-inducible Factor-1α

  • Li, Da-Wei;Dong, Pin;Wang, Fei;Chen, Xin-Wei;Xu, Cheng-Zhi;Zhou, Liang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4853-4858
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    • 2013
  • Objectives: To investigate whether hypoxia has an effect on regulation of multidrug resistance (MDR) to chemotherapeutic drugs in laryngeal carcinoma cells and explore the role of hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$). Methods: Laryngeal cancer cells were cultured under normoxic and hypoxic conditions. The sensitivity of the cells to multiple drugs and levels of apoptosis induced by paclitaxel were determined by MTT assay and annexin-V/propidium iodide staining analysis, respectively. HIF-$1{\alpha}$ expression was blocked by RNA interference. The expression of HIF-$1{\alpha}$ gene was detected by real-time quantitative RT-PCR and Western blotting. The value of fluorescence intensity of intracellular adriamycin accumulation and retention in cells was evaluated by flow cytometry. Results: The sensitivity to multiple chemotherapy agents and induction of apoptosis by paclitaxel could be reduced by hypoxia (P<0.05). A the same time, the adriamycin releasing index of cells was increased (P<0.05). However, resistance acquisition subject to hypoxia in vitro was suppressed by down-regulating HIF-$1{\alpha}$ expression. Conclusion: HIF-$1{\alpha}$ could be considered as a key regulator for mediating hypoxia-induced MDR in laryngeal cancer cells via inhibition of drug-induced apoptosis and decrease in intracellular drug accumulation.

Evidences that Suggest the Spread of Multiple-Antibiotic-Resistance (mar) Operon of Escherichia coli Mutants among Gram-Negative Bacilli (Mar (Multiple-Antibiotic-Resistance) Operon 돌연변이 대장균의 그람음성 세균들간 전파 가능성에 대한 근거)

  • Byung-Tae Park
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.17-26
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    • 1999
  • To evaluate the spreading possibilities of the marRAB mutation of E. coli Mar mutant among gram-negative bacilli, chromosomal marRAB mutations of Mar mutants were transduced by $\lambda$placMu9 into pUC19 (Lac$^{+}$, Ap$^{r}$) cloning site in another strains of E. coli or onto the chrmosome of S. typhimurium and P. aeruginosa, selected for transduction by Mar phenotype, Lac$^{-}$, or Ap$^{r}$, and tested for their antimicrobial resistance with or without addition of salicylate (SAL). Compared with wild type strains of JM109, NM522, harboring pUC19 or not, respectively, all strains of JM109 or NM522 carrying pUC19::marRAB mutation showed higher levels of antimicrobial resistance and SAL induction of Mar phenotype than those of wild type. However, in contrast to the original Mar mutants, there were some tendencies of decreased antimicrobial resistance of JM109 or NM522 harboring pUC19::marRAB mutation with SAL induction against chlorarnphenicol (Cm) and tetracycline (Tc), or Tc and ciprofloxacin (Cp), respectively. Almost the same results, as shown as the cases of E. coli JM109 or NM522, were obtained from all transductants of S. typhimurium and P. aeruginosa, except Cp, against which increased antimicrobial resistance with SAL induction was shown. This study, employed the methods of transformation or transduction among intercellular gene transfer methods between gram-negative bacteria, shows the evidences that suggest indirectly the spreading possibilities of marRAB mutation among gram-negative bacilli.

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Transgenic Tobacco Expressing the hrpNEP Gene from Erwinia pyrifoliae Triggers Defense Responses Against Botrytis cinerea

  • Sohn, Soo-In;Kim, Yul-Ho;Kim, Byung-Ryun;Lee, Sang-Yeob;Lim, Chun Keun;Hur, Jang Hyun;Lee, Jang-Yong
    • Molecules and Cells
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    • v.24 no.2
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    • pp.232-239
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    • 2007
  • $HrpN_{EP}$, from the gram-negative pathogen, Erwinia pyrifoliae, is a member of the harpin group of proteins, inducing pathogen resistance and hypersensitive cell death in plants. When the $hrpN_{EP}$ gene driven by the OsCc1 promoter was introduced into tobacco plants via Agrobacterium-mediated transformation, their resistance to the necrotrophic fungal pathogen, Botrytis cinerea, increased. Resistance to B. cinerea was correlated with enhanced induction of SA-dependent genes such as PR-1a, PR2, PR3 and Chia5, of JA-dependent genes such as PR-1b, and of genes related to ethylene production, such as NT-EFE26, NT-1A1C, DS321, NT-ACS1 and NT-ACS2. However the expression of NPR1, which is thought to be essential for multiple-resistance, did not increase. Since the pattern of expression of defense-related genes in $hrpN_{EP}$-expressing tobacco differed from that in plants expressing $hpaG_{Xoo}$ from Xanthomonas oryzae pv. Oryzae, these results suggest that different harpins can affect the expression of different defense-related genes, as well as resistance to different plant pathogens.

Ambient Air Waste Sorting Facilities Could Be a Source of Antibiotic Resistant Bacteria

  • Calheiros, Ana;Santos, Joana;Ramos, Carla;Vasconcelos, Marta;Fernandes, Paulo
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.367-373
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    • 2021
  • The antimicrobial resistance of Staphylococcus spp. and Gram negative strains present in air samples from waste sorting facilities was assessed. Phenotypic studies have revealed a high percentage of strains of Staphylococcus spp. resistant to methicillin. Genotypically and by RT-PCR, it was found that the mecA gene usually associated with methicillin resistance was present in 8% of the Staphylococcus strains isolated. About 30% of the Gram negative strains from the same samples also displayed resistance to meropenem and 79% of these were resistant to multiple antibiotics from different classes, namely cephalosporins and β-lactams. The results suggest that in professional activities with high levels of exposure to biological agents, the quantification and identification of the microbial flora in the work environment, with the determination of the presence of potential agents displaying multi-resistances is of relevance to the risk assessment. The personal protection of workers is particularly important relevance in these cases, since many of the strains that exhibit multi-resistance are potential opportunistic agents.

Functional Analysis of the marB gene of Escherichia coli K-12

  • Lee, Chang-Mi;Park, Byung-Tae
    • Biomedical Science Letters
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    • v.10 no.2
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    • pp.153-161
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    • 2004
  • Antibiotic resistance is often associated with the production of inner membrane proteins (for example, AcrAB/TolC efflux pump) that are capable to extrude antibiotics, detergents, dyes and organic solvents. In order to evaluate the unknown MarB function of Escherichia coli, especially focused on the function of OmpF porin, several mutants were construted by T4GT7 transduction. MarA plays a major roles in mar (multiple antibiotic resistance) phenotype with AcrAB/TolC efflux pump in E. coli K-12. Futhermore, MarA decreases OmpF porin expression via micF antisense RNA. Expression of acrAB is increased in strains containing mutation in marR, and in those carrying multicopy plasmid expressing marA. MarB protein of E. coli K-12 showed its activity at OmpF porin & TolC protein as target molecule. Some paper reported MarB positively regulates OmpF function. MarA shows mar phenotype, and MarB along with MarA show decreased MIC through OmpF function. By this experiment, MarB could decrease MIC through the OmpF porin & TolC protein as target.

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Proposing new models to predict pile set-up in cohesive soils

  • Sara Banaei Moghadam;Mohammadreza Khanmohammadi
    • Geomechanics and Engineering
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    • v.33 no.3
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    • pp.231-242
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    • 2023
  • This paper represents a comparative study in which Gene Expression Programming (GEP), Group Method of Data Handling (GMDH), and multiple linear regressions (MLR) were utilized to derive new equations for the prediction of time-dependent bearing capacity of pile foundations driven in cohesive soil, technically called pile set-up. This term means that many piles which are installed in cohesive soil experience a noticeable increase in bearing capacity after a specific time. Results of researches indicate that side resistance encounters more increase than toe resistance. The main reason leading to pile setup in saturated soil has been found to be the dissipation of excess pore water pressure generated in the process of pile installation, while in unsaturated conditions aging is the major justification. In this study, a comprehensive dataset containing information about 169 test piles was obtained from literature reviews used to develop the models. to prepare the data for further developments using intelligent algorithms, Data mining techniques were performed as a fundamental stage of the study. To verify the models, the data were randomly divided into training and testing datasets. The most striking difference between this study and the previous researches is that the dataset used in this study includes different piles driven in soil with varied geotechnical characterization; therefore, the proposed equations are more generalizable. According to the evaluation criteria, GEP was found to be the most effective method to predict set-up among the other approaches developed earlier for the pertinent research.

A Broad-Host-Range Promoter-Probe Vector, pKU20, and Its Use in Promoter Cloning and Expression of Bacillus thuringiensis Crystal Protein Gene in Pseudomonas putida

  • SHIN, BYUNG SIK;BON TAG KOO;SEUNG HWAN PARK;HO YONG PARK;JEONG IL KIM
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.240-245
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    • 1991
  • We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range plsmid RSF1010, as a base. The pKU20 contains structural gene for aminoglycoside phos-photransferase (aph), without promoter, and a multiple cloning site upstream the aph. Using this vector, a 412base pairs (bp) PstI fragment showing strong promoter activity both in Escherichia coli LE392 and Pseudomonas putida KCTC1644 has been cloned from Pseudomonas fluorescens chromosomal DNA on the basis of streptomycin resistance. The nucleotide sequence of the 412 bp fragment has been determined and the putative - 35 and -10 region was observed. Insecticidal protein gene of Bacillus thuringiensis subsp. kurstaki HD-73 inserted on downstream of the promoterlike DNA fragment was efficiently expressed in E. coli and P. putida. The toxin protein was efficiently synthesized in an insoluble form in both strains.

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OsAREB1, an ABRE-binding protein responding to ABA and glucose, has multiple functions in Arabidopsis

  • Jin, Xiao-Fen;Xiong, Ai-Sheng;Peng, Ri-He;Liu, Jin-Ge;Gao, Feng;Chen, Jian-Min;Yao, Quan-Hong
    • BMB Reports
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    • v.43 no.1
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    • pp.34-39
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    • 2010
  • Expression patterns of OsAREB1 revealed that expression of OsAREB1 gene can be induced by ABA, PEG and heat. Yeast one-hybrid assay demonstrated it can bind to ABA-responsive element (ABRE), which was found in most stress-induced genes. Transgenic Arabidopsis over-expressing OsAREB1 had different responses to ABA and glucose compared to wild-type plants, which suggest OsAREB1 might have a crucial role in these two signaling pathways. Further analysis indicate that OsAREB1 have multiple functions in Arabidopsis. First, OsAREB1 transgenic plants had higher resistance to drought and heat, and OsAREB1 up-regulated the ABA/stress related gene such as RD29A and RD29B. Second, it delayed plant flowering time by down-regulating the expression of flowering-related genes, such as FT, SOC1, LFY and AP1. Due to the dates, OsAREB1 may function as a positive regulator in drought/heat stresses response, but a negative regulator in flowering time in Arabidopsis.

Detection of Point Mutations in the rpoB Gene Related to Drug Susceptibility in Mycobacterium Tuberculosis using an Oligonucleotide Chip (올리고뉴클레오티드 칩(Oligonucleotide Chip)을 이용한 항결핵제 감수성과 관련된 Mycobacterium tuberculosis rpoB 유전자의 점돌연변이 판별 방법)

  • Kim, Hyun-Jung;Kim, Seong-Keun;Shim, Tae-Sun;Park, Yong-Doo;Park, Mi-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.29-41
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    • 2001
  • Background : The appearance of multiple-drug-resistant Mycobacterium tuberculosis strains has been seriously compromising successful control of tuberculosis. Rifampin-resistance, caused by mutations in the rpoB gene, can be indicative of multiple-drug-resistance, and its detection is of great importance. The present study aimed to develop an oligonucleotide chip for accurate and convenient screening of drug-resistance. Methods : In order to detect point mutations in the rpoB gene, an oligonucleotide chip was prepared by immobilizing specific probe DNA to a microscopic slide glass by a chemical reaction. The probe DNA that was selected from the 81 bp core region of the rpoB gene was designed to have mutation sites at the center. A total of 17 mutant probes related to rifampin-resistance including 8 rifabutin-sensitive mutant probes were used in this study. For accurate determination, wild type probes were prepared for each mutation position with an equal length, which enabled a direct comparison of the hybridization intensities between the mutant and wild type. Results : Mycobacterial genomic DNA from clinical samples was tested with the oligonucleotide chip and the results were compared with those of the drug-susceptibility test in addition to sequencing and INNO-LiPA Rif. TB kit test in some cases. Out of 15 samples, the oligonucleotide chip results of 13 samples showed good agreement with the rifabutin-sensitivity results. The two samples with conflicting result also showed a discrepancy between the other tests, suggesting such possibilities as existence of mixed strains and difference in drug-sensitivity. Further verification of these samples in addition to more case studies are required before the final evaluation of the oligonucleotide chip can be made. Conlcusion : An oligonucleotide chip was developed for the detection of rpoB gene mutations related to drugsusceptibility. The results to date show the potential for using the oligonucleotide chip for accurate and convenient screening of drug-resistance to provide useful information in antituberculosis drug therapy.

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