• Title/Summary/Keyword: Acid-resistant

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Mutagenesis of Streptomyces kasugaensis for Kasugamycin Production

  • Cho, Hoon;Choi, Du-Bok;Lim, Chae-Kyu
    • Journal of environmental and Sanitary engineering
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    • v.23 no.4
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    • pp.23-29
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    • 2008
  • This study was performed to develop mutant strain using a combination of UV irradiation procedures with protoplast mutagenesis in order to achieve an effective kasugamycin production from Streptomyceskasugaensis. Whenlessthan 1.0g/lof Linoleic acid was used, the cell growth was not inhibited. On the other hand, the cell growth was greatly inhibited when more than 1.6 g/l of linoleic acid was used. Among the various mutant strains, SK-12 was obtained in medium containing 1.6g/l of linoleic acid, showing the highest rate of both cell growth and kasugamycin production. In order to compare kasugamycin production with the SK-12 and the parent strain using soybean oil, cultures were performed in a flask. The production of kasugamycin was increased with the increase time. The maximum kasugamycin concentration was 1.2g/l after 6 days of culture. The product yield from soybean oil was 0.05g/l/g consumed carbon source, which was roughly 5.0 fold higher than the parent strain. These results show that it was effective method for obtaining a mutant resistant to linoleic acid for the effective production of kasugamycin from soybean oil.

Fungal Development, Respiration and Activity of Oxidative Enzymes in Rice Plants Inoculated with Pyricularia oryzae in Both Compatible and Incompatible Combinations (벼 도열병균에 감염된 친화 및 비친화 조합 벼에서의 균생장, 호흡 및 산화효소 활성)

  • Chung Bong-Koo;Chung Hoo-Sup
    • Korean journal of applied entomology
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    • v.21 no.3 s.52
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    • pp.113-122
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    • 1982
  • Appressorial formation of Pyricularia oryzae on leaves showed no marked difference between highly resistant Tongil and susceptible Norm No. 6. Race N-2 of the blast fungus penetrated directly into motor cells of susceptible cultivar Norm No. 6, later extensively spreading hyphae were developed, while in the cultivar Tongil, after penetration, no further hyphal extension resulted. In discoloration of infected tissues, the highly resistant cultivar Tongil not only discolored rapidly, but also the percentage of discolored cells was higher than the susceptible cultivars, Jinheung and Norm No. 6. The respiratory rate, was generally higher in infected tissue than in healthy tissue. No significant difference in the respiration rate of resistant Suwon No. 180 was not found between the infected and healthy leaf tissue, whereas, in susceptible Jinheung, a marked increase in respiratory rate was caused by blast infection. The respiratory rate increased at the appearance of the first visible symptom in all cultivars resistant or susceptible. Higher peroxidase activity was found in infected tissues as compared with healthy tissue. Peroxidase activity increased in resistant and susceptible reactions. Particularly, in resistant reaction, the increase of the activity was more pronounced. In highly resistant reaction, there was no difference in peroxidase activity between healthy and infected tissues. Ascorbic acid oxidase, hydroquinone oxidase and catechol oxidase had the same trend as the peroxidase. In contrast, activity of catalase rather decreased in leaf tissues infected with compatible races of the fungus.

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Bacteriological Identification of Strains Isolated from Diarrheal Patients at Ham Yang Gun, Kyeongsang-Namdo in July 1971 (1971년(年) 경남(慶南) 함양군(咸陽郡)에서 유행(流行)한 설사환자(患者)에서 분리(分離)한 Salmonella.Shigella 균(菌)의 세균학적(細菌學的) 연구(硏究))

  • Kim, Je-Soo;Ju, Byung-Chil;Park, Soo-Yeon;Ju, Jin-Woo;Yang, Hak-Do
    • The Journal of the Korean Society for Microbiology
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    • v.7 no.1
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    • pp.1-8
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    • 1972
  • The authors made a study on the bacteriological identification about unidentified strains which were roughly screened by local health center, which an epidemic diarrhea was outbreak at Ham Yang Gun, Kyeongsang-Namdo in July 1971. And the authors made an attempt to bacteriological Identification, serotyping with slide agglutination, and determination of the susceptibility of identified strains to various chemotherapeutic agents. The results Were obtained as follows: 1. The isolated strains identified Shigella flexneri type 2b(2 strains) and Salmonella paratyphi B(4 strains). 2. Sensitivity test using with three concentrations chemotherapeutic agents(Paper disk used; Eiken chemical Co., Tokyo, Japan): (1) Shigella flexneri type 2b isolated strains were sensitive to kanamycin, colistin(100%) and penicllin(50%) respectively, but resistant to erythromycin, oleandomycin, leucomycin, chloramphenicol, tetracycline, dihydrostreptomycin and sulfadimethoxin(100%) respectively. (2) Salmonella paratyphi B isolated strains were sensitive to chloramphenicol, tetracycline, dihydrostreptomycin, kanamycin, sulfadimethoxine(100%) and colistin(50%) respectively, but resistant to penicillin, erythromycin, oleandomycin and leucomycin(100%) respectively. 3. Sensitivity test using with multodisk urinary code(Paper disk used; Oxoid, London): (1) Shigella flexneri type 2b isolated strains were sensitive to Bactrim, ampicillin, nitrofurantoin, nalidixic add and gentamicin(100%) respectively, but resistant to chloramhpenicol, tetracycline, sulfadiazine and Fanasil(100%) respectively. (2) Salmonell paratyphi B isolated strains were sensitive to Bactrim, chloramphenicol, ampicillin, nitrofurantoin, tetracycline, nalidixic acid, gentamicin(100%) and sulfadiazine(50%) respectively, but resistant to Fanasil(100%) and sulfadiazine(50%) respectively.

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Vesicular Stomatitis Virus G Glycoprotein and ATRA Enhanced Bystander Killing of Chemoresistant Leukemic Cells by Herpes Simplex Virus Thymidine Kinase/Ganciclovir

  • Hu, Chenxi;Chen, Zheng;Zhao, Wenjun;Wei, Lirong;Zheng, Yanwen;He, Chao;Zeng, Yan;Yin, Bin
    • Biomolecules & Therapeutics
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    • v.22 no.2
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    • pp.114-121
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    • 2014
  • Refractoriness of acute myeloid leukemia (AML) cells to chemotherapeutics represents a major clinical barrier. Suicide gene therapy for cancer has been attractive but with limited clinical efficacy. In this study, we investigated the potential application of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) based system to inhibit chemoresistant AML cells. We first generated Ara-C resistant K562 cells and doxorubicin-resistant THP-1 cells. We found that the HSV-TK/GCV anticancer system suppressed drug resistant leukemic cells in culture. Chemoresistant AML cell lines displayed similar sensitivity to HSV-TK/GCV. Moreover, HSV-TK/GCV killing of leukemic cells was augmented to a mild but significant extent by all-trans retinoic acid (ATRA) with concomitant upregulation of Connexin 43, a major component of gap junctions. Interestingly, HSV-TK/GCV killing was enhanced by expression of vesicular stomatitis virus G glycoprotein (VSV-G), a fusogenic membrane protein, which also increased leukemic cell fusion. Co-culture resistant cells expressing HSV-TK and cells stably transduced with VSV-G showed that expression of VSV-G could promote the bystander killing effect of HSV-TK/GCV. Furthermore, combination of HSV-TK/GCV with VSV-G plus ATRA produced more pronounced antileukemia effect. These results suggest that the HSV-TK/GCV system in combination with fusogenic membrane proteins and/or ATRA could provide a strategy to mitigate the chemoresistance of AML.

Identification of Lactobacillus ruminus SPM0211 Isolated from Healthy Koreans and Its Antimicrobial Activity against Some Pathogens

  • Yun Ji-Hee;Yim Dong-sool;Kang Jin-Yang;Kang Byung-Yong;Shin Eun-ah;Chung Myung-Jun;Kim Soo-Dong;Baek Dae-Heoun;Kim Kyungjae;Ha Nam-Joo
    • Archives of Pharmacal Research
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    • v.28 no.6
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    • pp.660-666
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    • 2005
  • The intestinal microbiota are important to the host with regard to resistance they impart against bacterial infections and their involvement in mediating metabolic functions. Lactic acid producing bacteria such as Lactobacillus play an important physiological role in these matters. The aim of the present study was to isolate Lactobacillus sp. that inhibits enteric pathogens. Initially, 17 isolates from healthy Koreans were collected on Lactobacillus selective medium. Resistance of the isolates to antibiotics including rifampicin, streptomycin, clindamycin and vancomycin was measured. One of the isolate was identified as Lactobacillus ruminus on the basis of bacterial cell morphology, cultural characteristic and biochemical characteristics, 16S rRNA sequence analysis and PCR-RAPD. Antimicrobial activity of the bacterium against Vancomycin Intermediate Resistant Staphylococcus aureus (VISA) and Vancomycin-Resistant Enterococci (VRE) was measured. About $10^4$ cells of VISA or VRE were mixed with 1, 5, and 9 mL of L. ruminus SPM 0211 and the final volume was adjusted to 10 mL with brain heart infusion (BHI) broth. The cell suspension was incubated for 3, 6, 9, and 24 h, serially diluted and then plated on BHI agar plates. As numbers of L. ruminus SPM 0211 were increased, viable cell count of VISA and VRE decreased. The strongest antimicrobial activity of SPM 0211 was observed after 9 h incubation in any mixture, almost completely inhibiting the growth of these two bacteria. The results suggest that the freshly isolated L. ruminus SPM 0211 may be used as a pro-biotic microbe that prevents the colonization of enteric pathogens and can thereby promote good gastrointestinal health.

Genetic properties of R plasmids in Shigella isolates of swine origin in Korea (돈(豚)에서 분리(分離)한 Shigella균유래(菌由來) R plasmid의 유전적(遺傳的) 특성(特性)에 관한 연구(硏究))

  • Choi, Won-pil;Kwun, Hae-byeng;Jung, Suk-chan
    • Korean Journal of Veterinary Research
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    • v.29 no.1
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    • pp.37-44
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    • 1989
  • This paper dealt with the distribution of Shigella spp. on 5 piggeries in Taegu and Kyungpook during the period from August to October 1987. Isolated Shigella were examined for serogrouping, antimicrobial drug resistance and detection of R plasmid. Genetic properties of R plasmid in Shigella have examined to fertility inhibition (Fi) and gel electrophoresis was performed for the isolation of plasmid DNA. The results obtained were summarised as followings; 1. Of total 2,978 samples from 5 piggeries, 82 strains (2.8%) of Shigella spp. were isolated from 82 samples. The isolated strains were identified as S dysenteriae (60 strains), S flexneri (20 strains) and S sonnei (2 strains). 2. Of the 82 strains examined 67 (95.1%) were resistant to one or more antibiotics, such as ampicillin (Am), chloramphenicol (Cm), kanamycin (Km), nalidixic acid (Na), rifampicin (Rf), streptomycin (Sm), sulfademethoxine (Su), and tetracycline (Tc) and higher resistant to Su (90.2%), Sm (63.4%) and Tc (63.4%). 3. Of the 78 resistant Shigella strains 26 (33.3%) harbored conjugative R plasmids and the transfer frequency of Sm (50.0%), Cm(33.3%) resistance was much higher than that of the other drug resistance. 4. The most common resistant patterns were SmSuTc, Su and AmSmSuTc. 5. Out of the 26 Shigella R plasm ids examined for Fi, 14(53.8%) were $Fi^+$ and the remainder were $Fi^-$. 6. The plasmid DNA profiles in Shigella spp. (9 strains) isolated from pigs were confirmed as being 2 to 9 fragments by the gel electrophoresis. Their molecular size ranged 2.17 to 87.62 kilobase (Kb). All strains of Shigella spp. consisted in 15.4 Kb plasmids.

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Genetic Characterization of β-lactamase (VPA0477) in Vibrio parahaemolyticus (장염비브리오가 보유하는 β-lactamase (VPA0477)의 유전학적 특성)

  • Lee, Nam-Hyung;Song, Hyun-Jung;Park, Chang-Soo;Kim, Hee-Dai;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.6
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    • pp.597-604
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    • 2011
  • Using 108 strains of Vibrio parahaemolyticus isolated from seawater, we investigated ampicillin-resistance profiles and the genetic characterization of ${\beta}$-lactamase (VPA0477). All of the strains studied, except one strain, were resistant to ampicillin. However, the strain that was susceptible to ampicillin had the same ${\beta}$-lactamase gene as the ampicillin-resistant strains. We compared ${\beta}$-lactamase promoter region sequences among five strains, including both ampicillin-resistant and -susceptible strains. In the susceptible strain, a nucleotide at position -19 in the methionine initiation codon for ${\beta}$-lactamase was not present in the ampicillin-resistant strains. The genes in the region containing the gene VPA0477 were present in all of the tested strains, and LA-PCR analysis showed that the distance between VPA0474 and VPA0479 in all of the V. parahaemolyticus samples was precisely 5.7 kb. In V. parahaemolyticus ${\beta}$-lactamase, four important structural features that are conserved in Class A ${\beta}$-lactamases were present in the deduced amino acid sequences. Taken together, our study demonstrates that V. parahaemolyticus ${\beta}$-lactamase is included in the Class A ${\beta}$-lactamase group, and some nucleotides within the promoter region are of particular importance for ${\beta}$-lactamase activity.

Antimicrobial Resistance and the Presence of Virulence Genes in Escherichia coli Strains Isolated from Ruditapes philippinarum in Gomso Bay, Korea (곰소만 해역의 바지락(Ruditapes philippinarum)에서 분리한 대장균 (Escherichia coli)의 항균제 내성 및 병원성 유전자의 보유성)

  • Kim, Tae-Ok;Eom, In-Seon;Park, Kwang-Ho;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.6
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    • pp.800-806
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    • 2016
  • In total, 151 Escherichia coli isolates from Ruditapes philippinarum in Gomso Bay were analyzed for their susceptibility to 18 different antimicrobial agents and for genes associated with virulence. For virulence genes, each strain of the isolates was positive for the enterotoxigenic E. coli (ETEC)-specific heat-stable toxin (estA), enteroinvasive E. coli (EIEC)-specific invasion-associated locus (iaa) gene and enteropathogenic E. coli (EPEC)-specific attaching and effacing (eae) gene. According to a disk diffusion susceptibility test, resistance to ampicillin was most prevalent (23.2%), followed by resistance to amoxicillin (22.5%), ticarcillin (20.5%), tetracycline (18.5%), nalidixic acid (12.6%), ciprofloxacin (10.6%), streptomycin (9.9%), and chloramphenicol (6.6%). More than 35.8% of the isolates were resistant to at least one antimicrobial agent, and 19.9% were resistant to four or more classes of antimicrobials; these were consequently defined as multidrug resistant. Minimum inhibitory concentration (MIC) ranges for the antimicrobial resistance of the 15 different antimicrobial agents of 54 E. coli strains were confirmed by varying the concentrations from $32-2,048{\mu}g/mL$. Overall, these results not only provide novel insights into the necessity for seawater and R. philippinarum sanitation in Gomso Bay but they also help to reduce the risk of contamination by antimicrobial-resistant bacteria.

Outcomes and Use of Therapeutic Drug Monitoring in Multidrug-Resistant Tuberculosis Patients Treated in Virginia, 2009-2014

  • Heysell, Scott K.;Moore, Jane L.;Peloquin, Charles A.;Ashkin, David;Houpt, Eric R.
    • Tuberculosis and Respiratory Diseases
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    • v.78 no.2
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    • pp.78-84
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    • 2015
  • Background: Reports of therapeutic drug monitoring (TDM) for second-line medications to treat multidrug-resistant tuberculosis (MDR-TB) remain limited. Methods: A retrospective cohort from the Virginia state tuberculosis (TB) registry, 2009-2014, was analyzed for TDM usage in MDR-TB. Drug concentrations, measured at time of estimated peak ($C_{max}$), were compared to expected ranges. Results: Of 10 patients with MDR-TB, 8 (80%) had TDM for at least one drug (maximum 6 drugs). Second-line drugs tested were cycloserine in seven patients (mean $C_{2hr}$, $16.6{\pm}10.2{\mu}g/mL$; 4 [57%] below expected range); moxifloxacin in five (mean $C_{2hr}$, $3.2{\pm}1.5{\mu}g/mL$; 1 [20%] below); capreomycin in five (mean $C_{2hr}$, $21.5{\pm}14.0{\mu}g/mL$; 3 [60%] below); para-aminosalicylic acid in five (mean $C_{6hr}$, $65.0{\pm}29.1{\mu}g/mL$; all within or above); linezolid in three (mean $C_{2hr}$, $11.4{\pm}4.1{\mu}g/mL$, 1 [33%] below); amikacin in two (mean $C_{2hr}$, $35.3{\pm}3.7{\mu}g/mL$; 1 [50%] below); ethionamide in one ($C_{2hr}$, $1.49{\mu}g/mL$, within expected). Two patients died: a 38-year-old woman with human immunodeficiency virus/acquired immune deficiency syndrome and TB meningitis without TDM, and a 76-year-old man with fluoroquinolone-resistant (pre-extensively drug-resistant) pulmonary TB and low linezolid and capreomycin concentrations. Conclusion: Individual pharmacokinetic variability was common. A more standardized approach to TDM for MDR-TB may limit over-testing and maximize therapeutic gain.

Comparative Performance of Line Probe Assay (Version 2) and Xpert MTB/RIF Assay for Early Diagnosis of Rifampicin-Resistant Pulmonary Tuberculosis

  • Yadav, Raj Narayan;Singh, Binit Kumar;Sharma, Rohini;Chaubey, Jigyasa;Sinha, Sanjeev;Jorwal, Pankaj
    • Tuberculosis and Respiratory Diseases
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    • v.84 no.3
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    • pp.237-244
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    • 2021
  • Background: The emergence of drug-resistant tuberculosis (TB), is a major menace to cast off TB worldwide. Line probe assay (LPA; GenoType MTBDRplus ver. 2) and Xpert MTB/RIF assays are two rapid molecular TB detection/diagnostic tests. To compare the performance of LPA and Xpert MTB/RIF assay for early diagnosis of rifampicin-resistant (RR) TB in acid-fast bacillus (AFB) smear-positive and negative sputum samples. Methods: A total 576 presumptive AFB patients were selected and subjected to AFB microscopy, Xpert MTB/RIF assay and recent version of LPA (GenoType MTBDRplus assay version 2) tests directly on sputum samples. Results were compared with phenotypic culture and drug susceptibility testing (DST). DNA sequencing was performed with rpoB gene for samples with discordant rifampicin susceptibility results. Results: Among culture-positive samples, Xpert MTB/RIF assay detected Mycobacterium tuberculosis (Mtb) in 97.3% (364/374) of AFB smear-positive samples and 76.5% (13/17) among smear-negative samples, and the corresponding values for LPA test (valid results with Mtb control band) were 97.9% (366/374) and 58.8% (10/17), respectively. For detection of RR among Mtb positive molecular results, the sensitivity of Xpert MTB/RIF assay and LPA (after resolving discordant phenotypic DST results with DNA sequencing) were found to be 96% and 99%, respectively. Whereas, specificity of both test for detecting RR were found to be 99%. Conclusion: We conclude that although Xpert MTB/RIF assay is comparatively superior to LPA in detecting Mtb among AFB smear-negative pulmonary TB. However, both tests are equally efficient in early diagnosis of AFB smear-positive presumptive RR-TB patients.