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Molecular Characterization and Prevalence of 16S Ribosomal RNA Methylase Producing Bacteria in Amikacin Resistant Gram-negative Bacilli Isolated from Clinical Specimens

  • Shin, Kyung-A;Hwang, Seock-Yeon;Hong, Seung-Bok
    • Biomedical Science Letters
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    • v.18 no.3
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    • pp.299-306
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    • 2012
  • Recently, the prevalence of 16S rRNA methylase conferring high-level resistance to aminoglycosides has been increasing in Gram-negative bacilli globally. We determined the prevalence and genotype of these methylase-producing bacteria, and characterized the co-resistance to ${\beta}$-lactam antibiotics and quinolone in Gram-negative clinical isolates collected in 2010 at a hospital in Korea. Among 65 amikacin-resistant isolates screened from 864 Gram-negative bacilli (GNB), 16S rRNA methylase genes were detected from 49 isolates, including Acinetobacter baumannii (43), Klebsiella pneumoniae (2), Proteus mirabilis (2) and Serratia marcescens (1), Empedobacter brevis (1). All of the 16S rRNA methylase genotype was armA and no variant sequences of amplified PCR products for armA were noted. The 16S rRNA methylase producing bacteria showed much higher resistance to aminoglycoside for Enterobacteriaceae and glucose non-fermenting (NF)-GNB and to imipenem for glucose NF-GNB, than the non-producing isolates. All of the 16S rRNA methylase producing Enterobacteriaceae had the extended-spectrum-${\beta}$-lactamase. In addition, two K. pneumoniae concurrently produced both plasmid-mediated AmpC ${\beta}$-lactamase and qnrB gene. All of the amikacin-resistant A. baumannii (43) co-harbored armA 16S rRNA methylase and $bla_{OXA-23}$ carbapenemase. In conclusion, 16S rRNA methylase producing bacteria were very prevalent among GNB in South Korea, and were commonly associated with co-resistance, including carbapenem and quinolone.

Microorganisms Involved in Natural Fermentation of Asparagus cochinchinensis Roots and Changes in Efficacies after Fermentation (천문동 뿌리의 자연발효에 관여하는 미생물 및 발효 후 효능 변화)

  • Kim, Min-Jee;Shin, Na Rae;Lee, Myeong-Jong;Kim, Hojun
    • Journal of Korean Medicine for Obesity Research
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    • v.18 no.2
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    • pp.96-105
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    • 2018
  • Objectives: The aim of this study was to examine the effect of Asparagus cochinchinensis (AC) and fermented AC (fAC) on microorganisms and efficacies. Methods: AC was fermented for four weeks without using any bacterial strains. Then we investigated fermentation characteristics including potential of hydrogen (pH), total sugar, microbial profiling and antioxidant compound contents such as total polyphenol and total flavonoid. The anti-obesity effects of AC and fAC were evaluated by using Oil Red O staining in 3T3-L1 adipocyte. Also anti-diabetic effects of them were evaluated by using 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose (2-NBDG) uptake in C2C12 skeletal muscle cell. Results: Both pH and total sugar of fAC were decreased significantly compared to unfermented AC. And the abundance of total bacteria and lactic acid bacteria increased during fermentation, especially Lactobacillus plantarum. Also fermentation of AC increased the content of total polyphenol. On the metabolic aspects, we found that AC and fAC suppressed fat accumulation. Conclusions: After four weeks of fermentation, AC increased concentrations of active compounds, altered microbial composition, and inhibited fat accumulation such as triglyceride. These results indicate that fermentation of AC might be a beneficial therapeutic approach for obesity.

Analysis of Volatile Components of the Musk of Ondatra zibethicus by Gas Chromatography-Mass Spectrometry (가스크로마토그래피-질량분석기를 이용한 서사향의 향기성분 분석)

  • Jin, Yong-Xie;Choi, Se-Jin;Jung, Eui-Jin;Kim, Sun-Ju;Jeon, Young-Eun;Kang, Il-Jun;Chung, Cha-Kwon;Lim, Soon-Sung
    • Korean Journal of Pharmacognosy
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    • v.40 no.4
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    • pp.303-308
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    • 2009
  • Essential oils of the musk from wild muskrat (Ondatra zibethicus) were analyzed by gas chromatography/mass spectrometry. Fifty-six chemical components were separated, and 14 volatile compounds (78% of composition of total essential oil) of macrocyclic musk, mainly cyclopentadecanone, 12-hydroxy-14-methyl-oxa-cyclotetradec-6-en-2-one, cycloheptadecanone and 9-cycloheptadecen-1-one, were identified by the comparison of Wiley/NBS Library from the results of GC-MS. However, l-muscone, a compound reported by earlier workers in the musk from muskrat, was not detected using the spike test in this study. Interestingly, cyclohexadecanone, which has the same molecular weight (Mw 238) as l-muscone, was identified. In addition, it is considered that because of possessing much higher amounts of macrocyclic components compared to the original musk, muskrat has higher economic values in perfume and pharmaceutical industries.

In vitro Antimicrobial Activity in Combination of Antibacterials Against Fish-pathogenic Bacteria (병용 항균제의 어류질병 세균에 대한 시험관내 항균활성)

  • Jung, Sung-Hee;Kim, Jin-Woo
    • Journal of fish pathology
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    • v.13 no.1
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    • pp.45-51
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    • 2000
  • Bacterial diseases with mixed infection have recently occurred at land-based flounder farms in Korea. Thus, single antibacterial is not effective for therapy of mixed bacterial diseases of fish because of their different causative bacteria. The purpose of the present study was to obtain basic data for positive usefulness of a combination of antibacterials used for synergism to mixed bacterial diseases of fish. Snergistic interaction in combination of antibacterials was determined by in vitro antimicrobial activity against selected fish-pathogenic bacteria, Vibrio anguillarum, Edwardsiella tarda, Streptococcus sp., Staphylococcus epidermidis, on the basis of Checkerboard assay using fractional inhibitory concentration (FIC) indices. Synergistic interactions were observed in combinations of (oxytetracycline HCL+lincomycin), (tetracycline HCL+florfenicol), (oxytetracycline HCL+florfenicol) against V. anguillarum, (sodium nifurstyrenate+florfenicol), (tetracycline HCL+florfenicol), (sodium nifurstyrenate+oxolinic acid), (oxytetracycline HCL+florfenicol) against E. tarda, (ciprofloxacine+oleandomycin), (oxytetracycline HCL+oleandomycin), (tetracycline HCL+oleandomycin), (oxytetracycline HCL+lincomycin), (oxytetracycline HCL+spiramycin), (oxytetracycline HCL+erythromycin), (doxycycline HCL+oleandomycin), (tetracycline HCL+spiramycin) against Streptococcus sp., and (ciprofloxacine+erythromycin), (florfenicol+erythromycin), (doxycycline HCL+oleandomycin), (ciprofloxacine+oleandomycin) against S. epidermidis.

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The Effects of Galgunhwanggumhwangryun-tang on Glucose and Energy Metabolism in C2C12 Myotubes (C2C12 골격근 세포에서 갈근황금황련탕의 당 대사 및 에너지 조절 효과)

  • Jihong Oh;Song-Yi Han;Soo Kyoung Lim;Hojun Kim
    • Journal of Korean Medicine for Obesity Research
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    • v.22 no.2
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    • pp.93-101
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    • 2022
  • Objectives: This study aimed to observe the anti-diabetic effect and underlying mechanisms of Galgunhwanggumhwangryun-tang (GHH; Gegen-Qinlian-decoction) in the C2C12 myotubes. Methods: GHH (1.0 mg/ml) or metformin (0.75 mM) or insulin (100 nM) were treated in C2C12 myotubes after 4 days differentiation. The glucose uptake was assessed by 2-[N-(7-160 nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose uptake by C2C12 cells. The expression of adenosine monophosphate-activated protein kinase (AMPK) and phosphorylation AMPK (pAMPK) were measured by western blot. We also evaluated gene expression of glucose transporter type 4 (Slc2a4, formerly known as GLUT4), glucokinase (Gk), carnitine palmitoyltransferase IA (Cpt1a), nuclear respiratory factors 1 (Nrf1), mitochondrial transcription factor A (Tfam), and peroxisome proliferator-activated receptor γ coactivator 1α (Ppargc1a) by quantitative real-time polymerase chain reaction. Results: GHH promoted glucose uptake in C2C12 myotubes. The expression of AMPK protein, which plays an essential role in glucose metabolism, was increased by treatment with GHH. GHH treatment tended to increase gene expression of Slc2a4, Gk, and Nrf1 but was not statistically significant. However, GHH significantly improved Tfam and Ppargc1a gene expression in C2C12 myotubes. Conclusions: In summary, GHH treatment promoted glucose uptake in C2C12 myotubes. We suggest that these effects are associated with increased gene expression involved in mitochondrial biosynthesis and oxidative phosphorylation, such as Tfam and Ppargc1a, and increased expression of AMPK protein.

Phospholipase $A_2$ excreted from the cells of hyperthermophilic microbes (초호열성균이 생성하는 phospholipase $A_2$에 관한 연구)

  • Joh, Yong-Goe;Woo, Hyo-Kyeng;Kim, Yeon-Sim
    • Journal of the Korean Applied Science and Technology
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    • v.16 no.3
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    • pp.263-271
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    • 1999
  • We checked the presence of phospholipase $A_2(PLA)_2$ which could split the ester bond at the position 2 in the glycerol backbone of glycerophospholipids, in the cells of hyperthermophiles of Pyrococcus horikoshii and Sulfolobus acidocaldarius. The results obtained are as follows; (1). Pyrococcus horikoshii cells were grown in obligate anaerobic conditions at $95^{\circ}C$ and they needed sulfur as energy source instead of oxygen, while Sulfolobus acidocaldarius species grew well in the aerobic medium (pH 2.5) containing yeast and sucrose at $75^{\circ}C$. (2). Pyrococcus horikoshii cells produced phospholipase $A_2$ in the cell culture media although this species did not show lipase activity at least in the pH range of 1.5 ${\sim}$ 3.5. Sulfolobus acidocaldarius cells produced lipase hydrolyzing triacylglycerols such as triolein, but did not split any kind of phospholipids used as substates. (3). The compound of 1-decanoyl-2-(p-nitrophenylglutaryl) phosphatidylcholine was not suitable for a substrate in this experiment, though frequently used as a subtrate for checking presence of phospholipase $A_2$, for its decomposi-tion in this experiment. The L-${\alpha}$-phosphatidylcholine-${\beta}$-[N-7-nitrobenz-2-oxa-1, 3-diazol]aminohexanoyl-${\gamma}$-hexadecanoyl labelled with a fluorescent material, did not show any migration of acyl chains in the molecule during the reaction with phospholipase $A_2$ under a hot condition. (4). Phospholipase $A_2$ in the cells of Pyrococcus horikoshii, showed the optimum activity at $pH6.7{\sim}7.2$ and $95{\sim}105^{\circ}C$, respectively, and was activated by addition of calcium chloride solution. Andthe phospholipase $A_2$ specifically hydrolyzed glycero-phospholipids such as phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine and phosphatidyl inositol, but could not split phospholipid containing ether bonds in the molecule such as DL -${\alpha}$-phosphatidylcholine-${\beta}$-palmitoyl-${\gamma}$-O-hexadecyl, DL-${\alpha}$-phosphati- dylcholine-${\beta}$- oleoyl-${\gamma}$-O-hexadecyl, DL-phosphatidylcholine-dihexadecyl.

Comparison of Molecular Characteristics of Extended Spectrum ${\beta}$-lactamase Producing Escherichia coli Strains Isolated from Patients with Urinary Tract Infections between 2 Time Periods of 1989 and 2010 at Gangwon Province in Korea

  • Park, Min;Park, Soon Deok;Kim, Sa-Hyun;Lee, Gyusang;Woo, Hyun Jun;Kim, Hyun Woo;An, Byungrak;Jang, In Ho;Uh, Young;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.19 no.3
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    • pp.275-279
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    • 2013
  • Etiological agents of extended spectrum ${\beta}$-lactamase (ESBL) producing uropathogenic Escherichia coli (UPEC) have become a major problem in urinary tract infections. The purpose of this study was to compare the molecular characteristics of ESBL producing UPEC strains isolated from 1989 and 2010. A total of 301 strains of UPEC clinical isolates was collected from Korean healthcare facility in 1989 (126 strains) and in 2010 (175 strains). UPEC clinical isolates were analyzed by multiplex polymerase chain reaction method (ESBL related bla genes and phylogenetic groups) and amplified fragment length polymorphism (AFLP). Among 301 isolates, ESBL producing UPEC were 8 strains (6.3%) in 1989 isolates and 35 strains (20%) in 2010 isolates. The rate of bla genes in ESBL producing UPEC from 1989 isolates and 2010 isolates were $bla_{TEM}$ (75% and 85.7%), $bla_{CTX-M}$ (0% and 91.4%), $bla_{OXA}$ (25% and 20%), $bla_{PER}$ (0% and 2.9%). The distribution of phylogenetic groups in 1989 isolates and 2010 isolates were A (37.5% and 11.4%), B2 (12.5% and 51.4%), and D (50% and 37.1%). The most prevalent ESBL related bla gene and phylogenetic group were $bla_{CTX-M}$ (91.4%) and B2 (51.4%) in 2010 isolates, while $bla_{CTX-M}$ was not detected in 1989 isolates. Among 43 ESBL producing UPEC were grouped into 12 clusters up to 76% of genetic similarities by AFLP analysis. During past twenty one years, the rate of the ESBL producing UPEC strains in 2010 isolates was increased than that of in 1989 isolates. Also, the most prevalent ESBL related bla gene has been changed from $bla_{TEM}$ to $bla_{CTX-M}$.