• Title/Summary/Keyword: Clinical Microbiology

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In vitro and in vivo Evaluations of LB 10517, a Novel Parenteral Broad-Spectrum Cephalosporin

  • Song, Hye-Kyong;Nishino, Takeshi;Seo, Mi-Kyeong;Kim, Mu-Yong;Lee, Yong-Hee;Kim, In-Chull;Kwak, Jin-Hwan
    • Archives of Pharmacal Research
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    • v.19 no.1
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    • pp.46-51
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    • 1996
  • The in vitro activity of LB 10517, a new catechol-substituted cephalosporin, was compared with those of E-1077, cefpirome and ceftazidime 1034 clinical isolates collected in Japan. LB10517 showed a broad-spectrum antibacterial activity against a wide range of grampositive and gram-negative bacteria including non-glucose fermenting rods, Pseudomonas aeruginosa. Against the methicillin-susceptible strains of Staphylococcus aureus (MSSA) and Strptoccus pyogenes, the $MIC_{90}$ values of LB10517 which required to inhibit 90% of the strains wre $3.13\mug/ml\; and\; 0.1\mug/ml$, respectively. It was as active as E-1077 but more active than cefpirome and ceftazidime. Methicillin-resistant strains of S.aureus (MRSA) and Enterococcus spp. were highly resistant to all the test compunds. LB10517 was highly active against most members of the family Enterobacteriaceae, 90% of which were inhibited at a concentration of less than $0.78\mug/ml$, except for Enterobacter cloacae ($1.56\mug/ml$) and Serratia marcescens ($3.13\mug/ml$)Its activity was comparable to those of E-1077 and cefpirome but it was greater than that of ceftazidime. Against Pseudomonas aeruginosa, LB10517 showed the most potent antibacterial activity among the compounds tested. Ninety percent of P. aeruginosa isolates were susceptible at the concentration of $0.39\mug/ml$. Its activity was 32-to 128 fold higher than those of E-1077, cefpirome and ceftazidime. Against imipenem- or ofloxacin-resistant P. aeruginosa, LB10517 with $MIC_{90}\; of\; 6.25 \\mug/ml\; and\; 3.13\mug/ml$, respectively, showed 16-fold more potent activity than the other test compounds. LB10517 showed a relatively high plasma level and long plasma elimination half-life in rats $(t_{1/2}(\beta,\; 52 min)\; and\; dogs\; (t_{1/2}(\beta),\; 103 min)$.

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Inhibitory Effect of Extract of Teucrium veronicoides on the Production of Inflammatory Cytokines (곽향 추출물의 염증성세포활성물질 억제효과)

  • Bae, Gi-Sang;Park, Hee-Je;Kim, Do-Yun;Seo, Sang-Wan;Park, Kyung-Bae;Kim, Byung-Jin;Song, Je-Moon;Lee, Kyung-Yong;Na, Chul;Shin, Byung-Chul;Park, Sung-Joo;Song, Ho-Joon;Hwang, Sung-Yeon
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.119-125
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    • 2008
  • Objectives : The purpose of this paper was to investigate the anti-inflammatory effects of extract from Teucrium veronicoides (TV) on the RAW 264.7 cells. Methods : To evaluate of anti-inflammatory of TV, we examined the cytokine productions on lipopolysacchride (LPS)-induced RAW 264.7 cells and also inhibitory mechanisms using Western blot. Furthermore, We examined LPS-induced endotoxin shock. Results : 1. Extract from TV itself does not have any cytotoxic effect. 2. Extract from TV reduced LPS-induced Nitric oxide (NO),interleukin (IL)-1b, IL-6 and IL-10, tumor necrosis factor-a (TNF-a) production in RAW 264.7 cells. 3. TV inhibited the activation of mitogen-activated protein kinases (MAPKs) such as p38, extracelluar signal-regulated kinase (ERK 1/2) and c-Jun NH2-terminal kinase (JNK) and also the degradation of inhibitory kappa B a (Ik-Ba) in the LPS-stimulated RAW 264.7 cells. 3. TV slightly increased the duration of survival after LPS-induced endotoxin shock. Conclusions : TV down-regulated LPS-induced NO and cytokines production, which could provide a clinical basis for anti-inflammatory properties of TV.

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Characterization of Streptococcosis Occurrence and Molecular Identification of the Pathogens of Cultured Flounder in Jeju Island (제주지역 양식넙치의 연쇄구균증 발생동향 및 원인균에 대한 분자적 동정)

  • Jeong, Yong-Uk;Kang, Chul-Young;Kim, Min-Ju;Heo, Moon-Soo;Oh, Duck-Chul;Kang, Bong-Jo
    • Korean Journal of Microbiology
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    • v.42 no.3
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    • pp.199-204
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    • 2006
  • Streptococcosis of olive flounder(Paralichthys olivaceus) is an important bacterial disease in Jeju island. In this study, we investigated monthly infection pattern of this disease in different size of the flounder fish. Even though the disease occurred throughout the year, the infection ratio was relatively higher in the months with warm water season. The infection was more prevalent in adult flounder over 30 cm total length compare to these of small size fish. Two infectious species of streptococcosis pathogens were detected by multiplex PCR assay. Detection ratios of Streptococcus iniae and S. parauberis reached up to 46% and 54%, respectively, from June 2003 to May 2005 in Jeju island. S. iniae occurred intensively from September to October, whereas S. parauberis reported from March to May. S. iniae and S. parauberis infections of cultured flounder share some common features, but clinical findings showed considerable differences between two diseases. Distended abdomen, protruded anus and ascitic fluid in the peritoneal cavity are evident lesions detected in S. iniae infection, whereas, flounders infected by S. parauberis showed prominent lesions such as darkened surface and haemorrhaging in the non-ocular side. Both streptococcosis pathogens were sensitive to antibiotics, such as ampicillin and amoxicillin. However, S. iniae strains were more sensitive to doxycycline, erythromycin and oxytetracycline than S. parauberis strains.

Molecular Characterization of Clinically Isolated Staphylococcus aureus (인천지역에서 분리된 황색포도상구균의 분자생물학적 특성 분석)

  • Oh, Bo-Young;Kim, Jung-Hee;Gong, Young-Woo;Lee, Jae-Mann;Go, Jong-Myoung;Kim, Yong-Hee
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.305-310
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    • 2008
  • Staphylococcus aureus is one of the most significant pathogens and a causative agents of nosocomial infections. The emergence of methicillin resistant S. aureus (MRSA), in particular, has become a major clinical and epidemiological problems worldwide. In this study, we analyzed the toxin genes and investigated molecular epidemiological characteristics of S. aureus isolated from stools of diarrheal patients at the hospitals in Incheon. Of the 609 strains from 2,281 specimens, 173 strains retained enterotoxin; 68 isolates (39.30%), 100 isolates (57.80%) were classified to A and C type, respectively. In the antibiotic susceptibility, all of enterotoxin positive isolates were resistant to oxacillin. Eighty eight strains (50.86%) of 173 MRSA isolate possessed tsst gene, but eta and eth genes were not detected at all. In the detection of MRSA associated genes by PCR method, mecA genes were detected in 167 strains (96.53%). From the result of PFGE analysis, we classified tsst-positive MRSA to 10 types and 24 subtypes. Type A, H and F were the major strains comprised of 57.95% (51 strains), 10.22% (9 strains) and 9.09% (8 strains) respectively.

Antifungal Activity of Rheum undulatum on Candida albicans by the Changes in Membrane Permeability (막투과성 변화로 인한 대황의 Candida albicans에 대한 항진균 활성)

  • Lee, Heung-Shick;Kim, Younhee
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.360-367
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    • 2014
  • Candida albicans is an opportunistic and the most prevalent fungal pathogen that can cause superficial and systemic infections in immunocompromised patients. C. albicans can promote the transition from budding yeast to filamentous form, generating biofilms. Infections associated with C. albicans biofilms are frequently resistant to conventional antifungal therapy. Therefore, the development of more effective antifungal drugs related with biofilm formation is required urgently. The roots of Rheum undulatum have been used for medicinal purposes in Korea and China traditionally. The aim of present study was to evaluate the effect of R. undulatum extract upon preformed biofilms of 12 clinical C. albicans isolates and the antifungal activities. Its effect on preformed biofilms was evaluated using XTT reduction assay, and metabolic activity of all tested strains was reduced significantly ($49.4{\pm}6.0%$) at 0.098 mg/ml R. undulatum. The R. undulatum extract blocked the adhesion of C. albicans biofilms to polystyrene surfaces, and damaged the cell membrane integrity of C. albicans which was analyzed by CFDA, AM, and propidium iodide double staining. It caused cell lysis which was observed by Confocal laser scanning and phase contrast microscope after propidium iodide and neutral red staining, respectively. Membrane permeability was changed as evidenced by crystal violet uptake. The data suggest that R. undulatum inhibits biofilm formation by C. albicans, which can be associated with the damage of the cell membrane integrity, the changes in the membrane permeability and the cell lysis of C. albicans.

Fluoroquinolone Resistance and gyrA and parC Mutations of Escherichia coli Isolated from Chicken

  • Lee Young-Ju;Cho Jae-Keun;Kim Ki-Seuk;Tak Ryun-Bin;Kim Ae-Ran;Kim Jong-Wan;Im Suk-Kyoung;Kim Byoung-Han
    • Journal of Microbiology
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    • v.43 no.5
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    • pp.391-397
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    • 2005
  • Escherichia coli is a common inhabitant of the intestinal tracts of animals and humans. The intestines of animals also represent an ideal environment for the selection and transfer of antimicrobial resistance genes. The aim of this study was to investigate the resistance of E. coli isolated from chicken fecal samples to fluoroquinolones and to analyze the characterization of mutations in its gyrA and parC gene related resistance. One hundred and twenty-eight E. coil isolates showed a high resistance to ciprofloxacin (CIP; $60.2\%$), enrofloxacin (ENO; $73.4\%$) and norfloxacin (NOR; $60.2\%$). Missense mutation in gyrA was only found in the amino acid codons of Ser-83 or Asp-87. A high percentage of isolates ($60.2\%$) showed mutations at both amino acid codons. Missense mutation in parC was found in the amino acid codon of Ser-80 or Glu-84, and seven isolates showed mutations at both amino acid codons. Isolates with a single mutation in gyrA showed minimal inhibitory concentrations (MIC) for CIP (${\le}0.5\;to\;0.75{\mu}g/ml$), ENO (1 to $4{\mu}g/ml$) and NOR (0.75 to $4{\mu}g/ml$). These MIC were level compared to isolates with two mutations, one in gyrA and one in parC, and three mutations, one in gyrA and two in parC (CIP, ${\le}0.5\;to\;3{\mu}g/ml;\;ENO,\;2\;to\;32<{\mu}g/ml;\;NOR,\;1.5\;to\;6\;{\mu}g/ml$). However, the isolates with two mutation in gyrA regardless of whether there was a mutation in parC showed high MIC for the three fluoroquinolones (CIP, 0.75 to $32{\le}{\mu}g/ml;\;ENO,\;3\;to\;32{\le}{\mu}g/ml;\;NOR,\;3\;to\;32{\le}{\mu}g/ml$). Interestingly, although the E. coil used in this study was isolated from normal flora of chicken, not clinical specimens, a high percentage of isolates showed resistance to fluoroquinolones and possessed mutations at gyrA and parC associated with fluoroquinolone resistance.

Effect of Non-homologous Spacing in Target DNA Sequence on the Frequency of Cloning Based Homologous Recombination (Target DNA 염기서열 내에 존재하는 비상동성 간격이 상동성재조합을 이용한 클로닝 빈도에 미치는 영향)

  • Kim Jae-Woo;Do Eun-Ju;Yoon Se-Lyun;Jeong Yun-Hee;Yoon Young-Ho;Leem Sun-Hee;Sunwoo Yangil;Park In-Ho
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.239-245
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    • 2005
  • Transformation-Associated Recombination (TAR) cloning technique allows selective isolation of chromosomal regions and genes from complex genomes. The procedure requires knowledge of relatively small genomic sequences that reside adjacent to the chromosomal region of interest. This technique involves homologous recombination during yeast spheroplast transformation between genomic DNA and a TAR vector that has 5' and 3' gene targeting sequences. In this study, we examined the effect of non-homologous spacing sequence in target hooks on homologous recombination using a plasmid model system. The efficiency of homologous recombination between the modified his3-TRP1-his3 fragments and HlS3 gene on plasmid were analyzed by the characterization of $Ura^+$ transformants. The numbers of $Ura^+$ transformant showed same level when seven different modified his3-TRP1-his3 fragments were used. But the percentage of positive recombinants. $Trp^+His^-$, dramatically decreased when used the modified his3-TRP1-his3 fragments contained incorrect spacing of nonhomologous region. As a result, we suggest that incorrect spacing inhibits the homologous recombination between target hook and substrate DNA. Therefore, we should consider the correct spacing in target hook when the target hook are used for cloning of orthologue gene.

A Characteristics of Phagocytic Plaque on Staphylococcus aureus Layer Formed by Leukocytes of the Alcoholics (Staphylococcus aureus를 이용한 주정중독환자 백혈구의 Phagocytic Plaque 형성력)

  • 성희경;서병배;김용호
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.205-212
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    • 2002
  • This study was conducted to develop a method for direct determination of phagocytic activities in human circulatic systems and to measure the phagocytic activities in human leukocytes from the alcoholics, since phagocytic activity was considered to be very important in human immune mechanism at early stage for the health care of the alcoholics. The subjects for this study were 130 among which 95 males and 3 females were diagnosed as alcoholism and 32 was healthy blood donors. A thin layer of heat-killed Staphylococcus aureus Cowan I was placed on a plastic dish and reacted with whole blood to measure the phagocytic plaque formation by human leukocytes. In order to determine the health conditions of the subjects, some clinical laboratory tests, such as white blood cell counts, hemoglobin contents (Hgb), mean corpuscular volume of red blood cells(MCV), serum electrophoresis, B and T-lymphocytes, T-lymphocyte subtypes and phytohemagglutination test were also implemented. Compared to the non-alcoholism, new and old alcoholic inpatients showed statistically significant differences on levels of Hgb and MCV (p<0.05), but showed that T and B-lymphocyte numbers decreased and Helper T cell/Suppressor T cell ratio ($1.6{\pm}0.8$%) increased. Compared to non-alcoholism, phagocytic plaque activities of leukocytes from alcoholic patients decreased significantly and an unusual pattern in phagocytic plaque was observed, showing a strange body and chain shaped phagocytosis. Based upon these results, it is concluded that a phagocytic-plaques of Staphylococcus aureus Cowan I by leukocytes was very simple and useful method for the early immunological determination of phagocytic activities in alcoholic patients without requiring any special equipments.

Characteristics of Coagulase-negative Staphylococci Isolates from Dental Clinic Environments in Busan, Korea (부산지역 치과환경에서 분리된 coagulase-negative staphylococci의 특성)

  • Jung, Hye-In;Jung, So Young;Park, Indal;Bae, Il Kwon
    • Journal of Life Science
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    • v.26 no.2
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    • pp.220-225
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    • 2016
  • Coagulase-negative staphylococci (CNS) have recently become the bacteria most frequently found in clinical infections. The aim of this study was to investigate the prevalence, antimicrobial susceptibilities, and molecular characteristics of CNS isolates from dental clinic environments in Busan, Korea. One hundred and fifty-four samples were collected from 10 dental clinics and dental hospitals in Busan from December 2014 to January 2015. Species were identified by matrix-assisted laser desorption/ionization–time-of-flight. Antimicrobial susceptibility was determined by disk diffusion methods. A polymerase chain reaction was performed to detect mecA, mupA gene, and SCCmec types. Of the 154 samples, 10(6.5%) isolates were identified as CNS (5 Staphylococcus epidermidis, 2 Staphylococcus capitis, 2 Staphylococcus, and 1 Staphylococcus haemolyticus). Among the 10 isolates, 6 were resistant to penicillin, 5 were resistant to gentamicin, 3 were resistant to tetracycline, and 2 were resistant to cefoxitin and erythromycin. However, clindamycin, ciprofloxacin, teicoplanin, and trimethoprim-sulfamethoxazole resistant isolates were not present. Genes encoding mecA were detected in 4 (2 S. warneri and 2 S. haemolyticus) isolates, and mupA in 1 (S. epidermidis) isolate. One methicillin-resistant CNS (S. warneri) isolate was determined as being of the SCCmec type I. It is concluded that CNS resistant to various antimicrobial agents was widely distributed in dental clinic environments in Korea.

Biological Effect of Metronidazole Resistance in Helicobacter Pylori (Helicobacter pylori에서 metronidazole 내성이 미치는 생물학적 영향)

  • Lee Gin Hyug;Lee Sun-Mi;Nam Won-Hee;Kim Eun Sil;Kim Jin Ho;Kang Ho Young;Jeong Jin-Yong
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.955-960
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    • 2005
  • Resistance to metronidazole, a key component of therapies against Helicobacter pylori, is common in clinical isolates. Resistance generally requires inactivation of rdxA (HP0954), and sometimes also frxA (HP0642), two related nitroreductase genes. Here we studied the effect of resistance to metronidazole on fitness of the gastric pathogen H. pylori. The effect of metronidazole resistance for H. pylori in culture was assessed first by looking at colonies formed by freshly constructed mutant derivatives of H. pylori strain 26695. Mutations resulting in metronidazole resistance caused premature death of H.pylori in stationary phase, but had no significant effect on early exponential growth. The effect of nitroreductase deficiencies on fitness in vivo was tested by infecting C57BL/6 mice with 1:1 mixtures of SS1 wild type and its isogenic metronidazole resistant derivatives. Inactivation of rdxA caused an inability to colonize mice in SS1 H. pylori strain. Derivatives of a metronidazole resistant strain that survived better in stationary phase, although remaining metronidazole resistant, could again colonize mice. In conclusion, metronidazole resistance diminishes H. pylori's fitness, but their costs can be suppressed by additional mutation.