• Title/Summary/Keyword: ITS-PCR

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Antibiotic Resistance and Bacterial Biofilm Formation by Staphylococcus aureus Strains Isolated from Various Foods (식품에서 분리한 황색포도상구균의 항생제 내성 특징 및 균막 형성)

  • Lee, Joo-Young;Wang, Hae-Jin;Shin, Dong-Bin;Cho, Yong-Sun
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.96-104
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    • 2013
  • Staphylococcus aureus is a major human pathogen that produces a wide array of toxins, leading to a number of adverse symptoms. We examined 275 strains of Staphylococcus aureus isolated from various foods between 2006 and 2008 for antimicrobial susceptibility. At least 259 (94.2%) of the tested strains showed antibiotic resistant properties, and 106 (40.7%) of them showed multiple antibiotic resistance. Eleven of the tested strains were resistant to oxacillin and mec A-positive. Moreover, oxacillin-resistant strains were significantly more likely to be multi-drug resistant (p < 0.01). Of the 275 isolates tested, 24.4% were noted as being positive for slime production and 30.5% were positive for biofilm assay. Antibiotic resistance was not associated with a significantly higher prevalence of biofilm formation. Twenty strains were classified using the DiversiLab system. Most of the strains could be classified into 2 clusters and 4 unique types. All 10 mec A-positive strains (cluster I) were grouped together into the same sub-cluster. Cluster II (6 strains) was not found to be resistant to oxacillin in this study. Although the prevalence of methicillin-resistant S. aureus in food is currently low, the risk of its transmission through the food chain cannot be disregarded.

Microbial Floral Dynamics of Chinese Traditional Soybean Paste (Doujiang) and Commercial Soybean Paste

  • Gao, Xiuzhi;Liu, Hui;Yi, Xinxin;Liu, Yiqian;Wang, Xiaodong;Xu, Wensheng;Tong, Qigen;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.12
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    • pp.1717-1725
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    • 2013
  • Traditional soybean paste from Shandong Liangshan and Tianyuan Jiangyuan commercial soybean paste were chosen for analysis and comparison of their bacterial and fungal dynamics using denaturing gel gradient electrophoresis and 16S rRNA gene clone libraries. The bacterial diversity results showed that more than 20 types of bacteria were present in traditional Shandong soybean paste during its fermentation process, whereas only six types of bacteria were present in the commercial soybean paste. The predominant bacteria in the Shandong soybean paste were most closely related to Leuconostoc spp., an uncultured bacterium, Lactococcus lactis, Bacillus licheniformis, Bacillus spp., and Citrobacter freundii. The predominant bacteria in the Tianyuan Jiangyuan soybean paste were most closely related to an uncultured bacterium, Bacillus licheniformis, and an uncultured Leuconostoc spp. The fungal diversity results showed that 10 types of fungi were present in the Shandong soybean paste during the fermentation process, with the predominant fungi being most closely related to Geotrichum spp., an uncultured fungal clone, Aspergillus oryzae, and yeast species. The predominant fungus in the commercial soybean paste was Aspergillus oryzae.

Expression Analysis of Lily Type Lectin Isotypes in the Rock Bream, Oplegnathus fasciatus: in the Tissue, Developmental Stage and Viral Infection

  • Lee, Young Mee;Yang, In Jung;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Noh, Gyeong Eon;Kim, Woo-Jin;Kim, Kyung-Kil
    • Development and Reproduction
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    • v.20 no.4
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    • pp.297-304
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    • 2016
  • Lectins belong to the pattern-recognition receptors (PRRs) class and play important roles in the recognition and elimination of pathogens via the innate immune system. Recently, it was reported that lily-type lectin-1 is involved when a pathogen attacks in the early immune response of fish. However, this study is limited to information that the lectin is involved in the innate immune response against viral infection. In the present study, the lily-type lectin-2 and -3 of Oplegnathus fasciatus (OfLTL-2 and 3) have been presented to be included B-lectin domain and two D-mannose binding sites in the amino acid sequence that an important feature for the fundamental structure. To investigate the functional properties of OfLTLs, the tissue distribution in the healthy rock bream and temporal expression during early developmental stage analysis are performed using quantitative real-time PCR. OfLTL-2 and 3 are predominantly expressed in the liver and skin, but rarely expressed in other organ. Also, the transcripts of OfLTLs are not expressed during the early developmental stage but its transcripts are increased after immune-related organs which are fully formed. In the challenge experiment with RBIV (rock bream iridovirus), the expression of OfLTLs was increased much more strongly in the late response than the early, unlike previously known. These results suggest that OfLTLs are specifically expressed in the immune-related tissues when those organs are fully formed and it can be inferred that the more intensively involved in the second half to the virus infection.

Effects of 17β-estradiol, Interleukin-1β, and Human Chorionic Gonadotropin on Activity and mRNA Expression of Plasminogen Activators in Porcine Endometrial Cells

  • Hwangbo, Yong;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Development and Reproduction
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    • v.22 no.2
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    • pp.155-163
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    • 2018
  • This study aimed to investigate changes in the activity and mRNA expression of plasminogen activators (PAs) induced by $17{\beta}$-estradiol ($E_2$), human chorionic gonadotropin (hCG), and interleukin-$1{\beta}$ ($IL-1{\beta}$) in porcine endometrial cells. Endometrial cells were isolated from the epithelium and cultured to 80% confluence. They were then treated for 24 h with $E_2$ (0.2, 2, 20, and 200 ng/mL), $IL-1{\beta}$ (0.1, 1, 10, and 100 ng/mL), and hCG (0.5, 1, 1.5 and 2 IU/mL). mRNA expressions of urokinase-type (uPA) and tissue-type (tPA) PAs were analyzed using reverse transcription PCR, and activities were measured using a PA activity assay. mRNA expressions of uPA and tPA increased with $E_2$ treatment; however, this was not significant. Similarly, treatment with hCG did not influence the mRNA expressions of PAs. Interestingly, treatment with 0.1 ng/mL $IL-1{\beta}$ significantly reduced the mRNA expression of uPA, but did not affect that of tPA. Treatment with 2, 20, and 200 ng/mL $E_2$ increased PA activity compared with the control group; treatment with 0.1 and 1 ng/mL $IL-1{\beta}$ significantly increased PA activity compared with the other $IL-1{\beta}$ treatment groups, whereas treatment with 10 and 100 ng/mL $IL-1{\beta}$ decreased. Treatment with 2 IU/mL hCG increased PA activity compared with the other treatment groups, although there were no significant differences between the hCG and control groups. In conclusion, the activity and mRNA expression of PAs were differently regulated by the hormone/cytokine and its concentration in porcine endometrial cells. Therefore, understanding PA regulatory mechanisms may help to improve the reproductive potential of domestic animals.

Serologic Survey of Toxoplasmosis in Seoul and Jeju-do, and a Brief Review of Its Seroprevalence in Korea

  • Lim, Hyemi;Lee, Sang-Eun;Jung, Bong-Kwang;Kim, Min-Ki;Lee, Mi Youn;Nam, Ho-Woo;Shin, Jong-Gyun;Yun, Cheong-Ha;Cho, Han-Ik;Shin, Eun-Hee;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.50 no.4
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    • pp.287-293
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    • 2012
  • Knowledge of the prevalence of human Toxoplasma gondii infection is required in the Republic of Korea. In this study, we surveyed the seroprevalence of T. gondii infection and analyzed the risk factors associated with seropositivity among residents in 2 administrative districts; Seoul and the island of Jeju-do, which have contrasting epidemiologic characteristics. Sera and blood collected from 2,150 residents (1,114 in Seoul and 1,036 in Jeju-do) were checked for IgG antibody titers using ELISA and for the T. gondii B1 gene using PCR. In addition, participants completed a questionnaire that solicited information on gender, age, occupation, eating habits, history of contact with animals, and travel abroad. The T. gondii B1 gene was not detected in all residents examined. However, ELISA showed 8.0% (89 of 1,114 sera) positive for IgG antibodies against T. gondii in Seoul and 11.3% (117 of 1,036 sera) in Jeju-do. In both districts, the positive rates were higher in males than in females, and those 40-79 years of age showed higher rates than other ages. In Seoul, residents older than 70 years of age showed the highest positive rate, 14.9%, whereas in Jeju-do the highest prevalence, 15.6%, was in those in their sixties. The higher seropositive rate in Jeju-do than in Seoul may be related to eating habits and occupations. The present results and a review of related literature are indicative of an increased seroprevalence of T. gondii in Korea in recent years.

Effects of Chelidonii Herbal-acupuncture solution Anti-inflammatory in RAW 264.7 macrophages (백굴채약침액(白屈菜藥針液)이 LPS로 유도(誘導)된 RAW 264.7 대식세포(大食細胞)에서의 항염증효과(抗炎症效果))

  • Park, Dong-Cheon;Park, Ji-Hyeon;Lee, U-Kyung;Leek, Jin-Kyu;Seo, Il-Bok;Kim, Ho-Hyun;Kim, Jeong-Seon;Kim, Ee-Hwa
    • Korean Journal of Acupuncture
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    • v.21 no.2
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    • pp.125-137
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    • 2004
  • Objectives : Recently, Herbal-acupuncture therapeutics has been used for the treatment of inflammatory diseases such as rheumatoid arthritis. Especially, we have been interested in chemical mediators concerned with inflammation such as prostaglandin, cytokine, nitrous oxide. The purpose of this study is investigated that the effect of Chelidonii Herbal-acupuncture solution in lipopolysaccharide-stimulated RAW 264.7 macrophages, performed several expeimental items : those are prostaglandin $E_2$, nitric oxide and cyclooxygenase-2. Methods : The cytotoxicity of Chelidonii Herbal-acupuncture solution in RAW 264.7 macrophages were measured by MTT-based cytotoxicity assay. In order to observe cyclooxygenase-2 mRNA expression in lipopolysaccharide-stimulated RAW 264.7 macrophages, RT-PCR was used. Prostaglandin $E_2$ formation and nitric oxide production was measured by competitive enzyme immunoassay and Griess assay. Results : 1.The MTT assay demonstrated that cytotoxic effect of Chelidonii Herbal-acupuncture solution in RAW 264.7 macrophages were not appeared before concentration of 1mg/ml. 2.Chelidonii Herbal-acupuncture solution inhibited cyclooxygenase-2 mRNA expression in lipopolysaccharide-stimulated RAW 264.7 macrophages. 3. Chelidonii Herbal-acupuncture solution inhibited nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophages. 4. Chelidonii Herbal-acupuncture solution inhibited prostaglandin $E_2$ formation in lipopolysaccharide-stimulated RAW 264.7 macrophages. Conclusions : On the basis of these results, It was shown that Chelidonii Herbal-acupuncture solution is significantly able to inhibit the production of $PGE_2$ and NO, as well as COX-2 mRNA expression. Our results may provide new mechanism by which Chelidonii Herbal-acupuncture solution accounts for its beneficial effect on accelerating wound healing and anti-inflammation.

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Effect of Poria Cocos on the Scopolamine-induced Memory Impairment and Its Underlying Molecular Mechanism (스코폴라민으로 유도된 기억력 손상에 대한 복신의 보호 효과 및 작용기전 연구)

  • JeGal, Kyoung-Hwan;Park, Sung-Jun;Kim, Chang-Yul;Lee, Chan;Park, Jong-Hyun;Jang, Jung-Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.2
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    • pp.228-235
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    • 2010
  • This study was performed to investigate the memory enhancing effect of Poria cocos Wolf (Hoelen cum radix) against scopolamine-induced amnesia in Sprague-Dawley (SD) rats. To induce amnesia, scopolamine (0.75 mg/kg) was intraperitonically injected into SD rats 30 min before starting behavior tests. We have conducted Morris water-maze and Y-maze tests to monitor learning and memory functions. Poria cocos effectively reversed scopolamine-induced memory impairment in SD rats which was represented by an improvement of mean escape latency in water-maze test and spontaneous alterations in Y-maze test. To elucidate possible molecule mechanism, we have measured mRNA as well as protein expression of acetylcholine esterase (AchE), choline acetyltransferase (ChAT), muscarinic acetylcholine receptor (mAchR), and brain-derived neurotrophic factor (BDNF) using RT-PCR and Western blot analysis, respectively. Poria cocos increased mRNA levels of ChAT and mAchR in rat hippocampus compared with those in the scopolamine-injected amnesic group. In addition, protein expression of ChAT and BDNF was also elevated by Poria cocos intake. Furthermore, as an upstrem regulator, the activation of cAMP response element-binding protein (CREB) was assessed by immunohistochemistry. In this immunohistochemical analysis, the phosphorylation of CREB (p-CREB) was reduced by scopolamine injection, which was restored back to control levels by administration of Poria cocos. These results suggest that Poria cocos may improve memory and cognitive deficit in amnesia and have therapeutic potentials through up-regulation of ChAT, mAchR, and BDNF, which seemed to be mediated by activation of CREB.

Chondroprotective Effects of Wogonin in Experimental Models of Osteoarthritis in vitro and in vivo

  • Park, Jin Sung;Lee, Hyun Jae;Lee, Dong Yeong;Jo, Ho Seung;Jeong, Jin Hoon;Kim, Dong Hee;Nam, Dae Cheol;Lee, Choong Jae;Hwang, Sun-Chul
    • Biomolecules & Therapeutics
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    • v.23 no.5
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    • pp.442-448
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    • 2015
  • We evaluated the chondroprotective effects of wogonin by investigating its effects on the gene expression and production of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as on production of MMP-3 in the rat knee. Rabbit articular chondrocytes were cultured in a monolayer, and RT-PCR was used to measure interleukin-$1{\beta}$ (IL-$1{\beta}$)-induced expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), and type II collagen. In rabbit articular chondrocytes, the effects of wogonin on IL-$1{\beta}$-induced production and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of wogonin on MMP-3 protein production was also examined in vivo. In rabbit articular chondrocytes, wogonin inhibited the expression of MMP-3, MMP-1, MMP-13, and ADAMTS-4, but increased expression of type II collagen. Furthermore, wogonin inhibited the production and proteolytic activity of MMP-3 in vitro, and inhibited production of MMP-3 protein in vivo. These results suggest that wogonin can regulate the gene expression and production of MMP-3, by directly acting on articular chondrocytes.

Detection Rate of Periodontopathogens Associated with Cardiovascular Diseases in Denture. (의치 표면에서 심혈관질환과 관련된 치주질환 원인 세균의 검출)

  • Lim Mi-Young;Kim Hwa-Sook;Jeong Jae-Heon;Yang Ji-Youn;Oh Sang-Ho;Kook Joong-Ki
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.237-243
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    • 2004
  • The aim of this study is to investigate the detection rate of putative periodontopathogens, Porphyromonas gingivalis, Tannerella forsythia, and Actiobacillus actinomycetemcomitans, related to cardiovascular diseases(CVD). Plaques were sampled from 15 subjects (4 sites of denture base and/or tooth) with sterilized explorers and were transported in IX PBS. The detection of periodontopathogens was performed by polymerase chain reaction with species-specific primers based on 16S rDNA. The PCR products were cloned into pGEM-T easy vector and its nucleotides were sequenced in order to confirm the specificity. Our data showed that the detection rate of P. gingivalis and T forsythia in denture base of edentulous patients was 25% and 75%, respectively. And the detection rate of P. gingivalis and T.forsythia in denture base of patient having one more tooth was 91%. The results indicate that plaque of denture base may serve as reservoirs of oral bacteria related to CVD.

Comparative Genomics Profiling of Clinical Isolates of Helicobacter pylori in Chinese Populations Using DNA Microarray

  • Han, Yue-Hua;Liu, Wen-Zhong;Shi, Yao-Zhou;Lu, Li-Qiong;Xiao, Shudong;Zhang, Qing-Hua;Zhao, Guo-Ping
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.21-28
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    • 2007
  • In order to search for specific genotypes related to this unique phenotype, we used whole genomic DNA microarray to characterize the genomic diversity of Helicobacter pylori (H. pylori) strains isolated from clinical patients in China. The open reading frame (ORF) fragments on our microarray were generated by PCR using gene-specific primers. Genomic DNA of H. pylori 26695 and J99 were used as templates. Thirty-four H. pylori isolates were obtained from patients in Shanghai. Results were judged based on In(x) transformed and normalized Cy3/Cy5 ratios. Our microarray included 1882 DNA fragments corresponding to 1636 ORFs of both sequenced H. pylori strains. Cluster analysis, revealed two diverse regions in the H. pylori genome that were not present in other isolates. Among the 1636 genes, 1091 (66.7%) were common to all H. pylori strains, representing the functional core of the genome. Most of the genes found in the H. pylori functional core were responsible for metabolism, cellular processes, transcription and biosynthesis of amino acids, functions that are essential to H. pylori's growth and colonization in its host. In contrast, 522 (31.9%) genes were strain-specific genes that were missing from at least one strain of H. pylori. Strain-specific genes primarily included restriction modification system components, transposase genes, hypothetical proteins and outer membrane proteins. These strain-specific genes may aid the bacteria under specific circumstances during their long-term infection in genetically diverse hosts. Our results suggest 34 H. pylori clinical strains have extensive genomic diversity. Core genes and strain-specific genes both play essential roles in H. pylori propagation and pathogenesis. Our microarray experiment may help select relatively significant genes for further research on the pathogenicity of H. pylori and development of a vaccine for H. pylori.