• 제목/요약/키워드: Microbiological assay

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Effect of paternal folate deficiency on placental folate content and folate receptor ${\alpha}$ expression in rats

  • Kim, Hye-Won;Choi, Yun-Jung;Kim, Ki-Nam;Tamura, Tsunenobu;Chang, Nam-Soo
    • Nutrition Research and Practice
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    • v.5 no.2
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    • pp.112-116
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    • 2011
  • We investigated the effect of paternal folate status on folate content and expression of the folate transporter folate receptor ${\alpha}$ ($FR{\alpha}$) in rat placental tissues. Rats were mated after males were fed a diet containing 0 mg of folic acid/kg of diet (paternal folate-deficient, PD) or 8 mg folic acid/kg of diet (paternal folate-supplemented, PS) for 4 weeks. At 20 days of gestation, the litter size, placental weight, and fetal weight were measured, and placental folate content (n=8/group) and expression of $FR{\alpha}$ (n=10/group) were analyzed by microbiological assay and Western blot analysis, respectively. Although there was no difference observed in litter size or fetal weight, but significant reduction (10%) in the weight of the placenta was observed in the PD group compared to that in the PS group. In the PD group, placental folate content was significantly lower (by 35%), whereas $FR{\alpha}$ expression was higher (by 130%) compared to the PS group. Our results suggest that paternal folate status plays a critical role in regulating placental folate metabolism and transport.

Detection of Lymphotropic Herpesviruses by Multiplex Polymerase Chain Reaction

  • Park, Sang-Tae;Kim, Seung-Han;Lee, Dong-Gun;Park, Jung-Hyun;Shin, Wan-Shik;Kim, Tai-Gyu;Paik, Soon-Young;Kim, Chun-Choo
    • Journal of Microbiology
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    • v.39 no.3
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    • pp.226-228
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    • 2001
  • Human lymphotropic herpesvirus is known to be a major pathogen associated with various diseases in bone marrow transplantation (BMT) recipients. A multiplex nested-polymerase chain reaction (PCR) method was developed for the simultaneous detection of human lymphotropic herpesviruses, including Ebstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus 6 variants A and B (HHV6-A, HHV6-B). To demonstrate the usefulness of multiplex PCR for the analysis of clinical samples, peripheral blood mononuclear cells and serum from BMT recipients were analysed. The results skewed that a clear detection could be made between EBV, HCMV and HHV-6. This multiplex PCR assay is an efficient and cost-effective approach to the analysis of large numbers of samples to determine the epidemiological importance of EBV HCMV and HHV-6.

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Nitro oxide in human cytomegalovirus replication and gene expression

  • Lee, Jee-Yeon;Lee, Chan-Hee
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.152-157
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    • 1997
  • Infection of human fibroblast (HF) cells with human cytomegalovirus (HCMV) result in changes in the intracellular level of second messengers. Since nitric oxide (NO) production has been known to be related with other second messengers, it is probable that HCMV infection of HF cells may involve NO. To test this possibility, the amount of NO was measured following ogenous addition of NO generators such as sodium nitroprusside (SNP) or S-nitroso-N-a-cetylpenicillamine (SNAP) immediately after HCMV infection, however, inhibited virus multiplication. Furthermore, immunoblot experiment using monoclonal antibody to HCMV major immediate early (MIE) proteins or CAT assay using pCMVIE/CAT (plasmid containing CAT gene driven by HCMV MIE promoter) revealed that SNP or SNAP blocked the MIE gene expression. SNP was more effective than SNAP in hibiting HCMV multiplication or MIE gene expression. SNP produced more NO than SNAP in inhibiting HCMV multiplication or MIE gene expression. SNP produced more NO than SNAP. Although the mechanism for the inhibition of HCMV multiplication and MIE gene expression by NO is still elusive some correlation with NO-mediated inhibition of HCMV-induced increase in cytosolic free Ca$\^$2+/ concentration ([Ca$\^$2+/]) was observed. The increase of [Ca$\^$2+/] following HCMV infection was inhibited by SNP, and less effectively by SNAP. Raising [Ca$\^$2+/ with bromo-A23187 partially reversed the SNP block of MIE gene expression. Thus, there appear to e some relationships among NO. [Ca$\^$2+/], and HCMV MIE gene expression.

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Analytical Techniques for Vancomycin - A Review

  • Avinash P. Sattur;Lee, Je-Hyuk;Song, Ki-Bang;T. Panda;Kim, Chul-Ho;Rhee, Sang-Ki;B. Gokul
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.3
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    • pp.153-158
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    • 2000
  • Vancomycin belongs to the vancomycin-risocetin family of glycopeptides, and is a subclass of linear sugar containg peptides composed of seven amino acids. Its strochemical configuration forms the basic of a peptidoglycon monomer. The glycosylated hexapeptide chainconsists of chloro-$\beta$-hydroxytyrosines, p-hytidoglycines, N-anthylleucine and aspartic acid forms a rigid molecular frame work and gives the difficulty in the analysis. Voncomycin in the serum samples is usually estimated by liquid chromatography and the bacterial sensitivity was genereally tested by the microbiological assay. The pressent review deals with the qualitative, quantutative, microbioligical and immunological assays and the comparison of the quantitative methods. Clinical implications of vancomycin have also been cited in the review.

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Identification and Characterization of pH-Regulated Genes in Saccharomyces cerevisiae

  • Hong, Sung-Ki;Choi, Eui-Yul
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.327-333
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    • 1996
  • Yeast, like many other microbes, encounters large variations in ambient pH in their natural environments. Microorganisms capable of growing over a wide pH range require a versatile, efficient pH homeostatic mechanism protecting intracellular processes against extremes of pH. In several organisms, fusions to the bacterial lacZ gene have been extremely useful for the identification of genes expressed at different time during the life cycle or under different growth conditions. In this study, using the lacZ gene screening system, we surveyed a large number of yeast strains with lacZ insertion to identify genes regulated by pH. A yeast genomic library was constructed and inserted with lacZ by a shuttle mutagenesis procedure. The yeast transformants were individually picked up with a toothpick, replica-plated, and grown in alkaline pH medium. Among the 35,000 colonies screened, 10 candidate strains were identified initially by the $\beta$-gal assay. We finally confirmed two yeast strains carrying the genes whose expression are strictly dependent on pH of growth medium. One of the fusions showing a 10-fold induction in expression level in response to alkali pH was selected and further characterized. The pH-regulated gene was cloned by inverse PCR and a partial sequence of the gene was determined. Identification and characterization of the gene is currently under investigation.

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Molecular Cloning and Characterization of Catechol 2, 3-Dioxygenase Gene from Aniline-Degrading Psseudomonas acidovorans

  • Lee, Ji-Hyun;Bang, Sung-Ho;Park, Youn-Keun;Lee, Yung-Nok
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.316-321
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    • 1992
  • Catechol 2, 3-dioxygenase (C230) catalyses the oxidative ring cleavage of catechol to 2-hydroxymuconic semialdehyde. This is one of the key reactions in the metabolism of the widespresd pollutant aniline. We have cloned a gene encoding C230 from cells of the aniline degrading bacteria, Pseudomonas acidovorance KCTC2494 strain and expressed in E. coli, A 11.3-kilobase Sau3A partial digested DNA fragment from KCTC2494 was cloned into phagemid vector pBluescript and designated as pLP201. The C230 gene was mapped to a 2.8-kb region, and the derection of transcription was determined. The cloned C230 gene contains its own promoter which can be recognized and employed by E. coli transcriptional apparatus. C230 activities of subclones were identified by enzyme assay and activity staining. The T7 RNA promoter/polymerase system and maxicell analysis showed that a polypeptide with Mw of 35 kDa is the C230 gene product.

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Isolation of Cellulose-Degrading Myxobacteria Sorangium cellulosum (셀룰로오스 분해성 점액세균 Sorangium cellulosum의 분리)

  • Hyun, Hye-Sook;Chung, Jin-Woo;Lee, Han-Bit;Youn, Jin-Kwon;Lee, Cha-Yul;Kim, Do-Hee;Cho, Kyung-Yun
    • Korean Journal of Microbiology
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    • v.45 no.1
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    • pp.48-53
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    • 2009
  • 591 strains of myxobacteria that grew on cellulose as a sole carbon source were isolated from soils collected from various locations in Korea. Morphological characteristics such as the shape of cells, colonies and fruiting bodies and 16S rRNA sequences of the isolates suggested that all the isolates are Sorangium cellulosum. An antifungal assay has indicated that at least 20 strains among the 114 isolates tested produce antifungal substances inhibiting growth of Candida albicans.

Isolation, Identification , and Biodegradability of Phosphamidon-Degrading Bacteria (Phosphamidon 분해세균의 분리동정 및 생분해능)

  • 강양미;송홍규;안태석;허성남
    • Korean Journal of Microbiology
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    • v.35 no.1
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    • pp.61-64
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    • 1999
  • Organophosphorus inseclicide phosphamidon-degrading bacteria were isolated from agricultural soils and identified using Biolog microtiter assay. All Gram-positive degrading bacterial strains belong to genus Bacillus and many Gram-negative bacteria were rare soil species. Among them fast growing strains on phosphamidon-containing minimal medium were sclected and their biodegrading capability wcre measured. YD-17 which was identified as Capnocytophaga gingivalis showed the highest biodegradation rate. It could incrcase the removal of phosphamidon up to 52%. During the biodegradation continuous increase of amount of cell protein was observed, which indicated that phosphamidon was utilized as a carbon source for phosphamidon-degrading bacteria.

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Analysis of fusogenic activity of autographa californica nuclear polyhedrosis virus (Ac NPV) gp64 envelope glycoprotein

  • Kim, Hee-Jin;Yang, Jai-Myung
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.7-14
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    • 1996
  • Teh baculovirus gp64 glycoprotein is a major component of the envelope of budded virus (BV) and has been shown that it plays an essential role in the infection process, especially virus-cell membrane fusion. We have cloned Autographa californica Nuclear Polyhedrosis Virus (AcNPV) gp64 protein were examined for membrane fusion activity by using a synchtium formation assay under various conditions. The optimal conditions required for inducing membrane fusion are 1) form pH 4.0 to 4.8 2) 15 min exposure of cells to acidic pH 3) at least 1 .mu.g of gp64 cloned plasmid DNA per 3 * 10$^{6}$ cells 4) and an exposure of cells to acidic pH at 72 h post-transfection. In order to investigate the role of hydrophobicity of the gp64 glycoprotein for the membrane fusion, the two leucine residues (amino acid position at 229 and 230) within hydrophobic region I were substituted to alanine by PCR-derived site-directed mutagenisis and the membrane fusion activity of the mutant was anlaysed. The gp64 glycoprotein carrying double alamine substitution mutation showed no significant difference in fusion activity. This result suggested that minor changes in hydrophobicity at the amino acid position 229 and 230 does not affect the acid-induced membrane fusion activity of the gp64 glycoprotein.

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The Antimicrobial Activity of Essential Oil from Dracocephalum foetidum against Pathogenic Microorganisms

  • Lee, Saet-Byoul;Cha, Kwang-Hyun;Kim, Su-Nam;Altantsetseg, Shataryn;Shatar, Sanduin;Sarangerel, Oidovsambuu;Nho, Chu-Won
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.53-57
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    • 2007
  • A number of essential oils from Mongolian aromatic plants are claimed to have antimicrobial activities. The essential oil of Dracocephalum foetidum, a popular essential oil used in Mongolian traditional medicine, was examined for its antimicrobial activity. Eight human pathogenic microorganisms including B. subtilis, S. aureus, M. lutens, E. hirae, S. mutans, E. coli, C. albicans, and S. cerevisiae were examined. The essential oil of Dracocephalum foetidum exhibited strong antimicrobial activity against most of the pathogenic bacteria and yeast strains that were tested; by both the agar diffusion method and the minimum inhibitory concentration (MIC) assay ($MIC\;range\;was\;26-2592{\mu}g/ml$). Interestingly, Dracocephalum foetidum even showed antimicrobial activity against methicilin-resistant Staphylococcus aureus (MRSA) strains. We also analyzed the chemical composition of the oil by GC-MS and identified several major components, including n-Mentha-1,8-dien-10-al, limonene, geranial, and neral.