• Journal of Microbiology
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• 제44권1호
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• pp.92-97
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• 2006

We developed an mPCR assay for the simultaneous detection, in one tube, of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes using species-specific primers. The mPCR employed the E. coli O157:H7 specific primer Stx2A, Salmonella spp. specific primer Its, S. aureus specific primer Cap8A-B and L. monocytogenes specific primer Hly. Amplification with these primers produced products of 553, 312, 405 and 210 bp, respectively. All PCR products were easily detected by agarose gel electrophoresis, and the sequences of the specific amplicons assessed. Potential pathogenic bacteria, in laboratory-prepared and four commercially available kimchi products, were using this mPCR assay, and the amplicons cloned and sequenced. The results correlated exactly with sequences derived for amplicons obtained during preliminry tests with known organisms. The sensitivity of the assay was determined for the purified pathogen DNAs from four strains. The mPCR detected pathogen DNA at concentrations ranging from approximately 0.45 to $0.05\;pM/{\mu}l$. Thus, this mPCR assay may allow for the rapid, reliable and cost-effective identification of four potentially pathogens present in the mixed bacterial communities of commercially available kimchi.

• 미생물학회지
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• 제27권3호
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• pp.291-296
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• 1989

Campylobacter jejuni was studied for its disinfection by heat-and cold-treatment and UV-irradiation. When C. jejuni was treated by heat, no viable cell was found after 10 min treatment at $55^{\circ}C$, whereas small fraction of cell population was survived after 60 min treatment at $45^{\circ}C$ and $50^{\circ}C$. When they were treated by cold temperature for 30 days, no cell was survived at -$23^{\circ}C$ but about 4 log of the cells were survived at both temperature of $4^{\circ}C$ and -$40^{\circ}C$. When the organisms were UV-irradiated, thier survival rates were proportionally varied to the distance of irradiation. The scanning electron microscopic studies of C. jejuni cells treated by the disinfecting agents revealed that shapes of thecells were deformed from spiral rod into spherical form. The heat-treated cells showed rough and damaged surface on the scanning electron micrographs. In the heat-treated cells, some proteins of high molecular weight appeared to become accumulated in the electrophoretic analysis. The DNAs extracted from the cells treated with the physical agents showed some differences in agarose gel electrophoresis, comparing those of normal cells.

• BMB Reports
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• 제32권6호
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• pp.605-609
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• 1999

Thioltransferase, also called glutaredoxin, is a general GSH-disulfide reductase of importance for redox regulation. Previously, the protein thioltransferase, now called S-type thioltransferase, was purified and characterized from Arabidopsis thaliana seed. In the present study, a second thioltransferase, called L-type thioltransferase, was purified to homogeneity from Arabidopsis thaliana leaves. The purification procedures included DEAE-cellulose ion-exchange chromatography, Sephadex G-50 gel filtration, and glutathione-agarose affinity chromatography. The purified enzyme was confirmed to show a unique band on SDS-PAGE and its molecular weight was estimated to be 26.6 kDa, which appeared to be atypical compared with those of most other thioltransferase. It could utilize 2-hydroxyethyl disulfide, S-sulfocysteine, and insulin as substrates, and also contained dehydroascorbate reductase activity. Its optimum pH was 8.5 and its activity was greatly activated by L-cysteine. When it was kept for 30 min, it appeared to be very stable up to $70^{\circ}C$. It was activated by $MgCl_2$ and, on the contrary, inhibited by $ZnCl_2$, $MnCl_2$, and $AlCl_3$.

• Journal of Nutrition and Health
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• 제36권4호
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• pp.382-388
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• 2003

(-)-Epigallocatechin-3-gallate (EGCG) is a polyphenolic compound found in peen tea leaves, and has been known to be one of the most potent catechin species which inhibits cell growth most possibly through an apoptotic cell death. We investigated the apoptotic activity of (-)-EGCG on the human myeloid leukemia cell line, HL-60. Our results of MTT test indicated that (-)-EGCG had a significant antiproliferation effect in HL-60 cells with $IC_{50}$/ (50% inhibition concentration) value of 65 $\mu$M. Giemsa statining of HL-60 cells treated with (-)-EGCG (100 $\mu$M) for 6hrs showed a typical apoptosis-specific morphological change including shrinkage of the cytoplasm, membrane blobbing and compaction of the nuclear chromatin. The DNA fragmentation was observed from the agarose gel electrophoresis of cells treated with (-)-EGCG for 3hrs or longer, and was progressed to a greater degree as treatment time increases. Treatment of the cells with (-)-EGCG (100 $\mu$M) resulted in a rapid release of mitochondrial cytochrome c into the cytosol, and a subsequent cleavage of caspase-3 to an active form in a treatment-time dependent manner. (-)-EGCG (100 $\mu$M) also stimulated proteolytic cleavage of poly-(ADP-ribose) polymerase (PARP) to an active form in HL-60 cells. Tlken together, (-)-EGCG appears to induce the apoptosis in human myeloid leukemia cells via a caspase-dependent pathway. These results suggest the possible application of (-)-EGCG, the major active compound in green tea, as an antiproliferative agent for cancer prevention.

• The Plant Pathology Journal
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• 제25권3호
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• pp.291-293
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• 2009

The low virus titer in woody plant tissues and the presence of inhibitor compounds such as polyphenols, tannins and polysaccharides are common difficulties that compromise purification of plant viroids from their woody hosts. A simple, reliable method of RNA isolation using CF11 cellulose column on a microcentrifuge tube scale for detecting Apple scar skin viroid (ASSVd) in apple was developed. Total RNA extracted from leaf, woody bark and the fruit skin was used for reverse transcription. RT-PCR products could be detected from RNA prepared from dormant woody bark, fruit skin and fresh leaves with both the CF11 cellulose column method and NucliSens extractor in February, August and November. Meanwhile, with the RNeasy kit RT-PCR, products were detected only in leaves and not from bark or fruit skin. The PCR product, about 330 base pairs, was analyzed by agarose gel electrophoresis. The CF11 cellulose column method was effective for detecting ASSVd. The method enabled the processing of a large numbers of samples of dormant woody bark, leaf and fruit skin of apple.

• BMB Reports
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• 제31권4호
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• pp.345-349
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• 1998

Aspartase (EC 4.3.1.1) from Hafnia alvei was purified to homogeneity by a combination of DEAE-cellulose, Red A-agarose, and Sepharose 6B chromatography. The purified enzyme appeared homogeneous on denatured SDS-polyacrylamide gel electrophoresis. The purified enzyme was a tetrameric protein composed of identical subunits with a molecular weight of 55,000 daltons. The optimum pH for the enzymatic reaction was 8.5 and the optimum temperature for maximum activity was $45^{\circ}C$. The enzyme has an absolute requirement of divalent metal ions ($Mg^{2+}$, $Mn^{2+}$) at the alkaline pH. The enzyme, however, was inactivated in the presence of other divalent cations such as $Zn^{2+}$, $Ca^{2+}$. The helical content of the purified enzyme was estimated by CD spectropolarimetry to be 61%.

• BMB Reports
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• 제30권1호
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• pp.37-40
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• 1997

Earthworm extracts are known for anti-inflammatory, analgesic. antipyretic, and anticancer effects but can also influence blood circulation. It was previously shown that an earthworm, Lumbricus rubelius. contained a water-extractable anticoagulant which was a heat- and acid-stable molecule with hydrophilic property. In order to uncover the biochemical nature of this molecule, the anticoagulant was processed with various hydrolases such as trypsin, DNase, RNase. and lysozome. When the digested samples were analyzed with an in vitro coagulation test measuring activated partial thromboplastin time (APTT) and agarose gel electrophoresis, the anticoagulant proved to be a relatively homogeneous DNA fragment with relative molecular size around 72 base pairs. Interestingly, the activity was further stimulated with a trypsin digestion. RNA. on the other hand, did not prolong the APTT. It was also demonstrated that the DNA accelerated the antithrombin III (AT-III) inhibition of thrombin from $IC_{50}$ of 0.34 to 0.16 unit determined with S-2238 as a substrate, whereas heparin, a popular anticoagulant. shifted the value to 0.05. Therefore, it is suggested that the DNA could be considered as an alternative antithrombotic agent to heparin, which would exhibits bleeding side effects.

• 한국작물학회지
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• 제34권4호
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• pp.345-352
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• 1989

대두(Glycine max L.) 종자로부터 분리된 렉틴을 토끼에 면역화시켜 렉틴항혈청을 얻었다. 이를 이용한 면역화학적 방법을 사용하여 대두렉팅의 면역화학적 특성과 6개 대두 품종의 종자에서 렉틴의 존재를 조사하였다. Crossed immunoeiectrophoresis한 결과 종자는 4종의 침강선을 보임으로서 렉틴에 4종의 항원성분이 존재함을 보여 주었다. 1％ agarose gel을 이용하여 Immunodouble diffusion test한 결과 6개 대두 공시품종 모두 렉틴항체에 대하여 침강선을 나타내었고 푸종간 침강선은 모두 유사한 양상을 보였다. ELISA 방법에 의하여 6개 대두 품종간의 렉틴함량을 조사, 비교한 결과 렉틴함량은 장엽콩>광교>황금콩>백운콩>장백콩=팔달콩 순으로 나타났다. 공시품종의 조자로부터 분리한 렉틴은 모두 토끼 적혈구는 응집시켰으나 Fusarium sp., Alternaria sp., Cuvalaria sp. 등의 곰팡이 포자는 응집시키지 못하였다.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2003년도 춘계학술대회
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• pp.177-177
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• 2003

Adult periodontitis is a chronic inflammatory disease whose etiology is not well defined. Recent studies have shown that vitamin D receptor gene has been a candidate for the susceptibility of adult periodontitis. The purpose of this study is to investigate the frequency of Taq I restriction fragment length polymorphism (RELP) in the vitamin D receptor gene in 141 periodontically healthy controls and 32 adult periodontitis patients. Taq I RFLP in the vitamin D receptor gene were detected by PCR amplification, followed by restriction enzyme digestion and 2% agarose gel electrophoresis. There were no significant difference in the distribution of Taq I RFLP between healthy controls and adult periodontitis group (P > 0.05). Thus, Taq I RFLP in the vitamin D receptor gene may not confer the susceptibility to adult periodontitis in Korean population. However, t allele distributions of this RFLP showed various frequencies among ethnic groups studied. Further studies in other ethnic groups will be required.

• Food Science and Biotechnology
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• 제18권4호
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• pp.872-877
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• 2009

By-products of marine organisms including salmon, skate, flatfish, and yellow goosefish were investigated to search for new source of chondroitin sulfate (CS). Agarose gel electrophoresis with chondroitinase depolymerization showed that purified chondroitin sulfate did not contain any other glycosaminoglycans. 1H-nuclear magnetic resonance (NMR) spectra were acquired to confirm the structure and purity. The average molecular weight ranging from 22 to 64 kDa was determined by high performance size exclusion chromatography. Disaccharide compositions and purities were determined by strong anion exchange-high performance liquid chromatography (SAX-HPLC) after chondroitinase ABC depolymerization. SAX-HPLC data exhibited that the purity was from $81.7{\pm}1.3$ to $114.2{\pm}2.5%$ and the yield was from 1.3 to 12.5%. All analytical results indicate that salmon cartilage, skate cartilage, and yellow goosefish bone could be promising sources of CS to substitute shark cartilage CS in commercial neutraceuticals.