• Journal of Radiation Protection and Research
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• v.22 no.1
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• pp.15-21
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• 1997

In vitro somatic mutation induced by ${\gamma}$-radiation and pentachlorophenol(PCP) which is representative of chemical pollutant was measured at the hypoxanthine-guanine phosphoribosyl transferase(hprt) locus in human T- lymphocytes by a cell cloning assay. Mutant cells were selected by their ability to form a clone in the presence of purine analogue 6-thioguanine. The mutant frequencies by ${\gamma}$-irradiation to a dose of 1.0 Gy, 2.0 Gy and 3.0 Gy were 40%, 400% and 750% higher than those in controls. Significant changes were not observed in mutant frequencies in the 0.2 Gy and 0.5 Gy irradiated groups. When the doses of PCP were 15 ppm, 25 ppm and 50 ppm, the mutant frequencies increased by 30%, 90% and 520%, respectively. No changes were observed in the 10 ppm treated group. Similar types of dose-response relationship were shown in the two different mutagens. Reverse transcriptase/polymerase chain reaction technique(RT/PCR) was needed for the mutation spectrum to discriminate combined exposure to radiation and chemicals.

• Korean Journal of Medicinal Crop Science
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• v.17 no.4
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• pp.266-272
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• 2009

Cinnamyl alcohol dehydrogenase (CAD, EC 1.1.1.95), catalyzes the reduction of hydroxycinnamaldehydes to give hydroxycinnamyl alcohols, or "monolignols," the monomeric precursors of lignin. Lignins are important components of cell walls and lignified secondary cell walls play crucial roles in long distance transport of water and nutrients during plant growth and development and in plant defense against biotic and abiotic stresses. Here a cDNA clone containing a CAD gene, named as PgCAD, was isolated from a commercial medicinal plant Panax ginseng. PgCAD is predicted to encode a precursor protein of 177 amino acid residues, and its sequence shares high homology with a number of other plant CADS. The expression of PgCAD in adventitious roots and hairy roots of P. ginseng was analyzed using reverse transcriptase (RT)-PCR under various abiotic stresses such as salt, salicylic acid, wounding and chilling treatment that triggered a significant induction of PgCAD at different time points within 2-48 h post-treatment. This study revealed that PgCAD may help the plants to survive against various abiotic stresses.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.61-61
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• 2003

Aquaporins (AQPs)는 다양한 상피세포와 내피세포에 존재하며 다량의 물 수송을 촉진하는 막성단백질로 현재 11개의 AQP가 (AQP0-10) 발견되었으나, 아직 생리적, 기능적 분석은 불충분한 상태이다. 생쥐의 자궁내막은 발정주기 동안 호르몬의 자극에 따라 부풀어오르거나 수축하는 변화를 보이며 에스트로젠과 몇몇 혈관에 작용하는 매개체에 의해 자궁 혈관의 투수성이 증가한다는 보고는 있으나, 자궁액의 수송 메커니즘에 대해서는 뚜렷하게 밝혀진 바가 없다. 발정기의 생쥐 자궁은 자궁내막세포의 증식과 함께 수화되는 특징을 보이며 자궁내강으로 물이 수송되어 luminal fluid의 점성이 낮아지는 현상이 나타나는데, 이 때 AQP가 water channel로서 중요한 역할을 할 것으로 보고 본 실험에서는 면역조직화학법(immunohistochemistry)과 역전사중합효소연쇄반응(Reverse-transcriptase polymerase chain reaction)을 통해 발정기 자궁의 수화와 AQP 발현의 상관성에 대해 알아보고자 하였다. 면역조직화학법의 결과 발정주기의 다른 시기에 비해 발정기(estrus phase)에 자궁상피세포에 AQP4, 5, 8 protein이 다량 존재하는 것으로 밝혀졌고, 근육층(myometrium)에서의 발현은 발정주기 동안 차이가 없었다. Whole uterus로 RT-PCR을 수행한 결과 AQP4, 5, 8 mRNA는 luteal phase에 비해 follicular phase에 더 많이 발현하는 것으로 확인되었다. 또한 LCM(Laser Capture Microdissection) system을 이용하여 luminal epithelium과 stromal cell을 분리하여 RT-PCR을 수행한 결과 AQP4, 5, 8 mRNA는 stromal cell 보다는 luminal epithelium에 더 많이 발현하며, 이 역시 follicular phase에 발현량이 증가함을 확인하였다. 이러한 결과로 미루어 생쥐 자궁에서 AQP4, 5, 8은 발정주기 내막에 발현이 증가하며 이는 자궁내강 안으로 수분을 수송하는데 주요한 기작으로 사료되며 estrogen에 의한 조절 가능성을 암시한다.

• Tuberculosis and Respiratory Diseases
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• v.72 no.2
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• pp.156-162
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• 2012

Background: The main goal of this study was to evaluate the diagnostic efficacy of reverse transcription-nested polymerase chain reaction (RT-nested PCR) in bronchial washing fluid with MAGE A1-6 common primers for the detection of lung cancers invisible by bronchoscopy. Methods: To determine the expression of MAGE A1-6 gene in 189 lung cancers diagnosed by conventional fluoroscopy-guided lung biopsy and 89 cancer-free controls, RT-nested PCR was performed in bronchial washing specimens. We analyzed MAGE A1-6 RT-nested PCR data according to tumor histology, stage, size, and compared them with cytological data. Results: 189 patients (111 cases in adenocarcinoma, 47 cases in squamous cell carcinoma, 22 cases in small cell lung carcinoma, and 9 cases in other cancers) and 89 benign patients were investigated. The expression of MAGE was performed by nested RT-PCR using common MAGE primer. Among 189 cancer patients, the expression rate of MAGE was 49.2%, and the positive predictive value was 89.4%. However, the expression rate of MAGE in patients with benign lesions was 12.4%. In peripheral lung cancer, the positive rate of MAGE expression was 57.4% in squamous cell carcinoma, 44.1% in adenocarcinoma and 59.1% in small cell lung cancer. Whereas the expression rate of bronchial washing cytology in peripheral lung cancer was 9.0% (p=0.011). Conclusion: MAGE RT-PCR in bronchial washing fluid gave us promising data for the detection of peripheral lung cancer. It could be a useful method for selecting diagnostic tools for peripheral lesions.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.1
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• pp.7-12
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• 2010

The full-length cDNA of the porcine peroxisomal ${\Delta}^3$,${\Delta}^2$-enoyl-CoA isomerase (PECI) gene encodes a monofunctional peroxisomal ${\Delta}^3$,${\Delta}^2$-enoyl-CoA isomerase. Cloning and sequencing of the porcine PECI cDNA revealed the presence of an 1185-base pair open reading frame predicted to encode a 394-amino acid protein by the 5'rapid amplification of cDNA ends (5'RACE) and EST sequences. The porcine PECI gene was expressed in seven tissues (heart, liver, spleen, lung, kidney, skeletal muscle, fat) which was revealed by reverse transcriptase-polymerase chain reaction (RT-PCR). The porcine PECI was mapped to SSC71/2 p11-13 using the somatic cell hybrid panel (SCHP) and the radiation hybrid panel (RH) (LOD score 12.84). The data showed that PECI was closely linked to marker S0383. A C/T single nucleotide polymorphism in PECI exon 10 (3'UTR) was detected as a PvuII PCR-RFLP. Association analysis in our experimental pig population showed that different genotypes of PECI gene were significantly associated with the Average Backfat thickness (ABF) (p<0.05) and Buttock backfat thickness (p<0.01).

• Journal of fish pathology
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• v.10 no.1
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• pp.39-44
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• 1997

Metallothionein is an important and essential protein to control the intracellular concentration of heavy metals, which exist in all organisms from bacteria to vertebrates. Although the detailed functions and induction mechanisms of metallothionein gene have not been clearly characterized until yet, the structure of several metallothionein genes has been revealed. Especially, piscine metallothionein is regarded as an important protein because it is induced by several heavy metal pollutants and environmental stress and it could be determined the comparative amount of heavy metals and the extent of environmental stress by assaying the RNA transcript of metallothionein gene in the method of the quantitative RT-PCR(Reverse Transcriptase Polymerase Chain Reaction). In this study I have characterized the 450 bp PCR fragment of metallothionein gene amplified by using the mixture of internal specific primers and universal 3' end primer. The nucleotide sequence analysis of 450 bp PCR fragment amplified in cDNA library of channel catfish did not show strong homology to other piscine metallothionein genes.

• The Journal of Korean Society of Virology
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• v.29 no.3
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• pp.155-163
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• 1999

Hantaan virus (HTNV), the etiologic agent of hemorrhagic fever with renal syndrome (HFRS), belongs to the genus Hantavirus, and has three single negative stranded RNA genome segments. HTNV strain Howang isolated from the blood of severe case of Korean HFRS is more virulent than HTNV 76/118 and the M and S genome segments' nucleotide sequence of Howang strain showed 93.5% and 94% homology to each segment of HTNV 76/118. We have obtained 6533 nucleotides long sequence of the L genome segment of Howang strain using reverse transcriptase in conjunction with PCR amplification and compared to other hantaviruses. The messenger sense of the L segment contains one long single long open reading frame of 2151 amino acids, which encodes a deduced RNA dependent RNA polymerase of 246.4 kDa caculated molecular weight protein. The nucleotide sequence of the L segment of Howang strain shows 93%, 74%, 66%, 65% homology to HTNV 76/118, Seoul virus 80/39, Puumala virus $H{\ddot{a}}lln{\ddot{a}}s$ B1 and Sin Nombre virus, respectively. The amino acid sequence of the L segment of Howang strain shows 99%, 85%, 68%, 68% homology to HTNV 76/118, Seoul virus 80/39, Puumala virus $H{\ddot{a}}lln{\ddot{a}}s$ B1 and Sin Nombre virus, respectively.

• Toxicological Research
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• v.34 no.1
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• pp.7-12
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• 2018

Laboratory animal models have been developed to investigate preventive or therapeutic effect of medicinal products, or occurrence or progression mechanism of atopic dermatitis (AD), a pruritic and persistent inflammatory skin disease. The murine model with immunologic phenomena resembling human AD was introduced, which demonstrated skewedness toward predominance of type-2 helper T cell reactivity and pathophysiological changes similar as human AD following 2,4-dinitrochlorobenzene (DNCB) sensitization and challenge. Molecular mechanism on the DNCB-mediated AD was further evaluated. Skin tissues were collected from mice treated with DNCB, and each tissue was equally divided into two sections; one for protein and the other for mRNA analysis. Expression of filaggrin, an important protein for keratinocyte integrity, was evaluated through SDS-PAGE. Level of mRNA expression for cytokines was determined through semi-quantitative reverse transcriptase polymerase chain reaction. Expression of filaggrin protein was significantly enhanced in the mice treated with DNCB compared with the vehicle (acetone : olive oil = 4 : 1 mixture) treatment group or the normal group without any treatment. Level of tumor necrosis factor-alpha and interleukin-18 mRNA expression, cytokines involved in activity of type-1 helper T ($T_H1$) cell, was significantly downregulated in the AD group compared with other control groups. These results suggest that suppression of $T_H1$ cell-mediated immune response could be reflected into the skin tissue of mice treated with DNCB for AD induction, and disturbance of keratinocyte integrity might evoke a compensatory mechanism.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.4
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• pp.1129-1133
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• 2004

For the purpose of developing new anti-HIV agents from natural sources, the extracts of Kalopanax pictus were tested for their inhibitory effects on HIV-1 replication and its essential enzymes as the reverse transcriptase (RT). protease and α-glucosidase. In the assay of HIV-1-infected human T-cell line, water extracts of stem and leafstalk inhibited the HIV-1-induced cytopathic effects with Ie (inhibitory concentration) of 25 and 50㎍/㎖, respectively. Moreover water extracts (100㎍/㎖) of stem and leafstalk showed strong activity of 80% and 90% on anti-HIV-1 RT using Enzyme Linked Oligonucleotide Sorbent Assay (ELOSA) method. In the HIV-1 protease inhibition assay, aqueous stem extract inhibited the activity of the enzyme to cleave an oligopeptide, resembling one of the cleavage sites in the viral polyprotein which can only be processed by HIV-1 protease with 58%, but no glucosidase inhibitory activities. We found out this result, for these samples it is possible that the inhibition of the viral replication in vitro is due to the inhibition at least one of RT and protease. It would be of great interest to identify the compounds which are responsible for this inhibition, since all therapeutically useful agent up to date are RT, PR and α-glucosidase inhibitors.

• Korean Journal of Pharmacognosy
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• v.42 no.4
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• pp.348-353
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• 2011

The aim of the present study is to investigate the cytokine production inhibitory effect of a Dictamni Radicis Cortex (DRC). DRC has been commonly used as important medicinal herb in China and it used to control eczema, atopic dermatitis, fever and inflammatory diseases. Inflammation, such as a bacterial infection in vivo metabolites, such as external stimuli or internal stimuli to the defense mechanisms of the biological tissue a variety of intracellular regulatory factors deulin inflammatory TNF-${\alpha}$, IL-$1{\beta}$, IL-6, IL-8, such as proinflammatory cytokines, prostagrandin, lysosomal enzyme, free radicals are involved in a variety of mediators. The present study was designed to determine the effect of the DRC on proinflammatory factors such as NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 in lipopolysaccharide (LPS) - stimulated RAW264.7 cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of DRC, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, the DRC reduced NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 production without cytotoxicity. Our results suggest that the DRC may have an anti-inflammatory property through suppressing inflammatory mediator productions.