• Toxicological Research
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• 제24권1호
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• pp.29-36
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• 2008

The present study investigated the anti-proliferative and chemosensitizing effects of Crinum asiaticum var. japonicum against multi-drug resistant (MDR) cancer cells. The 80% methanol extract, chloroform ($CHCl_3$) fraction and butanol (BuOH) fraction of C. asiaticum inhibited the growth of mitoxantrone (MX) resistant HL-60 (HL-60/MX2) cells. When HL-60/MX2 cells were treated with the $CHCl_3$ and BuOH fractions, DNA ladder and sub-G1 hypodiploid cells were observed. Furthermore, the fractions reduced BcI-2 mRNA levels, whereas Bax mRNA levels were increased. These results suggest that the inhibitory effect of C. asiaticum on the growth of the HL-60/MX2 cells might arise from the induction of apoptosis. Treatment of HL-60/MX2 cells with the fractions markedly decreased the mRNA levels of the multi-drug resistance protein-1 and breast cancer resistance protein. The $CHCl_3$ fraction and hexane fraction increased MX accumulation in HL-60/MX2 cells. These results imply that the $CHCl_3$ fraction of C. asiaticum plays a pivotal role as a chemosensitizer. We suggest that components of C. asiaticum might have a therapeutic potential for the treatment of MDR leukemia.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2003년도 식물바이오벤처 페스티발
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• pp.65-71
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• 2003

Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorho-damine 123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to play a novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

• 한국토양비료학회지
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• 제41권6호
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• pp.415-418
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• 2008

폐기물 연용토양의 미생물 군집 분석을 위해 공단지역 하수 슬러지 처리등 16처리(3수준)에 대하여 가장 좋은 방법을 찾고자, RFLP, CLSU, PLFA, TGGE 의 4가지 방법을 이용하였으며, 이들 4가지 방법에 대하여 시기별 군집변동 조사를 통한 통계적 비교 분석을 하였다. 그 결과, 미생물의 경시적 군집 변동상 관찰은 PLFAs (Phospholipid Fatty Acids) 방법이 가장 효율적이었으며 RFLP는 많은 종류의 제한 효소의 사용이 필요하다. TGGE는 DNA 추출을 통해 비배양학적인 분석에서는 가장 좋은 방법이나 PCR조건 등 실험 과정에서 토양종류에 따라 민감하게 반응하여 비슷한 조성의 토양분석에 효율적이라고 생각한다.

• 한국식품영양과학회지
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• 제38권8호
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• pp.1003-1007
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• 2009

부추를 암 예방 식품 소재로 활용하기 위하여 부추로부터 주요 생리활성 물질인 함황화합물을 분리하여 인체 암세포성장 억제 및 간암세포의 사멸이 apoptosis에 의해 유도되는지를 조사하였다. 부추 함황화합물을 1, 5, 10, 20 및 30 $\mu g$/mL 농도로 24, 48 및 72시간별로 간암(HepG2) 및 폐암세포 (A549)에 처리하여 암세포 증식억제 효과를 측정한 결과 HepG2 및 A549 세포에서 농도 및 시간 의존적으로 그 성장을 억제하였으며, 20 $\mu g$/mL 농도 이상에서 암세포 성장이 60% 이상 억제되었다. 또한 부추 함황화합물 30 $\mu g$/mL 농도로 처리 시 대조군에 비하여 폐암 및 간암 세포수 감소 및 심한 형태학적 변화가 관찰되었다. 이들 암세포의 $IC_{50}$ 값을 측정한 결과 부추 함황화합물은 폐암세포(A549)보다 간암세포(HepG2)에 더 효과가 있었다. 한편 부추 함황화합물은 30 $\mu g$/mL 농도에서 핵 응축 및 apoptotic body를 나타내었으며, 농도 의존적으로 subG1 DNA 함량이 증가함으로써 HepG2 암세포 사멸이 apoptosis에 의해 유도되는 것을 확인할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제27권3호
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• pp.561-572
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• 2017

The global commercial cultivation of transgenic crops, including glyphosate-tolerant soybean, has increased widely in recent decades with potential impact on the environment. The bulk of previous studies showed different results on the effects of the release of transgenic plants on the soil microbial community, especially rhizosphere bacteria. In this study, comparative analyses of the bacterial communities in the rhizosphere soils and surrounding soils were performed between the glyphosate-tolerant soybean line NZL06-698 (or simply N698), containing a glyphosate-insensitive EPSPS gene, and its control cultivar Mengdou12 (or simply MD12), by a 16S ribosomal RNA gene (16S rDNA) amplicon sequencing-based Illumina MiSeq platform. No statistically significant difference was found in the overall alpha diversity of the rhizosphere bacterial communities, although the species richness and evenness of the bacteria increased in the rhizosphere of N698 compared with that of MD12. Some influence on phylogenetic diversity of the rhizosphere bacterial communities was found between N698 and MD12 by beta diversity analysis based on weighted UniFrac distance. Furthermore, the relative abundances of part rhizosphere bacterial phyla and genera, which included some nitrogen-fixing bacteria, were significantly different between N698 and MD12. Our present results indicate some impact of the glyphosate-tolerant soybean line N698 on the phylogenetic diversity of rhizosphere bacterial communities together with a significant difference in the relative abundances of part rhizosphere bacteria at different classification levels as compared with its control cultivar MD12, when a comparative analysis of surrounding soils between N698 and MD12 was used as a systematic contrast study.

• 미생물학회지
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• 제41권2호
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• pp.152-156
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• 2005

신생아의 분변에서 분리한 Pediococcus acidilacti GMB7330은 내산성(pH 2-3) 및 내담즙성$(1.0\%)$을 가지며 Helicobacter pylori에 대해 항균 활성을 나타내었다. P. acidilactici GMB7330의 H. pylori에 대한 저해 특성을 H. Pylori의 균수와 urease 활성 변화, 그리고 전자현미경을 이용한 세포구조 파괴를 관찰하므로써 조사하였다. P. acidilactici GMB 7330의 배양여액(pH 4.5)은 배양 24시간만에 H. pylori의 중식을 약 4 log 감소시켰다. 이때 P. acidilactici GMB7330의 배양여액의 pH를 중성으로 조절하였을 경우에는 pH 4.5의 경우보다 항균력이 다소 감소하기는 하였으나 완전히 소실되지는 않았다. 뿐만 아니라, P. acidilactici GMB7330과 유사한 농도의 젖산을 생성하는 유산균주 Lactobacillus sp. GM7311의 경우 H. pylori에 대해서 항균력을 나타내지 않는 것으로 미루어 P. acidilactici GMB7330의 항균 활성에는 배양 중 생성된 젖산 이외의 물질이 관여하는 것으로 관찰되었다. 또한 P. acidilactici GMB7330은 pH값에 관계없이 H. pylori의 urease 활성을 $50\%$이상 감소시켰으며, P. acidilactici GMB7330의 배양여액을 처리한 H. pylori 균주는 세포벽 구조의 파괴로 인한 세포 내부물질의 유출이 관찰되었다.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 국제심포지움
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• pp.88.2-98
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• 2002

A wide arry of naturally occurring substances particularly those present in dietary and medicinal plants, have been reported to possess substantial cancer chemopreventive properties. Certain phytochemicals retain strong antioxidative and anti-inflammatory properties which appear to contribute to their chemopreventive or chemoprotective activities. Inducible cyclooxygenase(COX-2) and nitric oxide synthase (iNOS) are important enzymes that mediate inflammatory processes. There is some evidence that expression of both COX-2 and iNOS is co-regulated by the eukaryotic transcription factor NF-$textsc{k}$B. Increased expression of COX-2 and/or iNOS has been associated with pathophysiology of certain types of human cancers as well as inflammatory diseases. Since inflammation is closely linked to tumor promotion, substances with potent anti-inflammatory activies are anticipated to exert chemopreventive effects on carcinogenesis, particularly in the promotion stage. An example is curcumin, a yellow pigment of turmeric (Curcuma longa L., Zingiberaceae), that strongly occurring diaryl heptanoids structurally related to curcumin have substantial anti-tumor promotional activities in two-stage mouse skin carcinogenesis. Thus, yakuchinone A [1-(4'-hydroxy-3'-methoxyphenyl)-7-phenyl-3heptanone] and yakuchinone B [1-(4'-hydroxy-3'methoxyphenyl)-7-phenylhept-1-en-3-one] present in Alpinia oxyphylla Miquel (Zingiberacease) attenuate phorbol ester-induced inflammation and papilloma formation in female ICR mice. These diarylheptanoids also suppressed phorbol ester-induced activation of epdermal ornithine decarboxylase and its mRNA expression when applied onto shaven backs of mice. Yakuchinone A and B as well as curcumin inhibited phorbol ester-induced expression of COX-2 and iNOS and their mRNA in mouse skin via inactivation of NF-$textsc{k}$B. Capsaicin, a major pungent ingredient of red pepper also attenuated phorbol ester-induced NF-$textsc{k}$B activation. Similar suppression of COX-2 and iNOS and down-regulation of NF-$textsc{k}$B activation for its DNA binding were observed with the ginsenosied Rg3 and the ethanol extract of Artemisia asiatica. We have also found that certain anti-inflammatory phytochemicals exert inhibitory effects on phorbol ester-induced COX-2 expression and NF-$textsc{k}$B activation in immortalized human breast epithelial (MCF-10A) cells in culture. One of the plausible mechanisms undelying inhibition by aforementioned phytochemicals of phorbol ester-induced NF-$textsc{k}$B activation involves interference with degragation of the inhibitory unit, I$textsc{k}$Ba, which blocks subsequent nuclear translocation of the functionally active p65 subunit of NF-$textsc{k}$B. the activation of epidermal NF-$textsc{k}$B by phorbol ester and subsequent induction of COX-2 hence appear to play an important role in intracellular signaling pathwasy leading to tumor promotion and targeted inhibition of NF-$textsc{k}$B may provide a new promising cancer chemopreventive strategy.

• Parasites, Hosts and Diseases
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• 제37권3호
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• pp.163-169
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• 1999

House dust mite allergens have been well established as sensitizing agents that are important in the induction of allergic diseases. In order to analyze epitopes of the allergen and to develop a quantitative method of the allergen exposure, monoclonal antibodies against a recombinant Der p 2 (rDer p 2), one of the major allergens of Dermatophogoides pteronyssinus, were produce. Four monoclonal antibodies produced wee species-specific and did not cross-react to the D. farinae crude extract. Two of the monoclonal antibodies were found to be IgG1 and the others were IgM. For the analysis of epitopes, a Der p 2 cDNA encoding 126 amino acids (aa) was dissected into three fragments with several overlapping peptides, A (aa residues 1-49), B (44-93), and C fragment (84-126). Three monoclonal antibodies showed reactivities to the recombinant B fragment and to the full-length rDer p 2, but one monoclonal antibody reacted only with the full-length rDer p 2. Two-site capture ELISA was developed using two different monoclonal antibodies for quantitating Der p2 in house dust. The sensitivity limit was 4ng/ml with rDer p2 and $8{\;}\mu\textrm{g}/ml$ with the d. pteronyssinus crude extract. The result suggested that the assay using monoclonal antibodies against rDer p2 could be useful for the environmental studies and for the standardization of mite allergen extracts.

• Journal of Forest and Environmental Science
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• 제11권1호
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• pp.81-101
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• 1995

지표-형질의 상관은 우량 개체 선발과 유전획득량의 증대를 위해 임목 육종에서 해결되어야 할 중요한 과제 중의 하나로 최근 분자유전학적 수준에서의 임의 증폭 다형 디엔에이 (RAPD) 기술의 발달로 이의 해결이 눈 앞에 다가왔다. 호주 퀸즈랜드산림청과 퀸즈랜드대 임목생물공학연구소가 공동 연구하고 있는 슬래쉬소나무, 카리비아소나무 및 그 교잡종에 있어 이 기술을 이용한 수피 두께에 대한 연구 및 육종 계획 전략을 소개한다. 1대 잡종에서 186개의 지표를 포함한 총 길이 1641cM의 16개 연관군의 유전적 지도가 작성되었고, 이 연관군 지도에 수피 두께를 지배하는 6개의 유전자좌가 추정되었다. 또한, 유전적 지표를 이용한 조기 선발을 위해 먼저 중요 형질을 지배하는 유전자들에 대한 종 특성 유전적 지표를 결정하고, 다음 여러가지 대립유전자형에 대한 지표-대립유전자 상관을 구명하는 2단계 전략이 제시되었다. 소나무류는 발아시 양료로 쓰이는 자성배우체는 모수에서 유래하나, 접합자인 배는 양친수로부터 유래하므로 이러한 이질적 유전 조성을 갖인 종자의 발달을 이용한 RAPD 지표와 형질의 상관 연구는 배 단계에서도 우량 개체의 선발을 가능하게 하여 소나무류 육종의 장래를 밝게 하고 있다.

• Journal of Forest and Environmental Science
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• 제32권2호
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• pp.158-163
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• 2016

Mushroom surveys and collections were conducted in the western and eastern forest areas in Cambodia, and then fungal biodiversity was analyzed by identifying mushrooms. One thousand and three hundreds eighty three specimens were identified by morphological and genetical characteristics, and were classified into 238 species, 160 genera, 52 families, 15 orders, and 3 phylums. The collected mushrooms were immersed in 70% ethyl alcohol for DNA extraction, and the rest of them were dried in the portable mushroom dryer for 12 hrs. Among these mushrooms, the genera Mycena (8.7%), Ganoderma (5.6%), Microporus (5.3%), Marasmius (4.2%), Marasmiellus (3.0%), Phellinus (2.5%), Trametes (2.5%), Hygrocybe (1.9%) and Pycnoporus (1.5%) were dominant. In the western Cambodia, 1,061 specimens were collected from Koh Kong forests, while 263 specimens were collected from the eastern Cambodia, Seima and Mondulkiri forests. Elevations of surveyed sites were ranged from 0 to 750 m above sea level. The number of species observed in the elevation of 251-500 m was the highest as compared to the other ranges of elevation. Daldinia concentrica, Microporus vernicipes, Microporus xanthopus, Pycnoporus coccineus, Stereum hirsutum, and Stereum ostrea were commonly distributed in all ranges of elevation, while the distribution of Ceratomyxa fruticulosa, Panus fulvus, Schizophyllum, Trametes versicolor, and Tyromyces chioneus were limited under 500 m. One hundred and forty one species including Amauroderma sp., Bjerkandera adusta, Trichaptum abietinum, and Tyromyces chioneus were collected only in Cardamom, while 20 species including Auricularia auricula-judae, Coriolopsis sanguinaria, Rigidoporus microporus, and Xylaria polymorpha were collected only in Seima. Ganoderma sp., Mycena sp., Marasmius sp., Microporus xanthopus, Phellinus sp., and Russula sp. were dominant species in both the western and eastern Cambodia. Species diversity indices in the eastern and western survey sites were 1.83 and 1.77, respectively, while evenness indices were 0.92 and 0.90. The species similarity index between two survey sites was 0.42.