• 대한의생명과학회지
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• 제2권1호
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• pp.121-126
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• 1996

Vinblastine을 포함하는 bis-indole alkaloids에 대한 단일클론 항체를 생산하여 Vinca alkaloids의 양을 측정할 수 있는 간편한 immunoassay체계를 확립하였다. Vinca alkaloids는 periwinkle식물체의 배양된 세포로부터 추출하여 BSA와 접합한 후 Balb/c생쥐에 면역시켜 얻은 비장세포와 골수종양세포의 융합을 유도하여 VBL-BSA에 반응하는 클론을 ELISA 방법으로 분석하였으며 이들 클론 중 bis-in-dole alkaloids와 특이적으로 반응하는 항체는 inhibition assay를 통하여 분리할 수 있었고 그 결과 두개의 단일클론 항체를 형성하는 세포주(KN-1과 KN-2)를 확립하였다. KN-1의 경우 dimeric bis-indole alkaloids 와는 상당한 교차반응을 나타낸 반면 monomeric bis-indole alkaloids 와는 교차반응을 나타내지 않았으며 이 클론의 항체를 이용하여 배양된 세포 추출물에 포함된 Vinca alkaloids의 양을 측정한 결과 0.05 nM정도의 dimeric Vinca alkoloids까지도 측정할 수 있었다.

• Applied Biological Chemistry
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• 제42권4호
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• pp.293-297
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• 1999

Escherichia coli의 중요한 sugar 흡수 system인 Phosphoenolpyruvate. carbohydrate phosphotransferase system(PTS)의 주요 구성 enzyme을 만드는 pts operon에는 여러 개의 promoter가 존재하여 어느 환경에서도 적절한 정도의 PTS 활성을 유지하도록 한다. E. coli pts operon의 P1 promoter transcription이 in vitro와 in vivo에서 차이가 나는 원인을 밝히기 위하여 pts promoter activity에 영향을 줄 수도 있는 pts P0 Promoter의 1kbp upstream에서부터 P0와 P1 promoter까지 transcription vector에 cloning하여 in vitro transcription assay를 한 결과, pts promoter의 upstream DNA가 pts P1 promoter의 in vitro transcription에 미치는 영향이 없음을 알 수 있었다. 여러 가지 PTS sugar들을 이용하여 in vivo에서 이들 sugar 들이 pts transcription에 미치는 영향을 cAMP농도 변화와 비교 조사한 바, glucose존재 하에 자랄 때보다 CAMP농도가 높은 mannose나 mannitol 존재 하에 bacteria가 자랄 때 P1b transcription은 증가하나 P0 transcription은 glucose존재 하에 자랄 때 더 높은 결과를 보였다. 이 결과는 P0에 glucose에 의해 induction되는 repressor가 존재하고, P1 에는 glucose. mannose, mannitol에 의해 공통적으로 induction되는 제 2의 repressor가 존재할 것이라는 가능성을 보여주는 것이다.

• 대한본초학회지
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• 제26권1호
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• pp.111-117
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• 2011

Objectives : The present study was designed to investigate effects of mixed extract from korean medicinal herbs (MIX) on oxidation/reduction reaction-related and aging-related enzyme in vitro. Methods : We performed MTT assay, collagenase inhibition assay, elastase inhibition assay, tyrosinase inhibition assay, DPPH free radical scavenging assay, SOD-like activity and xanthine oxidase inhibition assay. Results : Recently, many studies have reported that elastin is also involved in inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. The MIX showed 97% inhibition of collagenase activity, and 64% inhibition of elastase activity at 1 mg/ml concentration of MIX, next only to positive control, which indicate good efficacy for anti-wrinkle ingredient. Also it's treatment showed 34% inhibition of tyrosinase activity, to relate whitening effect, at the same dose of MIX. Antioxidant activities were determined by DPPH radical scavenging, xanthine oxidase (XO) inhibiting activity and SOD-like activity. Also these scavenging, XO-inhibiting and SOD-like activities were measured in 91%, 80%, and 63% inhibition, respectively, at a treated dose of 1 mg/ml, compare to control. Conclusions : These results suggest that possibility of mixed korean medicinal herbs as a functional ingredient for anti-wrinkle and whitening, anti-oxidation and anti-aging cosmetic formula.

• 생약학회지
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• 제47권3호
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• pp.264-272
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• 2016

This study was carried out to evaluate the preventive effect of the Corni Fructus (SSU) 50 % EtOH extract (SSU-E50) against bisphenol A (BPA) toxicity in Leydig cells and improving testosterone deficiency syndrome in orchidectomized Sprague-Dawly (SD) rats. Antioxidant properties were measured by radical scavenging activity of SSU-E50 in ABTS assay and DPPH assay. Also, real-time polymerase chain reaction(real-time PCR) was performed to quantify the mRNA expression levels of antioxidant enzyme. SD rats were divided into eight group: normal, sham operation (Sham), orchidectomized (ORX), ORX treated with testosterone 1 mg/kg (Tes. 1), ORX treated with SSU water extract 100 mg/kg (SSU-A 100) and 300 mg/kg (SSU-A 300), ORX treated with SSU 50 % EtOH extract 100 mg/kg (SSU-E 100) and 300 mg/kg (SSU-E 300). On a comparative basis, the SSU showed better activity quenching ABTS with an IC50 value of 0.29 mg/ml and DPPH with an IC50 value of 0.33 mg/ml. Cell viability was evaluated by MTS assay as described not cytotoxic at the highest concentration of $500{\mu}g/ml$. Cytotoxicity of BPA showed in $200{\mu}M$, but definitely survived by treatment with SSU in Leydig cells. In addition, SSU increased the mRNA expression levels of antioxidant enzyme in BPA induced Leydig cells. Superoxide dismutase (SOD) level was slightly increased and malondialdehyde (MDA) level was decreased with SSU-A 100 in in-vivo. These results suggest that Corni Fructus extracts have the greatest property as a natural anti-oxidative and improves testosterone deficiency syndrome source.

• 약학회지
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• 제44권4호
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• pp.340-346
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• 2000

Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV types 16 and 18) and cervical cancer has been well known. An extract of natural products, named as Somatid, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Somatid inhibited the proliferation of human cervical cancer cell lines (C-33A, SiHa, CaSki) and HaCaT keratinocytes in a dose response manner, In vitro binding assay and ELISA showed that Somatid inhibited the in vitro biding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Somatid inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by Somatid. Our data suggested that Somatid inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus can be used as a putative anti-HPV agent for the treatment of cervical carcinomas caused by HPV.

• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.445-450
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• 1996

Gelatinase A is a member of the matrix metalloproteinases that play an important role in cancer invasion and metastasis. In the course of screening gelatinase A inhibitors from microbial sources, a fungal strain PT-262 showed a strong inhibitory activity. The strain was identified as Chaunopycnis alba on the basis of its morphological characteristics. The inhibitor was isolated from acetone extract of mycelial cake by sequential chromatographies on MCI-gel, Sephadex LH-20, and a reverse-phase HPLC column. The purified inhibitor was identified as pyridoxatin by its physico-chemical properties and spectroscopic analysis. Pyridoxatin is not a peptide analog and has cyclic hydroxamic acid moiety. It inhibited activated gelatinase A with an $IC_{50}$ value of 15.2 ${\mu}M$ using fluorescent synthetic peptide. It also had a strong cytotoxicity against human cancer cell lines in vitro. Furthermore, this compound inhibited DNA synthesis with an $IC_{50}$ value of 2.92 ${\mu}M$ in PC-3 prostate cancer cells by [$^3H$]thymidine incorporation assay.

• 한국약용작물학회지
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• 제7권1호
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• pp.37-44
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• 1999

60종(種)의 약용식물과 식용작물을 대상으로 항종양활성등의 생리활성을 조사함으로서 약용식물과 식용작물의 유용적인 측면의 확인뿐만 아니라 나아가서 새로운 생리활성물질 탐색의 가능성을 검토하기 위해서 실험하였으며 결과는 다음과 같다. 약용식물 및 식용작물에 대한 80% EtOH 추출물을 이용하여 항종양효과를 보면 PKC법에서는 명아주(73.4%) 및 antibleb형성억제력검정에서는 검정콩이, PLC법에서는 검정콩(91.9%), MTT법에서 50%의 억제력을 나타내는 농도$(IC_{50})$가 검정콩과 쑥의 경우 각각 $4.7{\mu}g/ml$을 보였다.

• KSBB Journal
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• 제29권6호
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• pp.432-436
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• 2014

We investigated antioxidative activity of the ethanol extracts of leaves of Cornus walteri Wanger (CWE) by treated enzyme in human dermal fibroblast (HDFs) irradiated by UVB. We examined the in vitro chemical and cellular antioxidant activities of CWE in HDFs. We employed scavenging assay for the 1,1-diphenyl-2,5-picrylhydrazyl (DPPH) radicals and cellular antioxidative activity of CWE, and we was investigated in $H_2O_2$-treated or UVB-irradiated HDFs. The CWE effectively scavenged DPPH radicals ($IC_{50}$ $7.03{\pm}0.4{\mu}g/mL$) when compared to the scavenging activities of L-ascorbic acid ($IC_{50}$ $4.69{\pm}0.3{\mu}g/mL$). CWE reduced UVB-induced cellular damage in HS68 cells by MTT assay and inhibited intracellular ROS generation in dose-dependent manner. In addition, CWE also attenuated the elevated levels of 8-isoprostane resulting from UVB-mediated oxidative stress. Collectively, these results suggest that CWE could be a new potential candidate as antioxidant against UVB-induced oxidative stress in HDFs.

• 한국작물학회지
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• 제47권4호
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• pp.328-334
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• 2002

A Lindernia dubia (L.) Pennell var. dubia accession from Jeonnam province, Korea was tested for resistance to sulfonylurea (SU) herbicides, imazosulfuron and pyrazosulfuron-ethyl in whole-plant response bioassay. The accession was confirmed resistant to both herbicides. The $GR_{50}$ (herbicide concentration that reduced shoot dry weight by 50%) values of resistant accession were 264 and 19 times higher to imazosulfuron and pyrazosulfuronethyl, respectively, than that of the standard susceptible accession. The surviving resistant L. dubia after pyrazosulfuron-ethyl + molinate application can be controlled by sequential applications of soil-applied herbicides, butachlor, dithiopyr, pyrazolate, and thiobencarb and foliar herbicides, bentazon. Sulfonylurea-based mixtures such as mixtures of azimsulfuron + anilofos, bensulfuron-methyl + oxadiazon, pyrazosulfuron-ethyl + fentrazamide, and pyrazosulfuron-ethyl + anilofos + carfentrazon can also be used to control the surviving resistant L. dubia. However, use of these mixtures should be restricted to a special need basis. Thus, we suggest that sequential applications of non-SU-based mixtures such as butachlor + pyrazolate and MCPB + molinate + simetryne be used to control the surviving resistant L. dubia after SU herbicide applications. Rice yield was reduced 24 % by resistant L. dubia that survived after the pyrazosulfuron-ethyl + molinate application compared with pyrazolate + butachlor in transplanted rice culture. In vitro ALS activity of the resistant biotype was 40 and 30 times more resistant to imazosulfuron and pyrazosulfuron-ethyl, respectively, than the susceptible biotype. Result of in vitro ALS assay that the resistance mechanism of L. dubia to SU herbicides may be due, in part, to an alteration in the target enzyme, ALS.

• Journal of Microbiology and Biotechnology
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• 제21권1호
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• pp.62-70
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• 2011

The antibacterial effects of nine lactic acid bacteria (LAB) against Campylobacter jejuni were investigated by using agar gel diffusion and co-culture assays. Some differences were recorded between the inhibition effects measured with these two methods. Only two LAB, Lb. pentosus CWBI B78 and E. faecium THT, exhibited a clear anti- Campylobacter activity in co-culture assay with dehydrated poultry excreta mixed with ground straw (DPE/GS) as the only growth substrate source. It was observed that the supplementation of such medium with a cellulase A complex (Beldem S.A.) enhanced the antimicrobial effect of both LAB strains. The co-culture medium acidification and the C. jejuni were positively correlated with the cellulase A concentration. The antibacterial effect was characterized by the lactic acid production from the homofermentative E. faecium THT and the lactic and acetic acids production from the heterofermentative Lb. pentosus CWBI B78. The antagonistic properties of LAB strains and enzyme combination could be used in strategies aiming at the reduction of Campylobacter prevalence in the poultry production chain and consequently the risk of human infection.