• Archives of Pharmacal Research
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• 제29권8호
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• pp.624-626
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• 2006

Four known butenolides were isolated from the ethyl acetate extracts of the culture broth of the marine-derived bacterium, Streptoverticillium luteoverticillatum, by bioassay-guided fractionation. The structures were identified on the basis of spectral data. The absolute configuration of compound (1) was determined by CD spectrum for the first time. Compounds 1-4 showed in vitro cytotoxicity against the murine lymphoma P388 and human leukemia K562 cell lines. This is the first report on the isolation of butenolides from the marine bacterium, Streptoverticillium luteoverticillatum, and their cytotoxic activities.

• 한국식품영양과학회지
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• 제36권7호
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• pp.926-931
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• 2007

LC-MS/MS법에 의하여 설사성 패류독소 7종(okadaic acid, dinophysistoxin-1, -3, pectebotoxin-1, -2, -6, yessotoxin)의 동시 분석 조건을 확립하였고, 2006년 3월부터 9월까지 매월 5종의 이매패(굴, 진주담치, 반지락, 가리비, 개조개, 6월부터 9월까지는 가리비 대신 피조개)를 정기적으로 통영시장에서 구입하여 분석하였으며, 기존의 마우스 검사법과 비교하였다. 마우스 검사법으로는 총 35종 시료 중 3월의 굴과 진주담치에서 0.05${\sim}$0.1 MU/g의 독성을 나타내었으나, 이미 알려진 설사성 패류독소는 전혀 검출되지 않아 다른 성분에 의한 것으로 판단되었다. 한편, LC-MS/MS법에 의하여서는 6월 진주담치에서 dinophysistoxin-1(DTX1)이 0.05 ${\mu}g/g$ 검출되었으며, 8월의 모든 시료에서 okadaic acid(OA)가 0.01${\sim}$0.02 ${\mu}g/g$ 검출되었다. 이들은 외국의 기준치[0.05 MU/g(일본), OAs(OA+DTX1) 0.16 ${\mu}g/g$(EU)]보다 매우 낮은 함량으로 식품 위생상 안전하였다.

• 한국해양학회지:바다
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• 제13권2호
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• pp.129-139
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• 2008

해양생태독성평가를 위한 표준시험방법 개발을 위하여 해양생태계의 1차 소비자인 윤충류 및 저서 요각류를 이용한 시험방법을 정립하였다. 표준시험생물로 윤충류 Brachionus plicatilis와 저서 요각류 Tigriopus japonicus를 선정하였으며, endpoint는 윤충류 신생개체의 사망률(24 hr $LC_{50}$) 및 개체군성장률(48 hr $EC_{50}$)과 저서 요각류 유생($100{\sim}200\;{\mu}m$)의 사망률(48 hr $LC_{50}$)로 설정하였다. 표준시험방법은 미국재료시험협회 (ASTM)의 독성시험법을 참고하였으며, 표준시험생물은 국내 해양생태계의 특성 및 종의 유용성 등을 고려하여 재설정하였다. 윤충류 B. plicatilis와 저서 요각류 T. japonicus는 광염성으로 $5{\sim}35\;psu$ 구간에서 실험이 가능하고, 또한 배양이 용이한 점이 고려되었다. 상기 두 종을 이용한 독성시험은 시험생물 확보 및 배양이 쉽고 시험기간이 짧고 간단한 장점이 있는 반면 독성시험의 민감도가 무척추동물의 유생을 이용한 시험법보다 떨어지는 단점이 있다. 두 종 모두 해양유입 유해물질을 이용한 독성실험결과, 농도-반응의 선형관계가 뚜렷하였다. 카드뮴을 이용한 윤충류 개체군성장에 대한 실험실간 교차분석결과 $EC_{50}$이 각각 39.3 mg/L와 33.7 mg/L로 유사한 값을 보였다. 따라서 윤충류 및 저서 요각류는 독성시험생물로서 유용한 것으로 판단되며, 위의 두 종중 최소 1종은 해양생태 독성실험의 "battery test"에 포함할 것을 권장한다.

• 한국수산과학회지
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• 제29권6호
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• pp.856-861
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• 1996

The occurrence of TTX in ciliated protozoa was investigated in order to clarify tetrodotoxin (TTX) accumulation mechanisms in marine organisms. Tissue culture bioassay, HPLC, and GC-MS analyses confirmed the occurrence of TTX in Euplotes mutabilis and also in bacteria isolated from the culture medium. Fluorescently labeled bacteria (FLB) were prepared with those bacteria, and predation by E. mutabilis was observed. The results indicated that TTX in bacteria can be transferred to higher trophic levels through the food chain.

• 한국해양바이오학회지
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• 제1권4호
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• pp.260-267
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• 2006

재조합 Agrobacterium tumefaciens균주를 사용한 Liquid Culture Assay는 quorum sensing autoinducer를 검색하는 생물학적 분석 방법으로 개발되었다. 그러나 이 시스템에 사용되는 해양천연물 시료들이 일반적으로 낮은 수용액에 대한 용해도를 갖기 때문에 활성검색에 걸림돌이 되고 있다. 시료의 용해도는 유기용매 혹은 계면활성제의 첨가로 증가될 수 있으나, 유기용매나 계면활성제 자체가 검색결과의 정확한 해석을 방해할 수 있으므로 적절한 조건의 확립이 매우 중요하다. Methanol, ethanol, 1-propanol, DMSO와 DMF를 0~10% 농도 범위에서 재조합 A. tumefaciens균주에 대한 세포 성장에 미치는 영향과 ${\beta}$-galactosidase activity에 미치는 영향을 살펴본 결과 methanol 2%이하의 농도가 가장 적합하였으며, 계면활성제의 경우, Tween 20, Tween 80보다는 Triton X-100이 약 0.05% 농도에서 세포내로의 활성물질의 전달효율을 높이는데 효과적이었다.

• 한국수산과학회지
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• 제36권4호
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• pp.346-351
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• 2003

Single-cell gel electrophoresis (comet assay) was used to detect DNA single strand break in blood cells from several marine fish species. Three fish species were collected from Georgia coastal area. Mummichog, Fundulus heteroclitus showed higher DNA damage than sea bass, Lateolabrax japonicus and trout, Oncorhynchus masou masou under the same experimental conditions. Mummichogs had more alkaline-labile sites on their DNA than other fish species. The comet assay with mummichog blood cells at pH 12.5 showed a dose-response curve with the increasing concentrations of hydrogen peroxide. While the isolated leucocytes showed no increase of DNA damage after in vitro exposure to 2-methyl-1,4-naphthoquinone (MNQ), erythrocytes showed dose-dependent DNA damage. These results indicate that the comet assay can be applied successfully as a bioassay using erythrocyte for environmental monitoring.

• Journal of Microbiology and Biotechnology
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• 제16권4호
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• pp.637-638
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• 2006

Bioassay-guided fractionation of an organic extract of the culture broth from an unidentified marine-derived fungus led to the isolation of a new metabolite, N-[2-(4-hydroxyphenyl) acetyl]formamide (1), along with four known polyketides, 4-hydroxyphenyl acetamide (2), 4-hydroxyphenyl acetic acid (3), 3,4-dihydroxyphenyl acetic acid (4), and N-[2-(4-hydroxyphenyl)ethenyl]formamide (5). The structures of 1-5 were elucidated by spectral data analyses. Among them, compounds 1, 4, and 5 exhibited significant radical scavenging activity against 1, 1-diphenyl-2-picrylhydrazyl (DPPH) with $IC_{50}$ values of 8.4, 11.9, and $0.2{\mu}M$, respectively.

• Environmental Analysis Health and Toxicology
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• 제20권4호통권51호
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• pp.333-341
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• 2005

In order to know whether polychaetes could be used as an appropriate organism for the detection of genotoxicity, DNA strand breaks were evaluated in blood cells of a nereidae worm (Perinereis aibuhitensis) exposed to various aquatic chemical pollutants (e.g. Cd, Pb, Pyrene, Benaor[a]pyrene). Hydrogen peroxide increased DNA strand breaks up to the highest concentration (10 $\mu$M). Higher concentration than 0.1 $\mu$M showed a significantly more DNA damage than control. Cadmium and lead also showed higher DNA damage than control, over 1.0 and 1 $\mu$g/L, respectively. In case of pyrene, DNA damage was detected even at 0.001 $\mu$g/L. However, DNA damage decreased due to apoptosis at the highest concentration of pyrene and Pb. This study suggested that the polythaetous blood cells could be used effectively for screening genotoxic contaminants in the environment.

• Ocean and Polar Research
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• 제33권3호
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• pp.255-263
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• 2011

The aim of this investigation was to evaluate antioxidant activity of crude extracts from the halophyte Vitex rotundifolia, their solvent fractions, and isolated compounds (1-3). Antioxidant capacity was determined by measuring DPPH radical, and authentic $ONOO^-$ and $ONOO^-$ generated from 3- morpholinsydnonimine (SIN-1) in vitro as well as degree of occurrence of intracellular ROS, NO and GSH in mouse macrophage Raw 264.7 cells. From comparative analysis, MeOH extract, n-BuOH, and 85% aq. MeOH solvent fractions showed significant antioxidant effect in DPPH radical and $ONOO^-$ assay systems. Activity-guided purification of n-BuOH and 85% aq. MeOH fractions led to the isolation of flavonoids 1-3. Among them, compound 1 exhibited excellent antioxidant effect in all bioassay systems tested. On the other hand, compounds 2 and 3 revealed potent inhibitory effect against $ONOO^-$ generated from SIN-1, comparable with the positive control penicillamine.

• Biomolecules & Therapeutics
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• 제32권4호
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• pp.492-498
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• 2024

Bioassay and HPLC-UV guided fractionations of the crude extract of marine-derived Streptomyces sp. SNA-077 have led to the isolation of a red pigment, undecylprodigiosin (1). The chemical structure of undecylprodigiosin (1) was revealed by the interpretation of NMR and mass spectroscopic (MS) data. Further, anti-melanogenic effects of undecylprodigiosin (1) were investigated. First, the melanin contents of undecylprodigiosin (1)-treated B16 cells were evaluated. Furthermore, undecylprodigiosin (1) significantly inhibited the key enzymes involved in melanogenesis, including tyrosinase, tyrosinase related protein-1 (TYRP-1), and dopachrome tautomerase (DCT). The mRNA and protein expression levels of Microphthalmia-associated transcriptian factor (MiTF), a critical transcription factor for tyrosinase gene expression, were also suppressed by undecylprodigiosin (1) treatment in B16 analyses. Collectively, our results suggest for the first time that undecylprodigiosin (1), a potent component isolated from an extract of marine Streptomyces sp. SNA-077, critically exerts the anti-melanogenic ability for melanin synthesis.