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Japanese Medaka, Oryzias latipes as a Test Animal for Marine Ecotoxicological Evaluation (해양생태독성평가를 위한 표준시험생물로서의 송사리(Oryzias latipes)에 관한 연구)

  • Park Gyung Soo;Yoon Seong Jin;Lee Seung Min;Kim Ae Hyang;Park Soung Yun;Kang Duk Young
    • Korean Journal of Environmental Biology
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    • v.23 no.3 s.59
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    • pp.293-303
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    • 2005
  • Japanese medaka, Oryzias latipes is widely distributed in the North East Asia including Korea, Japan and east China, and commonly used for freshwater toxicity tests and cytotoxicological studies worldwide. In this study, a series of experiments were conducted to identify the potential of the fish as a standard test species for saltwater toxicity evaluation such as marine receiving waters, ocean-dumped materials and sediment pore waters etc. Hatching, growth and mortality rates of the fish were estimated with the wide ranges of salinity from freshwater to seawater (35 psu). Direct exposure of the fertilized eggs in freshwater to the wide ranges of salinity (from 0 to 35 psu) without pre- acclimation to the saltwater revealed no significant differences in hatching rates by salinities (p =0.24). On the other hand, medaka larvae hatched in freshwater and exposed to saltwater directly showed high mortality at > 25 psu treatment groups (p < 0.0001). However, there was no significant difference in mortality of medaka larvae hatched in 13.8 and 14.2 psu at the wide ranges of salinities ($0\~35$ psu). Growth rates of medaka larvae hatched in the above two salinities showed no differences in body length either from 0 to 35 psu treatment groups (p =0.64 for 13.8 psu group and p=0.32 for 14.2 psu group). The number of gill chloride cell in medaka larvae sharply increased when the larvae were exposed to high salinity. Reference tests with zinc chloride revealed 96h $LC_{50}=8.84(7.19\~10.87)mg\;L^{-1}$ using 7~10 day old medaka larvae. These were comparable or better sensitivity in comparison with the other standard test species such as North American sheepshead minnow Cyprinodon variegatus. Based on the results of these experiments, hatching rates and larvalmortality of medaka must be good toxicity parameters for seawater bioassay and the species seems to be a good standard species for both the freshwater and seawater toxicity test.

Mechanism Underlying a Proteasome Inhibitor, Lactacystin-Induced Apoptosis on SCC25 Human Tongue Squamous Cell Carcinoma Cells (사람혀편평상피세포암종세포에서 proteasome 억제제인 lactacystin에 의해 유도된 세포자멸사의 기전에 대한 연구)

  • Baek, Chul-Jung;Kim, Gyoo-Cheon;Kim, In-Ryoung;Lee, Seung-Eun;Kwak, Hyun-Ho;Park, Bong-Soo;Tae, Il-Ho;Ko, Myung-Yun;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.34 no.3
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    • pp.261-276
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    • 2009
  • Lactacystin, a microbial natural product synthesized by Streptomyces, has been commonly used as a selective proteasome inhibitor in many studies. Proteasome inhibitors is known to be preventing the proliferation of cancer cells in vivo as well as in vitro. Furthermore, proteasome inhibitors, as single or combined with other anticancer agents, are suggested as a new class of potential anticancer agents. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in SCC25 human tongue sqaumous cell carcinoma cell line treated with lactacystin. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingiva fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay respectively. The hoechst staining, hemacolor staining and TUNEL staining were conducted to observe SCC25 cells undergoing apoptosis. SCC25 cells were treated with lactacystin, and Western blotting, immunocytochemistry, confocal microscopy, FAScan flow cytometry, MMP activity, and proteasome activity were performed. Lactacystin treatment of SCC25 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Interestingly, lactacytin remarkably revealed cytotoxicity in SCC25 cells but not normal cells. And tested SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, the up-regulation of Bax, and the activation of caspase-7, caspase-3, PARP, lamin A/C and DFF45 (ICAD). Flow cytometric analysis revealed that lactacystin resulted in G1 arrest in cell cycle progression which was associated with up-regulation in the protein expression of CDK inhibitors, $p21^{WAF1/CIP1}$ and $p27^{KIP1}$. We presented data indicating that lactacystin induces G1 cell cycle arrest and apoptois via proteasome, mitochondria and caspase pathway in SCC25 cells. Therefore our data provide the possibility that lactacystin could be as a novel therapeutic strategy for human tongue squamous cell carcinoma.

Phytochemical compounds and quality characteristics of spray-dried powders with the blanching condition and selected forming agents from pressed extracts of Ligularia fischeri leaves (블랜칭 처리 및 부형제 종류에 따른 곰취 착즙액 분무건조 분말의 phytochemical 성분 및 품질특성)

  • Kim, Jae-Won;Park, In-Kyung;Youn, Kwang-Sup
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.659-667
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    • 2013
  • This study was performed to determine the effects of the blanching condition (immersion ratio 1:15 (w/v) for 3 min at $95^{\circ}C$, and solution containing 1% sodium chloride) and selected forming agents (dextrin DE=10, dextrin DE=20, ${\beta}$-cyclodextrin; each forming agents added 5%) on the phytochemical compounds and quality characteristics of Ligularia fischeri leaves. The moisture was not affected by the forming agent. The color of a, b and chroma values were low in the blanching treatment groups and were significantly lowest with ${\beta}$-cyclodextrin (CD). The polyphenol and flavonoid contents in the blanching treatment groups were higher than those in the non-blanching-treatment group. The ascorbic acid content was higher in the non-blanching-treatment group and was significantly highest in the group treated with dextrin (DE=10) whereas the blanching treatment groups showed lower dehydroascorbic acid content than the non-blanching-treatment group. The water absorption was higher in the non-blanching-treatment group and was significantly highest in the group treated with CD. The water solubility in the blanching treatment groups treated with dextrin (DE=20) and CD was higher than that in the blanching treatment group treated with DE=10. The total chlorophyll and chlorophyll a and b contents were high in the blanching treatment group treated with CD, and for the total carotenoid contents, the same tendency as that seen with the chlorophyll content was observed. With regard to the particle diameter, those in the blanching treatment groups were lower than that in the non-blanching-treatment group and was significantly lowest in the blanching treatment groups treated with DE=20 and CD. The result of SEM observation showed that the spray-dried powders in blanching treatment groups treated with the DE=20 and CD forming agents had uniform particle distribution.

Studies on the Tissue Culture of Some useful woody species (유용수종(有用樹種)의 조직배양(組織培養)에 관(關)한 연구(硏究))

  • Kim, Jai Saing;Lee, Sam Sup
    • Journal of Korean Society of Forest Science
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    • v.43 no.1
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    • pp.6-13
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    • 1979
  • In order to substract the time and cost of propagation for inducing the haploid plants per each species. 500 anthers of late uninucleate microspore on early binucleate microspore stage of Robinia pseudoacacia (Fuel tree) Punius granatum (Ornamental tree). Aleurites fordii (Faty tree) and Styrax japonica (Silvicultural tree) were cultured on the modified Murashige and Skoog's medium supplemented with Kinetin, 2.4-D and NAA as growth regulators. And I observed the samples of cultured anthers under the microscope which were made by Microtoming method and Paraffin method. The results were summarized as follows: 1) Among 500 cultured anthers per each species, anther numbers inducing the diploid callus were as follow: Styrax japonica 20 (4% for the species total); Aleurites fordii 10 (2% for the species, total) and Punica granatum 45 (9% for the species total) were showed. 2) 2n Callus were induced from anther wall. but haploid callus were induced from anther locule. 3) Haploid callus were induced only in 25 anthers (5% for the species total) of Robinia pseudoacacia. 4) These haploid callus were not originated from body cell of anther wall tissue, but from reduced microspores, 5) Since already reported many herbaceous haploid plants were induced from the callus which were originated from reduced microspores, I conclude that the anther of woody plant which induced the haploid callus also will be cultured haploid plant.

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Several causes of non virus-induced mosaic symptom on potato leaves and its induction by herbicides (감자 이상모자이크증상의 몇 가지 발생원인 및 제초제에 의한 증상 유기)

  • Kwon, Min;Hahm, Young-Il;Kim, Hyun-Jun;Yiem, Myoung-Soon
    • The Korean Journal of Pesticide Science
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    • v.5 no.2
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    • pp.45-50
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    • 2001
  • In recent, non virus-induced mosaic symptoms(NVMS) on potato leaves were observed in the seed potato fields, and its incidence rate was $5{\sim}20%$ nationwide. It made difficult to rogue out virus-infected plants, and caused much arguments between seed potato production farmers and seed potato inspectors. The objectives of these experiments were to find out the causes of NVMS, and also to induce mosaic symptom(phytotoxicity) on potato plants by treatment of several herbicides. No significant correlations were found between incidence rates of NVMS and values from soil analyses; soil pH, soil EC, organic matter content, and contents of inorganic constituents($P_2O_5,\;NO_3$, Ca, Mg, K) in the soil around the potato planted. The examinations by ELISA, virus indicator plants, and TEM showed that NVMS on potato leaves was not caused by the viruses infection. But, the use of herbicides could induced the NVMS on potato leaves. The incidence rates of potato treated with pendimethalin linuron of 400 mL/10 a, pendimethalin of 200 mL/10 a, pendimethalin.oxadiazon of 300 mL/10 a, and control were 61.1%, 47.2%, 19.4%, and 1.4%, respectively. Based on these results, we confirmed that the treatment of pendimethalin alone and in mixture with other herbicides were the reason of NVMS on potato leaves. The yields among test plots were similar except dicamba treated plot, which decreased by about 23% compared to control plot. When their progenies harvested in 1999 were planted in the following season, no symptoms of mosaic were observed.

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