• Title/Summary/Keyword: Growth inhibition test

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Marine Ecotoxicological Evaluation on HNS Spill Accident : Nitric Acid Spill Case Study (HNS 유출사고가 해양생물에 미치는 생물독성 영향평가 : HNO3 유출사고 대상)

  • Kim, Tae-Won;Kim, Young Ryun;Jo, So Eun;Son, Min Ho;Lee, Moonjin;Oh, Sangwoo
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.21 no.6
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    • pp.655-661
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    • 2015
  • This study intends to evaluate the effect of nitric acid($HNO_3$) spill accidents on the marine ecosystem, while $HNO_3$ is known as one of the typical HNS. For this purpose, we performed (1) the growth inhibition test by using phytoplankton(Skeletonema costatum), (2) acute and chronic toxicity test by using invertebrate(Brachionus plicatilis and Monocorphium acherusicum), (3) fish(Cyprinodon variegatus) and (4) bacteria(Vibrio fischeri). In these tests, we observed the (1) pH changes induced by the nitric acid spill and (2) changes in nitrate($NO_3$) concentration disassociated from nitric acid after the accident, respectively. The toxicity test result on pH changes induced by $HNO_3$ shows that the no observed effect concentration(NOEC), lowest observed effect concentration(LOEC) and 50 % effect concentration($72h-EC_{50}$) values of M. acherusicum are pH 7 (0.3 mM), pH 5(1.1 mM) and pH 5.2(1.4 mM), respectively, indicating that M. acherusicum is the most sensitive species. The chronic toxicity test (population growth rate test) on $NO_3{^-}$ of B. plicatilis show that the NOEC, LOEC and $96h-EC_{50}$ are 5.9 mM, 11.8 mM and 32.6 mM, respectively, indicating that B. plicatilis is the most sensitive species. In conclusion, toxic effecst on the marine organism caused by the nitric acid spill accident is determined to be so slightly except for the most adjacent area of the ship in pH scale and such concentration of nitrate, to the extent of directly influencing the survival and reproduction of the marine organism, is determined practically not to be applicable in the typical accidents in the sea.

BCR/ABL mRNA Targeting Small Interfering RNA Effects on Proliferation and Apoptosis in Chronic Myeloid Leukemia

  • Zhu, Xi-Shan;Lin, Zi-Ying;Du, Jing;Cao, Guang-Xin;Liu, Gang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.4773-4780
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    • 2014
  • Background: To investigate the effects of small interference RNA (siRNA) targeting BCR/ABL mRNA on proliferation and apoptosis in the K562 human chronic myeloid leukemia (CML) cell line and to provide a theoretical rationale and experimental evidence for its potential clinical application for anti-CML treatment. Materials and Methods: The gene sequence for BCR/ABL mRNA was found from the GeneBank. The target gene site on the BCR/ABL mRNA were selected according to Max-Planck-Institute (MPI) and rational siRNA design rules, the secondary structure of the candidate targeted mRNA was predicted, the relevant thermodynamic parameters were analyzed, and the targeted gene sequences were compared with BLAST to eliminate any sequences with significant homology. Inhibition of proliferation was evaluated by MTT assay and colony-formation inhibiting test. Apoptosis was determined by flow cytometry (FCM) and the morphology of apoptotic cells was identified by Giemsa-Wright staining. Western blotting was used to analyze the expression of BCR/ABL fusion protein in K562 cells after siRNA treatment. Results: The mRNA local secondary structure calculated by RNA structure software, and the optimal design of specific siRNA were contributed by bioinformatics rules. Five sequences of BCR/ABL siRNAs were designed and synthesized in vitro. Three sequences, siRNA1384, siRNA1276 and siRNA1786, which showed the most effective inhibition of K562 cell growth, were identified among the five candidate siRNAs, with a cell proliferative inhibitory rate nearly 50% after exposure to 12.5nmol/L~50nmol/L siRNA1384 for 24,48 and 72 hours. The 50% inhibitory concentrations ($IC_{50}$) of siRNA1384, siRNA1276 and siRNA1786 for 24hours were 46.6 nmol/L, 59.3 nmol/L and 62.6 nmol/L, respectively, and 65.668 nmol/L, 76.6 nmol/L, 74.4 nmol/L for 72 hours. The colony-formation inhibiting test also indicated that, compared with control, cell growth of siRNA treated group was inhibited. FCM results showed that the rate of cell apoptosis increased 24 hours after transfecting siRNA. The results of annexinV/PI staining indicated that the rate of apoptosis imcreased (1.53%, 15.3%, 64.5%, 57.5% and 21.5%) following treamtne with siRNAs (siRNA34, siRNA372, siRNA1384, siRNA1276 and siRNA1786). Morphological analysis showed td typical morphologic changes of apoptosis such as shrunken, fragmentation nucleus as well as "apoptotic bodies" after K562 cell exposure to siRNA. Western blot analysis showed that BCR/ABL protein was reduced sharply after a single dose of 50nmol/L siRNA transfection. Conclusions: Proliferation of K562 cells was remarkbly inhibited by siRNAs (siRNA1384, siRNA1276 and siRNA1786) in a concentration-dependent manner in vitro, with effective induction of apoptosis at a concentration of 50 nmol/L. One anti-leukemia mechanism in K562 cells appeared that BCR/ABL targeted protein was highly down-regulated. The siRNAs (siRNA1384, siRNA1276 and siRNA1786) may prove valuable in the treatment of CML.

Acute Toxicity Studies of Octachlorostyrenes for Environment Organisms (Octachlorostyrenes의 환경생물에 대한 급성독성 연구)

  • Kim, Yong-Seok;Jeon, Yong-Bae;Lee, Dae-Yong;Lee, Jue-Hwan;Lee, Kyu-Seung;Sung, Ha-Jung
    • The Korean Journal of Pesticide Science
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    • v.13 no.2
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    • pp.87-97
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    • 2009
  • Octachlorostyrene (OCS) is a persistant and bioaccumulative toxic subtance (PBTs). In this study, acute toxicity tests on algae, daphnia and fish for octachlorostyrene and its isomers were done to determine effective concentration ($EC_{50}$), Lethal concentration ($LC_{50}$), no observed effect concentration (NOEC) or lowest observed effect concentration (LOEC). As a result, NOEC on algae growth inhibition test for octacholorostyrene and 2-, 3-chlorostyrene was determined as $0.50\;mg\;L^{-1}$, and NOEC for 4-chlorostyrene was determined as $0.13\;mg\;L^{-1}$. NOEC on daphnia, acute immobilisation test for octachlorostyrene and 2-, 3-chlorostyrene was determined as $5.00\;mg\;L^{-1}$ and $EC_{50}$ for 4-chlorostyrene was determined as $2.128\;mg\;L^{-1}$. NOEC on Oryzias Latipes, acute toxicity test for octachlorostyrene was determined as $80.0\;mg\;L^{-1}$ and NOEC for 2-, 3-chlorostyrene was determined as $60.0\;mg\;L^{-1}$. $LC_{50}$ for 4-chlorostyrene was determined as $39.0\;mg\;L^{-1}$ (48h) and $22.6\;mg\;L^{-1}$ (96h).

Antimutagenic and Cytotoxicity Effects of Extracts of Eleutherococcus senticosus Maxim fruits (가시오갈피 열매 추출물의 항돌연변이원성 및 세포독성 효과)

  • 전윤영;최승필;이효진;문선영;이득식;함승시
    • Food Science and Preservation
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    • v.10 no.3
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    • pp.394-400
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    • 2003
  • This study was earned out to investigate the antimutagenic and cytotoxic effects of Eleutherococcus senticosus Maxim fruits ethanol extract on Salmonella typhimurium TA98, TA100 and cancer cell lines using Ames test and SRB assay, respectively. They were extracted with ethanol and then fractionated with hexane, chloroform, ethyl acetate, butanol and water to get active fractions. In the Ames test, most of the extracts had strong antimutagenic effects against the mutagenesis induced by MNNG, 4NQO, B(${\alpha}$)P and Trp-P-l. The ethanol extract (200 $\mu\textrm{g}$/plate) of Eleutherococcus senticosus fruits showed 87.2% inhibitory effect on the mutagenesis induced by MNNG against TA100. And also, The suppression ratio against B(${\alpha}$)P and Trp-P-l in the TA100 showed 96.1% and 95.5%, respectively. In the cytotoxic effects against human cancer cell lines (A549, AGS, MCF-7, Hep3B), the value of inhibition were mostly above 60% for each fraction (1 mg/mL). Hexane fraction (1 mg/mL) against showed the strongest cytotoxic effects of 92.7% compared to those of other fraction and butano fraetion against Hep3B was relatively high growth inhibitory effect of 82%.

Some Physiological Activity of Phenolic Substances in Plant Foods (식물성 식품중 페놀성 물질의 몇가지 생리활성)

  • Lee, Jung-Hi;Lee, Su-Rae
    • Korean Journal of Food Science and Technology
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    • v.26 no.3
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    • pp.317-323
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    • 1994
  • Nine plant foods (persimmon leaf, perilla seed, Chinese quince, ginger root, walnut, mugwort leaf, arrowroot, buckwheat and sorghum) rich in phenolic substances were examined for their effects on the digestive enzymes, food-poisoning bacteria and mutagenicity/antimutagenicity by Ames test. Among tested samples, Chinese quince significantly inhibited the $\alpha-amylase$ activity (97%), exhibiting an uncompetitive inhibition type. Protease activity was inhibited by Chinese quince (86%), persimmon leaf (51%) and mugwort leaf (20%), in which mugwort extract exhibited a noncompetitive type. Lipase was activated >50% by all samples. The inhibition of $\alpha-amylase$ was highly correlated with the content of condensed tannin (r=0.89) and the inhibition of protease, with total phenolic content (r=0.84). Total phenolies fraction of tested samples showed the growth inhibition toward E. coli. Streptococcus faecalis and Salmonella enteritidis, in which the effect of perilla, sorghum and arrowroot was the highest for E. coli. Standard phenolics and food samples did not show any mutagenicity toward Salmonella typhimurium TA98 and TA100. Tannic acid inhibited the mutation of the two strains by benzo[a]pyrene whereas total phenolics fractions of Chinese quince and walnut exhibited antimutagenicity to a lesser extent.

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A Comparison of Sensitivity of Four Freshwater Algae to Five Pesticides (5종 농약에 대한 4종 담수조류(freshwater algae)의 감수성 비교)

  • Park, Yeon-Ki;Kim, Byung-Seok;Bae, Chul-Han;Kim, Yeon-Sik;Park, Kyung-Hoon;Lee, Jea-Bong;Shin, Jin-Sup;Hong, Soon-Sung;Lee, Kyu-Seung;Lee, Jung-Joon
    • The Korean Journal of Pesticide Science
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    • v.12 no.1
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    • pp.50-56
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    • 2008
  • The purpose of the study was to determine the effects of isoprothiolane, diazinon, butachlor, dimethametryn and molinate in Selenastrum capriconurtum, Scenedesmus subspicatus, Chlorella vufgaris and Nitzschia palea during an exposure period of 72 hours. The study was carried out in according with the OECD Guidelines for Testing of Chemicals No 201 Alga, Growth Inhibition Test. The toxicological responses of isoprothiolane to S. capriconutum, S. subspicatus, C. vulgaris and N. palea expressed as in individual $ErC_{50}$ (Median Effective Concentration by growth rate) value, were 5.87, 9.91, $18.55 and 38.79\;mg\;L^{-1}$, respectively. Diazinone to S. capriconutum, S. subspicatus, C. vulgaris and palea expressed as in individual $ErC_{50}$ value, were 10.31, 11.44, >32 and $14.32\;mg\;L^{-1}$, respectively. Butachlor to S. capriconutum, S. subspicatus, C. vulgaris and N. plaea expressed as in individual $ErC_{50}$ value, were 0.002, 0.019, 8.67 and $4.94\;mg\;L^{-1}$, respectively. Dimethamelryn to S. subspicatus, C. vulgaris and N. palea expressed as in individual $ErC_{50}$ value, were 0.0071, 0.011, 0.0065 and $0.009\;mg\;L^{-1}$, respectively. Molinate to S. capriconutum, S. subspicatus, C. vulgaris and N. palea expressed as in individual $ErC_{50}$ value, were 0.44, 1.26, 48.84 and $28.52\;mg\;L^{-1}$, respectively. The sensitivities of five pesticides were different depending on freshwater alga in the order of S. capriconutum > S. subspicatus > C. vulgaris, N. palea. Highly significant correlation (r=0.9677) based on $ErC_{50}$ were found between S. capriconutum and S. subspicatus.

An Improved Method to Determine Corn (Zea mays L.) Plant Response to Glyphosate (Glyphosate에 대한 옥수수 반응의 개선된 검정방법)

  • Kim, Jin-Seog;Lee, Byung-Hoi;Kim, So-Hee;Min, Suk-Ki;Choi, Jung-Sup
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.57-62
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    • 2006
  • Several methods for determining the response of corn to glyphosate were investigated to provide a fast and reliable method for identifying glyphosate-resistant corn in vivo. Two bioassays were developed. One assay is named 'whole plant / leaf growth assay', in which the herbicide glyphosate is applied on the upper part of 3rd leaf and the growth of herbicide-untreated 4th leaf is measured at 3 day after treatment. in this assay, the leaf growth of conventional corn was inhibited in a dose dependent from 50 to $1600{\mu}g/mL$ of glyphosate and growth inhibition at $1600{\mu}g/mL$ was 55% of untreated control. The assay has the potential to be used especially in the case that the primary cause of glyphosate resistance is related with a reduction of the herbicide translocation. Another assay is named 'leaf segment / shikimate accumulation assay', in which the four excised leaf segments ($4{\times}4mm$) are placed in each well of a 48-well microtiter plate containing $200{\mu}L$ test solution and the amount of shikimate is determined after incubation for 24 h in continuous light at $25^{\circ}C$. In this assay, 0.33% sucrose added to basic test solution enhanced a shikimate accumulation by 3 to 4 times and the shikimate accumulation was linearly occurred from 2 to $8{\mu}g/mL$ of glyphosate, showing an improved response to the method described by Shaner et al. (2005). The leaf segment / shikimate accumulation assay is simple and robust and has the potential to be used as a high throughput assay in the case that the primary cause of glyphosate resistance is related with EPSPS, target site of the herbicide. Taken together, these two assays would be highly useful to initially select the lines obtained after transformation, to investigate the migration of glyphosate-resistant gene into other weeds and to detect a weedy glyphosate-resistant corn in field.

Effects of Soil Amendments on the Early Growth and Heavy Metal Accumulation of Brassica campestris ssp. Chinensis Jusl. in Heavy Metal-contaminated Soil (중금속 오염 토양에서 안정화제가 청경채의 초기 생육과 중금속 흡수량에 미치는 영향)

  • Kim, Min-Suk;Koo, Namin;Kim, Jeong-Gyu;Yang, Jae-E.;Lee, Jin-Su;Bak, Gwan-In
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.6
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    • pp.961-967
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    • 2012
  • There have been many studies about efficiency of amendments for heavy metal stabilization through chemical assessment. The objective of this study was to evaluate the efficiency of several soil amendments (lime, agric-lime, dolomite, steel slag, fly ash and acid mine drainage sludge) on heavy metals stabilization through not only chemical but also biological assessments (phytotoxicity test) in abandoned mining area soil. In order to achieve the goal, we conducted preliminary screening experiment targeting 12 types of crop plants such as radish, young radish, chinese cabbage, winter grown cabbage, cabbage, bok choy, chicory, crown daisy, carrot, chives, spinach, and spring onion. The results of inhibition rates of early plant growth in metal-contaminated soil against non-contaminated soil and the correlations between inhibitions items showed that the bok choy was appropriate specie with respect to confirm the effect of several amendments. Several amendment treatments on contaminated soil brought about the changes in the root and shoot elongation of bok choy after 1 week. Agric-lime, dolomite and steel slag treatments showed the great efficiency of reducing on mobility of heavy metals using chemical assessment. But in contrary, these treatments resulted in the reduction of root and shoot elongation and only AMD sludge increased that of elongation, significantly. When considering both chemical and biological assessments, AMD sludge could be recommended the compatible amendment for target contaminated soil. In conclusion, biological assessment was also important aspect of decision of successful soil remediation.

Antagonistic inhibitory effects of probiotics against pathogenic microorganisms in vitro (Probiotics의 병원성미생물에 대한 길항적 억제효과)

  • Yuk, Young Sam;Lee, Young ki;Kim, Ga-Yeon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.12
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    • pp.110-116
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    • 2019
  • To investigate the antagonistic inhibitory effects in a mixed culture between probiotics and various pathogenic microorganisms, 140 probiotics were identified using a 16 rRNA sequencing phylogenetic analysis method, and various probiotics strains were isolated from Korean kimchi from January to December 2016. The antagonistic inhibition test of a mixed culture of four probiotics (Enterococcus faecalis, Lactobacillus plantarum, Lactobacillus rhamnosus, and Lactobacillus reuteri) with excellent antimicrobial activity and six pathogenic microorganisms (Candida albicans, Salmonella Enteritidis, E. coli O157:H7, Shigella flexneri, Staphylococcus aureus, and Pseudomonas aeruginosa)showed that the growth of most probiotics strains increased normally after culture, but growth was inhibited almost completely in most pathogenic microorganisms, except for S. Enteritidis. This antagonistic inhibitory effect in vitro was attributed to the low pH of the lactic acid and organic acid produced during fermentation. As a result, four probiotics strains isolated from Korean Kimchi are very likely to be developed as therapeutic agents for female yeast infections and colon and skin care. In the future, these therapeutic agents will help improve public health related to probiotics.

Inhibition of Starch Biosynthesis by Antisense Expression of cDNAs Encoding ADP-Glucose Pyrophosphorylase Small Subunit in Sweetpotato (고구마에서 ADP-Glucose Pyrophosphorylase Small Subunit cDNA의 Antisense 발현에 의한 전분생합성 저해)

  • Min, Sung-Ran;Bae, Jung-Myung;Harn, Chee-Hark;Jeong, Won-Joong;Lee, Young-Bok;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.277-283
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    • 2007
  • Embryogenic calluses derived from shoot apical meristem explants of sweetpotato were subjected to particle bombardment to generate transgenic plants for antisense expression of cDNAs encoding two different AGPase small subunit (ibAGP1 and ibAGP2). Plants were generated via somatic embryogenesis. PCR and Southern analysis demonstrated that the incorporation of ibAGP1 and ibAGP2 into the genome in an antisense orientation. Immunoblot analysis confirmed reduced levels of AGPase small subunit in transgenic plant leaves. Plants with both ibAGP1 and ibAGP2 produced a lower level of the protein than plants with ibAGP1 alone. iodine test demonstrated that transgenic plant leaves and storage root accumulated reduced amounts of starch. Iodine staining of leaf tissues indicated that transgenic plants accumulated less amount of starch than control. In accordance with western blot analysis, plants with both ibAGP1 and ibAGP2 accumulated a lower amount of starch than plants with ibAGP1 alone. Both transgenic plants exhibited a severely retarded growth, resulting in bare survival. It is suggested that disrupted expression of the gene encoding AGPase small subunit is lethal to the growth of sweetpotato contrast to other species including potato.