• Title/Summary/Keyword: In Vitro Development

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In Vitro Antitumor Activity and Nephrotoxicity of the Novel Platinum(II) Coordination Complex Containing Cis-dach/Diphosphine (새로운 Platinum(II)Complex ([Pt(II)(cis-dach)(DPPP)].$(NO_3)_2$의 항암효과 및 신독성)

  • Jung, Jee-Chang;Yim, Sung-Vin;Park, Seung-Joon;Chung, Joo-Ho;Ko, Kye-Chang;Chang, Sung-Goo;Rho, Young-Soo
    • The Korean Journal of Pharmacology
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    • v.32 no.1
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    • pp.93-102
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    • 1996
  • Platinum coordination complexes are currently one of the most compounds used in the treatment of solid tumors. However, its use is limited by severe side effects such as nephrotoxicity. Our platinum-based drug discovery program is aimed at developing drugs capable of diminishing toxicity and broadening the clinical spectrum of activity of cisplatin. We synthesized new Pt(II) complex analogue containing 1,2-diaminocyclohexane (dach) as carrier ligand and 1,3-bis(diphenyl phosphino)propane (DPPP) as a leaving group. Furthermore, nitrate was added to improve the solubility. A new series of PC-1 [Pt(cis-dach) (DPPP)]. $2NO_3_2$ was synthesized and characterized by their elemental analysis and by various spectroscopic techniques [infrared (IR), $^{13}carbon$ nuclear magnetic resonance (NMR)]. PC-1 was demonstrated acceptable antitumor activity aganist SKOV -3, OVCAR-3 human ovarian adenocarcinomacells and significant activity as compared with that of cisplatin. The toxicity of PC-1 was found quite less than that of cisplatin using MTT, $[^3H]thymidine$ uptake and glucose consumption tests in rabbit proximal tubule cells, human kidney cortical cells and human renal cortical tissues. Based on these results, this novel platinum compound represent a valuable lead in the development of a new anticancer chemotherapeutic agent capable of improving antitumor activity and low toxicity.

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MiR-29a and MiR-140 Protect Chondrocytes against the Anti-Proliferation and Cell Matrix Signaling Changes by IL-1β

  • Li, Xianghui;Zhen, Zhilei;Tang, Guodong;Zheng, Chong;Yang, Guofu
    • Molecules and Cells
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    • v.39 no.2
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    • pp.103-110
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    • 2016
  • As a degenerative joint disease, osteoarthritis (OA) constitutes a major cause of disability that seriously affects the quality of life of a large population of people worldwide. However, effective treatment that can successfully reverse OA progression is lacking until now. The present study aimed to determine whether two small non-coding RNAs miR-29a and miR-140, which are significantly down-regulated in OA, can be applied together as potential therapeutic targets for OA treatment. MiRNA synergy score was used to screen the miRNA pairs that potentially synergistically regulate OA. An in vitro model of OA was established by treating murine chondrocytes with IL-$1{\beta}$. Transfection of miR-29a and miR-140 via plasmids was investigated on chondrocyte proliferation and expression of nine genes such as ADAMTS4, ADAMTS5, ACAN, COL2A1, COL10A1, MMP1, MMP3, MMP13 and TIMP metallopeptidase inhibitor 1 (TIMP1). Western blotting was used to determine the protein expression level of MMP13 and TIMP1, and ELISA was used to detect the content of type II collagen. Combined use of miR-29a and miR-140 successfully reversed the destructive effect of IL-$1{\beta}$ on chondrocyte proliferation, and notably affected the MMP13 and TIMP1 gene expression that regulates extracellular matrix. Although co-transfection of miR-29a and miR-140 did not show a synergistic effect on MMP13 protein expression and type II collagen release, but both of them can significantly suppress the protein abundance of MMP13 and restore the type II collagen release in IL-$1{\beta}$ treated chondrocytes. Compared with single miRNA transfection, cotransfection of both miRNAs exceedingly abrogated the suppressed the protein production of TIMP1 caused by IL-$1{\beta}$, thereby suggesting potent synergistic action. These results provided1novel insights into the important function of miRNAs' collaboration in OA pathological development. The reduced MMP13, and enhanced TIMP1 protein production and type II collagen release also implies that miR-29a and miR-140 combination treatment may be a possible treatment for OA.

Effect of Mepanipyrim on the Resistant Isolates of Gray Mold fungus, Botrytis Cinerea to the Fungicides of Benzimidazole and Dicarboximide (Benzimidazole과 dicarboximide계 살균제 저항성 잿빛곰팡이병원균(Botrytis cinerea)에 대한 mepanipyrim의 효과)

  • Koo, Han-Mo;An, Seung-Joon;Shin, Ho-Chul;Do, Eun-Soo;Shin, Mi-Ho;Kim, You-Seok;Kim, Jin-Hee;Chun, Se-Chul
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.259-265
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    • 2006
  • Effect of the fungicide mepanipyrim on the resistant and sensitive isolates of Botrytis cinerea was studied in vitro and also tested to control Botrytis rot of strawberry, cucumber and grape in the field. These isolates were selected by relative mycelial growth and spore germination on potato dextrose agar(PDA) incorporated with $100{\mu}g\;a.i./ml$ of benomyl and procymidone, respectively, compared to the unamended PDA. Mycelial growth of the selected resistant isolates was significantly inhibited by mepanipyrim but the inhibition rate was similar to other fungicides belong to benzimidazole or dicarboximide, although spore germination was not inhibited even by the higher concentration of mepanipyrim. When the benomyl and procymidone resistant isolates were inoculated to cucumber leaves, lesion development was significantly inhibited with application of $250{\mu}g\;a.i./ml$ of mepanipyrim but not with that of benomyl and procymidone. In addition, when $250{\mu}g\;a.i./ml$ of mepanipyrim was applied to strawberry, cucumber, and grape in the field, the control of Botrytis rot was significantly different from that of the untreated control(Duncan's multiple range test, p<0.05). The results suggested that mepanipyrim might be an alternative fungicide for the control of benomyl- and procymidone-resistant pathogens of Botrytis rot.

Effect of cultivar and ascorbic acid on in vitro shoot regeneration and development of bombardment-mediated plastid transformation of tomato (Lycopersicon esculentum) (토마토 재분화 효율 향상 및 엽록체 형질전환 조건)

  • Roh, Kyung-Hee;Lee, Ki-Jong;Park, Jong-Sug;Kim, Jong-Bum;Lee, Seung-Bum;Suh, Seok-Cheol
    • Journal of Plant Biotechnology
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    • v.37 no.1
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    • pp.77-83
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    • 2010
  • Eighteen cultivars of tomato were tested for their regeneration response. Lycopersicon esculentum cv. 2001-58 showed a very high frequency of regeneration (93%). We evaluated the effect of two compounds with known antioxidant activity (ascorbic acid and cystein). The use of ascorbic acid ($200\;-\;300\;{\mu}M/L$) has a positive effect on shoot regeneration. To develope a system for plastid transformation in tomato via homologous recombination, we constructed the tomato plastid expression vector (pKRT22-AG) harboring 2.2 kb flanking sequences cloned from intact plastid genome and gfp gene. To investigate the factors affecting the delivery system of the pKRT22-AG into chloroplast using bombardment, We assessed the optimal DNA concentration, gold particle volume and target distance. Expression of the GFP protein was observed within chloroplast on protoplast of cotyledon explant by confocal laser scanning microscopy, which indicates that the protocol developed in this study be useful for the production of plastid transgenic plants in tomato.

Antioxidative and Anti-diabetic Activity of C3GHi, Novel Black Rice Breed (Cyanidine-3-glucoside (C3G) 색소의 함유량이 증대된 쌀 신품종(C3GHi)의 항산화 및 항당뇨 활성)

  • Kim, Hwa-Young;Kim, Joong-Hark;Lee, Sung-Ae;Ryu, Su-Noh;Han, Sang-Jun;Hong, Seong-Gil
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.55 no.1
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    • pp.38-46
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    • 2010
  • C3GHi, a novel black rice variety developed by conventional breeding has more high contents of cyanidine-3-glucoside (C3G) and a more strong antioxidant than normal black rice. In this study, we investigate the antioxidative potential and anti-diabetic activity of C3GHi and extract of it for the purpose of development of functional materials or replacement meal for diabetic patients. Using in vitro antioxidative activity assay system such as DPPH radical quenching assay, superoxide anion radical scavenging assay, inhibition of lipid peroxidation and DNA breakage assay, C3GHi extract was observed to have more high antioxidative activity than normal black rice breed. The glycemic index of a freeze dried powder of C3GHi brown rice showed $43.7\pm18.8$, which belonged to low GI food (GI of 55 or less). Using in vivo diabetic model such as db/db mice model and streptozotocininduced diabetic model, C3GHi rice extract decrease blood glucose level and inhibit oxidative stress in blood. From the results, we think that the C3GHi varieties have a potential for the functional materials or ingredient of meal replacement for diabetic patients.

Evaluation of Acetylcholinesterase Inhibitory and Antioxidative Activities of Rhus javanica (오배자 추출물의 Acetylcholinesterase 저해 및 항산화 활성 평가)

  • Bae, Jong-Sup;Lee, Hyun-Shik;Lee, Ha Yeong;Yoo, Byung-Hyuk;Kim, Taewan;Kim, Yong Han;Kim, Tae Hoon
    • Food Science and Preservation
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    • v.19 no.5
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    • pp.751-756
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    • 2012
  • Antioxidant capacity and acetylcholinesterase (AChE) inhibitory activity of the aqueous methanolic extract of Rhus javanica were investigated in vitro. The antioxidant properties was measured by radical scavenging assays using DPPH and $ABTS^+$ radicals. The AChE inhibitory efficacy of R. javanica was tested by Ellman's assay and the total phenolic content was determined using a spectrophotometric method. All tested samples showed a dose-dependent AChE inhibitory and radical scavenging activities. In particularly, ethyl acetate (EtOAc)-soluble portion from the methanolic extract of R. javanica showed significantly higher inhibitory activity than other organic solvent soluble-portions in an AChE and radical scavenging assay systems. These results suggest that R. javanica may be possess potential benefits which might be useful in development of antioxidant and anti-alzheimer's disease ingredient.

Development of Porous Cellulose Hydrogel for Enhanced Transdermal Delivery of Liquiritin and Liquiritigenin as Licorice Flavonoids (감초 플라보노이드 Liquiritin 및 Liquiritigenin을 담지한 피부전달체인 셀룰로오스 다공성 하이드로젤 제형 개발)

  • Kim, Su Ji;Kwon, Soon Sik;Yu, Eun Ryeong;Park, Soo Nam
    • Polymer(Korea)
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    • v.38 no.5
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    • pp.676-681
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    • 2014
  • Licorice, widely used as a herbal medicine, has flavonoids such as liquiritin and its aglycone, liquiritigenin that show anti-oxidant and anti-inflammatory properties. Licorice flavonoid-loaded cellulose hydrogels were prepared as carriers for skin drug delivery, and their properties were investigated. The porous cellulose hydrogel was made by reacting cellulose with epichlorohydrin as a cross-linking agent in NaOH/urea(1~10%) solutions. Through studies on the rheological properties and water uptake of the hydrogel, a NaOH/urea(6%) solution was established as being optimum for the synthesis of the cellulose hydrogel containing liquiritin and liquiritigenin. Scanning electron microscopy (SEM) observations of a cross-section of the prepared hydrogel indicated its porosity. In particular, in skin permeation experiments using a Franz diffusion cell, hydrogel containing the licorice flavonoids showed remarkable transdermal permeation compared to the control group. These results indicate that porous cellulose hydrogel is a potential drug delivery system to enhance the skin permeation of licorice flavonoids.

[ α ]-Amylase Inhibitory Activity of Flower and Leaf Extracts from Buckwheat (Fagopyrum esculentum) (메밀(Fagopyrum esculentum) 꽃, 잎 추출건조물의 α-Amylase 효소활성 저해)

  • Lee, Myung-Heon;Lee, Jung-Sun;Yang, Hee-Chul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.1
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    • pp.42-47
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    • 2008
  • Prevention of postprandial hyperglycemia is important, as it is implicated in the development of macro- and microvascular complications associated with diabetes. An inhibitor of ${\alpha}$-amylase which acts in the first step of carbohydrate digestion, is expected to be a suppressor of postprandial hyperglycemia. This study investigated the porcine pancreatic ${\alpha}$-amylase inhibitory activity of the extracts from buckwheat (Fagopyrum esculentum) flower, leaf, stem and grain. Flower, leaf, stem and grain of buckwheat were extracted by water and ethanol (40%, 70%, 100%), respectively. Flower and leaf extracts were more effective ${\alpha}$-amylase inhibitors than stem and grain extracts in all tested solutions. Ethanol extracts were more effective than water extracts or powders on the ${\alpha}$-amylase inhibitory activities. At concentrations of $0.5%{\sim}10%$ (w/w, starch basis), the flower extracts of 40%, 70% and 100% ethanol lowered the enzyme activity by about 90% and the results were similar to the values of acarbose. At the same concentrations, the leaf extracts of 100% ethanol lowered the enzyme activity by about 90%. These results suggest that buckwheat flower and leaf ethanol extracts may delay carbohydrate digestion and lower postprandial hyperglycemia.

Fertilization and in vitro Development of Bovine Oocytes Following Round Spermatid Injection (원형정자 미세주입에 의한 소 난자의 수정과 체외 배 발달)

  • Um, J.H.;Do, J.T.;Kim, N.H.;Park, H.D.;Lee, H.T.;Chung, K.S.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.193-201
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    • 1999
  • In this study we determined fertilization processes and developmental ability of porcine oocytes following injection of round spermatid in the presence of artificial activation. Electrical stimulation at 3 h before spermatid injection significantly increased the incidence of normal fertilization as compared to those following injection without stimulation or with stimulation immediately after injection. The incidences of two pronuclear formation and apposition were not different in oocytes between following intracytoplasmic spermatid and spermatid nucleus injection. Indirect immunocytochemistry and laser scanning confocal microscopy study revealed that micro tubules were organized from the oocyte cortex following round spermatid injection, and this seemed to move both male and female pronuclei into the oocyte center. Paternal mitochondria which are introduced with spermatid have been observed up to 4-cell. Our study indicated that either round spermatid or it's nucleus can be used to produce viable bovine embryos by injection into unfertilized oocytes.

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Suppressive Effect of Organic Farming Materials on the Development of Tomato Gray Mold (토마토 잿빛곰팡이병에 대한 유기농업자재의 억제효과)

  • Hong, Sung-Jun;Kim, Yong-Ki;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jung;Jee, Hyeong-Jin;Kim, Suk-Chul
    • Korean Journal of Organic Agriculture
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    • v.23 no.3
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    • pp.567-582
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    • 2015
  • Botrytis cinerea infects stems, leaves and fruits of greenhouse tomato and can cause serious economic losses. This study was conducted to develop organic farming control method against tomato gray mold. Twenty two organic farming materials including mineral and plant extracts were screened for the suppressive activity against Botrytis cinerea, in vitro and in vivo. Among the organic farming materials, sulfur, copper, Chinese twinleaf extract and rhubarb extract decreased by 51.7-90% of the spore germination of Botrytis cinerea. Also, gray mold incidence was reduced more than 90% on tomato stems by treating sulfur, seaweed extracts, rhubarb root extracts and Chinese twinleaf extract. After the selected four organic farming materials were applied on tomato cultivated in greenhouse, their control effects against the tomato gray mold were tested. When the water soluble sulfur was foliar-sprayed on the tomato leaves infected by artificial inoculation with spore suspension of Botrytis cinerea, it showed 87.9% of control value. Also, control activity of the water soluble sulfur was paralleled with chemical fungicide, diethofencarb+carbendazim. The above mentioned results indicate the sulfur formulation can be used as chemical fungicide alternatives for controlling tomato gray mold in the greenhouse.