• Title/Summary/Keyword: In-vitro study

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Effects of Cooling Rate and Equilibration Time on the Survival and Development of Frozen-thawed Bovine Immature Oocytes (동결속도와 평형시간이 소 미성숙 난자의 동결 융해후 생존율에 미치는 영향)

  • 양병철;양보석;성환후;임기순;최선호;장원경;진동일;임경순
    • Korean Journal of Animal Reproduction
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    • v.25 no.1
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    • pp.19-28
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    • 2001
  • The present study was undertaken to investigate the effects of cooling rate and equilibration time on the survival, in vitro maturation and development to embryos of frozen-thawed bovine immature oocytes(Germinal Vesicle Stage). The cryoprotectants are used 10% ethylene glycol(EG) as permeating cryoprotectant and 0.05M soc.ose(S) or trehalose(T) as low molecular weight nonpermeating cryoprotectants and 5% ficoll(F) or polyvinylpyrrolidone(PVP) as high molecular weight nonpermeating cryoprotectants. Four freezing solution were uysed in this experiment(EFT: 10% EG + 5% F + 0.05M T, EFS: 10% EG + 5% F + 0.05M S, EPT: 10% EG + 5% P + 0.05M T, EPS: 10% EG + 5% P + 0.05M S). The best equilibration time and freezing solution was 15 min in EPT(83% survival rate of frozen-thawed bovine immature oocytes). When frozen-thawed bovine oocytes were cultured following IVM and IVF, there was no significant difference in cleavage and development rates among the EFT, EFS, EPT and EPS solutions. When 9 blastocysts derived from frozen bovine oocytes were transferred to 6 recipients, two recipients were pregnant. And one was aborted at 45 days of pregnancy and the other had a stillbirth.

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Effects of 3,5-di-O-Caffeoylquinic acid from Artemisia scoparia Waldstein et Kitamura on the Function of HPV 16 Oncoproteins (인진에서 분리한 3,5-di-O-Caffeoylquinic acid가 자궁경부암 바이러스 발암단백질의 기능에 미치는 영향)

  • Baek, Tae-Woong;Lee, Kyung-Ae;Ahn, Min-Jung;Joo, Hae-Kyung;Cho, Min-Chul;Kang, Jung-Woo;Kim, Hee-Seo;Shim, Jung-Hyun;Lee, Hee-Gu;Oh, Hyun-Cheol;Ahn, Jong-Seok;Cho, Yong-Kwen;Myung, Pyung-Keun;Yoon, Do-Young
    • Korean Journal of Pharmacognosy
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    • v.35 no.4 s.139
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    • pp.368-374
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    • 2004
  • Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human papillomavirus) type 16. The HPV is cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Artemisia scoparia Waldstein et Kitamura has inhibitory effects on the binding between E6 oncoprotein and tumor suppressor p53, or the binding between E6 and E6 associated protein (E6AP), an E3 ubiquitin-protein ligase. HPV oncoprotein inhibitors from Artemisia scoparia W. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and the inhibitory compounds were finally purified by HPLC using an ELISA screening system based on the binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on the functions of E6 oncoprotein. We investigated whether 3,5-di-O-caffeoylquinic acid (DCQA) extracted from Artemisia scoparia W. Could inhibit the function of E6 oncoprutein. DCQA inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that DCQA inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.

Control of Pepper Anthracnose Caused by Colletotrichum acutatum using Alternate Application of Agricultural Organic Materials and Iminoctadine tris + thiram (유기농업자재와 유기합성 살균제(Iminoctadine tris + thiram) 교호살포에 따른 고추 탄저병 방제 효과)

  • Hong, Sung-Jun;Kim, Yong-Ki;Jee, Hyeong-Jin;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jung;Kim, Jung-Hyun;Kim, Seok-Cheol
    • The Korean Journal of Pesticide Science
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    • v.19 no.4
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    • pp.428-439
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    • 2015
  • Pepper anthracnose caused by Collectotrichum acutaum has been known as one of the most damaging diseases of pepper, which has reduced not only yield but also quality of pepper produce almost every year. This study was conducted to develop control strategy against pepper anthracnose by alternate application of agricultural organic materials and chemical fungicides. The alternate application effect of agricultural organic materials and chemical fungicides for controlling pepper anthracnose were examined in vitro and in the field. First, thirteen microbial agents and twenty two agricultural organic materials were screened for antifungal activity against C. acutatum through the dual culture method and bioassay. As a result, one microbial agent (Bacillus subtilis QST-713) and three agricultural organic materials (sulfur, bordeaux mixture, marine algae extracts) were found to show high inhibition effect against C. acutatum. In the field test, when Iminoctadine tris+thiram, a chemical fungicide for controlling pepper anthracnose, was sprayed, it reduced disease incidence by 89.5%. Meanwhile Sulfur, bordeaux mixture, copper, marine algae extracts and Bacillus subtilis QST-713 showed low disease incidence at the range of 33.1~81.0%. However, when Iminoctadine tris+thiram and agricultural organic materials(bordeaux mixture, marine algae extracts) were applied to pepper fruits alternately two times at 7 days interval, there was a 81.7 and 87.1% reduction in disease, respectively. Consequently, the alternate spray of chemical fungicide (Iminoctadine tris+thiram) and agricultural organic materials (bordeaux mixture, marine algae extracts) could be recommended as a control method to reduce the using amount of chemical fungicide.

Inhibitory Effects of Lentinus edodes and Rice with Lentinus edodes mycelium on Diabetes and Obesity (In vitro 및 In vivo에서 표고버섯, 표고버섯균사체배양쌀 추출물의 항당뇨와 항비만 작용)

  • Kim, Haeseop;You, Jeheon;Jo, Yeongcheol;Lee, Youngjae;Park, Inbae;Park, Jeongwook;Jung, Myung-A;Kim, Young-Suk;Kim, Sunoh
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.2
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    • pp.175-181
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    • 2013
  • This study investigated the ability of extracts from Lentinus edodes (LE) and rice with Lentinus edodes mycelium (LEM) to inhibit diabetes and obesity. Lipid accumulation significantly decreased by 78% and 74% upon treatment with 300 ${\mu}g/mL$ of LE and LEM, respectively (p<0.01). Cholesteryl ester transfer protein (CETP) inhibition activity increased by 94% and 99% upon treatment with 300 ${\mu}g/mL$ of LE and LEM, respectively. In order to investigate the effect of LE and LEM on diabetes, the inhibition of protein tyrosine phosphate 1B (PTP1B) activity from the LE and LEM extracts at various concentrations (1, 3, 10, 30, 100, 300 ${\mu}g/mL$) was assessed. PTP1B activity by treatment with 10, 30, and 100 ${\mu}g/mL$ of LE, was inhibited at a rate of 7, 9, and 7% respectively. Also, PTP1B activity from treatment with increasing concentration of LEM led to a significant concentration-dependent inhibition of PTP1B activity (p<0.01). LE and LEM were orally administered for 28 days after a high fat diet (HFD). LE and LEM significantly reduced triglyceride, cholesterol, HDL-cholesterol and LDL-cholesterol levels. GOT and GPT were not significantly effected. These results indicate that extracts of LE and rice with LEM have potent activities useful in the treatment of obesity and diabetes mellitus.

Antioxidative and Antiallergic Effect of Persimmon Leaf Extracts (감잎(Diospyros kaki Thunb) 추출물의 항산화 및 항알레르기 효과)

  • Yoo, Ki-Hwan;Jeong, Jong-Moon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.12
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    • pp.1691-1698
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    • 2009
  • The purpose of this study was to investigate the antioxidative and antiallergic effects of persimmon leaf extract. Antioxidative and anti-inflammatory effects of the crude persimmon leaf extract (PLE) and the partially purified persimmon leaf extract (PPLE) were determined in in vitro assays by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide anion radicals, and 5-lipoxygenase (5-LO) and cyclooxygenase (COX). Total phenols and total flavonoid levels of PLE and PPLE were $230.0{\pm}19.6$ mg/g and $475.5{\pm}38.7$ mg/g, and $34.8{\pm}6.5$ mg/g and $78.8{\pm}3.6$ mg/g, respectively. DPPH and superoxide radical-scavenging activities ($SC_{50}$) of the PLE and PPLE were $23.8{\pm}3.2$ ppm and $10.0{\pm}1.3$ ppm, and $47.6{\pm}3.4$ ppm and $22.4{\pm}3.3$ ppm, respectively. Inhibitory activities ($IC_{50}$) of PLE and PPLE against 5-LO, COX-1 and COX-2 were $77.1{\pm}11.7$, $38.6{\pm}7.0$ ppm, $47.4{\pm}7.7$, $25.3{\pm}6.3$ ppm, and $129.5{\pm}5.5$, $84.5{\pm}2.3$ ppm, respectively. Moreover, two extracts inhibited dose-dependently NO production in LPS-stimulated RAW 264.7 cells, and also effectively inhibited the cutaneous anaphylaxis reaction activated by anti-dinitrophenyl IgE antibody in mice. These results suggest that PLE and PPLE may be useful for phytochemical materials for prevention and treatment of radical-mediated pathological and allergy diseases.

Antioxidant, Tyrosinase Inhibitory, and Anti-proliferative Activities of Gochujang Added with Cheonggukjang Powder Made from Sword Bean (작두콩 청국장 첨가 고추장의 항산화, tyrosinase 저해 및 암세포 증식 억제 효과)

  • Chang, Moon-Ik;Kim, Jae-Young;Kim, Un-Sung;Baek, Seung-Hwa
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.221-226
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    • 2013
  • This study aimed to examine the antioxidant, tyrosinase inhibitory, and anti-proliferative activities (A549, G361, HT-29, and MDA-MB-231) of fermented gochujang (made from sword bean cheonggukjang powder (SBC) for 90 days. Gochujang was prepared by adding 0 (SBC 0), 2 (SBC 2), 5 (SBC 5), 8 (SBC 8) and 10% (SBC 10) levels with SBC, and all experiments were measured at diluted levels of 20, 50 and 100 times. The antioxidant activity and tyrosinase inhibitory effect demonstrated that SBC 10 increased approximately 1.2 and 1.1 times compared with SBC 0, respectively, at diluted levels of 50 and 100 times. The anti-proliferative effects of A549, G361, and HT-29 presented that SBC 10 were 2.8, 1.1, and 8.9 times higher compared with SBC 0, respectively, at diluted levels of 50, 20, and 100 times. In the case of MDA-MB-231, SBC 10 was 3.7 times higher compared with SBC 5 at diluted level of 20 times. As a result, we confirmed that SBC gochujang was improved for physiological activities and anti-proliferative effects.

REMINERALIZATION EFFECT OF COMMERCIALLY AVAILABLE ANTICARIOGENIC PRODUCTS ON ARTIFICIAL CARIES LESION USING QLF (QLF를 이용한 항우식 제품의 인공우식 재광화 효과에 대한 연구)

  • Song, Ju-Hyun;Kim, Young-Jae;Kim, Jung-Wook;Jang, Ki-Taeg;Kim, Chong-Chul;Hahn, Se-Hyun;Lee, Sang-Hoon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.35 no.2
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    • pp.287-296
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    • 2008
  • The purpose of this in vitro study was to compare the remineralization effect of commercially available anticariogenic products, exactly low level fluoride mouthrinse(500 ppm NaF), tooth cream with Casein phosphopeptide-amorphous calcium phosphate(CPP-ACP) and fluoride mouthrinse plus tooth cream on artificial caries lesion. Artificial caries lesion was induced at the buccal surface of permanent third molar and the specimens were then divided(16 specimens/group) into four group. Twice a day during 28 days specimens of each group were treated with a commercially anticariogenic product as follows and applied to the pH cycling system. Group 1: control group (No treatment) Group 2: Tooth $Mousse^{(R)}$ (GC Co. Japan) Group 3: $chikachika^{(R)}$ (Samil Co. Korea) Group 4: $chikachika^{(R)}$+Tooth Mousse$^{(R)}$ The long-term change of mineral loss(${\Delta}Q$) was evaluated by quantitative light-induced fluorescence (QLF) and the following results were obtained: 1. ${\Delta}Q$ of Group 1 was not noticed statistically significant during 28 days comparing that prior to treatment. There was a statistically significant increase in ${\Delta}Q$ of Group 2 and 3 since 14 days. So was in ${\Delta}Q$ of Group 4 since 7 days. 2. ${\Delta}Q$ was increased as follows: Group 1< Group 2, 3< Group 4. 3. Comparing with Group 1, Group 2 was a statistically significant increase since 7 days and Group 3 and 4 were since 3 days. Comparing Group 2 with 3, there was not noticed statistically significant during whole duration. Group 4 was significantly higher than Group 2 and 3 after 28 days. 4. All groups demonstrated a decrease in the rate of remineralization as time goes on.

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Screening of Extracts from Marine Green and Brown Algae in Jeju for Potential Marine Angiotensin-I Converting Enzyme (ACE) Inhibitory Activity (제주 자생 해양 녹조류와 갈조류 추출물로부터의 항고혈압 활성)

  • Cha, Seon-Heui;Ahn, Gin-Nae;Heo, Soo-Jin;Kim, Kil-Nam;Lee, Ki-Wan;Song, Choon-Bok;K.Cho, So-Mi;Jeon, You-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.3
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    • pp.307-314
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    • 2006
  • This study was conducted to screen in vitro angiotensin converting enzyme (ACE) inhibitory activities of methanol (MeOH) and aqueous extracts which were prepared by four different extractions-80% methanol extracts(ME) at $20^{\circ}C\;and\;70^{\circ}C$, respectively and aqueous extracts (AE) at both temperatures with the residue of the MEs-of ten marine green algae and nineteen brown algae collected along Jeju coast of Korea. Most marine brown algae extracts showed higher capacities than those of marine green algae in ACE inhibitory activity. Particularly, $70^{\circ}C$ MeOH extract (70ME) of Hizikia fusiforme showed the strongest inhibition activity (about 87%) among all the extracts. Also, 70 MEs of Enteromorpha linza, Ishige sinicola, Laminaria ochotensis, Petrospongium rugosum, Sagrassum horneri, Undaria pinnatifida and $20^{\circ}C$ MeOH extracts (20ME) of Myagropsis myagroides, Petrospongium rugosum, $20^{\circ}C$ aqueous extracts (20AE) of Codium contractum, Enteromorpha compressa, and $70^{\circ}C$ aqueous extracts (70AE) of Ecklonia cava, Petrospongium rugosum showed moderate ACE inhibitory activities more than 50% and the other extracts exhibited weak activities. On tile other hand, E. cava had the best ACE inhibitory activity among 70AEs. This indicates that 70AE of E. cava contains potential anti-ACE macromolecular. We tried to proteolytic digest 70AE of E. cava to induce production of anti-ACE peptides from E. cava 70AE. The enzymes used are five pretenses including Kojizyme, Flavourzyme, Neutrase, Alcalase, and Protamex, which are food grade-commercial enzymes from Novo Co. Flavourzyme-digest of E. cava 70AE showed the highest inhibitory activity about 90%. And the five different enzymatic digests of the E. cava 70AE ranged from 2.33 to 3.56 ${\mu}g/mL$, respectively in $IC_{50}$ values of anti-ACE activity.

Physiological Interactions Between the Herbicide Pretilachlor and the Safener Fenclorim on Rice (제초제(除草劑) Pretilachlor와 해독제(害毒劑) Fenclorim의 수도(水稻)에 대한 생리적(生理的) 상호작용(相互作用))

  • Han, S.S.;Hatzios, K.K.
    • Korean Journal of Weed Science
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    • v.10 no.4
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    • pp.328-337
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    • 1990
  • The incividual and combined effects of the chloroacetanilide herbicide pretilachlor and of the safener fenclorim on the growth and selected physiological processes of rice (Oryza sativa L., var 'Lemont')were evaluated under greenhouse and laboratory conditions. Fenclorim applied at rates ranging from 50 to 300 g a.i./ha antagonized the injurious effects caused by 150 to 900 g a.i./ha of pretilachlor on 15-day old wet-sown rice grown under greenhouse conditions. When used rates of 150 g/ha or higher, fenclorim reversed completely the effects of all doses of pretilachlor on rice. When the two compounds were given simultaneously, fenclorim enhanced the uptake of $^{14}C$pretilachlor into rice leaf mesophyll protoplasts measured for 1 hr, indicating that competition for uptake at the protoplast level is not involved in the protective action of this safener. The safener-induced stimulation of pretilachlor uptake was particularly evident when fenclorim was used at concentrations of 10, 20 and $40{\mu}M$. Following 4 hr of incubation, individual treatments with pretilachlor inhibited the in vitro incorporation of radiolabeled precursors into proteins, DNA, and lipids of rice leaf protoplasts only when used at the high concentration of $100{\mu}M$M. Individual treatments with high concentrations (10 or $100{\mu}M$) of the safener fenclorim inhibited the incorporation of radiolabeled precursors into proteins and lipids of rice protoplasts, but had no DNA synthesis. The combined effects of pretilachlor and fenclorim on the incorporation of radiolabeled precursors into these macromolecules of isolated rice mesophyll protoplasts appeared to be additive or slightly synergistic rather than antagonistic. Fenclorim at $1{\mu}M$ antagonized the effects of pretilachlor on total lipids of rice leaf protoplasts. In addition, individual and combined treat-menu with pretilachlor and fenclorim influenced the incoroporation of$^{14}C$acetate into polar lipids, triglycerides and steryl esters of rice leaf protoplas causing a redistribution of carbon in these lipid fractions. However, these effects were not large enough to explain the herbicidal activity of pretilachlor or to account for the protective action of the safener fenclorim. Overall, the uesults of the present study idnicate that the safener fenclorim does not seem to protect rice against pretilachlor injury by antagonizing its effects on protein, DNA, or lipid syntheses.

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Herbicidal Properties of 5,8-dihydroxy-1,4-naphthoquinone and Their Possible Mode of Action (천연물 유래 5,8-dihydroxy-1,4-naphthoquinone의 살초특성과 작용기구)

  • Choi, Jung-Sup;Kim, Ji-Yeon;Seo, Bo-Ram;Ko, Young-Kwan;Cha, Mi-Ran;Kim, Young-Sup;Ryu, Shi-Yong;Hwang, In-Taek
    • Korean Journal of Weed Science
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    • v.31 no.3
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    • pp.250-259
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    • 2011
  • This study was conducted to assess the possibility of 5,8-dihydroxy-1,4-naphthoquinone (DHNQ) as a environmental friendly herbicide candidate. Foliar application of DHNQ showed excellent herbicidal effect to the 3 grasses and 5 broad-leaved weeds. Among them, Digitaria sanguinalis and Solanum nigrum were completely controlled by $250{\mu}g\;mL^{-1}$ of DHNQ with main symptoms of desiccation or burndown within 24 hours. Aeschynomene indica was also sensitive to DHNQ treatment. All of the eight weed species were controlled by 90~100% at a concentration of $1000{\mu}g\;mL^{-1}$. However, soil application of DHNQ to Digitaria sanguinalis did not show any herbicidal symptoms. DHNQ strongly inhibited KAPAS activities in vitro and the $IC_{50}$ was $4.4{\mu}M$. Cellular leakage from cucumber leaf squares treated with DHNQ increased depending on the concentrations increased from 6.25 to $100{\mu}M$ after 24 hours incubation with or without light. However, chlorophyll loss in cucumber leaf squares was negligible. Biotin supplements significantly rescued the inhibition of germination rate of Arabidopsis thaliana seeds previously inhibited by the DHNQ. According to above results, DHNQ is a good natural herbicide candidate having a new target KAPAS, which is involved in biotin biosynthesis pathway, with environmental friendly.