• Title/Summary/Keyword: 생물제제

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Expression and Role of Trypsin-Like Enzyme Involved in Hatching of Preimplantation Mouse Embryos (생쥐 배아의 부화에 관여하는 Trypsin 유사 효소의 발현과 역할)

  • Kim, Soo-Kyung;Kang, Hee-Kyoo;Jun, Jin-Hyun;Choi, Kyoo-Wan;Kim, Moon-Kyoo
    • Development and Reproduction
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    • v.5 no.1
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    • pp.17-21
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    • 2001
  • This study was conducted to investigate the expression pattern of Trypsin-like enzyme and the effect of a trypsin inhibitor(benzimidine) on hatching process during in-vitro culture of mouse preimplantation embryos. The Trypsin-like enzyme was identified by rhodamine-conjugated Trypsin substrate probe. The expression of trypsin-like enzyme was firstly detected at the late morula stage, and the enzyme was uniformly localized in the trophectoderm of late blastocysts. Especially, intense fluorescence was observed in the blebbing area of hatching blastocysts. Bisbenzamidine, contained in culture media, did not alter embryonic development from 4-cell stage to the expanded blastocyst but decrease the hatching rate in ImM concentration (15.8% vs 89.7%, p<0.02). In the treatment of bisbenzimidine (5mM) for 12 hours according to the embryonic stage of mouse, the hatching rate of control (83.0%) and treatment in late blastocysts (8.7%) were significantly (p<0.01) different. From these results, we suggested that the hatching enzyme having trypsin-like activity was localized from the late morula stage, and the hatching process by this enzyme was activated in the late blastocyst stage of mouse embryos.

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The Growth Promotion Effect of Useful Enterobacteria Bifidobacterium aolescentis KCTC 3216 by Combination of Natural Products Bearing Antioxioative Capacity (장내 유용세근 Bifidobacterium adolescentis KCTC 3216의 성장을 촉진시키는 항산화 천연산물의 조합구성)

  • 김종덕;김민용;안창범;서효진;김봉조;서재관;김점순;공재열
    • KSBB Journal
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    • v.17 no.4
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    • pp.388-395
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    • 2002
  • The growth of enterobacteria, Bifidobacteriurn adolescentis KCTC 3216 was promoted by natural products bearing antioxidative capacity and combined two, three and four kinds of them. B. adolescentis was showed a good growth by Sophorae flos, Phellodendri cofex, Mori cortex radicis, Aurantii nobilis pericarpium, Angelicae gigantis radix, alone, and two mixed combinations were composed of Paeonia japonica and Theae foiium, Epimedii herba and Angelicae gigantis radix, Paeonia japonica and epimedii herba, Atractylodis rhizoma alba and angelicae gigantis radix, and three mixed combinations were oraganized with Theae folium, Paeonia japonica and epimedii herba, Theae folium, Beiamcanda chinensis and Paeonia japonica, Theae foiium, Astragaii radix and Mori cortex radicis, and four mixed combinations were formed with Seiamcanda chinensis, Angelicae gigantis radix, Epimedii herba, Theae folium, and Angeiicae gigantis radix, Epimedii herba, Paeonia japonica, Theae folium, and Epimedii herba, Paeonia japonica, Sophorae flos, Theae folium. The best four mixed combination for the growth of B. adolescentis was mixture of Epimedii herba, Paeonia iaponica, Sophorae flow and Theae foiium, which promoted 2.6 times than that of control, and its antioxidative capacity was also 5.6 times higher, and the ratio of elimination of hydroxyl radical was more than 80% in each dilution rate. As these combinations of natural products will activate some parts of body, they may be applied to pharmaceuitcal applications, functional foods, antiaging tea, also expected to promote useful entero bacterial growth for fermentative beverage bearing multifunction.

Anti-angiogenic and Anti-cell Adhesion Effects and Their Mechanism with the Extract of Camellia japonica Leaf (동백잎 추출물의 신생혈관 및 세포부착 억제작용과 그 기전)

  • Song, Min-Gyu;Seo, Hyo-Jin;Moon, Je-Hak;Park, Keun-Hyung;Kim, Jong-Deog
    • KSBB Journal
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    • v.22 no.4
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    • pp.249-254
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    • 2007
  • Anti-angiogenesis and anti-cell adhesion effects were investigated with different dose of Camellia japonica leaf (CJL) extract for applying anti-cancer, anti-metastasis and anti-obesity. Cytotoxicity on HUVECs was very low at 200 ug/mL of CJL-extract. Anti-angiogenic ratio at increasing dose of 1.5 ug/mL, 3.0 ug/mL, 15 ug/mL and 30 ug/mL was showed 30.7%, 38.5%, 53.8%, and 70.0%, respectively. Also, anti-cell adhesion effect at concentration of $50{\mu}g{/well},\;100{\mu}g{/well}\;and\;200{\mu}g{/well}$ was expressed on E-selectin by 46.7%, 66.7%, and 86.76%, on VCAM-1, 23.0%, 61.5%, and 84.6%, and on ICAM-1, 11%, 55.5%, and 88.8%, respectively. For inquiring anti-angiogenesis mechanism, when western blot was performed with different dose of CJL extract, signal molecules of VEGFR-2, $\beta$-catenin and PI3-K were suppressed. As the signal transduction from VEGFR-2, $\beta$-catenin and PI3-K to NF-${\kappa}$B was interupted, angiogenesis could not be occurred causing not activated NF-kB. C. japonica leaf (CJL) is a useful herb for developing therapeutics of angiogenesis related diseases such as cancer, metastasis, rheumathioid arthritis and obesity.

Establishment and Validation of Gold Amalgamation Method for the Quantitation of Thimerosal in Biological Products (생물학적제제의 치메로살 함량 정량을 위한 가열기화 아말감 흡광도법의 확립 및 검증)

  • Kim, Byung-Chul;Kim, Do-Keun;Hong, Sung-Hwa;Kim, Yeon-Hee;Lim, Jong-Mi;Won, Yun-Jung;Kim, Seok-Hwan;Hong, Ji-Young;Yun, Young-Min;Kim, Jae-Ok
    • YAKHAK HOEJI
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    • v.55 no.4
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    • pp.284-288
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    • 2011
  • The test method for biologics of lot release system is based on 'Test procedure and specification for biological products,' generally, thimerosal content is measured by chemical analysis using O.D. In this study, the comparative analysis was carried out using the gold amalgamation method for thimerosal content was compared to the existing methods, which are described above. The gold amalgamation method, which uses atomic absorption spectrophotometry, was meets all the method validation acceptance criteria. It is considered to be proper as the assay and identification test for thimerosal. In this study, the comparative analysis was performed three times. As a result, gold amalgamation method is more convenient and easy to perform as this assay doesn't have pre-treatment procedure. Also this assay showed good precision and reproducibility compared to the conventional method. Therefore, it is appropriate to alternate the assay method of thimerosal from the conventional chemical analysis to gold amalgamation method to improve the credibility of lot release system and the quality control of biologics, by standardizing test method.

Effects of Carbon Substrates on Exopolysaccharide Production by Enterobacter sp. (Enterobacter sp. 의 다당 생산에 미치는 탄소원 기질의 영향)

  • Lee Ju-Ha;Lee Shin-Young
    • KSBB Journal
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    • v.20 no.1 s.90
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    • pp.26-33
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    • 2005
  • The effects of carbon sources for exopolysaccharide production during batch cultivation of an Enterobacter sp. isolated from the composter were investigated. The highest amount of exopolysaccharide was obtained when lactose was used as carbon source. Lactose in medium was converted into glucose and galactose. Glucose was metabolized fast and was completely consumed, but about $20\%$ of lactose was accumulated as galactose. On the other hand, enzyme activity was about $350\~450$ unit with the increase of lactose concentration. Thus, it was considered that the exopolysaccharide might be produced in the course of that lactose was hydrolyzed into glucose and galactose by $\beta-galactosidase$ with respect to that enzyme activity on lactose hydrolysis was accorded to the exopolysaccharide production. When glucose and galactose were added to lactose medium, respectively, it could be considered that glucose was as a repressor and galactose was as a inducer for $\beta-galactosidase$ synthesis even though the mechanisms were not elucidated. The increase of lactose concentration was almost ineffective to the specific growth rate $(0.133\~0.151\;hr^[-1})$ but showed the difference in the biomass content. The higher carbon source concentration, the more residual sugar remained. It was assumed that the optimum lactose concentration for exopolysaccharide production was $30\~70g/L.$ On the other hand, it was considered that the nitrogen acted as growth limiting nutrients to the cell growth. In the cases of 30 and 70 g/L of the fixed carbon concentrations, the increase of the nitrogen sources concentration caused a remarkable increase within the range of $0.059\~0.225\;hr^{-1}$ and $0.141\~0.237hr^{-1}$ of the specific growth rate, respectively, while there was no significant difference in biomass.

Bioequivalence of Enteric-coated Omeprazole Products (오메프라졸 장용성제제에 대한 생물학적 동등성 평가)

  • Kim, Chong-Kook;Jeong, Eun-Ju;Lee, Eun-Jin;Shin, Hee-Jong;Lee, Won-Keun
    • Journal of Pharmaceutical Investigation
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    • v.23 no.1
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    • pp.41-49
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    • 1993
  • The bioequivalence of two omeprazole enteric-coated products was evaluated in 16 normal male volunteers (age 26-32 yr, body weight 57-75 kg) following single oral administration. Test product was enteric-coated KD-182 tablet (Chong Kun Dang Corp., Korea) and reference product was $Rosec^{\circledR}$ capsule containing enteric-coated pellets of omeprazole (Yuhan Corp., Korea). Both products contain 20 mg of omeprazole. One tablet or capsule of the test or the reference product was administered to the volunteers, respectively, by randomized two period cross-over study ($2\;{\times}\;2$ Latin square method). Average drug concetrations at each sampling time and pharmacokinetic parameters calculated were not significantly different between two products(p>0.05); the area under the concentrationtime curve to last sampling time (8 hr) $(AUC_{0-8hr})$ $(1946.5{\pm}675.3\;vs\;2018.3{\pm}761.6\;ng{\cdot}hr/ml)$, AUC from time zero to infinite $(AUC_{o-\infty})$ $(2288.6{\pm}1212.8\;vs\;2264.9{\pm}1001.3\;ng{\cdot}hr/ml)$, maximum plasma concentration $(C_{max})$ $(772.5{\pm}283.3\;vs\;925.8{\pm}187.7\;ng/ml)$, time to maximum plasma concentration $(T_{max})$ $(2.38{\pm}1.06\;vs\;2.34{\pm}1.09\;hr)$, apparent elimination rate constant $(k_{\ell})$ $(0.5339{\pm}0.2687\;vs\;0.5769 {\pm}0.2184\;hr^{-I})$, apparent absorption rate constant $(k_a)$ $(1.1536{\pm}0.5278\;vs\;0.9739{\pm}0.9507 hr^{-1})$ and mean residence time (MRT) $(3.13{\pm}0.73\;vs \;3.41{\pm}1.04\;hr)$. The differences of mean $(AUC_{0-8hr})$, $C_{max}$, $T_{max}$ and MRT between the two products (3.69, 19.83, 1.32 and 8.99%, respectively) were less than 20%. The power $(1-{\beta})$ and treatment difference $(\triangle)$ for $AUC_{o-8hr}$ $C_{max}$ and MRT were more than 0.8 and less than 0.2, respectively. Although the power for $T_{max}$ was under 0.8, $T_{max}$ of the two products was not significantly different each other(p>0.05). These results suggest that the bioavailability of KD-182 tablet is not significantly different from that of $Rosec^{\circledR}$ capsule. Therefore, two products are bioequivalent based on the current results.

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Purification Nematicidal Substance and Nematicidal Activity from Ginkgo biloba L. Outer Seedcoat (은행 외종피로부터 살선충 물질의 순수 분리와 활성)

  • Jang, Yu Ju;Hwang, Hyeon Jeong;Kim, Keun Ki
    • Korean Journal of Organic Agriculture
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    • v.29 no.1
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    • pp.97-109
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    • 2021
  • Plant parasitic nematodes are causing significant damage in crop production. There is a need to develop eco-friendly nematicide that reduces the damage of nematode and has little effect on the environment and human. In this study, we have isolated a substance having nematicidal activity from Ginkgo biloba L. outer seedcoat. Studies of G. biloba L. outer seedcoat are insufficient compared to the seed and leaves due to their odor and toxicity. The dried G. biloba L. outer seedcoat was extracted with dichloromethane:methanol (1:1) and fractionated into hexane, ethyl acetate and H2O. Four steps TLC were performed from EtOAc fraction to purely isolate GB4-3 with nematicidal activity. To compare nematicidal activity, G. biloba L. seedcoat methanol extract and purified GB4-3 were investigated in terms of treatment concentration and time. As a result, the nematicidal activity increased with concentration and time. In the place treated with 20 ㎍/mL of crude G. biloba L. seedcoat MeOH extract, strong activity appeared after 12 hours, and 46% nematicidal activity shown after 18 hours. About 69% of nematicidal activity was confirmed in the place where GB4-3 purified from outer seedcoat was treated with 20 ㎍/mL, and the possibility of development as nematicide was very high. This study could be used as a basic data for the development of a nematode preparation from G. biloba L. outer seedcoat.

Isolation and Characterization of Indole-3-acetic acid- and 1-aminocylopropane-1-carboxylyic Acid Deaminase-producing Bacteria Related to Environmental Stress (환경스트레스와 관련된 indole-3-acetic acid 및 1-aminocylopropane-1-carboxylyic acid deaminase 활성을 갖는 박테리아의 분리와 특성 연구)

  • Kim, Hee Sook;Kim, Ji-Youn;Lee, Song Min;Park, Hye-Jung;Lee, Sang-Hyeon;Jang, Jeong Su;Lee, Mun Hyon
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.390-400
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    • 2019
  • In this study, strains isolated from soil samples collected from Busan, Changwon, and Jeju Island were examined to verify their abilities of phosphate solubilization and nitrogen fixation, production of indole-3-acetic acid (IAA), siderophore, and 1-aminocylopropane-1-carboxylyic acid (ACC) deaminase in order to select strains that promote plant growth and play a role in biocontrol of pests or pathogens. According to the results of this study, most of the isolated strains were found to have ability of phosphate solubilization, nitrogen fixation, IAA production, siderophore production, and production of ACC deaminase. These isolated strains might help plant growth by directly improving absorption of nutrients essential for phosphate solubilization and nitrogen fixation. In addition, they can promote plant growth and control resistance to plant diseases through extracellular enzyme activity and antifungal activity. In addition, most of the selected strains were found to survive in various environmental conditions such as temperature, salinity, and pH. Therefore, Pseudomonas plecoglossicida ANG14, Pseudarthrobacter equi ANG28, Beijerinckia fluminensis ANG34, and Acinetobacter calcoaceticus ANG35 were finally selected through a comparative advantage analysis to suggest their potential as novel biological agents. Further studies are necessary in order to prove their efficacy as novel biological agents through formulation and optimization of effective microorganisms, their preservation period, and crop cultivation tests.

Mass Cultivation of Rhodococcus sp. 3-2, a Carbendazim-Degrading Microorganism, and Development of Microbial Agents (카벤다짐 분해 미생물인 Rhodococcus sp. 3-2의 대량 배양 및 미생물 제제 개발)

  • Jun-Kyung Park;Seonghun Im;Jeong Won Kim;Jung-Hwan Ji;Kong-Min Kim;Haeseong Park;Yeong-Seok Yoon;Hang-Yeon Weon;Gui Hwan Han
    • Korean Journal of Environmental Agriculture
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    • v.42 no.4
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    • pp.259-268
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    • 2023
  • Rhodococcus sp. 3-2 strain has been reported to degrade benzimidazole-based pesticides, such as benomyl and carbendazim. Therefore, this study aimed to optimize culture medium composition and culture conditions to achieve cost-effective and efficient large-scale production of the Rhodococcus sp. 3-2 strain. The study identified that the optimal media composition for mass culture comprised 0.5% glucose, 0.5% yeast extract, 0.15% NaCl, 0.5% K2HPO4, 0.5% sodium succinate, and 0.1% MgSO4. Additionally, a microbial agent was developed using a 1.5-ton fermenter, with skim milk (20%), monosodium glutamate (15%), and vitamin C (2%) as key components. The storage stability of the microbial agent has been confirmed, with advantages of low temperature conservation, which helps to sustain efficacy for at least six months. We also assessed the benomyl degradation activity of the microbial agent within field soil. The results revealed an over 90% degradation rate when the concentration of viable cells exceeded 2.65 × 106 CFU/g after a minimum of five weeks had elapsed. Based on these findings, Rhodococcus sp. 3-2 strain can be considered a cost-effective microbial agent with diverse agricultural applications.

In vitro Effect of Ecklonia stolonifera Okamura Extract on the Cell Growth in CCD-986sk Human Fibroblast and Melanin Formation Inhibition in Clone M-3 Mouse Melanocyte Cell Line (청정해역 곰피추출물의 세포생리활성 연구)

  • Whang, Eun-Kyoung;Cho, Myung-Hwan;Park, Chan-Sun;Kim, Myung-Hee;Park, Kap-Joo
    • Korean Journal of Environmental Biology
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    • v.26 no.1
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    • pp.30-35
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    • 2008
  • In order to investigate whether or not CCD-986sk cell line can be affected by Korean Ecklonia stolonifera Okamura, we examined the MTT assay when we treated Korean Ecklonia stolonifera Okamura extract in CCD-986sk human fibroblast cell line. The sample were tested for cell proliferation activity by means of a modification of the MTT assay. Ecklonia stolonifera Okamura extract showed significantly strong cell proliferation activity at the range of from 6.25 mg $mL^{-1}$ to 1.56 mg $mL^{-1}$ compared with control group. And in order to search for inhibition agents of skin melanin formation, we tested for inhibition effect of melanin pigmentation of Korean Ecklonia stolonifera Okamura using Clone M-3 mouse melanocyte cell lines. when we treated the extracts of Ecklonia stolonifera Okamura to the mouse melanocyte cell lines, the sample showed a significantly little formation of melanin pigments compared with control group at the only range of 200 mg $mL^{-1}$. These results suggest that extract of Korean Ecklonia stolonifera Okamura may represents an excellent candidate for inhibition of melanin pigmentation and for protection of human skin aging at in vitro level.