• Title/Summary/Keyword: In vitro screening

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Frequency of Micronuclei in Lymphocytes Following Gamma and Fast-neutron Irradiations (방사선 조사량에 따른 인체 정상 림파구의 미세핵 발생빈도)

  • Kim Sung-Ho;Cho Chul-Koo;Kim Tae-Hwan;Chung In-Yong;Yoo Seong-Yul;Koh Kyoung-Hwan;Yun Hyong-Geun
    • Radiation Oncology Journal
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    • v.11 no.1
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    • pp.35-42
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    • 1993
  • The dose response of the number of micronuclei in cytokinesis-blocked (CB) lymphocytes after in vitro irradiation with $\gamma$-rays and neutrons in the 5 dose ranges was studied for a heterogeneous population of 4 donors. One thousand binucleated cells were systematically scored for micronuclei. Measurements performed after irradiation showed a dose-dependent increase in micronuclei (MN) frequency in each of the donors studied. The dose-response curves were analyzed by a linear-quadratic model, frequencies per 1000 CB cells were ($0.31{\pm}0.049$)D+($0.0022{\pm}0.0002)D^2+(13.19{\pm}1.854) (r^2=1.000,\;X^2=0.7074,\;p=0.95$) following $\gamma$ irradiation, and ($0.99{\pm}0.528$)\;D+(0.0093{\pm}0.0047)\;D^2+(13.31{\pm}7.309)\;(r^2=0.996,\;X^2=7.6834,\;p=0.11) following neutrons irradiation (D is irradiation dose in cGy). The relative biological effectiveness (RBE) of neutrons compared with $\gamma$-rays was estimated by best fitting linear-quadratic model. In the micronuclei frequency between 0.05 and 0.8 per cell, the RBE of neutrons was $2.37{\pm}0.17$. Since the MN assay is simple and rapid, it may be a good tool for evaluating the $\gamma$-ray and neutron response.

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Investigation on Antioxidant Activity in Plant resources (식물자원의 항산화활성 탐색)

  • Lee, Seung-Eun;Sung, Jung-Sook;Jang, In-Bok;Kim, Geum-Sook;Ahn, Tae-Jin;Han, Hee-Sun;Kim, Ji-Eun;Kim, Young-Ock;Park, Chung-Berm;Cha, Sun-Woo;Ahn, Young-Sup;Park, Ho-Ki;Bang, Jin-Ki;Seong, Nak-Sul
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.5
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    • pp.356-370
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    • 2008
  • This study was conducted for screening on antioxidant activity of 429 plants and selecting new potential antioxidant candidates. In vitro test models such as scavenging activity on DPPH radical and inhibitory activity on linoleic acid oxidation were used in the preliminary study. Flower of Sanguisorba officinalis, flower of Sedum kamtschaticum, flower of Rumex obtusifolius, and root of Sedum kamtschaticum showed very effective antioxidant activity on DPPH radical and linoleic acid oxidation. Those plants showed 8.1, 9.4, 9.9, $11{\mu}g/ml$ in DPPH radical scavenging activity as $SC_{50}$ and did 80.4, 80.1, 84.5, 88.0% in inhibition activity on linoleic acid oxidation, respectively. Root of Sedum middendorfianum M. showed positive effects in superoxide radical scavenging activity ($38.4{\mu}g/ml$) and inhibitory effect on $CuSO_4$-induced LDL oxidation (53.8% at final concentration of $1{\mu}g/ml$). Gleditsia japonica Mig. showed high antioxidant activity on LDL oxidation as 71.6% at final concentration of $1{\mu}g/ml$ and total phenol content of 958.5 mg% as tannic acid equivalent. In conclusion, we think that these plants having potent antioxidant activity might be studied further and could be used as new resources for many purposes including healthy food, functional cosmetics and drug development etc.

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.

Screening of Plant Extracts with Cholinesterase Inhibition Activity (콜린 에스테라제 저해효과 보유 식물 추출물 탐색)

  • Park, Saet-Byul;Lee, Jeong-Hoon;Kim, Hyung-Don;Soe, Kyung-Hae;Jeong, Hyeon-Soo;Kim, Dong-Hwi;Lee, Seung-Eun
    • Korean Journal of Plant Resources
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    • v.31 no.5
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    • pp.433-452
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    • 2018
  • This study was conducted to select candidates from plant resources for the purpose of improving or treating Alzheimer's disease, a type of dementia. One hundred and eighty-four plant extracts at a final concentration of $100{\mu}g/ml$ were screened to determine their capacity to inhibit acetylcholinesterase (AChE) by in vitro assay. From this AChE assay, seven plant extracts - including methanol ext. and water ext. of Phellaodendron amurense Rupr. (bark), methanol ext. of Nelumbo nucifera Gaertn (stamen/ovary), methanol ext. of Persicaria tinctoria H. GROSS (flower), methanol ext. of Coptis chinensis (rhizome), ethanol ext. of Cinnamomum cassia Blume(bark) and ethanol ext. of Carthamus tinctorius L. (fruit) - showed effective inhibition activity ranging from 18.7% to 63.1%. The selected extracts were testified their inhibition activities on AChE and BuChE (butyrylcholinesterase) at concentrations of 25, 50, 100, $200{\mu}g/ml$. In the AChE assay, five extracts including methanol ext. of Nelumbo nucifera Gaertn. (stamen/ovary), methanol ext. of Persicaria tinctoria H. GROSS (flower), methanol ext. of Coptis chinensis (rhizome), methanol ext. and water ext. of Phellaodendron amurense Rupr. (bark) showed inhibition activity of 15.0%~73.5%, 19.5%~63.5%, 81.6%~58.5%, 69.9%~80.5%, and 54.8%~78.3%, respectively, at concentrations of 25, 50, 100, $200{\mu}g/ml$. In the BuChE assay, the extracts of Nelumbo nucifera Gaertn. (stamen/ovary), Persicaria tinctoria H. GROSS (flower), and Coptis chinensis (rhizome) showed inhibitory capacities of 58.9~81.6%, 45.8%~72.4%, and 33.1%~55.4% at concentrations of 25, 50, 100, $200{\mu}g/ml$, respectively. In conclusion, it is suggested that Nelumbo nucifera Gaertn. (stamen/ovary), Persicaria tinctoria H. GROSS, Coptis chinensis (rhizome) and Phellaodendron amurense Rupr. (bark) could be selected as candidate materials for improving or treating Alzheimer's disease on the basis of further study.

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.

In vitro Screening of Jeju Island Plants for Customerized Cosmetics (맞춤형화장품 소재 개발을 위한 제주 식물 탐색)

  • Yoon, Kyung-Sup;Kim, Mi Jin;Kim, Moo-Han
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.4
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    • pp.1487-1495
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    • 2018
  • In this study, we investigated collagen production and hyaluronic acid production effects for wrinkle improvement test on 50 kinds of land plants and 10 kinds of marine plants native to Jeju Island as a part of developing customized cosmetic materials. Collagen and hyaluronic acid are recognized as major factors affecting skin aging. Cerastium holosteoides var. hallaisanense Mizushima extract ($100{\mu}g/mL$) produced more than 190% of collagen in the extracts of 50 kinds of land plants. Vicia angustifolia var. segetilis K. Koch. extract ($100{\mu}g/mL$) produced more than 160% of collagen. Ftsia japonica Decne. et Planch. extract ($100{\mu}g/mL$), Euonymus japonica Thunb. extract ($100{\mu}g/mL$), Suaeda malacosperma H.Hara extract ($100{\mu}g/mL$), Elaeagnus umbelellata Thunb. extract ($100{\mu}g/mL$), Sedum oryzifolium Makino extract ($100{\mu}g/mL$), Vicia unijuga A. Br. extract ($100{\mu}g/mL$), and Brassica juncea var. integrifolia Sinsk. extract ($100{\mu}g/mL$) showed more than 140% collagen production effect. Among the 10 species of marine plants, Sargassum macrocarpum C. Agardh extract ($50{\mu}g/mL$) produced more than 190% of collagen, and Carpopeltis angusta (Harvey) Okamura extract ($100{\mu}g/mL$), Codiumcoactum Okamura extract ($100{\mu}g/mL$), and Codium tenuifolium S. Shimada, T. Tadano & J. Tanaka extract ($100{\mu}g/mL$) showed more than 140% collagen production. Suaeda malacosperma H.Hara extract ($100{\mu}g/mL$) showed the effect of producing hyaluronic acid more than 140%, and Ftsia japonica Decne. et Planch. extract ($20{\mu}g/mL$) and Wistaria floribunda A.P. DC extract ($100{\mu}g/mL$) showed more than 130% hyalunonic acid production effect. Among the 10 species of marine plants, Peyssonnelia capensis Montagne extract ($100{\mu}g/mL$) was the most effective. Carpopeltis angusta (Harvey) Okamura extract ($100{\mu}g/mL$), Codiumcoactum Okamura extract ($100{\mu}g/mL$), and Codium tenuifolium S. Shimada, T. Tadano & J. Tanaka extract ($100{\mu}g/mL$) showed more than 120% hyalunonic acid production. Jeju resources, which have good collagen and hyaluronic acid production, showed the potential to be applied to solve the skin troubles of customized cosmetics in the future.

A Study of Guidelines for Genetic Counseling in Preimplantation Genetic Diagnosis (PGD) (착상전 유전진단을 위한 유전상담 현황과 지침개발을 위한 기초 연구)

  • Kim, Min-Jee;Lee, Hyoung-Song;Kang, Inn-Soo;Jeong, Seon-Yong;Kim, Hyon-J.
    • Journal of Genetic Medicine
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    • v.7 no.2
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    • pp.125-132
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    • 2010
  • Purpose: Preimplantation genetic diagnosis (PGD), also known as embryo screening, is a pre-pregnancy technique used to identify genetic defects in embryos created through in vitro fertilization. PGD is considered a means of prenatal diagnosis of genetic abnormalities. PGD is used when one or both genetic parents has a known genetic abnormality; testing is performed on an embryo to determine if it also carries the genetic abnormality. The main advantage of PGD is the avoidance of selective pregnancy termination as it imparts a high likelihood that the baby will be free of the disease under consideration. The application of PGD to genetic practices, reproductive medicine, and genetic counseling is becoming the key component of fertility practice because of the need to develop a custom PGD design for each couple. Materials and Methods: In this study, a survey on the contents of genetic counseling in PGD was carried out via direct contact or e-mail with the patients and specialists who had experienced PGD during the three months from February to April 2010. Results: A total of 91 persons including 60 patients, 49 of whom had a chromosomal disorder and 11 of whom had a single gene disorder, and 31 PGD specialists responded to the survey. Analysis of the survey results revealed that all respondents were well aware of the importance of genetic counseling in all steps of PGD including planning, operation, and follow-up. The patient group responded that the possibility of unexpected results (51.7%), genetic risk assessment and recurrence risk (46.7%), the reproduction options (46.7%), the procedure and limitation of PGD (43.3%) and the information of PGD technology (35.0%) should be included as a genetic counseling information. In detail, 51.7% of patients wanted to be counseled for the possibility of unexpected results and the recurrence risk, while 46.7% wanted to know their reproduction options (46.7%). Approximately 96.7% of specialists replied that a non-M.D. genetic counselor is necessary for effective and systematic genetic counseling in PGD because it is difficult for physicians to offer satisfying information to patients due to lack of counseling time and specific knowledge of the disorders. Conclusions: The information from the survey provides important insight into the overall present situation of genetic counseling for PGD in Korea. The survey results demonstrated that there is a general awareness that genetic counseling is essential for PGD, suggesting that appropriate genetic counseling may play a important role in the success of PGD. The establishment of genetic counseling guidelines for PGD may contribute to better planning and management strategies for PGD.

Experimental Studies on the Antitumor Effects of Jinryungtang Gagambang Extract (진령탕가감방의 항종양효과(抗腫瘍效果)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Jeong, Jun-Tak;Moon, Goo;Moon, Suk-Jae
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.4 no.1
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    • pp.37-53
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    • 1998
  • The sprig of Jinryungtang Gagambang has been used for curing as a traditional medicine without any experimental evidence to support the rational basis for their clinical use. This experiment was carried out to evaluate the possible therapeutic or antitumoral effects of Jinryungtang Gagambang extract against cancer, and to study some mechanisms responsible for its effect. The cytotoxic and antitumor effects were evaluated on human cell liens (A549, hep3B, Caki-1, Sarcoma 180) after exposure to Jinryungtang Gagambang extract using in ILS, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. As a result of exposure to Jinryungtang Gagambang extract, the proliferation of A549, hep3B, Caki-1, good correlations were shown from the results of SRB assay and those of clogenetic assay. 2. The oral administration of Jinryungtang Gagambang extract showed significant effects of increase of MST(mean survival time) and ILS(increased life span) depending on the increasing concentration. 3. Against squamous cell carcinoma induced by MCA, Jinryungtang Gagambang decreased not only the frequency of tumor production but also the number and weight of tumors per tumor bearing mice(TBM). Jinryungtang Gagambang also significantly suppressed the development of 3LL cell-implanted tumors by frequency and their size, and some developed tumors were regressed by the continuous treatment of Jinryungtang Gagambang extract into TBM. 4. Jinryungtang Gagambang extract also increased NK cell activities. According to the above results, it could be suggested that Jinryungtang Gagambang extract has prominent antiutmor effect.

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