• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 1998년도 춘계학술대회 및 워크숍
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• pp.12-28
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• 1998

The technology of creating transgenic animals has a potential value in improving productivity and disease resistance of animals, gene therapy, drug pharming and production of model animals for certain diseases. Up to date, fairly low success rate of production of transgenic animals and a pronounced variability with respect to the expression of transgenes have been much observed. The mechanisms how to integrate the injected genes with a certain part of the genomes are unknown yet. Many techniques in gene transfer, beside microinjection, have been introduced and explored thus to improve the production efficiency of transgenic animals. In this article, the methods and efficiency of gene-transfer techniques, the detection and preselection of transgenes in embryos by PCR- and GFP-screenings and cloning of preselected transgenic embryos by nuclear transplantation are described and discussed. Some experimental results showed that the early screening and selection of integration of the injected gene with embryonic genome by polymerase chain reaction(PCR) and green fluorecence protein(GFP) were promising methods. Further, the application of nuclear transplantation technology to cloning and multiplication of the positively integrated genes in the cleaving embryos and embryonic cells will be beneficially used for the mass production of transgenic embryos and consequently improving the production efficiency in transgenic animals.

• Journal of Pharmaceutical Investigation
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• 제35권1호
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• pp.17-23
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• 2005

Polyethylenimine (PEI) has been used as cationic polymers for efficient gene transfer without the need for endosomolytic agents. Various kinds of PEIs with different molecular weight were tested in order to investigate the effects of the molecular weight of PEI on the transfection efficiency and cell cytotoxicity. The ${\beta}-galactosidase$ expression $(pCMV-{\beta}-gal)$ plasmid was used as a model DNA. Complex formation between PEI and pDNA was assessed by 1% agarose gel electrophoresis method. Particle size and zeta-potential of complexes were determined by electrophoretic light scattering spectrometer. In vitro transfection efficiency was assayed by measuring ${\beta}-galactosidase$ activity. Cell cytotoxicity was determined by MTT assay. Particle sizes of the complexes became smaller on increasing molecular weights of PEI and N/P ratios. Surface potential of complexes was increased as the molecular weight of PEI increased. Transfection efficiency of $pCMV-{\beta}-ga1$ on the HEK 293 cells was greatest with PEI 25 K system but having the lowest cell viability. PEI with high molecular weight showed higher transfection efficiency and cell viability than PEI with low molecular weight.

• Archives of Pharmacal Research
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• 제20권4호
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• pp.297-305
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• 1997

A new serum-free defined medium was developed that supports the growth of normal rat mammary epithelial cells. Mammary organoids from the glands of female F344 rats were cultured in a serum-free medium. Monolayer culture colonies developed within a week and remained viable for months in culture. Upon subculture of one-week-old primary colonies, almost the same morphology of colonies was developed. The scrape loading/dye transfer technique showed that most of colonies that developed in a serum-free medium containing EGF, human transferrin, insulin, and hydrocortisone (basal serum-free medium, BSFM) failed to show cell-cell communication. However, colonies cultured in BSFM supplemented with prolactin, $E_2$, and progesterone (complete hormone serum-free medium, CHSFM) showed cell-cell communication at 14 days of primary culture or of subculture. By flow cytometry with FITCPNA and PE-anti-Thy-1.1 monoclonal antibody, we distinguished four RMEC subpopulations in cultures in both media: Thy-1.1+ cells, PNA+ cells, cells negative to both reagents and cells positive to both reagents. It is likely that combined prolactin, cortisol, and insulin in CHSFM stimulate terminal differentiation of clonogenic cells.

• 약학회지
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• 제49권3호
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• pp.217-224
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• 2005

Human Immunodeficiency Virus type 1 (HIV-l) based lentivirus vector has demonstrated great potential as gene therapy vectors mediating efficient gene delivery and long-term transgene expression in both dividing and nondividing cells. However, for clinical studies it must be confirmed that vector preparations are safe and not contaminated by replication competent lentivirus (RCL) related to the parental pathogenic virus, HIV-l. In this study, we would like to establish the method for titration and RCL detection of lentivirus vector. The titration was determined by vector expression containing the green fluorescent protein, GFP in transduced cells. The titer was $1{\times}10^7$ Transducing Unit/ml in the GFP expression assay and $8.9{\times}10^7$ molecules/ml in the real-time PCR. Also, for the detection of RCL, we have used a combination method of PCR and p24 antigen detection. First, PBS/psi and VSV-G region in the genomic DNA of transduced cells was detected by PCR assay. Second, transfer and expression of the HIV-1 gag gene was detected by p24 ELISA. In an attempt to amplify any RCL, the transduced cells were cultured for 3 weeks (amplification phase) and the supernatant of amplified transduced cell was used for the second transduction to determine whether a true RCL was present (indicator phase). Analysis of cells and supernatant at day 6 in indicator phase were negative for PBS/psi, VSV-G, and p24 antigen. These results suggest that they are not mobilized and therefore there are no RCL in amplification phase. Thus, real-time PCR is a reliable and sensitive method for titration and RCL detection of lentivirus vector.

• 약학회지
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• 제34권1호
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• pp.54-63
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• 1990

These studies were made to assess the present stage of resistance of Shigella species to antibiotics and to find characteristics of R-plasmid of these bacteria. From 1986 to 1988, 125 strains of Shigella species were isolated from patients specimens collected in Chung Cheong-do Hospital, Kyunghee Medical Center, city an provincial health & environmental institutes. These specimens were tested for resistance to 12 kinds of antimicrobial agents by agar dilution method. Using Muller-Hinton agar for the assay of drug resistance and Trypticane Soy Broth as propagating medium for conjugation. All the strains (100%) were resistant to one or more antibiotics. Drug resistance patterns of isolated strains were found as the highest resistance to ampicillin (98%) in 1986, to tetracycline (98%) in 1987, to tetracycline (100%) in 1988, all strains were sensitive to gentamicin, amikacin, tobramycin. Chronologically, resistance decreased gradually as it was shown in relation to kanamycin, rifampicin in 1986, 1987 and 1988, (4%, 2%) (4%, 2%) (0%, 0%) respectively. But, resistance was increased year by year as it was shown in relation to tetracycline, nalidixic acid, streptomycin in 1986, 1987, 1988 (89%, 19%, 45%) (98%, 46%, 71%) and (100%, 58%, 88%). The resistance in correlation to more than 5 drugs, which was 13 strains among 47 strains in 1986, 38 strains among 87 strains in 1987, 23 strains among 26 strains in 1988, was increased gradually. In the transfer test of drug resistance by conjugation methods, the rate which was 3 strains (50%) in 1986, 8 strains (62%) in 1987, 3 strains (100%) in 1988, was increased gradually. When the donor strains were conjugated with the recipient strains, the conjugation rate was high in the multiple resistant strains. The relationships of transferring patterns of drug resistance and molecular weight of R-plasmid were variable. However, only a plasmid which has more than 35 Mgd was transferred.

• 한국동물위생학회지
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• 제15권2호
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• pp.134-143
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• 1992

The present study was conducted to investigate the biochemical characteristics and anti-biotic resistance of Escherichia coli(E. coli) isolated from piglets with diarrhea in Kyongbuk province during the Period from February to November 1991. 368 E. coli strains were isolated from 382 piglets with diarrhea and the biochemical and cultural reaction were compared with the classification criteria of Edwards and Ewing. Tetracycline and sulfadimethoxine were found to be highly ineffective at in vitro inhibition of the E. coli of piglets origin. The majority of E. coli were susceptible to amikacin, chloramphenicol and gentamicine. 89 (89.0%) of 100 strains of E. coil were resistant to one or more drugs. The organisms resistant to 20 or 3 drugs were 54(60.6%) of 89 strains, whereas 16(17.9%) strains were found to be resistant to one drug. 55(61.8%) out of 89 drug resistance strains carried R factors($R^+$) which were transfer-able to the recipients by conjugation.

• 한국추진공학회:학술대회논문집
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• 한국추진공학회 2008년 영문 학술대회
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• pp.566-569
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• 2008

Recently, the biolistic process is emerging as an effective needle-free drug delivery technique to transfer adequate concentrations of pharmacologic agents to soft living tissues with minimum side effects. We have started developing an effective method for delivering drug coated particles using laser ablation. A thin metal foil with deposited micro-particles on one side is irradiated with laser beam on the opposite side so that a shock wave is generated. This shock wave travels through the foil and is reflected, which causes and instantaneous deformation of the foil. Due to such a sudden deformation, the micro-particles are ejected at a very high speed. Here we present the experimental results of direct and confined laser ablation, which correspond to the initial stage of the whole experiment.

• 한국벤처창업학회:학술대회논문집
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• 한국벤처창업학회 2022년도 춘계학술대회
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• pp.25-30
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• 2022

Despite the importance of partnership for commercialization of innovations in startups, it is not easy for startups to persuade an established firm to collaborate on a completely novel idea. If information transfer about the innovations is too costly, startups may avoid pursuing radically new projects. Our paper examines the impact of policy signals on the novelty of the innovations pursued by startups. In the context of the Orphan Drug Act(ODA), we find that startups develop more radical therapies when policy signals help them to convince potential partners of the value of prospective therapies. While the likelihood of partnership increases, the timing of partnership is delayed in ODA-affected fields.

• Tuberculosis and Respiratory Diseases
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• 제42권3호
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• pp.302-313
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• 1995

연구배경: 종양괴사인자(Tumor necrosis factor; TNF)는 다양한 생물학적인 작용을 가지며 종양 세포에 대한 세포 독성은 그 대표적인 기능중의 하나이다. TNF-$\alpha$는 생체외에서(in vitro) 몇몇 종양 세포주에 대하여 항암제, 특히 topoisomerase II targeted chemotherapeutic agent의 세포 독성 효과를 상승적으로 증가시키는 것이 알려져 있다. 최근 암세포에 대한 cytokine 유전자 요법에서 TNF는 중요한 대상으로 여겨지고 있으며, 유전자 이입에 의해 암조직이 TNF를 생성하게 될 경우 암 증식 억제 효과가 있음이 보고되고 있다. 연구자는 암세포에 TNF-$\alpha$ 유전자를 이입하여 자신이 TNF-$\alpha$를 생성하도록 형질을 변환시킨 암세포는 topoisomerase II 억제 항암제에 대한 김수성에 변화가 있을 것이라는 가설을 수립하였고 이를 검증하고자 본 연구를 수행하였다. 본 연구에서는 생체외로(in vitro) TNF-$\alpha$ 유전자를 이입하여 TNF-$\alpha$를 생성하는 암세포주에서 topoisomerase II targeted drug에 대한 항암제 감수성 효과가 모세포주에 비하여 증대될 수 있는지를 알아 보고자하였다. 방법: TNF-$\alpha$에 감수성을 보이는 것으로 알려진 인체 중피종 세포주인 NCI-H2058 세포주 및 생쥐의 섬유육종 세포주인 WEHI164 세포주와 인체 비소세포 폐암 세포주인 A549 세포주를 배양하여, 먼저 임상에서 흔히 폐암의 항암 화학 요법 치료에 널리 쓰이는 대표적인 topoisomerase II targeted chemotherapeutic drug인 etoposide(VP-16)와 doxorubicin(adriamycin)을 가하였을 때 관찰된 세포 독성을 MTT assay로 측정하고, 각 모세포주(parenta1 cell line)에 TNF-$\alpha$의 유전자를 이입시켜서 형절 변환한 세포주(transformed cell line)에 대하여 각각 동일한 항암제를 가하였을 때 관찰된 세포 독성의 정도를 같은 방법으로 측정하여, 그 결과를 비교 분석하였다. 또한 모세포주에 외부에서 TNF를 가하여 전처치한 후 동일한 항암제를 가하였을 때의 세포독성을 관찰하여 비교 분석하였다. 결과: H2058 세포주에서는 TNF-$\alpha$ 유전자를 이입한 세포주 topoisomerase II targeted drug을 가하였을 때, 항암제 감수성이 모세포주에 같은 항암제를 가하였을 때에 비하여 의미있게 증가함을 관찰할 수 있었으나(p<0.05), WEHI 세포주와 A549 세포주에 있어서는 TNF-$\alpha$ 유전자를 이입한 세포주에서 모세포주에 비하여 항암제 감수성이 증가하지는 않았다. 결론: TNF-$\alpha$ 유전자의 이입이 topoisomerase II targeted chemotherapeutic drug에 대한 항암제 감수성을 증가시키는 효과는 세포주에 따라 다양한 결과를 보이는 것을 알 수 있었으며, 적어도 선택된 특정 종류의 호흡기계 암세포에 있어서는 TNF-$\alpha$ 유전자의 이입으로 항암제 감수성(chemosensitivity)을 증가시킬 수 있을 것으로 사료된다.

• 약학회지
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• 제43권4호
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• pp.437-441
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• 1999

Enoxacin[1-ethyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-\piperazinyl)-1,8-naphthyridine-3-carboxylic acid, ENX] is a new quinolone antibacterial agent. The method is based on the highly colored charge-transfer complex formation of this drug as a $\pi$-electron donor with 7,7,8,8-tetracyanoquinodimethane(TCNQ) or chloranil(CL) as $\pi$-acceptors. The colored products were measured spectrophotometrically at 842 nm and 552 nm for TCNQ and CL, respectively. The different experimental conditions are optimized. The linearities for TCNQ and CL were $1.6{\;}\mu\textrm{g}/mL~32{\;}\mu\textrm{g}/mL$ and $6.4{\;}\mu\textrm{g}/mL~160{\;}\mu\textrm{g}/mL$, respectively and colors were produced in non-aqueous media. This report describes a simple and ra\pid method for the analysis of enoxacin.