• Title/Summary/Keyword: ln vitro

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Inhibitory Effect of Linolenic Acid on the Mutagens-Induced Mutagenicities in Ames Assay System and SOS Chromotest (Ames 실혐계 및 SOS Chromotest에서 Linolenic acid의 돌연변이유발 억제효과)

  • 임선영;이슥희;박건영
    • Journal of Life Science
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    • v.5 no.3
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    • pp.121-125
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    • 1995
  • To determine whether the omega 3 family, linolenic acis(LnA) is effective to inhibit carcinogens/mutagens-induced mutagenesis, we employed the Ames test using Salmonella typhimurium strain of TA100 and the SOS chromotest using Escherichia coli PQ37 strain. The inhibitory effect of LnA shown in the Ames assaying system was 95%, 78% and 73% when the mutagenicities were mediated by AFB$_{1}$, MNNG and 4-NQO, respectively. LnA shows a strong antimutagenic activity against indirect mutagen of AFB$_{1}$, whereas the same concentration of LnA exhibited weaker inhibitory effects on the direct mutagen of MNNG and 4-NQO than that of AFB$_{1}$. However. LnA reduced more than 80% of SOS responses induced by MNNG and 4-NQO when the adding concentration increased to 5%. We conclude that LnA contains in vitro antimutagenic properties and that this finding warrants further investigation both in vitro and in vivo to assess its possible chemotherapeutic potential.

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Establishment of an In Vitro TCD (Testosterone Compound Detection) System (테스토스테론 물질 검출을 위한 in vitro TCD 시스템 구축)

  • Lee, Dong-Geun;Jo, Jung-Kwon;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.29 no.10
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    • pp.1159-1163
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    • 2019
  • Although there is a growing interest in male menopause, a phenomenon associated with male hormone depletion, current kits using antibodies to quantify male hormones are expensive. In this study, we constructed an in vitro system for verifying the activity or concentration of male steroid hormones using a transcriptional activity test. A reporter plasmid, pGL2-Neo-ARE-AdE1BTATA, which reacts to testosterone, was constructed. In this plasmid, the ARE-AdE1bTATA sequences can be bounded by the testosterone - androgen receptor complex to express luciferase as a reporter. Then, a stable transfection was performed on the human prostate cancer cell line, LNcap-LN3. The constructed LNcap-LN3/pGL2-Neo-ARE-AdE1BTATA testosterone compound detection (TCD) system showed quantitatively proportional luciferase activities to concentrations of $10^{-13}$ to $10^{-8}M$ of standard testosterone. The established in vitro TCD system will contribute to the development of materials for health/functional foods and drugs as it will be possible to search en masse for testosterone-like or testosterone-inhibiting substances derived from natural materials.

Effect of Gal-Geun-Tang on Antigen-Specific Immune Response (갈근탕(葛根湯)이 면역조절작용에 미치는 영향)

  • Cho, Dae-Yeoun;Yun, Young-Gab;Jung, Myung;Lee, Eun-Hye;Bok, Young-Ok;Jung, Chang-Ohk;Lim, Kyu-Sang
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.29 no.3
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    • pp.134-149
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    • 2016
  • Objectives : Gal-Geun-Tang (GT) has been described from SANGHAN in Korean traditional medicine and known to act against cold, fever, hypertension, and nasal catarrh. However, little has yet been learned about the effect of GT on immune function. In the current study, in vitro and in vivo immunomodulatory activity of GT (water extract) was investigated.Methods : Water extract of GT induced in vitro proliferation of spleen cells and significantly increased their proliferative responses during anti-CD3 activation. Using purified splenic T and B cells, it was revealed that GT has a mitogenic activity to B cells and promotes their proliferation induced by lipopolysaccharide, whereas T cell proliferation was not triggered and GT was rather inhibitory to T cell activation caused by anti-CD3 antibody. In the presence of antigen presenting cells (APC), GT addition resulted in a significant increase of IFNγ and IL-4, but not IL-2, production. However, addition of high concentration (1,000㎍/㎖) of GT led to a marked reduction in T cell cytokine production and under such condition, GT facilitated apoptosis of T cells when examined by flow cytometry with propidium iodide staining.Results : In vivo immunomdulation of GT was also investigated using a mouse model. Following keyhole limpet hemocyanin (KLH) immunization, GT (1 ㎎/day) was orally administered for 9 days. Cell numbers in thymus, spleen and peripheral blood were not altered by GT administration, indicating that such dose is not immunotoxic. Cell numbers in draining lymph nodes (LN) and ex vivo Ag-specific proliferation of LN cells were significantly elevated by GT administration. However, any preferential stimulation of T or B and CD4+ or CD8+ T cell subpopulations was not observed in a flow cytometric analysis of LN cells. This result shows that GT does not promote in vivo B cell proliferation while GT enhances Ag-specific proliferation of LN cells, unlike what was observed in vitro.Conclusions : For a further understanding of in vivo immunomodulatory activity of GT, ex vivo cytokine production of LN cells obtained from KLH-immunized mice was evaluated. Ag-specific IFNγ production was significantly higher in GT-treated mice when compared to PBS-treated control mice. In contrast, IL-4 production in GT-treated group was comparable to control group unlike to in vitro data. In addition, GT administration did not result in any significant differences in serum levels of Ig (IgM, IgG1 and IgG2a) between GT-treated and control groups. Taken together, these data strongly support that GT promotes immune response, more profoundly type 1 helper T cell (Th1) activity and GT may be applicable for treatment of intracellular parasite infection such as viral diseases.

Lipid nanodispersion for parenteral drug delivery: in vitro characterization

  • Lee, Jung-Min;Choi, Sung-Up;Lee, Byoung-Moo;Lee, Sung-Jae;Choi, Young-Wook
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.295.2-295.2
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    • 2003
  • Lipid nanodispersion (LN) composed of biocompatible lipids and surfactants is an alternative parenteral drug delivery system especially for lipophilic drugs. It has been studied for versatile applications such as oral, parenteral, topical, ocular, vaccine, and peptide drug delivery. The purpose of this study was to produce a novel LN system for intravenous injection using the high pressure homogenization. (omitted)

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In vitro Anticancer Activity of Paclitaxel Incorporated in Low-melting Solid Lipid Nanoparticles

  • Lee, Mi-Kyung;Yang, Jae-Heon
    • Journal of Pharmaceutical Investigation
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    • v.39 no.3
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    • pp.201-205
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    • 2009
  • Triglyceride solid lipid with medium chain fatty acid, tricaprin (TC), was used as a core matrix of lipid nanoparticles (LN) to solubilize water-insoluble paclitaxel and enhance the stability of nanoparticles by immobilization of incorporated drug in the solid core during storage at low temperature. In the present study, TC-LN containing paclitaxel was prepared by hot melt homogenization method using TC as a core lipid and phospholipids as stabilizers. The particle size of TC-LN containing paclitaxel was less than 200 nm and its zeta potential was around -40 mV. Calorimetric analysis showed TC core could be solidified by freezing and thawing in the manufacturing process in which the hot dispersion should be prepared at elevated temperature and subsequently cooled to obtain solid lipid nanoparticles. The melting transition of TC core was observed at $27.5^{\circ}C$, which was lower than melting point of TC bulk. The particle size of TC-LN remained unchanged when kept at $4^{\circ}C$. Paclitaxel containing TC-LN showed comparable anticancer activity to the Cremophore ELbased paclitaxel formulation against human ovarian (OVCAR-3) and breast (MCF-7) cancer cell lines. Thus, lipid nanoparticles with medium chain solid lipid may have a potential as alternative delivery system for parenteral administration of paclitaxel.

Pharmacokinetic Behavior and Biodistribution of Paclitaxel-Loaded Lipid Nanosuspension

  • Choi, Sung-Up;Park, Jung-Min;Choi, Woo-Sik;Lee, Jae-Hwi;Choi, Young-Wook
    • Journal of Pharmaceutical Investigation
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    • v.39 no.5
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    • pp.359-366
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    • 2009
  • In this study, paclitaxel-loaded lipid nanosuspension (PxLN) was prepared and the in vivo profiles after intravenous administration in rats were investigated. We compared the manufacturing processes depending on the temperature: PxLN-H for a hot homogenization process and PxLN-C for solidification of lipid-drug mixtures by liquid nitrogen. Both formulations showed submicron size distribution and the similar drug loading efficiency of about 70%. In vitro release of PxLNs and Taxol$^{(R)}$ performed by a dialysis diffusion method showed similar pattern for PxLN-H and Taxol$^{(R)}$, but the reduced release profile for PxLN-C. PxLN or Taxol$^{(R)}$ was intravenously administered to the rats at a dose of 5 mg/kg as paclitaxel. The drug in blood samples were assayed by the HPLC/MS/MS method. The AUC$_t$ of PxLN-H was 3.4-fold greater than that of Taxol$^{(R)}$. PxLN-H gave higher biodistribution in all tissues than did Taxol$^{(R)}$. In addition, it maintained the higher drug concentration for 12 h. This lipid nanosuspension might be a promising candidate for an alternative formulation for the parenteral delivery of poorly water-soluble paclitaxel.

In Vitro Effect of Yuza (Citrus junos SIEB ex TANAKA) Extracts on Proliferation of Human Prostate Cancer Cells and Antioxidant Activity (In vitro 상에서 유자(Citrus junos SIEB ex TANAKA) 용매 추출물의 암세포 억제효과 및 항산화성)

  • Yoo, Kyung-Mi;Hwang, In-Kyeong
    • Korean Journal of Food Science and Technology
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    • v.36 no.2
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    • pp.339-344
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    • 2004
  • Effects of 18 kinds of yuza extracts on viability of prostate cancer cells, DU 145 and LN-CaP, were investigated. Chloroform and methanolic extracts of yuza peel exhibited moderate cytotoxicity against both cancer cell lines dose-dependently and also showed antioxidant activity matching on inhibition of cell viability (author's intension not clear). Chloroform extract of yuza peel exhibited highest radical-scavenging activity and cytotoxicity against prostate cancer cells in vitro.

Regeneration of Cryopreserved Pear Shoot Tips Grown in Vitro by Encapsulation-Dehydration

  • Yi, JungYoon;Lee, YoungYi;Lee, GiAn;Son, EunHo;Park, HongJae
    • Korean Journal of Plant Resources
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    • v.30 no.6
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    • pp.612-617
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    • 2017
  • The preservation of pear germplasm, like that of other clonal germplasms, is difficult because it requires conservation of whole plants or their tissues. Among the currently available methods for long-term conservation of clonal germplasm, cryopreservation of shoot tips is the most reliable and cost- and space-effective option. Alginate-coated axillary shoot tips from in vitro-grown pear were conserved successfully in liquid nitrogen (LN) following dehydration. Shoot recovery from cryopreserved shoot tips was improved greatly after 8 weeks of cold acclimation, but recovery decreased slightly after then. The highest regeneration rate was observed when in vitro shoot tips were preincubated in MS (Murashige and Skoog) medium with 0.3 M sucrose for 48 h, and when alginate-coated shoot tips were precultured in MS medium with increasing sucrose concentrations (0.5 M and 0.7 M) for 8 and 16 h, respectively. When the encapsulated beads were dehydrated for up to 7 h [25% water content (fresh weight basis)] under laminar flow, the highest regeneration rate was observed in "BaeYun No. 3" (55.7%) and "Whanggeum" (43.3%) after warming from LN. This technique is useful as a practical procedure to cryopreserve plant material that is sensitive to freezing of the surrounding cryoprotectant medium. Therefore, this technique appears to be promising for the cryopreservation of shoot tips from in vitro-grown plantlets of pear germplasm.

Effects of cryoprotectants and sucrose concentrations on the viability of aggregated mouse embryos frozen rapidly in liquid nitrogen vapour (동결보호제 및 Sucrose 농도가 급속동결한 마우스 집합배의 생존성에 미치는 영향)

  • Shin, Sang-tae
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.523-527
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    • 1991
  • The effects of ethylene glycol, DMSO and glycerol as cryoprotectants and the effect of concentrations(0, 0.25, 0.5 and 1.0M) of sucrose in the diluent on the viability of the aggregated morulae frozen rapidly in liquid nitrogen$(LN_2)$ vapour were examined. The morulae were produced by aggregation of ICR and CBA mice embryos at 8-cell stage. Before freezing the embryos were equilibrated in 1.5M cryoprotectants+0.25M sucrose in oae-step or in 3.0M cryoprotectants+0.25M sucrose in two-steps. The embryos were pipetted into the freezing medium fraction of 0.25ml plastic straws. The straws were frozeu by directly transfer into $LN_2$ vapour(about lcm above $LN_2$) for 2 minutes, and then plunged into $LN_2$. After thawing the cryoprotectants were diluted with 0, 0.25, 0.5 or 1.0M sucrose solution. The post-thawed in vitro viability of the aggregated embryos was significantly dependent on the type and concentration of cryoprotectants in the freezing medium and also on the concentration of sucrose in the diluent. When the aggregated embryos were equilibrated in 1.5M cryoprotectants +0.25M sucrose in one-step and diluted with 0.5M sucrose after thawing, the survival rate of the embryo5 was significantly(p<0.05) higher in DMSO(62.5%) or ethylene glycol(52.2%) than in glycerol(33.3 %). In the case that the concentration of the cryoprotectants was raised to 3.0M in two-steps, thc higher survival rate of the embryos was obtained in ethylene glycol or glycerol than in DMSO followed by diluting them with 0.5 or 1.0M sucrose after thawing(p<0.01).

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Synthesis and evaluation of 64Cu-labeled avidin for lymph node imaging

  • Kang, Choong Mo;Kim, Hyunjung;Lee, Yong Jin;Choe, Yearn Seong
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.5 no.1
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    • pp.54-60
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    • 2019
  • Sentinel lymph node (SLN) imaging plays an important role in surgery of patients with breast cancer and melanoma. In this study, avidin (Av), a tetrameric protein glycosylated with mannose and N-acetylglucosamine molecules, was labeled with $^{64}Cu$ and then evaluated for LN imaging. $^{64}Cu$-Labeled $NeutrAvidin^{TM}$ (NAv), a non-glycosylated form of Av, was used for comparison. 1,4,7,10-Tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA)-conjugated Av and NAv were prepared from the corresponding proteins and DOTA-NHS ester, which were then labeled with copper-64 and purified using PD-10 columns. The numbers of DOTA molecules conjugated to Av and NAv were 4.9 and 3.3, respectively. [$^{64}Cu$]Cu-DOTA-conjugated Av and NAv were prepared in 93% and 73% radiochemical yields, respectively. In vitro serum stability study showed that copper-64 remained stable on all radiotracers for 24 h (>97%). MicroPET/CT images showed that high radioactivity was accumulated in LNs within 15 min after footpad-injection of radiotracers. Tissue distribution data of mice demonstrated significantly higher uptake in the popliteal (PO) LN than lumbar (LU) LN for $^{64}Cu$-labeled Av (relative % ID/g excluding the injection sites: 66.2% and 26.0%, respectively) compared with those of $^{64}Cu$-labeled NAv (43.0% and 49.2%, respectively). The results of this study suggest that mannose molecules on Av enabled the radiotracer to retain in the first LN after mouse footpad-injection.