• Title/Summary/Keyword: In-vitro study

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Impact of Sodium Copper Chlorophyllin on Mercury Absorption Using an in Vitro Digestion with Human Intestinal Cell Model

  • Hwang, Han-Joon;Shim, Soon-Mi
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.564-568
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    • 2008
  • The effects of sodium copper chlorophyllin (SCC) on bioaccessibility and uptake of mercury from fish were investigated using an in vitro digestion coupled with a Caco-2 cell. Fish along with SCC was subjected to a simulated in vitro digestion, which simulates both the gastric and small intestinal phase in vivo. Mercury bioaccessibility, the amount of mercury released from fish to aqueous phase following a digestion, was measured. Various amounts of SCC (0.1-25 mg) significantly reduced mercury bioaccessibility in a dose dependent manner by 49-89% compared to the negative control (fish without SCC) (p<0.05). Mercury bioaccessibility in varying molar ratios of mercury to positive control, 2,3-dimercapto-1-propane sulfonate (DMPS) was between 24 and 52%. Mercury uptake by Caco-2 cells from test media containing aqueous phase following in vitro digestion was measured after 6 hr incubation at $37^{\circ}C$. Cellular mercury uptake with increasing amount of SCC ranged from 0.352 to $0.052\;{\mu}g$ mercury/mg protein, while those in DMPS treatment were between 0.14 and $0.27\;{\mu}g$ mercury/mg protein. Our study suggests that SCC can reduce mercury absorption following fish consumption and may be efficient as a synthetic chelating agent for long term chronic mercury exposure in fish eating populations.

Effects of Pretense Inhibitors and Antioxidants on In Vitro Survival of Porcine Primordial Germ Cells

  • Lee, Chang-Kyu;Jorge A. Piedrahita
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2001.10a
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    • pp.47.1-47
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    • 2001
  • One of the problems associated with in vitro culture of primordial gern cells (PGCs) is the large loss of cells during the initial period of culture. This study characterized the initial loss and determined the effectiveness of two classes of apoptosis inhibitors, protease inhibitors and antioxidants, on the ability of the porcine PGCs to survive in culture. Results from electron microscopic analysis and in situ DNA fragmentation assay indicated that porcine PGCs rapidly undergo apoptosis when placed in culture. Additionally, \ulcorner2-macroglobulin, a protease inhibitor and cytokine carrier, and N-acetylcysteine, an antioxidant, increased the survival of PGCs in vitro. While other protease inhibitors tested did not affect survival of PGCs, all antioxidants tested improved survival of PGCs (p<0.05). Further results indicated that the beneficial effect of the antioxidants was critical only during the initial period of culture. Finally, it was determined that in short-term culture, in the absence of feeder layer, antioxidants could partially replace the effect(s) of growth factors and reduce apoptosis. Collectively, these results indicate that the addition of \ulcorner2-macroglobulin and antioxidatns can increase the number of PGCs in vitro by suppressing apoptosis.

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In Vitro Production of Pig Embryos using Intracytoplasmic Injection of Flow Cytometry Sorted Boar Spermatozoa

  • Kim, Dae-Young;Hyun, Sang-Hwan;Lee, Eun-Song
    • Journal of Embryo Transfer
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    • v.23 no.4
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    • pp.275-281
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    • 2008
  • The ability to preselect the sex of piglets is advantageous in the pig industry. The objective of this study was to examine the feasibility of using intracytoplasmic sperm injection (ICSI) with sorted spermatozoa to produce piglets with a preselected sex. Pig embryos were produced by ICSI of frozen X- and Y-sperm that had been separated by flow cytometry. The developmental competence of the embryos was investigated in vitro and in vivo. The populations of X- and Y-spermatozoa were 52.7% and 47.3%, respectively in our samples. The in vitro development of ICSI embryos was enhanced by longer of in vitro maturation of oocytes ($44{\sim}48\;h$ vs. $40{\sim}43\;h$). Their cleavage ($65{\sim}70%$) and blastocyst formation ($9{\sim}12%$) rates were not significantly different between male and female ICSI embryos, or between sorted and unsorted sperm-derived embryos. One pregnancy was established in a recipient that was transferred with 110 female ICSI embryos, but the pregnancy was terminated on Day 89 of gestation. Our results suggest that the separation X- and Y-spermatozoa by flow cytometric sorting can be a useful tool in combination with ICSI for the production of pig embryos and piglets of preselected sex.

Instability in in vitro fruiting of Cordyceps militaris (In vitro에서 큰번데기 동충하초 자실체형성의 불안정성)

  • Shrestha, Bhushan;Park, Young-Jin;Han, Sang-Kuk;Choi, Sung-Keun;Sung, Jae-Mo
    • Journal of Mushroom
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    • v.2 no.3
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    • pp.140-144
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    • 2004
  • Traditionally, Cordyceps species have been used as a part of herbal medicine in Oriental countries, including Korea for internal health, vigor and to cure different diseases related to heart, lung etc. In recent years, research on artificial fruiting of some species of the genus Cordyceps including C. militaris has been carried out extensively because of their medicinal value. Instability observed in the in vitro fruiting of C. militaris is reported in the present study.

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Effects of Protease Inhibitors and Antioxidants on In Vitro Survival of Porcine Primordial Germ Cells

  • Lee, Chang-Kyu;Jorge A. Piedrahita
    • Proceedings of the KSAR Conference
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    • 2001.10a
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    • pp.47-47
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    • 2001
  • One of the problems associated with in vitro culture of primordial germ cells (PGCs) is the large loss of cells during the initial period of culture. This study characterized the initial loss and determined the effectiveness of two classes of apoptosis inhibitors, protease inhibitors and antioxidants, on the ability of porcine PGCs to survive in culture. Results from electron microscopic analysis and in situ DNA fragmentation assay indicated that porcine PGCs rapidly undergo apoptosis when placed in culture. Additionally,? 2-macroglobulin, a protease inhibitor and cytokine carrier, and N-acetylcysteine, an antioxidant, increased the survival of PGCs in vitro. While other protease inhibitors tested did not affect survival of PGCs, all antioxidants tested improved survival of PGCs (p〈0.05). Further results indicated that the beneficial effect of the antioxidants was critical only during the initial period of culture. Finally, it was determined that in short-term culture, in the absence of feeder layers, antioxidants could partially replace the effect(s) of growth factors and reduce apoptosis. Collectively, these results indicate that the addition of ?2-macroglobulin and antioxidants can increase the number of PGCs in vitro by suppressing apoptosis.

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In vitro Nasal Cell Culture Systems for Drug Transport Studies

  • Cho, Hyun-Jong;Termsarasab, Ubonvan;Kim, Jung-Sun;Kim, Dae-Duk
    • Journal of Pharmaceutical Investigation
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    • v.40 no.6
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    • pp.321-332
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    • 2010
  • Growing interest in the nasal route as a drug delivery system calls for a reliable in vitro model which is crucial for efficiently evaluating drug transport through the nasal cells. Various in vitro cell culture systems has thus been developed to displace the ex vivo excised nasal tissue and in vivo animal models. Due to species difference, results from animal studies are not sufficient for estimating the drug absorption kinetics in humans. However, the difficulty in obtaining reliable human tissue source limits the use of primary culture of human nasal epithelial cells. This shortage of human nasal tissue has therefore prompted studies on the "passage" culture of nasal epithelial cells. A serially passaged primary human nasal epithelial cell monolayer system developed by the air-liquid interface (ALI) culture is known to promote the differentiation of cilia and mucin gene and maintain high TEER values. Recent studies on the in vitro nasal cell culture systems for drug transport studies are reviewed in this article.

Sex Determination of In Vitro Fertilized Bovine Embryos by Fluorescence In Situ Hybridization Technique

  • Han, M.S.;Cho, E.J.;Ha, H.B.;Park, H.S.;Sohn, S.H.
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.133-137
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    • 2004
  • Sexing from bovine embryos which were fertilized in vitro implicate a possibility of the sex-controlled cattle production. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. To evaluate Y-chromosome specificity of the FISH probe, metaphase spreads of whole embryos and lymphocytes were prepared and tested. A male-specific signal was detected on 100% of Y chromosome bearing metaphase specimens. Using the FISH technique with a bovine Y-specific probe, 232 whole embryos of 8 cell- to blastocyst-stage were analyzed. Observing the presence of the Y-probe signal on blastomeres, 102 embryos were predicted as male, and 130 embryos as female. The determining rate of embryo sex by FISH technique was about 93% regardless of embryonic stages. In conclusion, the FISH using a bovine Y-specific DNA probe is an accurate, reliable and quick method for determining the sex of bovine embryos.

In vitro Screening of Dietary Factors on Buckwheat(Fagopyrum esculentum Moench) Influencing the Regulation of Blood Pressure, Glucose and Cholesterol Level (혈압, 혈당 및 콜레스테롤농도 조절에 영향을 주는 식이 메밀인자의 in vitro 검정)

  • 최용순;김복란;김려화;이병훈;심태흠;이상영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.2
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    • pp.280-287
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    • 2000
  • Generally, buckwheat has been regarded as a crop of secondary importance in many countries. In vitro functionalities of buckwheats as a food were evaluated in this study. Five of buckwheat cultivars were extracted with methanol, and the extractant were dried and lyophilized, separately. Or water soluble buckwheat components were digested with the commercial enzymes and the obtained protein hydrolysate was again fractionated by acid precipitation. The antioxidant capacity of the methanol extracts determined using Fe2+-ascorbic acid system was dependent ont the cultivars: The extract of Suwon 4 showed 3.3 times stronger activity than ascorbic acid in terms of IC50. Also, the extracts of buckwheats inhibited efficiently the activities of $\alpha$-amylase and lens aldose reductase. Buckwheat soluble protein or rutin suppressed the in vitro activities of angiotensin-converting enzyme, and the inhibitory degree depended largely on the cultivars. Buckwheat proteins exerted higher hydrophobicity being related to the sterol binding capacity than casein. The results suggested that buckwheat seeds may be desirable and functional food resources in human living in current society.

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Comparison of in Vivo, in Vitro 3T MR Spectroscopy and Proton NMR Spectroscopy for the Fluid from Cystic Tumor: Preliminary Study (낭성 종양의 체액에 대한 생체내, 생체외 3T 양성자 자기공명분 광법과 양성자 핵자기공명기법의 비교: Preliminary Study)

  • Lee, Hui-Joong;Kim, Jong-Yeol;Chang, Yong-Min
    • Investigative Magnetic Resonance Imaging
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    • v.12 no.2
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    • pp.107-114
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    • 2008
  • Purpose : The aim of this study is to determine possibility of application of in vivo proton ($^1H$) magnetic resonance spectroscopy (MRS) in distinguishing cystic mass arising around pancreas by comparison of in vivo MRS, in vitro MRS using 3T MR machine, based on nuclear magnetic resonance (NMR). Materials and Methods : We obtained spectra of in vivo MRS, in vitro MRS and NMR from abdominal mass arising around pancreas (mucinous cystic neoplasm=5, intraductal papillary mucin producing tumor=5, pseudocyst=1, and lymphangioma=1). We estimated existence of peak of in vivo MRS, and in vitro MRS concordant to that of NMR. We also evaluated differential peak for predicting specific disease. Results : Correlation of presence of peak with NMR showed showed sensitivity of 29.6%, specificity of 82.6% and accuracy of 67.7% on in vivo MRS (p = 0.096, McNemar test), sensitivity of 57.1% and specificity of 92.6% and accuracy of 82.3% on in vitro MRS (p = 0.362, McNemar test). The spectra of NMR for IPMT showed more frequent peaks at 3.5-4.0 ppm (p=0.026). Conclusion : Although chemical analysis, using NMR could be regarded as possible tool to differentiate cystic masses, in vivo and in vitro MRS need further technical evolution for clinical application.

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In vitro experimental study on flow characteristics of abdominal aorta aneurysm (복부대동맥 동맥류의 유동특성에 관한 in vitro 실험적 연구)

  • Lee, J.P.;Kim, D.S.;Lee, S.J.
    • 한국전산유체공학회:학술대회논문집
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    • 2008.03b
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    • pp.10-12
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    • 2008
  • Hemodynamic features of blood flow in the abdominal aorta aneurysm (AAA) are very important, because they are closely related with the rupture of aneurysm to death. It has been considered that the wall shear stress of blood flows influences the formation, growth, and rupture of AAA. On this account, it is important to understand the flow structure of blood in the aneurysm. In this study, the whole velocity field information inside a typical AAA was measured using an in vitro AAA model under the pulsatile flow condition. The vessel geometry was reconstructed based on the computerized tomography (CT) data of a patient. The AAA model was made by using a rapid prototyping (RP) method, based on the reconstructed vessel geometry. Velocity fields in the AAA model were measured at different pulsatile phases using a PIV (particle image velocimetry) system. As experimental results, a large-scale vortex is formed inside the AAA model and the vortices located near the AAA wall are supposed to increase the local pressure and wall shear stress. In this study, the AAA wall stress found to be was one of the most important governing parameters giving rise to the ruptured aneurysm.

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