• Title/Summary/Keyword: In-vitro study

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A Comparative Study of in vitro Methods on the Phototoxicity of Phenothiazines (Phenothiazines의 광독성에 대한 in vitro 실험법의 비교 연구)

  • 김종예;김현진;김봉희
    • Environmental Analysis Health and Toxicology
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    • v.15 no.1_2
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    • pp.13-18
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    • 2000
  • A few in vitro methods were developed to compare the result on the phototoxicity of phenothiazines. By the MTT assay, the Candida test, and the RBC photohemolysis, the phototoxicities of UVA and UVB irradiation were measured. This paper presents the comparisons of methods which are effective to measure the phototoxicities of the chemicals causing phototoxicity and photoallergy. The tested chemicals of phenothiazines include Chlorpromazine, Promethazine, Perphenazine, Chlorprothixene, Trifluoperazine and Thioridazine. Each chemical represented variable results according to the test methods. MTT assay shows the most sensitive method.

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Fermentation-Mediated Enhancement of Ginseng's Anti-Allergic Activity against IgE-Mediated Passive Cutaneous Anaphylaxis In Vivo and In Vitro

  • Hwang, Seon-Weon;Sun, Xiao;Han, Jun-Hyuk;Kim, Tae-Yeon;Koppula, Sushruta;Kang, Tae-Bong;Hwang, Jae-Kwan;Lee, Kwang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1626-1634
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    • 2018
  • Ginseng (the root of Panax ginseng Meyer) fermented by Lactobacillus plantarum has been found to attenuate allergic responses in in vitro and in vivo experimental models. Ginseng has been reported to also possess various biological functions including anti-inflammatory activity. The present study was aimed at comparing the anti-allergic effect of ginseng and fermented ginseng extracts on IgE-mediated passive cutaneous anaphylaxis in vitro in a murine cell line and in vivo in mice. Fermented ginseng extract (FPG) showed higher inhibitory effect against in vitro and in vivo allergic responses when compared with ginseng extract (PG). The secretion of ${\beta}$-hexosaminidase and interleukin (IL)-4 from the IgE-DNP-stimulated RBH-2H3 mast cells were significantly (p < 0.05) inhibited by FPG treatment, and this effect was concentration-dependent. Further, MKK4 activation and subsequent JNK phosphorylation were attenuated by FPG treatment. The inhibitory effect of FPG on the in vitro allergic response was verified in vivo against IgE-DNP-induced passive cutaneous anaphylaxis in a mouse model. These data indicated that the fermentation of ginseng with L. plantarum enhanced its anti-allergic effects both in vitro and in vivo. We predict that compositional changes in the ginsenosides caused by the fermentation may contribute to the change in the anti-allergic effects of ginseng. The results of our study highlight the potential of the use of FPG as a potential anti-allergic agent.

Effects of In Vitro Fertilization Conditions of In Vitro Matured Cumulus-Intact Pig Oocytes on Embryo Development

  • Kim, Jae-Young;Lee, Eun-Ji;Park, Jin-Mo;Lee, Hong-Cheol;Park, Hum-Dai;Kim, Jae-Myeong
    • Journal of Embryo Transfer
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    • v.26 no.2
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    • pp.117-122
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    • 2011
  • In this study, we examined the effectiveness of in vitro fertilization of porcine immature oocytes on the embryo development of blastocysts or hatched blastocysts and the number of cells according to the in vitro fertilization conditions. In the in vitro fertilization of in vitro matured porcine oocytes, there were no significant differences between treatment groups regarding fertilization rate, blastocyst rate, and embryo development of hatched blastocysts according to the storage periods of liquid sperm of 24, 48, and 72 hours. The embryo development rate of hatched blastocysts after the fertilization according to different spermatozoa concentrations ($0.4{\times}10^5$, $1.2{\times}10^5$, and $3.6{\times}10^5$ cells/ml) showed the highest rate in the group with a spermatozoa concentration of $1.2{\times}10^5$ cells/ml; in particular, this rate was significantly higher than that in the $0.4{\times}10^5$ cells/ml group (p<0.05). The total number of blastocysts cells as well as trophectoderms (TE) that developed in each treatment group were also significantly higher in the $1.2{\times}10^5$ cells/ml group than in any other groups (p<0.05). In contrast, the embryo development rate of blastocysts according to different co-incubation periods of sperm and oocyte (1, 3, and 6 hr) was high in the 6-hour group; in particular, the rate was significantly higher than that of the I-hour group (p<0.05). Furthermore, the total number of oocytes cells and TEs that developed was significantly higher in the 6-hour group than any other group (p<0.05). In this study, the most effective treatment conditions for porcine embryo development and high cell number were found to be as follows: a sperm storage period of less than 72 hours, a spermatozoa concentration of $1.2{\times}10^5$ cells/ml, and a 6-hour co-incubation period for sperm and ooocyte.

In vitro and in vivo studies on theophylline mucoadhesive drug delivery system

  • Bandyopadhyay, AK;Perumal, P
    • Advances in Traditional Medicine
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    • v.7 no.1
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    • pp.51-64
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    • 2007
  • Mucus is an aqueous gel complex with a constitution of about 95% water, high molecular weight glycoprotein (mucin), lipid, salts etc. Mucus appears to represent a significant barrier to the absorption of some compounds. Natural mucoadhesive agent was isolated and purified from the aqueous extract of the seeds of prosopis pallida (PP). Formulated tablet with the isolated material by wet granulation method. Some natural edible substances are in consideration for candidates as mucoadhesive agents to claim more effective controlled drug delivery as an alternative to the currently used synthetic mucoadhesive polymers. Subjected the materials obtained from natural source i.e. PP and standard synthetic substance, sodium carboxymethyl cellulose for evaluation of mucoadhesive property by various in vitro and in vivo methods. Through standard dissolution test and a model developed with rabbit, evaluated in vitro controlled release and bioadhesive property of theophylline formulation. Mucoadhesive agent obtained from PP showed good mucoadhesive potential in the demonstrated in vitro and in viνo models. The results suggest that the mucoadhesive agent showed controlled release properties by their application, substantially. In order to assess the gastrointestinal transit time in vivo, a radio opaque X-ray study performed in healthy rabbit testing the same controlled release formulation with and without bioadhesive polymer. Plasma levels of theophylline determined by the HPLC method and those allowed correlations to the in vitro mucoadhesive study results. Better correlation found between the results in different models. PP may acts as a better natural mucoadhesive agent in the extended drug delivery system.

Effect of supplement of SCM in culture medium for in vitro development of bovine in vitro fertilized oocytes

  • Sang Jun Uhm
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.3
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    • pp.143-150
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    • 2023
  • Background: The successful production of superior or transgenic offspring from in vitro produced embryos in cattle relies heavily on the quality of blastocyst stage embryos. In order to enhance the developmental competency of these embryos, a novel culture method was devised. Methods: This study utilized stem cell culture medium (SCM) from hESCs as a supplement within the culture medium for bovine in vitro produced embryos. To gauge the efficacy of this approach, in vitro fertilized embryos were subjected to culture in CR1aa medium enriched with one of three supplements: 0.3% BSA, 10% FBS, or 10% SCM. Results: The blastocyst development and hatching rates of one-cell zygotes cultured in CR1aa medium supplemented with SCM (23.9% and 10.2%) surpassed those cultured in CR1aa medium supplemented with BSA (9.3% and 0.0%) or FBS (3.1% and 0.0%) (p < 0.05). Furthermore, post-zygotic gene activation, cleaved embryos cultured in CR1aa medium supplemented with SCM (57.8% and 34.5%) exhibited notably higher rates (p < 0.05) compared to those cultured with BSA (12.9% and 0.0%) or FBS (45.7% and 22.5%) supplementation. Furthermore, the microinjection of SCM into the cytoplasm or pronucleus of fertilized zygotes resulted in elevated blastocyst development and hatching rates, particularly when the microinjected embryos were subsequently cultured in CR1aa medium supplemented with SCM from the 8-cell embryo stage onwards (p < 0.05), in contrast to those cultured with FBS supplementation. Conclusions: In conclusion, this study conclusively demonstrated that the incorporation of SCM into the culture medium significantly enhances the developmental progress of preimplantation embryos.

Nutritional Evaluation of Rice with Different Processing Treatments on in vitro Rumen Fermentation Characteristics and in situ Degradation (재고미의 가공처리에 따른 in vitro, in situ 소화율 및 발효성상 평가)

  • Yang, Sung-Jae;Jung, Eun-Sang;Kim, Han-Been;Shin, Taek-Soon;Cho, Byung-Wook;Cho, Seong-Keun;Kim, Byeong-Woo;Seo, Ja-Kyeom
    • Korean Journal of Organic Agriculture
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    • v.26 no.2
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    • pp.281-296
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    • 2018
  • This study was conducted to evaluate the effect of different processing of rice on rumen fermentation in in vitro and in situ experiments. Different processing treatments (extruding, roasting, and steaming) were used in this study and all treatments were ground through a cyclone mill (Foss, Hillerød, Denmark) fitted with a 1 mm screen. Non-treated rice was considered to a control substrate. Then, all treatments were used in in vitro and in situ experiments. Total gas production and dry matter digestibility in control were lower than any other treatment at all incubation times (P<0.01). The lowest ammonia nitrogen ($NH_3-N$) concentration was observed in control among treatments at 6, 12, and 24 h incubation (P<0.01). Extruding had a highest total volatile fatty acids (VFA) concentration at 6, 12 h incubation (P<0.01) and Steaming exhibited a highest total VFA at 24 h (P<0.01). The lowest total VFA concentration was observed in control at 6, 12, and 24 h (P<0.01). In an in situ, The highest value of soluble fraction, degradation rates, effective degradability was observed in extruding (P<0.01). It was considered that feed processing increased dry matter digestibility, total VFA concentration, and decreased pH as well as $NH_3-N$ concentration indicating that processing may increase nutrient degradation of rice in the rumen.

Effect of Culture Media and Co-culture with Bovine and Rabbit Oviductal Epithelial Cells on In Vitro Development of Rabbit Embryos (토끼 수정란의 체외발달에 미치는 배양액 및 소와 토끼의 난관상피세포들과의 공배양 효과)

  • 노규진;이효종;송상현;윤희준;박충생
    • Korean Journal of Animal Reproduction
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    • v.18 no.1
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    • pp.39-46
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    • 1994
  • This experiment was carried out to develop an in vitro culture system for rabbit embryos. The zygotes or 2-cell embryos were collected from the oviducts of the superovulated and mated does with D-PBS/10% FCS at 24 hours after hCG injection. The in vitro developmental rate of blastocyst formation and the number of nuclei in the embryos were examined under the following treatments; 1) TCM-199 with 10% FCS, 2) EBSS with 10% FCS, 3) rabbit vitreous humor(VH), 4) TCM-199 with 10% FCS+BOEC, 5) TCM-199 with 10% FCS+ROEC, 6) EBSS with 10% FCS+BOEC and 7) EBSS with 10% FCS+ROEC. For a comparative study of in vivo and in vitro development, the fresh blastocysts, which were developed in vivo for 96 hours after hCG injection, were collected from the uterus and their numbers of nuclei were counted. 1. The zygotes or 2-cell embryos developed to the blastocyst stage in TCM-199, EBSS and VH at the rates of 93, 92 and 89%, respectively. 2. The higher developmental rates 95~98% of blastocyst formation was achieved when the embryos were co-cultured with a monolayer of bovine or rabbit oviductal epithelial cells in TCM-199 or EBSS. No significant difference in developmental rates was shown between bovine and rabbit oviductal epithelial cells. 3. In a comparative study of in vivo and in vitro development, the total numbers of nuclei were significantly less in the in vitro cultured embryos(104~224) than the in vivo developed embryos(1, 0090 at 96 hours after hCG injectin. 4. The mean cell cycle numbers in the embryos cultured for 72 hours in TCM-199 with 10% FCS, EBSS with 10% FCS, TCM-199 with 10% FCS+BOEC, TCM-199 with 10% FCS+ROEC, EBSS with 10% FCS+BOEC and in vivo was 7.38, 6.63, 7.76, 7.69, 7.01 and 9.92, respectively. From these results, it can be suggested the optimal culture system for in vitro culture of rabbit embryos is a co-culture system with bovine or rabbit oviductal epithelial cells in TCM-199 with 10% FCS. Considering the significant reduction in total numbers of nuclei in the in vitro cultured embryos, the advanced research on development of in vitro culture system for rabbit embryos is expected.

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In-Vitro Model Design of Mitral Valve Regurgitation and Comparative Study of Quantification between PISA and 4D flow MRI (승모판 역류 In-Vitro 모델을 활용한 초음파 및 4D flow MRI 기반 혈류 정량화 비교연구)

  • Juyeon Lee;Minseong Kwon;Hyungkyu Huh
    • Journal of the Korean Society of Visualization
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    • v.22 no.1
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    • pp.40-48
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    • 2024
  • This study presents an in-vitro model designed to simulate mitral valve regurgitation, aiming to compare the quantification results between Proximal Isovelocity Surface Area(PISA) and 4D Flow MRI on both fixed and valve annulus tracking(VAT) views. The in-vitro model replicates the dynamic conditions of the mitral valve in a pulsatile environment, utilizing a piston pump set at 60 bpm. Through systematic experiments and analysis, the study evaluates the accuracy and effectiveness of PISA and 4D Flow MRI in assessing regurgitation severity, considering both fixed and valve annulus tracking. The displacement length measured in echo closely resembled that of optical measurements, making it advantageous for structural analysis. VAT-4D flow MRI exhibited the smallest deviation from actual flow rate values, establishing it as most accurate method for quantitative regurgitation assessment.

Effects of IGF-I and EGF Supplemented to PZM3 Culture Medium on the Development of Porcine Embryos In vitro

  • Kim, J.Y.;Park, M.C.;Kim, S.B.;Park, H.D.;Lee, J.H.;Kim, Jae-Myeoung
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.8
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    • pp.1117-1123
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    • 2009
  • This study investigated the effects of IGF-I and EGF on the development of blastocysts or hatched blastocysts during the in vitro culture of embryos from immature porcine oocytes. After the in vitro maturation and fertilization of cumulus-oocyte complexes (COCs) and their culture in vitro in PZM3 medium, we examined the embryo development rate for 168 h. When different concentrations of IGF-I (0, 1, 10, 20 ng/ml) were supplemented to fertilized porcine embryos in vitro, there were no significant differences in cleavage rate, blastocyst development rate or blastocyst hatching rate among the treated groups. On the other hand, when different concentrations of EGF (0, 1, 10, 20 ng/ml) were supplemented to the in vitro culture medium, blastocyst development rate was highest in the group in which EGF was not supplemented and, specifically, it was higher than in the 20 ng/ml treatment group (p<0.05). When 10 ng/ml IGF-I and 1 ng/ml EGF were supplemented separately or simultaneously, there were no significant differences among the treated groups in blastocyst hatching rate and the number of cells in each condition. This study demonstrated that the addition of IGF-I and EGF into PZM3 medium did not enhance development of the blastocyst stage and total cell number in blastocysts.

Development of Antifreezing Agent for Bisected Embryo in Mouse (생쥐 분할배의 동결보호물질 개발)

  • 이병오
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.233-238
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    • 1996
  • This study was conducted to investigate the effects of cryoprotective agents and thawing temperature on the survival rate of the bisected embryos of mouse. The results of this study were summairzed as follows: 1. In in vitro culture after frozen with DMSO, glycerol, ethylene glycol or propanediol and thawed, the rates of normally developed bisected morula which was denuded were 31.7, 39.1, 28.0 and 23.1%, respectively. 2. The survival rates of bisected morula encased into the zona pellucida in in vitro culture after frozen with DMSO, glycerol, ethylene glycol or propanediol and thawed were 72.4, 68.7, 64.0 and 59.5%, respectively. 3. The survival rates of bisected denuded blastura in in vitro culture after frozen with DMSO, glycerol, ethylene glycol or propanediol and thawed were 48.3, 44.8, 32.1 and 28.6%, respectively. 4. The survival rates of bisected blasturae encased into the zona pellucida in in vitro culture after frozen with DMSO, glycerol, ethylene glycol or propanediol and thawed were 73.6, 67.4, 53.0 and 49.1%, respectively. 5. In in vitro culture after frozen with DMSO, glycerol, ethylene glycol or propanediol and thawed at room temperature, the rates of the normally developed bisected morula which was encased into the zona pellucida were 67.1, 62.3, 57.7 and 53.0%, respectively, and those of the bisected blasturae encased into the zona pellucida were 70.8, 65.4, 56.6 and 52.1%, respectively. 6. The survival rates of bisected morula which was encased into the zona pellucida in in vitro culture after forzen with DMSO, glycerol, ethylene glycerol or propanediol and thawed at 37$^{\circ}C$ were 74.3, 71.3, 63.9 and 57.4%, respectively, and those of bisected blastura encased into zona pellucida were 76.0, 69.1, 61.1 and 56.1%, respectively.

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