• Reproductive and Developmental Biology
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• 제29권4호
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• pp.223-227
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• 2005

In vitro maturation of porcine immature cumulus-enclosed oocytes can be enhanced by co-incubation with spermatozoa even before fertilization. The aim of this study was to determine whether the addition of spermatozoa into the culture medium can stimulate the meiosis resumption of porcine cumulus-enclosed oocytes arrested at germinal vesicle (GV). Cumulus-enclosed oocytes (CEOs) were collected from follicles of 3 to 5mm diameter. Porcine CEOs were cultured in tissue culture medium containing various meiosis inhibitors and spermatozoa. Oocytes were examined for evidence of GV and GV breakdown after 24 h culture. After 24 h culture $43.8\%$ of oocytes cultured in only TCM 199 remained at GV stage whereas $56.2\%$ of oocytes were able to resume meiosis. When porcine CEOs were cultured in the medium with meiosis inhibitor such as, dibutyryl cAMP (dbcAMP) and forskolin (Fo), more than $90\%$ of oocytes were not able to resume meiosis. However, co-culture of porcine CEOs with spermatozoa was able to overcome the inhibitory effect of dbcAMP and Fo. Irrespective of the presence of 3-isobutyl-1-methylxanthine (IBMX), no difference was observed in the proportion of oocyte reached germinal vesicle breakdown (GVBD). The present study suggests that dbcAMP and Fo prevent the spontaneous maturation of competent oocyte in culture after isolation from follicles and that mammalian spermatozoa contain a substance(s) that improves meiosis resumption in vitro of porcine cumulus-enclosed oocytes.

• Journal of Applied Biological Chemistry
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• 제64권1호
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• pp.49-55
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• 2021

The diet plays a fundamental role in the formation of the gut microbiota, determining the interrelationship between the gut microbiota and the host. The current study investigated the effect of Chlorella vulgaris on the gut microbiota by using simulated in vitro digestion and colonic fermentation. Bioaccessibility was measured after in vitro digestion, and SCFAs and microbial profiling were analyzed after colonic fermentation. The bioaccessibility of C. vulgaris was 0.24 g/g. The three major SCFAs (acetate, propionate, and butyrate) increased significantly when compared to the control group. In microbial profiling analysis, microorganisms such as Faecalibacterium, Dialister, Megasphaera, Dorea, Odoribacter, Roseburia, Bifidobacterium, Butyricmonas, and Veillonella were high in C. vulgaris group. Among them, Faecalibacterium, Dialister, Megasphaera, Roseburia, and Veillonella were thought to be closely associated with the increased level of SCFAs. Finally, it can be expected to help improve gut microbiota and health through ingestion of C. vulgaris. However, further studies are vital to confirm the changes in the gut microbiota in in vivo, when C. vulgaris is ingested.

• 대한수의학회지
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• 제6권1호
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• pp.45-52
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• 1966

Hexachlorophene, Bithionol and Dithiazanine iodide were used to examine the vermicidal effect on Fasciola hepatica in vitro. A kymographic study was performed to investigate the motility of Fasciola hepatica under the influence of these drugs. Following conclusions were made: 1) With 10,000 : 1 solution; exhibited the vermicidal effect on Fasciola hepatica, Hexachlorophene in 5 minutes, Bithionol in 10 minutes, and Dithiazanine, iodide in 40 minutes, respectively. 2) With 100,000 : 1 solution; exhibited the vermicidal effect on Fasciola hepatica, Hexachlorophene in 20 minutes and Bithionol in 30minutes. In the case of Dithiazanine iodide the stimulation upon the fluke continued for 120 minutes. 3) With 1,000,000 : 1 solution; exhibited the vermicidal effect on Fasciola hepatica, Hexachlorophene in 50 minutes and Bithionol in 80 minutes. In the case of Dithiazanine iodide the stimulation upon the fluke continued for 120 minutes. 4) With 10,000,000 : 1 solution; the only Hexachlorophene showed the vermicidal effect on Fasciola hepatica. In the case of Bithionol and Dithiazanine iodide the slight stimulation upon the fluke continued for 120 minutes.

• Fisheries and Aquatic Sciences
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• 제20권12호
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• pp.32.1-32.7
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• 2017

Recent in vitro studies have demonstrated that extract of soft coral Dendronephthya gigantea (SCDE) had strong anti-inflammatory activities. However, the direct effects of SCDE on anti-inflammatory activities in vivo model remained to be determined. Therefore, the present study was designed to assess in vivo anti-inflammatory effect of SCDE using lipopolysaccharide (LPS)-stimulated zebrafish model. We also investigated whether SCDE has toxic effects in zebrafish model. The survival, heart beat rate, and developmental abnormalities were no significant change in the zebrafish embryos exposed to at a concentration below $100{\mu}g/ml$ of SCDE. However, lethal toxicity was caused after exposure to 200 and $400{\mu}g/ml$ of SCDE. Treating zebrafish model with LPS treatment significantly increased the reactive oxygen species (ROS) and nitric oxide (NO) generation. However, SCDE inhibited this LPS-stimulated ROS and NO generation in a dose-dependent manner. These results show that SCDE alleviated inflammation by inhibiting the ROS and NO generation induced by LPS treatment. In addition, SCDE has a protective effect against the cell damage induced by LPS exposure in zebrafish embryos. This outcome could explain the profound anti-inflammatory effect of SCDE both in vitro as well as in vivo, suggesting that the SCDE might be a strong anti-inflammatory agent.

• 산업식품공학
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• 제14권4호
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• pp.342-345
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• 2010

The purpose of current study was to examine bioaccessibility of antioxidant activity and total phenolic content in each part of water spinach (Ipomoea aquatic Forsk.). In vitro biomimicking system simulated human digestive fluid was employed in order to measure bioavailable anti-oxidative effect and phenolic content. Antioxidant activity and total phenolic content was measured by using the DPPH method and the Folin-Ciocalteu assay, respectively. Stem of water spinach had a higher DPPH free radical scavenging effect (5.43 mg/mL for $IC_{50}$) than leaf (5.95 mg/mL for $IC_{50}$), while leaf had a greater level of total phenolic content (287.45 ${\mu}g$ GAE/mL) than stem (216.45 ${\mu}g$ GAE/mL). Bioaccessible antioxidant capacity and digestive stability of total phenolic content showed a similar pattern to what found in raw materials. Our result also indicated that total phenolic content was not found to be a major marker for prediction of antioxidant activity. It is plausible that other constituents such as vitamin E and C in water spinach could be contributors for antioxidant activities.

• 한국수정란이식학회지
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• 제10권2호
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• pp.121-129
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• 1995

To examine the critical effect of oxygen concentration on embryonic development, in vitro fertilized embryos were cultured in media(TCM199 vs. SOF) supplemented sera(1O% FCS vs. 10% HS) with and without bovine oviduct epithelial cells under two gas atmosphere (5% $CO_2$ in air vs. 5% $CO_2$, 5% $O_2$, 90% $N_2$). Oocytes, obtained from abattoir ovaries, were matured in EGF containing TCM199 medium co-cultured with BOEC for 24 hours, followed by exposure to frozen-thawed, heparin4reated spermatozoa in TALP for 30 hours. And then early embryos(1~2 cell) were cultured in both TCM199 and SOF supplemented with 10% FCS or 10% RS under 5% $CO_2$ in air or 5% COi, 5% $O_2$, 90% $N_2$. Development to morulae and blastocysts was recorded on days 7, after the start of in vitro fertilization. The developmental rates of in vitro fertilized embryos to morulae and blastocysts cultured in SOF with BOEC under 5% $CO_2$, 5% $O_2$, 90% $N_2$(24.4%) were significantly(p<0.05) higher than cultured in SOF with BOEC under 5% $CO_2$ in air(14.1%) at seven days after in vitro fertilization. When early bovine embryos were cultured in TCM 199 and SOF under two different gas atmosphere, there were no significant differences in the developmental rates to morulae and blastocysts between supplements of 10% FCS and 10% HS. The rates of development to morulae and blastocysts were significantly(p<0.01) higher in TCM 199 with BOEC(24.7%) than TCM199 without BOEC(10.9%) under 5% $CO_2$ in air, otherwise SOF without BOEC(36.4%) were significantly (p<0.05) higher than in SOF with BOEC (24.4%) under 5% $CO_2$, 5% $O_2$, 90% $N_2$. In summary, these experiments have proved that the culture system in SOF supplemented 10% ES is effective on in vitro development of early bovine embryos under 5% $CO_2$, 5% $O_2$, 90% $N_2$. In addition, it is effective to development of bovine embryos that TCM 199 should be co-cultured with BOEC and SOF should be cultured without somatic cells under two different gas atmosphere.

• 한국수정란이식학회지
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• 제25권3호
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• pp.165-169
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• 2010

This study was carried out to assess the effect of vitamin E against the reactive oxygen species (ROS) on chemical activation of in vitro matured oocytes. Bovine oocytes were aspirated from slaughtered ovaries and transferred to maturation medium with or without vitamin E ($100\;{\mu}M$). After 22 hours of culture, oocytes with polar bodies were selected and submitted to activation treatments with or without vitamin E. After activation, oocytes were cultured in mSOF medium and rate of development was monitored. For ROS ($H_2O_2$) detection, in vitro matured and activated oocytes were selected and stained with DCFDA and observed under fluorescence microscope. The ROS contents were not significant differences in IVM rate, activation process and embryonic development to blastocysts with or without vitamin E. The cell number of blastocyst showed significant difference (p<0.05) in embryos matured and activated with vitamin E. The results of the present study demonstrated that the exposure of vitamin E in IVM and activation process improved the quality of embryos evaluated by the cell number of blastocysts.

• 한국가축번식학회지
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• 제21권1호
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• pp.53-59
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• 1997

This study was conducted to investigate the effect of hormones on in vitro maturation of porcine follicular oocytes. The results obtained were as follows : 1. The maturation rates of oocytes cultured in medium containing FSH 1, 10, 50 and 100$\mu\textrm{g}$/ml were 44.6, 58.5, 42.6 and 37.9%, respectively. And those were no difference to maturation rates among the medium containing FSH. However, the maturation rate(13.7%) of oocytes cultured without FSH was significantly(P<0.05) lower than those of oocytes cultured with FSH. 2. The maturation rates of oocytes cultured in medium containing hCG 0, 1, 10, 50 and 100IU/ml were 12.2, 11.9, 17.9, 21.9 and 45.6%, respectively. The maturation rate of oocytes cultured with 100IU/ml hCG was significantly(P<0.05) higher than those of oocytes cultured with 0~50IU/ml hCG concentrations. 3. The maturation rates of oocytes cultured in medium containing E2 0, 1, 10, 50 and 100$\mu\textrm{g}$/ml were 13.7, 10.5, 14.9, 11.4 and 5.9%, respectively. There were no differences to maturation rates among the E2 concentrations. 4. The FSH＋hCG treatment was the highest maturation rate in medium containing different combination of FSH, hCG and E2.

• Archives of Plastic Surgery
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• 제34권4호
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• pp.420-425
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• 2007

Purpose:The mechanism of scar formation is not fully understood. Fibroblast is an important cell in wound healing process. We experienced a patient who was taking progesterone orally. Upper blepharoplasty was performed on her but, wound healing was delayed. We hypothesized that progesterone was the cause of delayed wound healing and fibroblast proliferation inhibition. We investigated the effect of progesterone in vitro on human dermal fibroblasts to study the effects on fibroblast proliferation. Methods: Human dermal fibroblasts from four persons were cultured initially. Progesterone is mixed to them at various concentrations, and fibroblast cell count was measured by MTT assay method at 570 nm. We confirmed that progesterone has some inhibitory effect on fibroblast proliferation and maximal inhibitory concentration of progesterone was determined. Then fibroblasts from a total of nineteen persons were cultured and the effects of progesterone were studied. Results: The initial study showed the maximal inhibitory concentration of progesterone to be $50{\mu}g/ml$. The main study showed that progesterone had 70.9% inhibitory effect on human dermal fibroblast in vitro. Conclusion: Progesterone has inhibitory effect on cultured human dermal fibroblast proliferation in vitro.

• Archives of Pharmacal Research
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• 제17권5호
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• pp.287-293
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• 1994

The effect of several adsorbents on paraquat poisohing was investigated (1) by measuring the saturatd amount of the poison adhered on the adsorbents in vitro and (2) by assaying the blood level of paraquat in the rat in in situ intestinal absorption experiments. Activated charcoal powder, natural aluminum silifonate) were used as adorbents. The steady-state blood level of paraquat in its absorption experiment with the cationic exchange resins was markedly lower than those without the resins or with other adsorbents. A good relationship was achieved between the calculated AUC or adsorptioin rate (in situ) and the saturated adsorption amount (in vitro). The rank order of the effect was sodium polystyene sulfonate > calcium polystyene sulfonate>natural aluminum silicate>activated charcoal powder. The effect of sodium polystyrene sulfonate after intestinal washing with physiological saline ws also measured, and a synergistic effect (marked decrease in blood paraquat level0 was found as compared with the intestinal washing alone. The simultaneous use of G.I. weshing and powerful adsorbent was scientifically proven to be most benefical.