• Journal of Embryo Transfer
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• v.15 no.2
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• pp.191-196
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• 2000

the present study was carried out to develop a completely defined culture system and determine if high NaCl concentrations in defined (PVA added) or semi-defined (BSA added) medium is toxic to bovine embryos. Oocytes from slaughterhouse ovaries were matured and fertilized in vitro. After 30 h of insemination, only 2-cell stage embryos were selected and cultured for this experiment. The culture media used were as follows : TLP(114 mM of NaCl) + BSA (3 mg/ml), TLP + PVA (1 mg/ml), mTLP(96 mM of NaCl) + BSA, mTLP + PVA. Six to ten embryos were placed into a 30$\mu$1 drop of each medium and the embryos were examined at 10 day post-insemination without medium renewal. The experiment was replicated 4 times. All data were analyzed by chi-square. There were no significant differences among TLP-BSA, mTLP-BSA and mTLP-PVA in blastocyst development (21.6, 17.2 and 20.2%), respectively. Also, no differences were obtained in hatching rates (11.7, 9.9 and 12.2%), respecitively. However, there were significant differences between TLP-PVA (1.7% and 0.6%) and other group in blastocyst formation and hatching rates, respectively (p<0.01). Development of in vitro produced embryos cultured in BSA containing medium was not affected by high NaCl concentration, but in the completely defined medium, embryonic development was highly affected by NaCl. This study shows that reduced NaCl concentration in completely defined medium is beneficial for development of bovine pre-implantation embryos in vitro.

• Journal of Veterinary Science
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• v.24 no.1
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• pp.10.1-10.10
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• 2023

Background: The collection of ovaries from slaughterhouses is an important source of oocytes for in vitro embryo production. On the other hand, the physiological stage of slaughtered females varies and influences embryo production. Objectives: The study examined the in vitro efficiency of embryos and demi-embryos from young, non-pregnant adult, and pregnant adult ewes from a local slaughterhouse. Methods: One thousand three hundred ovaries were collected from August to October 2020. The recovered oocytes were matured, fertilized, and cultured at 5% CO2, 38.5℃, and 100% humidity. Embryo bisection was performed in 96 blastocysts (n = 32 per treatment). The demiembryo pairs were incubated for their reconstitution for 12 h. SAS was used for data analysis. Results: The number of oocytes collected from the experimental group of non-pregnant adult ewes was higher (p ≤ 0.007) than those collected from the group of pregnant adult ewes (2.67 ± 0.19 vs. 2.18 ± 0.15 oocytes/group, respectively). The blastocyst rate was higher (p ≤ 0.0001) in the non-pregnant adult group (36.39%) than in the young (17.96%). The ratio of demi-embryos that recovered the blastocoelic cavity was higher (p < 0.05) in the young group (81.25%) than in the pregnant adult group (59.38%). The diameter of the demi-embryos was higher (p < 0.05) in the non-pregnant adult group (186.54 ± 8.70 ㎛) than those in the young and pregnant adult groups. Conclusions: In conclusion, the in vitro embryo production efficiency was highest when using oocytes from non-pregnant adult ewes under the conditions of this study.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.15 no.4
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• pp.263-270
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• 1982

In an attempt to evaluate the nutritional quality of seaweed protein, the effects of heat treatment on the in vitro digestibility and trypsin inhibitor content in seaweed were determined. In this study, the nitrogen-to-protein conversion factors were also calculated on the basis of quantitative amino acid data. The results are as follows : 1. The in vitro protein digestbilty of red seaweeds (P. teoera anc P. suborbiculata) were ranged from 78.5 to 82.2, and green seawerd (E. linza) and brown seaweeds showed value under 80 in vitro digestibility. In general, trypsin inhibitor contents in brown seaweed were higher (0.33-0.54 mg/g) than those of red seaweeds (0.26-0.39 mg/g). And it is noted that the lowest trypsin inhibitor content was shown in green seaweed (E. linza) in spite of lowest in spite digestibility (78.5). 2. The in vitro protein digestibility of sun dried laver (P. tenera) was increased with cooling time (microwave heating), but it was not significant. Hot plate cooking raised the in vitro digestibility from 81. 1 to 84.5. The influence pot cooking time on trypsin inhibitor content was inversely proportional to in vitro digestibility. 3. Computed nitrogen factor, based on amino acid content (Factor method) and Kjeldahl nitrogen content (Kjeldahl mettled), were 5.83 (H. fusiforme)- 6.52 (P. tencra) as Factor method and 5.40 (U. pinnatifida)-6.29 (P. tenera) as Kjeldahl method. Individual value for each nitrogen conversion factor differed by species, especially in brown seaweeds. The best estimate of the protein content of seaweed can be calculated, from multiplying the summed amino acid content by conversion factor (Factor method).

• Translational and Clinical Pharmacology
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• v.26 no.3
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• pp.142-142
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• 2018

• Proceedings of the Korean Society of Food Hygiene and Safety Conference
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• 2002.05a
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• pp.137-137
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• 2002

Several epidemiologidal studies suggested that arsenic exposure was strongly correlated with the development of cardiovascular disease such as hypertension. In order to examine whether arsenic affects vasomotor tone in blood vessels, we investigated the effect of arsenic on agonist-induced vasorelaxation using the isolated rat aortic ring in in vitro organ bath system. Treatment with arsenite inhibited acetylcholine-induced relaxation of aortic rings in a concentration- dependent manner. The inhibitory effects by arsenic were also observed in the relaxation induced by sodium nitroprusside, a NO-donor. Consistent with these findings, the cGMP levels stimulated by acetylcholine in blood vessels were reduced significantly by arsenite treatment. In addition, higher concentration of arsenite decreased the relaxation by 8-Br-cGMP, a cGMP analog, in aortic rings without endothelium. These in vitro results indicated that arsenite that arsenite was capable of suppressing acetylcholine-induced relaxation in blood vessels by inhibiting production of nitric oxide in endothelial cells and by impairing the relaxation machinary in smooth muscle cells. In vivo studies revealed that the reduction of blood pressure by acetylcholine infusion was signigicantly suppressed after arsenite was administered intravenously to rate. These data suggest that vasomotor tone impaired by arsenite exposure may be one of the contrbuting factors in development of cardiovascular disease.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1808-1812
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• 2002

The present study was carried out to examine the efficiency of microcapsules and a stability of lactase in vitro in the simulated gastric and intestinal conditions. As a coating materials, medium-chain triacylglycerol (MCT) and polyglycerol monostearate (PGMS) were used. The highest efficiency of microencapsulation was found in the ratio of 15:1 as coating to core material with both MCT (91.5%) and PGMS (75.4%). In a subsequent experiment, lactose content was measured to study a microcapsule stability. Lysis of microcapsules made by MCT in simulated gastric fluid was proportionally increased such as 3% in pH 5 and 11% in pH 2 for 20 min incubation. In the case of PGMS microcapsulation, 11-13% of lactose was hydrolyzed at 20 min in all pHs and also very little amount (less than 3%) of lactose was hydrolyzed after 20 min in all pHs. The highest percentages of lactose hydrolysis in MCT and PGMS microcapsules were 68.8 and 60.8% in pHs 7 and 8 during 60 min, respectively. Based on our data, the lactase microcapsules seemed to be stable when they stay in the stomach, and hydrolyzed rapidly in small intestine where the bile acid was excreted.

• Journal of Life Science
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• v.9 no.4
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• pp.382-388
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• 1999

The effects of citrus flavonoids, hesperidin and naringin, on the lipid metabolism were investigated in cultured human hepatocyte HePG2 cells. HepG2 cells were cultured for 6 h and 24 h to the control medium or the media containing hespridin and narigin, which concentrations were 0.5 and 5.0 mg/$m\ell$. There were no significant effects on cell proliferation and cellular protein content, except for increased in these parameters by adding both citrus flavonoids (0.5 mg/$m\ell$). The cellular content of triacylglycerol after 6 h incubation with 0.5 mg/$m\ell$ hesperidin and naringin was markedly increased, and after 24 h incubation that was decreased in both citrus flavonoids supplementation. The supplementation of 5.0 mg/$m\ell$ hesperidin caused a marked decrease in the cellular cholesterol content following 6 h incubation, and that was also reduced markdly, in a dose-dependent manner, during incubation for 24 h. However, there was no significant difference in the cellular cholesterol content in medium supplemented with naringin. The effect of hesperidin and naringin on acyl-CoA: cholesterol acyltransferase (ACAT) activity was studied in vivo and in vitro. The data confirmed that hesperidin inhibit ACAT activity in vivo and in vitro, whereas naringin had no such effect on ACAT activity in vivo but not in vitro. The present study suggests that hesperidin reduces the cellular triacyglycerol and cholesterol contents in human hepatocyte HepG2 cells.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.6
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• pp.812-817
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• 2012

Nitrate can serve as a terminal electron acceptor in place of carbon dioxide and inhibit methane emission in the rumen and nitrate reducing bacteria might help enhance the reduction of nitrate/nitrite, which depends on the type of feed offered to animals. In this study the effects of three levels of sodium nitrate (0, 5, 10 mM) on fermentation of three diets varying in their wheat straw to concentrate ratio (700:300, low concentrate, LC; 500:500, medium concentrate, MC and 300:700, high concentrate, HC diet) were investigated in vitro using buffalo rumen liquor as inoculum. Nitrate reducing bacteria, isolated from the rumen of buffalo were tested as a probiotic to study if it could help in enhancing methane inhibition in vitro. Inclusion of sodium nitrate at 5 or 10 mM reduced (p<0.01) methane production (9.56, 7.93 vs. 21.76 ml/g DM; 12.20, 10.42 vs. 25.76 ml/g DM; 15.49, 12.33 vs. 26.86 ml/g DM) in LC, MC and HC diets, respectively. Inclusion of nitrate at both 5 and 10 mM also reduced (p<0.01) gas production in all the diets, but in vitro true digestibility (IVTD) of feed reduced (p<0.05) only in LC and MC diets. In the medium at 10 mM sodium nitrate level, there was 0.76 to 1.18 mM of residual nitrate and nitrite (p<0.01) also accumulated. In an attempt to eliminate residual nitrate and nitrite in the medium, the nitrate reducing bacteria were isolated from buffalo adapted to nitrate feeding and introduced individually (3 ml containing 1.2 to $2.3{\times}10^6$ cfu/ml) into in vitro incubations containing the MC diet with 10 mM sodium nitrate. Addition of live culture of NRBB 57 resulted in complete removal of nitrate and nitrite from the medium with a further reduction in methane and no effect on IVTD compared to the control treatments containing nitrate with autoclaved cultures or nitrate without any culture. The data revealed that nitrate reducing bacteria can be used as probiotic to prevent the accumulation of nitrite when sodium nitrate is used to reduce in vitro methane emissions.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.3
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• pp.1163-1169
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• 2014

Astragalus, a commonly used traditional Chinese medicine, has exhibited antitumor actions in patients. In this study, in vitro and in vivo antitumor effects of astragalus and synergistic antitumor efficacy in combination with pterostilbene were investigated. Melanoma cells were treated with pterostilbene (Pt), graduated doses of astragalus injection (AI), or these in combination. Cell viability was measured using a MTT assay. Released nucleosomes and caspase activity were measured using enzyme-linked immunosorbent assay. Growth inhibition in vitro and in vivo was also assessed. Analysis of variance and t tests were used for statistical analysis. Significant reduction (p<0.05) in cellular proliferation were observed with AI and AI-Pt in a time- and concentration-dependent manner. Apoptosis and caspase-3/7 activity were significantly increased by AI and AI-Pt treatment (p<0.05). In vivo, AI inhibited melanoma tumor growth, with inhibition rates ranging from 36.5 to 62.3%, by inducing apoptosis via up-regulation Bax expression and the Bax/Bcl-2 ratio and down-regulating Bcl-2 expression. AI significantly inhibits the growth of melanoma in vitro and in vivo by inducing apoptosis. These data suggest that combined treatment of astragalus with pterostilbene enhances antitumor efficacy.

• Journal of Life Science
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• v.16 no.3 s.76
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• pp.409-414
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• 2006

Molecular chaperones and folding enzymes in the endoplasmic reticulum (ER) associate with the newly synthesized proteins to prevent their aggregation and help them fold and assemble correctly. Chaperone function of BiP, which is a Hsp70 homologue in ER, is controlled by the N-terminal ATPase domain. The ATPase activity of the ATPase domain is affected by regulatory factors. BAP was identified as a nucleotide exchange factor of BiP (Grp78), which exchanges ADP with ATP in the ATPase domain of BiP This study presents whether BAP can influence folding of a protein, immunoglobulin heavy chain that is bound to BiP tightly. We first examined which nucleotide of ADP and ATP affects on BAP binding to BiP The data showed that endogenous BAP of HEK293 cells prefers ADP for binding to BiP in vitro, suggesting that BAP first releases ADP from the ATPase domain in order to exchange with ATP. Immunoglobulin heavy chain, an unfolded protein substrate, was released from BiP in the presence of BAP but not in the presence of ERdj3, which is another regulatory factor for BiP accelerating the rate of ATP hydrolysis of BiP The ADP-releasing function of BAP was, therefore, believed to be responsible for immunoglobulin heavy chain release from BiP. Grp170, another Hsp70 homologue in ER, did not co-precipited with BAP from $[^{35}S]$-metabolic labeled HEK293 lysate containing both overexpressed Grp170 and BAP. These data suggested that BAP has no specificity to Grp170 although the ATPase domains of Grp170 and BiP are homologous each other.