• Asian-Australasian Journal of Animal Sciences
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• v.29 no.3
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• pp.343-351
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• 2016

Although the chemical, physical, and nutritional properties of bovine milk have been extensively studied, only a few studies have attempted to characterize milk-synthesizing genes using RNA-seq data. RNA-seq data was collected from 21 Holstein samples, along with group information about milk production ability; milk yield; and protein, fat, and solid contents. Meta-analysis was employed in order to generally characterize genes related to milk production. In addition, we attempted to investigate the relationship between milk related traits, parity, and lactation period. We observed that milk fat is highly correlated with lactation period; this result indicates that this effect should be considered in the model in order to accurately detect milk production related genes. By employing our developed model, 271 genes were significantly (false discovery rate [FDR] adjusted p-value<0.1) detected as milk production related differentially expressed genes. Of these genes, five (albumin, nitric oxide synthase 3, RNA-binding region (RNP1, RRM) containing 3, secreted and transmembrane 1, and serine palmitoyltransferase, small subunit B) were technically validated using quantitative real-time polymerase chain reaction (qRT-PCR) in order to check the accuracy of RNA-seq analysis. Finally, 83 gene ontology biological processes including several blood vessel and mammary gland development related terms, were significantly detected using DAVID gene-set enrichment analysis. From these results, we observed that detected milk production related genes are highly enriched in the circulation system process and mammary gland related biological functions. In addition, we observed that detected genes including caveolin 1, mammary serum amyloid A3.2, lingual antimicrobial peptide, cathelicidin 4 (CATHL4), cathelicidin 6 (CATHL6) have been reported in other species as milk production related gene. For this reason, we concluded that our detected 271 genes would be strong candidates for determining milk production.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.4
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• pp.586-590
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• 2003

The pharmacological effect of Korean-Chinese traditional herbal medicine, Hawangyeonhaedoktang (HT) on experimentally induced-triglyceride accumulation in cultured human hepatocyte HepG2 cells was studied. HePG2 cells were cultured in the Dulbecco's modified Eagle's (DME) medium without (Control medium) or with HT (0.5 mg/mL and 5.0 mg/mL) containing 1 mM oleate, 0.2% bovine serum albumin (BSA), and glucose 4.5 mg/mL for 6 and 24 hours in experiment I and 2 mM oleate, 0.5% BSA, and glucose 4.5 mg/mL for 6, 24 and in hours in Experiment II or 1 and 3 hours in Experiment III. Oleate ［$^{14}$ C］(0.5 $\mu$Ci/mL medium) added as a radioactive lipid precursor in the experiment I. In the experiment I, the intracellular triglyceride concentration was decreased remarkably during incubation for 6 and 24 hours, in a dose-dependent manner. At the same time, HT caused a decrease in the incorporation of ［$^{14}$ C］ oleate into intracellular triglyceride fraction and the secretion of triglyceride labeled with ［$^{14}$ C］ oleate into medium. In the experiment II and III compared to experiment I, the triglyceride accumulation in HepG2 cells was occurred, and HT prevented the accumulation of triglyceride during incubation for 24 and 48 hours. This result suggest that HT prevent the triglyceride accumulation in human hepatocytes by its inhibiting action on the intercellular triglyceride biosynthesis.

• KSBB Journal
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• v.16 no.1
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• pp.48-53
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• 2001

Bovine serum albumin(BSA) was solubilized into the reverse micellar phase consisting of sodium bis(2-ethylhexyl) sulfosuccinate(AOT) and isooctane using the phase transfer method. Of particular interest in this study were the effects of pH and the added salt type and concentration on the solubilization efficiency. When univalent or divalent salts such as KCl, NaCl, $MgCl_2$, or $CaCl_2$ were added to the aqueous phase at a concentration of 0.1 M, maximum solubilization efficiency was attained at a pH ranging from 5 to 7, depending on the added salt type. Increased salt concentration up to 1 M resulted in an increased solubilization efficiency for $CaCl_2$ and NaCl, while the addition of $MgCl_2$ beyond 0.1 M showed an anomalous trend. Further, it was noteworthy that too a large extent the protein precipitated in the interface between the organic and aqueous phases at lower pHs and lower salt concentrations. The size of the reverse micelle water pool was estimated by measuring the molar ratio of the surfactant to the water, $W_0$. Irrespective of pH in the aqueous phase, the resulting value of $W_0$ was almost constant, eg., 20 for $MgCl_2$ . However, the value of $W_0$ decreased with increased salt concentration in the cases of KCl and $CaCl_2$.

• Journal of Life Science
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• v.24 no.11
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• pp.1252-1257
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• 2014

While dimethyl sulfoxide (DMSO) is the most commonly used cryoprotectant agent in the cryopreservation of cultured mammalian cells, it has been reported to cause differentiation of some cell lines by DNA methylation and associated histone modifications. To avoid the side effects of DMSO in cryopreservation, other agents might be more appropriate for maintaining the stable differentiation of cultured cell phenotypes through cryopreservation. All cryoprotectants should be highly soluble in water and display low cell toxicity. Cryoprotective agents have been shown to be effective in animal sperm preservation, and eight types of amides were examined in the cryopreservation of cultured mouse endothelial cells. Among the amides examined, acetamide and lactamide were effective cryoprotectants for cultured mammalian cells. The most effective concentration of lactamide, 1.5 M, had an even lower cryoprotective ability than 1M DMSO. Because successful cryopreservation of cultured cells is hampered by osmotic stress, the adequate ionic concentration was determined by diluting phosphate-buffered saline (PBS) in the 1.5M lactamide solution. The most effective concentration was $0.4{\times}PBS$, which minimized osmotic stress during the cryopreservation of cultured cells. As the addition of high molecular weight materials in cryopreservation media improves the viability of cells, the effects of bovine serum albumin (BSA), hydroxyethyl-starch (HES), and dextran were examined. The best combination of lactamide-based media for cryopreservation was found to be 1.5 M lactamide in $0.4{\times}PBS$ with 1% BSA.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.131-137
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• 1997

This study was carried out to investigate the in vivo development rates of vitrified-thawed mouse expanding, hatching and hatched blastoc ysts(BL). In vitro fertilization produced blastocysts were vitrified in EFS40(40% ethylene glycol, 30% Ficoll and 0.3 M sucrose in phosphate buffer saline containing 10% FBS). Expanding a and hatching blastocysts were equilibrated in 20% ethylene glyco](EG) for 5 min. before exposure to EFS40 at 25°C for 1 min., they were then vitrified in liquid nitrogen. Hatched blastocysts which cultured in m-CR1 medium supple mented 0.4% bovine serum albumin on day 5. were equilibrated in 10% EG for 5 min. and then vitrified in EFS40 for 30 sec. After thawing, re-expanding blastocysts were transferred to recipients(3 day of pseudopregnant) on one or both uterine horns(6-8 embryos per a horn). Preg¬n nancy rates of recipients and implantation were a assccessed by autopsy on 15 gestation. The res¬u ults obtained in these experiments were summar¬1 ized as follows; 1) The pregnancy and live fetus rates, for vitrified expanding BL(77.8 and 25.0%) and hatching BL(77.8 and 26.4%)n vitro were not significantly difference in those of control BL (66.7 and 42.9%: 83.3 and 40.4%), respectively, 2) in vitro development of vitrified hatched BL was 34.0%. and 3) in vivo developmental rate of vitrified hatched BL was only 33.3%. These results suggested that proposed rapid vitrification p procedures used EFS40 cryoprotectant can be effectively performed in mouse expanding Ihatching blastocysts and that mouse blastocysts a after being hatched from zona pellucida can be successfully cryopreserved.

• Journal of Embryo Transfer
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• v.17 no.3
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• pp.187-193
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• 2002

This study was conducted to determine the effect of extract of Cervi pantotrichum cornu on human sperm motility, Four different types of media were prepared such as plain Ham's F-10 medium(control medium), control medium containing 0.3% bovine serum albumin(BSA)(medium A), control medium containing the extract of Cervi pantotrichum cornu aqua-acupuncture medium(medium B) and medium B containing 0.3% BSA(medium C). Human semen were washed and divided into 4 fractions and sperm were cultured in those medium for up to 72 hours at 37$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$ in air. A total twenty eight semen samples including 14 normozoospermia and 14 asthenospermia were used for this study. In normozoospermia group, motility of control medium and medium A, B and C were 4.1%, 1.3%, 64.5%, and 77.1%, respectively after 24 hours of incubation, and were 0.0%, 0.0%. 8.8% and 44.9%, respectively after 48 hours of incubation. In asthenospermia group, motility of control medium and medium A, B and C were 2.0%, 2.2%, 58.3% and 85.1%, respectively after 24 hours of incubation, and decreased to 0.0%, 0.2%, 5.8% and 29.6%, respectively after 48 hours of incubation. In both groups, highest sperm motility was observed in medium C group when compared with other media. Furthermore motile sperm were found in medium C after 72 hours of incubation while no motile sperm was observed in the other media. Therefore it could be concluded that the extract of Cervi pantotrichum rornu affects on the human sperm motility.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.78-82
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• 2013

A novel Chryseobacterium sp. JK1 strain isolated from soil had been reported that this isolate produced large amount of extracellular protease at mesophilic temperature in previous study. The optimal temperature and pH of extracellular protease were $40^{\circ}C$ and 7.0, respectively, showing narrow range of optimal temperature and relatively broad activity from pH 6.0 to 9.0. In addition, the protease showed greatest activity against skim milk and lowest against bovine serum albumin (BSA). The protease strongly inhibited by ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA) or phenylmethylsulfonyl fluoride (PMSF), and addition of cation $Ag^+$ or $Cu^{2+}$, and slightly inhibited by $Al^{3+}$. No significant inhibition was found with pepstatin, and addition of cation, $K^+$, $Ca^{2+}$, $Na^+$, $Fe^{2+}$ or $Mg^{2+}$. On the contrary, protease was enhanced by addition of divalent cation $Mn^{2+}$ (5 mM). Zymography analysis of concentrated culture supernatant revealed two major bands at 67 and 145 kDa. These results suggest that Chryseobacterium sp. JK1 strain produced extracellular neutral serine proteases which could apply in food industry.

• BMB Reports
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• v.29 no.2
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• pp.105-110
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• 1996

Polyclonal antiserum R101 against aflatoxin $B_1$ ($AFB_1$) was raised in New Zealand white rabbits after injection of bovine serum albumin-$AFB_1$ conjugate. Competitive ELISA (enzyme linked immuno-sorbent assay) demonstrated that antiserum R101 has the highest binding for $AFB_1$ (50% inhibition at 170 fmol) and aflatoxicol II (50% inhibition at 112 fmol). It also reacts with other aflatoxins such as $AFB_2$, $AFG_1$, $AFG_2$, and aflatoxin metabolites ($AFM_1$, $AFM_2$, $AFP_1$, and $AFQ_1$), but it does not cross-react with $AFG_2a$. Using this antiserum, aflatoxins were quantitated in 100 urine samples of undergraduate students at the College of Pharmacy, Sung Kyun Kwan University, Republic of Korea. By ELISA, $AFB_1$ and its metabolites were detected in human urine samples (N=100, male=89, female=11, ages=20~31 yrs) with a range of 1.4~200.6 ng/kg/day (mean$\pm$SD=$18.11{\pm}33.01\;ng\;AFB_1/kg/day$ in males, $3.82{\pm}2.65\;ng/kg/day$ in females). Assuming that urinary excretion is about 7.6% of $AFB_1$ intake (Groopman et al., 1992), we estimated that Koreans were daily exposed to a total dietary $AFB_1$ of $240.20{\pm}438.67\;ng/kg/day$ in males and $50.35{\pm}29.88\;ng/kg/day$ in females, respectively. When the human monitoring data was applied to a linear regression model of Y=21.67X-10.04 {Y=liver cancer incidence per 100,000, X=Log $AFB_1$ intake (ng/kg/day), r=0.99} developed from previously reported epidemiological data, calculated liver cancer incidences attributed to $AFB_1$ exposure were 41.56/100,000 in males and 26.84/100,000 in females. The incidences were similarly correlated with liver cancer mortality rates of 43.43/100,000 in males and 11.23/100,000 in females in Korea. These results suggest that aflatoxin exposure may be an important risk factor for the high incidence of liver cancer in Korea.

• Membrane Journal
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• v.17 no.3
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• pp.219-232
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• 2007

The polyethersulfone(PES)-titanium oxide($TiO_2$) hybrid membranes were prepared by immersion precipitation phase inversion method. The casting solution for the preparation of $PES-TiO_2$ hybrid membrane was provided by adding $TiO_2$ nano particles into the basis polymer solution of 14 wt% and 20 wt% PES/N-methyl-2-pyrrolidone(NMP). The $TiO_2$ loading [wt% ($TiO_2/NMP$)] in eating solution was varied from 0 to 60 wt%. Membrane performance and morphological change of the resulting $PES-TiO_2$ hybrid membranes were discussed in aspect of $TiO_2$ loading, by viscosity, coagulation value and light transmittance of the casting solution, measurement of tensile strength, pore size and contact angle, surface and cross sectional SEM images of the hybrid membrane, and ultrafiltration experiments using the hybrid membrane. According as increase of $TiO_2$ loading in the casting solution, viscosity is increased and coagulation value becomes lower, therefore the thermodynamic instability of the casting solution is increased. It is found that when $TiO_2$ loading is increased, 1) precipitation rate becomes faster while instantaneous demixing is maintained, 2) pure water flux, membrane pore size and compaction stability of the resulting membranes are increased, 3) tensile strength and contact angle are decreased. Dead-end ultrafiltration of bovine serum albumin(BSA) solution using the hybrid membrane shows that membrane performance(flux of BSA solution) enhanced up to 7 times compared with the results obtained using the pure PES membrane(not containing $TiO_2$ particle), due to the increase of hydrophilicity.

• Proceedings of the Korea Environmental Mutagen Society Conference
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• 2002.11a
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• pp.80-88
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• 2002

Recent studies demonstrate that collagen IV selectively pro-motes the repair of physiological processes in sublethally injured renal proximal tubular ceils (RPTC). We sought to further define the mechanisms of cell repair by measuring the effects of toxicant injury and stimulation of repair by L-ascorbic acid-2-phosphate (AscP), exogenous collagen IV, or function-stimulating integrin antibodies on the expression and subcellular localization of collagen-binding integrins (CBI) in RPTC. Expression of CBI subunits ${\alpha}_1$, ${\alpha}_2$, and ${\beta}_1$ in RPTC was not altered on day 1 after sublethal injury by S-(1,2-dichlorovinyl)-L-cysteine (DCVC). On day 6, expression of ${\alpha}_1$ and ${\beta}_1$ subunits remained unchanged, whereas a 2.2-fold increase in ${\alpha}_2$ expression was evident in injured RPTC. CBI localization in control RPTC was limited exclusively to the basal membrane. On day 1 after injury, RPTC exhibited a marked inhibition of active $Na^+$ transport and a loss of cell polarity characterized by a decrease in basal CBI localization and the appearance of CBI on the apical membrane. On day 6 after injury, RPTC still exhibited marked inhibition of active $Na^+$ transport and localization of CBI to the apical membrane. However, DCVC-injured RPTC cultured in pharmacological concentrations of AscP (500 ${\mu}$M)or exogenous collagen IV (50 ${\mu}$g/ml) exhibited an increase inactive $Na^+$ transport, relocalization of CBI to the basal membrane, and the disappearance of CBI from the apical membrane on day 6. Function-stimulating antibodies to CBI ${\beta}_1$ did not promote basal relocalization of CBI despite stimulating the repair of $Na^+$/$K^+$-ATPase activity on day 6 after injury. These data demonstrate that DCVC disrupts integrin localization and that physiological repair stimulated by AscP or collagen IV is associated with the basal relocalization of CBI in DCVC-injured RPTC. These data also suggest that CBI-mediated repair of physiological functions may occur independently of integrin relocalization.