• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.95-95
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• 2002

Human embryonic stem (hES) cells can be induced to differentiate into tyrosine hydroxylase expressing (TH+) cells that may serve as an alternative for cell replacement therapy for Parkinson's disease (PD). To examine in vitro differentiation of hES (MB03, registered in NIH) cells into TH+ cells, hES cells were induced to differentiate according to the 4-/4+ protocol using retinoic acid (RA), ascorbic acid (AA), and/or lithium chloride (LiCl) followed by culture in N2 medium for 14 days, during which time the differentiation occurs. Immunocytochemical stainings of the cells revealed that approximately 21.1% of cells treated with RA plus AA expressed TH protein that is higher than the ratio of TH+ cells seen in any other treatment groups (RA, RA+LiCl or RA+AA+LiCl). In order to see the differentiation pattern in vivo and the ability of in vitro differentiation-induced cells in easing symptomatic motor function of PD animal model, cells (2 $\times$ 10$^{5}$ cells/2${mu}ell$) undergone 4-/4+ protocol using RA plus AA without any further treatment were transplanted into unilateral striatum of MPTP-lesioned PD animal model (C57BL/6). Following the surgery, motor behavior of the animals was examined by measuring the retention time on an accelerating rotar-rod far next 10 weeks. No significant differences in retention time of the animals were noticed until 2 weeks post-graft; however, it increased markedly at 6 weeks and 10 weeks time point after the surgery. Immunohistochemical studies confirmed that a reasonable number of TH+ cells were found at the graft site as well as other remote sites, showing the migrating nature of embryonic stem cells. These results suggest that in viかo differentiated hES cells relieve symptomatic motor behavior of PD animal model and should be considered as a promising alternative for the treatment of PD.

• Biomolecules & Therapeutics
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• v.18 no.1
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• pp.32-38
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• 2010

Activation of JNK has long been associated with the apoptotic response induced by various anti-cancer drugs including doxorubicin, vinblastine, and etoposide. In this study, we examined and compared patterns of apoptosis and JNK activation according to three different anti-cancer drugs (daunorubicin, vinblastine, and etoposide) and two different sources of HL60 cells (Jackson Laboratory and ATCC). HL60 cells from Jackson Laboratory (HL60/RPMI) were maintained in RPMI 1640 containing 5% fetal bovine serum and those from ATCC (HL60/IMDM) in IMDM containing 20% fetal bovine serum as to each manufacture's guideline. In general, HL60/RPMI cells were more sensitive to anti-cancer drugs compared to HL60/IMDM cells, demonstrated by the XTT and flow cytometric analyses. Apoptotic pathways after treatment with anti-cancer drugs seemed to be different between HL60/RPMI (daunorubicin and etoposide, caspase 3 dependent, but caspase 8 or 9 independent; vinblastine, caspase 3 independent) and HL60/IMDM (caspase 3 and caspase 9 dependent). The expression of apoptotic protein, BID, was consistent with caspase 3 activation. Immunoblotting of phospho-JNK and JNK kinase assay showed JNK activation by all three anti-cancer drugs in HL60/RPMI, while JNK activation was observed only in vinblastine-treated cells in HL60/IMDM. Our study results suggest that in vitro environmental conditions have a significant influence on JNK mediated apoptosis of HL60 cells by anti-cancer drugs and in vitro culture conditions are important factors in JNK or possibly other MAPK related studies.

• The Korean Journal of Food And Nutrition
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• v.23 no.1
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• pp.88-93
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• 2010

The principal objective of this study was to evaluate the physicochemical characteristics of brown rice Dasik prepared with different 5 levels(0, 25, 50, 75, 100%) of modified potato starch powder. We assessed the general compositions, Hunter's color values, the mechanical characteristics, and conducted a sensory evaluation analysis of brown rice Dasik at room temperature($20^{\circ}C$). The more modified potato starch powder composition was increased, the content of ash, protein and fat were significantly decreased(p<0.05). We noted that the luminance of samples were increased the more modified potato starch powder composition was increased, but in Hunter's a and b values were decreased. With regard to the mechanical properties of the Dasik samples, we noted significant differences in hardness, gumminess, chewiness but no differences in springiness, cohesiveness and adhesiveness were detected. The more modified potato starch powder composition was increased, the more the score of hardness, gumminess and chewiness were higher. The results of sensory evaluation showed that there were significant differences on the color, sweetness, mouth-feel, chewiness, overall quality of the Dasik samples(p<0.05). The score of brown rice Dasik with 50%(w/w) modified potato starch(BM2) in color, mouth-feel, chewiness, overall quality were significantly increased than those of other samples.

• Food Science of Animal Resources
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• v.33 no.6
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• pp.708-714
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• 2013

This study was performed to investigate the influence of age on the physicochemical properties of strip loin (m. longissimus lumborum), chuck tender (m. supraspinatus), and eye of round (m. semitendinosus) of Hanwoo cows. Hanwoo cows (n=126; 24-194 mon; live weight, 270-500 kg) were slaughtered and three muscles were obtained according to 3 age groups (G1, < 5 years old; G2, 6-8 years old; G3, > 9 years old). The chuck tender had significantly higher protein contents in G3 than in G1 or G2 (p<0.05). For strip loin and chuck tender, G1 had significantly higher intramuscular fat contents than G3 (p<0.05). The chuck tender had significantly higher $b^*$ (yellowness) values for G2 than for G1 (p<0.05). The three muscles had significantly higher cooking loss (%) and lower WHC (%) in G3 than in G1 or G2 (p<0.05). WBS values of strip loin were significantly higher in G3 than in the other groups (p<0.05). The three muscles had higher C18:1n9 in G1 than that in the other groups. The total content of saturated fatty acids was significantly higher in G3 than in G1 for all 3 muscles (p<0.05). Regarding free amino acid contents, G1 had significantly higher contents of threonine, alanine, valine, methionine, phenylalanine, leucine, and lysine in the strip loin than G2 or G3 (p<0.05). In conclusion, young cow beef were higher in the WHC, intramuscular fat and free amino acids contents, whereas old cow beef had higher cooking loss and WBS (p<0.05).

• Journal of Aquaculture
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• v.19 no.2
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• pp.140-146
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• 2006

This study was conducted to investigate the effect of dietary oxidized oil and ${\alpha}$-tocopherol level on growth and body composition of juvenile flounder. To prepare oxidized diets, squid liver oil was oxidized by aeration at $25^{\circ}C$ for 30 days. The six diets were prepared to contain 6% fresh or oxidized squid liver oil as the lipid sources in combination with three levels of ${\alpha}$-tocopheryl acetate at 0, 80 and 800 mg/kg diet. Triplicate groups of fish ($3.9{\pm}0.1$) were fed to apparent satiation twice a day for 8 weeks. Survival was not significantly different among treatments. Weight gain, feed efficiency, daily feed intake, protein efficiency ratio and condition factor of fish fed the fresh oil diets were significantly higher than those of fish fed the oxidized oil diets (P<0.05). The increase of the vitamin E level in diets did not result in any significant improvement on growth performance of fish fed both oil diets. The vitamin E content of the liver and dorsal muscle increased with increasing dietary vitamin E level at both oil diet groups. A decreasing trend in vitamin E content of the tissues was observed in fish fed the oxidized oil diets at the same dietary vitamin E level. Significantly higher moisture content and lower crude lipid content were observed in the whole body of fish fed the oxidized oil diets than fish fed the fresh oil diets (P<0.05). Dietary lipid source affected the fatty acid content of the whole body; higher contents of saturated and monoenoic fatty acids, and lower n-3 HUFA contents such as 20:5n-3 and 22:6n-3 were observed in fish fed the oxidized oil diets than those of fish fed fresh oil diets. The results of this study suggest that the dietary oxidized oil may impair the growth performance, and an increase in ${\alpha}$-tocopheryl acetate supplementation have no beneficial effect on growth and feed efficiency of juvenile flounder.

• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1295-1300
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• 2009

A 2,172-bp full-length gene (aga-F78), encoding a protease-resistant $\alpha$-galactosidase, was cloned from Rhizopus sp. F78 and expressed in Escherichia coli. The deduced amino acid sequence shared highest identity (45.0%) with an $\alpha$-galactosidase of glycoside hydrolase family 36 from Absidia corymbifera. After one-step purification with a Ni-NTA chelating column, the recombinant Aga-F78 migrated as a single band of ~82 and ~210 kDa on SDS-PAGE and nondenaturing gradient PAGE, respectively, indicating that the native structure of the recombinant Aga-F78 was a trimer. Exhibiting the similar properties as the authentic protein, purified recombinant Aga-F78 was optimally active at $50^{\circ}C$ and pH 4.8, highly pH stable over the pH range 5.0-10.0, more resistant to some cations and proteases, and had wide substrate specificity (pNPG, melidiose, raffinose, and stachyose). The recombinant enzyme also showed good hydrolytic ability to soybean meal, releasing galactose of $415.58\;{\mu}g/g$ soybean meal. When combined with trypsin, the enzyme retained over 90% degradability to soybean meal. These favorable properties make Aga-F78 a potential candidate for applications in the food and feed industries.

• Journal of Microbiology and Biotechnology
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• v.27 no.11
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• pp.2044-2051
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• 2017

The main pathological hallmark of Alzheimer's disease is the deposition of amyloid-beta ($A{\beta}$) peptides in the brain. $A{\beta}$ has been widely used to mimic several aspects of Alzheimer's disease. However, several characteristics of amyloid-induced Alzheimer's disease pathology are not well established, especially in mice. The present study aimed to develop a new Alzheimer's disease model by investigating how $A{\beta}$ can be effectively aggregated using prokaryotes and eukaryotes. To express the $A{\beta}42$ complex in HEK293 cells, we cloned the $A{\beta}42$ region in a tandem repeat and incorporated the resulting construct into a eukaryotic expression vector. Following transfection into HEK293 cells via lipofection, cell viability assay and western blotting analysis revealed that exogenous $A{\beta}42$ can induce cell death and apoptosis. In addition, recombinant His-tagged $A{\beta}42$ was successfully expressed in Escherichia coli BL21 (DE3) and not only readily formed $A{\beta}$ complexes, but also inhibited the proliferation of SH-SY5Y cells and E. coli. For in vivo testing, recombinant His-tagged $A{\beta}42$ solution ($3{\mu}g/{\mu}l$ in $1{\times}PBS$ containing $1mM\;Ni^{2+}$) was injected stereotaxically into the left and right lateral ventricles of the brains of C57BL/6J mice (n = 8). Control mice were injected with $1{\times}PBS$ containing $1mM\;Ni^{2+}$ following the same procedure. Ten days after the sample injection, the Morris water maze test confirmed that exogenous $A{\beta}$ caused an increase in memory loss. These findings demonstrated that $Ni^{2+}$ is capable of complexing the 50-kDa amyloid and that intracerebroventricular injection of $A{\beta}42$ can lead to cognitive impairment, thereby providing improved Alzheimer's disease models.

• Biomolecules & Therapeutics
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• v.22 no.1
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• pp.17-26
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• 2014

${\alpha}$-Asarone exhibits a number of pharmacological actions including neuroprotective, anti-oxidative, anticonvulsive, and cognitive enhancing action. The present study investigated the effects of ${\alpha}$-asarone on pro-inflammatory cytokines mRNA, microglial activation, and neuronal damage in the hippocampus and on learning and memory deficits in systemic lipopolysaccharide (LPS)-treated C57BL/6 mice. Varying doses of ${\alpha}$-asarone was orally administered (7.5, 15, or 30 mg/kg) once a day for 3 days before the LPS (3 mg/kg) injection. ${\alpha}$-Asarone significantly reduced TNF-${\alpha}$ and IL-$1{\beta}$ mRNA at 4 and 24 hours after the LPS injection at dose of 30 mg/kg. At 24 hours after the LPS injection, the loss of CA1 neurons, the increase of TUNEL-labeled cells, and the up-regulation of BACE1 expression in the hippocampus were attenuated by 30 mg/kg of ${\alpha}$-asarone treatment. ${\alpha}$-Asarone significantly reduced Iba1 protein expression in the hippocampal tissue at a dose of 30 mg/kg. ${\alpha}$-Asarone did not reduce the number of Iba1-expressing microglia on immunohistochemistry but the average cell size and percentage areas of Iba1-expressing microglia in the hippocampus were significantly decreased by 30 mg/kg of ${\alpha}$-asarone treatment. In the Morris water maze test, ${\alpha}$-asarone significantly prolonged the swimming time spent in the target and peri-target zones. ${\alpha}$-Asarone also significantly increased the number of target heading and memory score in the Morris water maze. The results suggest that inhibition of pro-inflammatory cytokines and microglial activation in the hippocampus by ${\alpha}$-asarone may be one of the mechanisms for the ${\alpha}$-asarone-mediated ameliorating effect on memory deficits.

• Childhood Kidney Diseases
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• v.12 no.1
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• pp.88-92
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• 2008

Infection of Epstein-Barr virus(EBV) gives rise to a broad spectrum of clinical manifestations in children. Although renal involvement is rare, diverse renal manifestations are known from hematuria to acute renal failure. Secondary membranous nephropathy(MN) associated with systemic EBV infection is an uncommon renal pathology and only two cases have been reported. We are adding another case of MN associated with EBV infection in a child. An 8-year-old girl was admitted for renal biopsy. She had been followed up for microscopic hematuria and intermittent proteinuria for 5 months. There had been no specific findings in serology and radiology. Tonsil biopsy had been done due to exudative tonsillar hypertrophy and enlarged multiple cervical lymph nodes. And it showed EBV-associated lymphoproliferative findings. Serologic tests for EBV showed positive evidence of recent infection; viral capsid antigen(VCA) IgM was borderline positive, VCA IgG and early antigen IgG were positive, and EB nuclear antigen IgG was negative. In Situ Hybridization of tonsil for EBV mRNA was positive. Because her proteinuria and hematuria were aggravated at that time(protein 3 +, RBC >60/HPF), renal biopsy was done. Renal biopsy showed the findings of MN, characterized by thickened capillary walls with epimembranous spikes on light microscopy and subepithelial, mesangial and subendothelial electron dense deposits on electron microscopy. On immunofluorescence microscopy, IgG, C1q, kappa and lambda chains were positive. After steroid administration, proteinuria and hematuria resolved gradually within 6 months.

• Food Science of Animal Resources
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• v.31 no.1
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• pp.129-138
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• 2011

Hydrolyzed wheat gluten (HWG) and low glutamic acid (Glu) hydrolyzed wheat gluten with different quantities of NaCl were reacted with several precursors to develop natural meat flavor based on Maillard reaction products (MRP). The MRP based flavors were analyzed for their pH, browning index, DPPH radical scavenging effect, and sensory properties. Synthetic meat flavor from low Glu hydrolyzed wheat gluten with 7% NaCl and ribose, cysteine, methionine, thiamin, lecithin, and garlic powder reacted at $140^{\circ}C$ for 30 min and were most favorable for a roasted meat flavor. Based on an omission test, cysteine was selected as the most important precursor for producing meat flavor compared to methionine, thiamine, and lecithin. Natural precursors including mushroom powder and fat medium were applied to compensate for the synthetic precursors. The optimum formula for meat flavor was 5% ribose, 7.7% cysteine, 6.9% garlic juice powder, 2.1% Lentinusedodes powder digested with protease, and 1% lard. The sulfuric pungent, oily, and salty attributes of the formula decreased and a mild roasted meat flavor was expressed.