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Park, Yeo-Reum,Park, Hye-Bin,Kim, Mi-Jeong,Jung, Bae-Dong,Lee, Seunghyung,Park, Choon-Keun,Cheong, Hee-Tae The Korean Society of Developmental Biology 2019 발생과 생식 Vol.23 No.1
We examined the effects of endoplasmic reticulum (ER) stress inhibitor treatment during the micromanipulation of porcine somatic cell nuclear transfer (SCNT) on the in vitro development of SCNT embryos. ER stress inhibitors such as salubrinal (200 nM) and tauroursodeoxycholic acid (TUDCA; $100{\mu}M$) were added to the micromanipulation medium and holding medium. The expression of X-box binding protein 1 (Xbp1), ER-stress-associated genes, and apoptotic genes in SCNT embryos was confirmed at the one-cell and blastocyst stages. Levels of Xbp1 splicing and expression of ER-stress-associated genes in SCNT embryos at the one-cell stage decreased significantly with TUDCA treatment (p<0.05). The expression of ER-stress-associated genes also decreased slightly with the addition of both salubrinal and TUDCA (Sal+TUD). The expression levels of caspase-3 and Bcl2-associated X protein (Bax) mRNA were also significantly lower in the TUDCA and Sal+TUD treatments (p<0.05). At the blastocyst stage, there were no differences in levels of Xbp1 splicing, and transcription of ER-stress-associated genes and apoptosis genes between control and treatment groups. However, the blastocyst formation rate (20.2%) and mean blastocyst cell number ($63.0{\pm}7.2$) were significantly higher (p<0.05) for embryos in the TUDCA treatment compared with those for control (12.6% and $41.7{\pm}3.1$, respectively). These results indicate that the addition of ER-stress inhibitors, especially TUDCA, during micromanipulation can inhibit cellular damage and enhance in vitro development of SCNT embryos by reducing stress levels in the ER.
Park, A Reum,Kim, Jung Sub,Kim, Kwang Su,Zhang, Kan,Park, Juhyun,Park, Jong Hyeok,Lee, Joong Kee,Yoo, Pil J. American Chemical Society 2014 ACS APPLIED MATERIALS & INTERFACES Vol.6 No.3
<P>Although Si is a promising high-capacity anode material for Li-ion batteries (LIB), it suffers from capacity fading due to excessively large volumetric changes upon Li insertion. Nanocarbon materials have been used to enhance the cyclic stability of LIB anodes, but they have an inherently low specific capacity. To address these issues, we present a novel ternary nanocomposite of Si, Mn, and reduced graphene oxide (rGO) for LIB anodes, in which the Si–Mn alloy offers high capacity characteristics and embedded rGO nanosheets confer structural stability. Si–Mn/rGO ternary nanocomposites were synthesized by mechanical complexation and subsequent thermal reduction of mixtures of Si nanoparticles, MnO<SUB>2</SUB> nanorods, and rGO nanosheets. Resulting ternary nanocomposite anodes displayed a specific capacity of 600 mAh/g with ∼90% capacity retention after 50 cycles at a current density of 100 mA/g. The enhanced performance is attributed to facilitated Li-ion reactions with the MnSi alloy phase and the formation of a structurally reinforced electroconductive matrix of rGO nanosheets. The ternary nanocomposite design paradigm presented in this study can be exploited for the development of high-capacity and long-life anode materials for versatile LIB applications.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/aamick/2014/aamick.2014.6.issue-3/am404608d/production/images/medium/am-2013-04608d_0008.gif'></P>
Yeo-Reum Park,Hye-Bin Park,Hwa-Yeon Lee,Hyo-Kyung Bae,Hui-Yeon Shin,Seunghyung Lee,Choon-Keun Park,Boo-Keun Yang,Hee-Tae Cheong 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
This study was conducted to examine the effects of activation methods on the ER stress induction and subsequent apoptosis and in vitro development of porcine parthenogenetic embryos. Porcine in vitro matured oocytes were activated by four activation methods; 1) electric stimulus(ES) with two DC pulses of 1.25 kV/cm, for 30 ㎲ (E), 2) ES + 10 μM Ca-ionophore (A23187) treatment for 5 min (EC), 3) ES + 2 mM 6-dimethylaminopurine treatment for 3 h (ED), or 4) ES + A23187 + 6-DMAP (ECD). After activation, parthenogenetic embryos were in vitro cultured in PZM-3 medium and sampled to analyze the x-box binding protein 1 (Xbp1) mRNA, ER stress-associated genes and apoptotic genes at 3 h post ES and the 1-cell and blastocyst stages. The un-spliced and spliced x-box binding protein 1 (Xbp1) mRNA were confirmed by RT-PCR. Also ER stress-associated genes, such as the C/EBP homologous protein (CHOP), binding protein (BiP), activating transcription factor 4 (ATF4) and glucose-regulated protein 94 (GRP94), and apoptotic genes were analyzed by real-time quantitative RT-PCR (RT-qPCR). The band intensities of spliced Xbp1 (Xbp1s) mRNA was higher in the EC group than other three groups at 3 h and the 1-cell stage, while it was higher in the ED groups compared with E group at the blastocyst stage. Four ER stress-associated genes were showed the highest expression in the EC group and weakly expressed in the ED group at 3 h. However, most of those genes were highly expressed in EC and ECD groups at the 1-cell and blastocyst stages with some variation. The expressions of Bcl-2-associated X protein (Bax) and caspase-3 mRNAs were significantly higher in EC group than other three groups at all stages. The developmental rate to the blastocyst stage was higher (p<0.05) in ED and ECD groups (32.1±3.8 to 34.6±2.2%) than that of E group (26.1±3.9%). These results suggest that the intracellular ER stress of parthenogenetic porcine embryos is affected by activation method and subsequently lead to the apoptosis of embryos.
Park, Yoorim,Jung, Min Kyung,Yoon, Sun Young,Lee, Ha-Reum,Hur, Dae Young,Kim, Daejin,Yang, Yoolhee,Kim, Tae Sung,Kim, Seonghan,Yoon, Suk Ran,Park, Hyun Jeong,Bang, Sa Ik,Cho, Dae Ho Published for the International Union of Biochemis 2013 Biotechnology and applied biochemistry Vol.60 No.3
<P>Adipose stem cells (ASCs) are pluripotent cells that can generate pure fat tissue for regeneration. Differentiated adipose cells have been generated by a common inducer cocktail composed of dexamethasone, insulin, and isobutylmethylxanthine (DIM). The major drawbacks of adipose cells are their tendency to float on the culture media and their cost. To overcome some of these disadvantages, a new inducer cocktail that includes insulin, dehydroepiandrosterone, and histamine (DH IH) was tested. As a result, lipid accumulation was elevated more than twofold with DH IH than with DIM. Cell adhesion and viability, which are important factors for stable differentiation, were increased with DH IH and were proven through measurement of mRNA expression levels of adhesion marker genes, N-cadherin and vascular cell adhesion molecule, as well as through an alamar blue assay. The expression of adipogenesis-related genes, adiponectin, and glucose transporter type 4 lasted for a long time. To improve the efficiency of grafting, cell adhesion and neovascularization need to be increased. Neovascularization was observed around the transplanted adipose cells, which showed a higher number of vessel formation in DH IH than in DIM. The above results suggest that DH IH can produce pure differentiated adipose cells effectively and enhance their adhesion onto the target location when these differentiated adipose cells were applied as a clinical resource.</P>
Park, Ah Reum,Park, Jeong-Hoon,Ahn, Hye-Jin,Jang, Ji Yeon,Yu, Byung Jo,Um, Byung-Hwan,Yoon, Jeong-Jun The Korean Society of Mycology 2015 Mycobiology Vol.43 No.1
${\beta}$-Glucosidase, which hydrolyzes cellobiose into two glucoses, plays an important role in the process of saccharification of the lignocellulosic biomass. In this study, we optimized the activity of ${\beta}$-glucosidase of brown-rot fungus Fomitopsis pinicola KCTC 6208 using the response surface methodology (RSM) with various concentrations of glucose, yeast extract and ascorbic acid, which are the most significant nutrients for activity of ${\beta}$-glucosidase. The highest activity of ${\beta}$-glucosidase was achieved 3.02% of glucose, 4.35% of yeast extract, and 7.41% ascorbic acid where ascorbic acid was most effective. The maximum activity of ${\beta}$-glucosidase predicted by the RSM was 15.34 U/mg, which was similar to the experimental value 14.90 U/mg at the 16th day of incubation. This optimized activity of ${\beta}$-glucosidase was 23.6 times higher than the preliminary activity value, 0.63 U/mg, and was also much higher than previous values reported in other fungi strains. Therefore, a simplified medium supplemented with a cheap vitamin source, such as ascorbic acid, could be a cost effective mean of increasing ${\beta}$-glucosidase activity.
Discovery of parallel pathways of kanamycin biosynthesis allows antibiotic manipulation
Park, Je Won,Park, Sung Ryeol,Nepal, Keshav Kumar,Han, Ah Reum,Ban, Yeon Hee,Yoo, Young Ji,Kim, Eun Ji,Kim, Eui Min,Kim, Dooil,Sohng, Jae Kyung,Yoon, Yeo Joon Nature Publishing Group, a division of Macmillan P 2011 Nature chemical biology Vol.7 No.11
Kanamycin is one of the most widely used antibiotics, yet its biosynthetic pathway remains unclear. Current proposals suggest that the kanamycin biosynthetic products are linearly related via single enzymatic transformations. To explore this system, we have reconstructed the entire biosynthetic pathway through the heterologous expression of combinations of putative biosynthetic genes from Streptomyces kanamyceticus in the non??aminoglycoside-producing Streptomyces venezuelae. Unexpectedly, we discovered that the biosynthetic pathway contains an early branch point, governed by the substrate promiscuity of a glycosyltransferase, that leads to the formation of two parallel pathways in which early intermediates are further modified. Glycosyltransferase exchange can alter flux through these two parallel pathways, and the addition of other biosynthetic enzymes can be used to synthesize known and new highly active antibiotics. These results complete our understanding of kanamycin biosynthesis and demonstrate the potential of pathway engineering for direct in vivo production of clinically useful antibiotics and more robust aminoglycosides.
Park, Hye-Bin,Park, Yeo-Reum,Lee, Hwa-Yeon,Bae, Hyo-Kyung,Lee, Seunghyung,Park, Choon-Keun,Yang, Boo-Keun,Cheong, Hee-Tae The Korean Society of Embryo Transfer 2017 한국동물생명공학회지 Vol.32 No.1
This study was conducted to investigate the effect of activation method on the endoplasmic reticulum (ER) stress induction, apoptosis and in vitro development of porcine parthenogenetic embryos. Porcine in vitro matured oocytes were activated by four activation methods; 1) electric stimulus (ES) (E), 2) $ES+10{\mu}M$ Ca-ionophore (A23187) treatment (EC), 3) ES+2 mM 6-dimethylaminopurine (6-DMAP) treatment (ED), or 4) ES+A23187 and 6-DMAP treatments (ECD). Parthenogenetic embryos were sampled to analyze x-box binding protein 1 (Xbp1) mRNA, ER stress-associated genes and apoptosis genes at 3 h after ES and the 1-cell and blastocyst stages. In the EC group, the band intensity of spliced Xbp1 (Xbp1s) mRNA was higher than those of the other groups at the 3 h and 1-cell stage, and higher than that of the E group at the blastocyst stage. Four ER stress-associated genes were expressed at the highest level in the EC group and weakly expressed in the ED group at 3 h after activation. However, most of the genes were highly expressed at the 1-cell and blastocyst stages with some variation in the EC and ECD groups. Expression of Bcl-2-associated X protein (Bax) and caspase-3 mRNA was significantly higher in the EC group than in the other groups at all development stages. The developmental rates to the blastocyst stage were higher in the ED and ECD groups than in the E and EC groups. These results suggest that the intracellular ER stress of parthenogenetic porcine embryos is affected by the activation method and subsequently lead to the apoptosis of embryos.