• Journal of Embryo Transfer
• /
• v.23 no.2
• /
• pp.101-108
• /
• 2008

This study investigated the developmental ability and gene expression of somatic cell nuclear transfer embryos using ear skin fibroblast cells derived from miniature pig. When miniature pig (m) and landrace pig (p) were used as donor cells, there were no differences in cleavage (79.2 vs. 78.2%) and blastocyst rates (27.4 vs. 29.7%). However, mNT blastocysts showed significantly higher apoptosis rate than that of pNT blastocysts (6.1 vs. 1.7%) (p<0.05). The number of nuclei in pNT blastosysts was significantly higher than that of mNT (35.8 vs. 29.3) (p<0.05). Blastocysts were analyzed using Realtime RT-PCR to determine the expression of Bax-${\alpha}$, Bcl-xl, H19, IGF2, IGF2r and Xist. Bax-${\alpha}$ was higher in mNT blastocyst than pNT blastocyst (p<0.05). There was no difference in Bcl-xl between two NT groups. Bax-${\alpha}$/Bcl-xl was, however, significantly higher in mNT blastocyst compared to pNT. The expression of imprinting genes were aberrant in blastocysts derived from NT compared to in vivo blastocysts. H19 and IGF2r were significantly lower in mNT blastocysts (p<0.05). The expression of IGF2 and Xist was similar in two NT groups. However, imprinting genes were expressed aberrantly in mNT compared to pNT blastocysts. The present results suggest that the NT between donor cells derived from miniature pig and recipient oocytes derived from crossbred pig might affect reprogramming of donor cell, resulting in high apoptosis and aberrant expression patterns of imprinting genes.

• Proceedings of the Plant Resources Society of Korea Conference
• /
• 2018.04a
• /
• pp.59-59
• /
• 2018

Panax ginseng Meyer (Korean ginseng) is in the spotlight of Oriental medicine and is proclaimed as the king of medicinal plants owing to its adaptogenic characteristics. Ginseng root rot is a devastating disease caused by the fungus, Ilyonectria mors-panacis that generally attacks younger roots (~2 years), leading to defects in root quality, ginsenoside accumulation and also life cycle of the plant. Hence, there is an indispensable need to develop strategies resulting in tolerance against ginseng root rot. In the present study, we evaluated the effect of silica nanoparticles(N-SiO2) in Panax ginseng during I. mors-panacis infection. Long term analysis (30 dpi) revealed a striking 50% reduction in disease severity index upon 1mM and 2mM treatment of N-SiO2. However, N-SiO2 did not have any direct antifungal activity against I. mors-panacis. Membrane bound sugar efflux transporter, SWEET (Sugars Will Eventually be Exported Transporters) was identified in ginseng and as expected, its expression was suppressed upon N-SiO2 treatment in the root rot pathosystem. Furthermore, the total and reducing sugars in the apoplastic fluid clearly revealed that N-SiO2 regulates sugar efflux into apoplast. In a nut shell, N-SiO2 administration induces transcriptional reprogramming in ginseng roots, leading to regulated sugar efflux into apoplast resulting in enhanced tolerance against I. mors-panacis.

• Journal of Embryo Transfer
• /
• v.29 no.4
• /
• pp.345-350
• /
• 2014

We investigate the effect of L-glutathione (GSH), an antioxidant, treatment during the somatic cell nuclear transfer (SCNT) procedures on the in vitro development and DNA methylation status of bovine SCNT embryos. Bovine in vitro matured (IVM) oocytes were enucleated and electrofused with a donor cell, then activated by a combination of Ca-ionophore and 6-dimethylaminopurine. The recipient oocytes or reconstituted oocytes were treated with $50{\mu}M$ GSH during these SCNT procedures from enucleation to activation treatment. The SCNT embryos were cultured for 7 days to evaluate the in vitro development, apoptosis and DNA methylation in blastocysts. The apoptosis was measured by TUNEL assay and caspase-3 activity assay. Methylated DNA of SCNT embryos at the blastocyst stages was detected using a 5-methylcytidine (5-MeC) antibody. The developmental rate to the blastocyst stage was significantly higher (P<0.05) in GSH treatment group ($32.5{\pm}1.2%$, 78/235) than that of non-treated control SCNT embryos ($22.3{\pm}1.8%$, 50/224). TUNEL assay revealed that the numbers of apoptotic cells in GSH treatment group ($2.3{\pm}0.4%$) were significantly lower (P<0.05) than that of control ($3.8{\pm}0.6%$). Relative caspase-3 activity of GSH treated group was $0.8{\pm}0.06$ fold compared to that of control. DNA methylation status of blastocysts in GSH treatment group ($13.1{\pm}0.5$, pixels/embryo) was significantly lower (P<0.05) than that of control ($17.4{\pm}0.9$, pixels/embryo). These results suggest that antioxidant GSH treatment during SCNT procedures can improve the embryonic development and reduce the apoptosis and DNA methylation level of bovine SCNT embryos, which may enhance the nuclear reprogramming of bovine SCNT embryos.

• Journal of Embryo Transfer
• /
• v.33 no.4
• /
• pp.255-264
• /
• 2018

This experiment was conducted to analyse the effects of flavone supplementation on the preimplantation development of in-vitro produced porcine embryos. During in-vitro development, immature oocytes and early embryos were exposed to different concentrations of flavone (0, $1{\mu}M$, $25{\mu}M$, $50{\mu}M$, and $100{\mu}M$ respectively). Results showed that $100{\mu}M$ of flavone significantly reduced the intracellular ROS levels of oocytes accompanied with a significant rise in GSH level. In parthenogenesis, no significant change was observed in the cleavage rates whether flavone was supplemented in IVM or IVC media. In IVM supplemented group, the blastocyst development rate was significantly enhanced by $1{\mu}M$ concentration than other groups (51.5% vs. 41.3%, 44.0%, 36.3%, 31.7%; P<0.05) respectively. However, in IVC group $1{\mu}M$ concentration significantly improved the blastocysts production than $50{\mu}M$ and control groups (50.0% vs. 40.5%, 38.0%; P<0.05) respectively. Following nuclear transfer, the cleavage rate of IVM group was significantly more in $1{\mu}M$ than $50{\mu}M$ and $100{\mu}M$ groups (92.9% vs. 89.7%, 87.8%; P<0.05), followed by similar pattern of cloned blastocysts production being significantly higher in $1{\mu}M$ group than $50{\mu}M$, $100{\mu}M$ and control groups (16.8% vs. 9.0%, 7.1%, 12.8%; P<0.05) respectively. In IVC group, $1{\mu}M$ concentration resulted in significantly higher cleavage rate than $25{\mu}M$ and $50{\mu}M$ groups (91.7% vs. 87.8%, 88.8%; P<0.05) respectively. However, the blastocysts production was significantly higher in $100{\mu}M$ group than others (26.2% vs. 13.6%, 14.0%, 18.2%; P<0.05) respectively. The optimal concentrations of flavone significantly enhanced the percentages of ICM:TE than control group (43.8% vs. 37.6%; P<0.05) accompanied with significantly higher expression levels of reprogramming related genes. In conclusion, the optimal concentrations of $1{\mu}M$ during IVM and $100{\mu}M$ during IVC can significantly improve the production of porcine in-vitro embryos.

• Journal of Life Science
• /
• v.33 no.9
• /
• pp.754-765
• /
• 2023

Exosomes are a type of extracellular vesicle containing proteins and messenger and microRNAs; they are secreted by all cell types. Once released, exosomes are selectively taken up by other cells adjacent or at a distance, releasing their contents and reprogramming the target cells. Since exosomes are natural vesicles produced by cells as small sizes, it is generally accepted that exosomes have a non-toxic nature and non-immunogenic behaviors. Recently, exosomes have elicited scientific attention as drug delivery vehicles to the central nervous system. The central nervous system has a blood-brain barrier that makes it difficult for drugs to penetrate. Thus, the blood-brain barrier has been a major obstacle to the development of drugs for treating neurodegenerative diseases. However, accumulating evidence suggests that exosomes can cross the blood-brain barrier primarily through transcytosis. Consequently, exosomes are expected to become a new delivery vehicle that can cross the blood-brain barrier and deliver drugs into the brain parenchyma. In addition, since different types of exosomes are secreted depending on the cell type and disease state, exosomes can also be utilized as biomarkers for the diagnosis of diseases in the central nervous system. In this review, we summarized recent research trends on exosomes, including clinical trials as biomarkers and treatment options for diseases in the central nervous system.

• IMMUNE NETWORK
• /
• v.20 no.5
• /
• pp.40.1-40.15
• /
• 2020

The protein encoded by the Gene Associated with Retinoid-Interferon-Induced Mortality-19 (GRIM-19) is located in the mitochondrial inner membrane and is homologous to the NADH dehydrogenase 1-alpha subcomplex subunit 13 of the electron transport chain. Multiple sclerosis (MS) is a demyelinating disease that damages the brain and spinal cord. Although both the cause and mechanism of MS progression remain unclear, it is accepted that an immune disorder is involved. We explored whether GRIM-19 ameliorated MS by increasing the levels of inflammatory cytokines and immune cells; we used a mouse model of experimental autoimmune encephalomyelitis (EAE) to this end. Six-to-eight-week-old male C57BL/6, IFNγ-knockout (KO), and GRIM-19 transgenic mice were used; EAE was induced in all strains. A GRIM-19 overexpression vector (GRIM19 OVN) was electrophoretically injected intravenously. The levels of Th1 and Th17 cells were measured via flow cytometry, immunofluorescence, and immunohistochemical analysis. IL-17A and IFNγ expression levels were assessed via ELISA and quantitative PCR. IL-17A expression decreased and IFNγ expression increased in EAE mice that received injections of the GRIM-19 OVN. GRIM19 transgenic mice expressed more IFNγ than did wild-type mice; this inhibited EAE development. However, the effect of GRIM-19 overexpression on the EAE of IFNγ-KO mice did not differ from that of the empty vector. GRIM-19 expression was therapeutic for EAE mice, elevating the IFNγ level. GRIM-19 regulated the Th17/Treg cell balance.

• Journal of Science and Technology Studies
• /
• v.5 no.1 s.9
• /
• pp.55-92
• /
• 2005

The factors contributing to the formation of an important scientific concept in South Korea and its circulation in the society are the scientific knowledge that had been already formed, matured, and established in the U.S.A, Europe and Japan and has been introduced into Korea, and the institutions that have been formed during the recent modernization in South Korea. The concept of 'genetic information' cannot be an exception in this context. The concept of genetic information is the one that has been extended and intensified by the genomics and bioinformatics formed and matured through the Human Genome Projects from the former concept of inheritance or heredity within the framework of classical and molecular genetics. The purpose of this study was to find out 'how the production structure of genetic information in South Korea has been formed', under the perspective of the conceptual, epistemic, and institutional holisticity or integratedness in the concept and knowledge production structure idealized in Western advanced nations. The discourse of genetic engineering popular in the mid 1980's in South Korea has catalyzed the development of molecular biology. However, the institutional balance that had been established for the biochemistry departments in Natural Science College and Medical College was not formed between the genetic engineering and genetics departments in South Korea. Therefore, they were unable to achieve the more integrative and macro-level disciplinary impact on life sciences, largely due to institutional lack of the capable (human) genetics departments in some leading Korean colleges of Medicine. In genomics, the cutting-edge reprogramming and restructuring of the traditional genetics in the West, South Korea has not invested, even meagerly, in the infrastructure, fund, and research and development (R & D) for the Basic or First Phase of the research trajectory in the Human Genome Project. Without a minimal Basic Phase, the genomics research and development in Korea has been running more or less for the Advanced or Second Phase. Bioinformatics has started developing in Korea under a narrow perspective which regards it as a mere sub-discipline of information technology (IT). Having developed itself in parallel with genomics, bioinformatics contains its own unique logics and contents that can be both directly and indirectly connected to the information science and technology. As a result, bioinformatics reveals a defect in respect of being synergistically integrated into genetics and life sciences in Korea. Owing to the structural problem in the production, genetic information appears to be produced in a fragmented pattern in the Korean society since its fundamental base is weak and thin. A good example of the conceptual and institutional fragmentedness is that 'the genetics of individual identification' is not a normal integrated part of the Korean genetics, but a scientific practice exercised in the departments of legal medicine in a few Medical Colleges. And the environment contributing to the production structure of genetic information in South Korea today comprises 'sangmyung gonghak'(or life engineering) discourse and non-governmental organization movement.

• Molecules and Cells
• /
• v.38 no.3
• /
• pp.210-220
• /
• 2015

Athletic performance is an important criteria used for the selection of superior horses. However, little is known about exercise-related epigenetic processes in the horse. DNA methylation is a key mechanism for regulating gene expression in response to environmental changes. We carried out comparative genomic analysis of genome-wide DNA methylation profiles in the blood samples of two different thoroughbred horses before and after exercise by methylated-DNA immunoprecipitation sequencing (MeDIP-Seq). Differentially methylated regions (DMRs) in the pre-and post-exercise blood samples of superior and inferior horses were identified. Exercise altered the methylation patterns. After 30 min of exercise, 596 genes were hypomethy-lated and 715 genes were hypermethylated in the superior horse, whereas in the inferior horse, 868 genes were hypomethylated and 794 genes were hypermethylated. These genes were analyzed based on gene ontology (GO) annotations and the exercise-related pathway patterns in the two horses were compared. After exercise, gene regions related to cell division and adhesion were hypermethylated in the superior horse, whereas regions related to cell signaling and transport were hypermethylated in the inferior horse. Analysis of the distribution of methylated CpG islands confirmed the hypomethylation in the gene-body methylation regions after exercise. The methylation patterns of transposable elements also changed after exercise. Long interspersed nuclear elements (LINEs) showed abundance of DMRs. Collectively, our results serve as a basis to study exercise-based reprogramming of epigenetic traits.

• Journal of Life Science
• /
• v.29 no.8
• /
• pp.895-903
• /
• 2019

The aim of the present study was to improve the cell viability of human dental papilla derived single-induced pluripotent stem cells (iPSCs) using a Rho-associated protein kinase (ROCK) inhibitor, Y-27632. The iPSCs were produced using an episomal plasmid-based reprogramming method. After cell separation using trypsin, the iPSCs were treated with 0, 0.5, 1, 2.5, 5, 7.5, or $10{\mu}M$ Y-27632 for 5 d. Cell viability increased significantly following the $5{\mu}M$ Y-27632 treatment (p<0.05). When the iPSCs were exposed to medium containing $10{\mu}M$ Y-27632 for 0, 1, 2, 3, 4, and 5 d, the cell viability rate increased significantly in accordance with the cell viability rate (p<0.05). To evaluate the effect of the Y-27632 treatment on stemness characteristics, the expression of stem cell-specific transcripts and telomerase activity were investigated in the iPSCs treated with $10{\mu}M$ Y-27632 for 5 d. The expression levels of stem cell-specific transcripts, such as OCT-4, NONOG, and SOX-2, and telomerase activity were not significantly different in the iPSCs treated with $10{\mu}M$ Y-27632 as compared with those of untreated control iPSCs (p>0.05). Taken together, the results demonstrated that cell viability can be improved by treatment with the ROCK inhibitor Y-27632, without losing iPSC stemness characteristics.