• Journal of Animal Reproduction and Biotechnology
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• v.36 no.4
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• pp.299-306
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• 2021

Vitrification methods are commonly used for mammalian reproduction through the long-term storage of blastocyst produced in vitro. However, the survival and quality of embryos following vitrification are significantly low compared with blastocyst from in vitro production (IVP). This study evaluates that the survival of frozen-thawed bovine embryos was relevant to mitochondrial superoxide derived mitochondrial activity. Here we present supplementation of the cryopreservation medium with Mito-TEMPO (0.1 µM) induced a significant (p < 0.001; non-treated group: 56.8 ± 8.7%, reexpanded at 24 h vs Mito-TEMPO treated group: 77.5 ± 8.9%, re-expanded at 24 h) improvement in survival rate of cryopreserved-thawed bovine blastocyst. To confirm the quality of vitrified blastocyst after thawing, DNA fragmentation of survived embryos was examined by TUNEL assay. As a result, TUNEL positive cells rates of frozen-thawed embryos were lower in the Mito-TEMPO treated group (4.2 ± 1.4%) than the non-treated group (7.1 ± 3.5%). In addition, we investigated the intracellular ROS and mitochondrial specific superoxide production using DCF-DA and Mito-SOX staining in survived bovine embryos following vitrification depending on Mito-TEMPO treatment. As expected, intracellular ROS levels and superoxide production of vitrified blastocysts after cryopreservation were significantly reduced (p < 0.05) according to Mito-TEMPO supplement in freezing medium. Also, mitochondrial activity measured by MitoTracker Orange staining increased in the frozen-thawed embryos with Mito-TEMPO compared with non-treated group. These results indicate that the treatment of Mito-TEMPO during cryopreservation might induce reduction in DNA fragmentation and apoptosis-related ROS production, consequently increasing mitochondrial activation for developmental capacity of frozen-thawed embryos.

• The Korean Journal of Physiology and Pharmacology
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• v.26 no.5
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• pp.367-375
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• 2022

Gastric cancer stem cells (GCSCs) are a major cause of radioresistance and chemoresistance in gastric cancer (GC). Therefore, targeting GCSCs is regarded as a powerful strategy for the effective treatment of GC. Atorvastatin is a widely prescribed cholesterol-lowering drug that inhibits 3-hydroxy-3-methylglutaryl-coenzyme A reductase, a rate-limiting enzyme in the mevalonate pathway. The anticancer activity of atorvastatin, a repurposed drug, is being investigated; however, its therapeutic effect and molecular mechanism of action against GCSCs remain unknown. In this study, we evaluated the anticancer effects of atorvastatin on MKN45-derived GCSCs. Atorvastatin significantly inhibited the proliferative and tumorsphere-forming abilities of MKN45 GCSCs in a mevalonate pathway-independent manner. Atorvastatin induced cell cycle arrest at the G0/G1 phase and promoted apoptosis by activating the caspase cascade. Furthermore, atorvastatin exerted an antiproliferative effect against MKN45 GCSCs by inhibiting the expression of cancer stemness markers, such as CD133, CD44, integrin α6, aldehyde dehydrogenase 1A1, Oct4, Sox2, and Nanog, through the downregulation of β-catenin, signal transducer and activator of transcription 3, and protein kinase B activities. Additionally, the combined treatment of atorvastatin and sorafenib, a multi-kinase targeted anticancer drug, synergistically suppressed not only the proliferation and tumorsphere formation of MKN45 GCSCs but also the in vivo tumor growth in a chick chorioallantoic membrane model implanted with MKN45 GCSCs. These findings suggest that atorvastatin can therapeutically eliminate GCSCs.

• Molecules and Cells
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• v.46 no.4
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• pp.209-218
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• 2023

In induced pluripotent stem cells (iPSCs), pluripotency is induced artificially by introducing the transcription factors Oct4, Sox2, Klf4, and c-Myc. When a transgene is introduced using a viral vector, the transgene may be integrated into the host genome and cause a mutation and cancer. No integration occurs when an episomal vector is used, but this method has a limitation in that remnants of the virus or vector remain in the cell, which limits the use of such iPSCs in therapeutic applications. Chemical reprogramming, which relies on treatment with small-molecule compounds to induce pluripotency, can overcome this problem. In this method, reprogramming is induced according to the gene expression pattern of extra-embryonic endoderm (XEN) cells, which are used as an intermediate stage in pluripotency induction. Therefore, iPSCs can be induced only from established XEN cells. We induced XEN cells using small molecules that modulate a signaling pathway and affect epigenetic modifications, and devised a culture method which can produce homogeneous XEN cells. At least 4 passages were required to establish morphologically homogeneous chemically induced XEN (CiXEN) cells, whose properties were similar to those of XEN cells, as revealed through cellular and molecular characterization. Chemically iPSCs derived from CiXEN cells showed characteristics similar to those of mouse embryonic stem cells. Our results show that the homogeneity of CiXEN cells is critical for the efficient induction of pluripotency by chemicals.

• International Journal of Stem Cells
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• v.16 no.1
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• pp.36-43
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• 2023

Background and Objectives: Lymphoblastoid cell lines (LCLs) deposited from disease-affected individuals could be a valuable donor cell source for generating disease-specific induced pluripotent stem cells (iPSCs). However, generation of iPSCs from the LCLs is still challenging, as yet no effective gene delivery strategy has been developed. Methods and Results: Here, we reveal an effective gene delivery method specifically for LCLs. We found that LCLs appear to be refractory toward retroviral and lentiviral transduction. Consequently, lentiviral and retroviral transduction of OCT4, SOX2, KFL4 and c-MYC into LCLs does not elicit iPSC colony formation. Interestingly, however we found that transfection of oriP/EBNA-1-based episomal vectors by electroporation is an efficient gene delivery system into LCLs, enabling iPSC generation from LCLs. These iPSCs expressed pluripotency makers (OCT4, NANOG, SSEA4, SALL4) and could form embryoid bodies. Conclusions: Our data show that electroporation is an effective gene delivery method with which LCLs can be efficiently reprogrammed into iPSCs.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.3
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• pp.153-163
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• 2024

Background: The clinical application of canine mesenchymal stem cells (MSCs) necessitates efficient and safe culture methods to produce large quantities of cells. Traditionally, fetal bovine serum (FBS) has been used for MSC expansion, but it carries risks such as contamination and adverse immune responses. Methods: In this study, we investigate the efficacy and efficiency of canine allogeneic serum as an effective alternative to FBS for the in vitro culture of canine MSCs. We measured the population doubling time of canine MSCs in allogeneic serum conditions and utilized qRT-PCR, flowcytometric analysis, and cellular staining/color-metric assay for investigating its effects on cellular senescence during long-term culture and the expression of key pluripotency-related transcriptomes. Results: Our findings demonstrate that canine MSCs cultured with allogeneic serum exhibited enhanced proliferation rates, reduced cellular senescence, and lower apoptosis levels compared to those cultured with FBS. Additionally, the expression of key pluripotency-related transcription factors, including Oct4, Sox2, and Nanog, was increased in canine MSCs cultured with allogeneic serum. Conclusions: These results highlight the potential of canine allogeneic serum to provide a safer and more effective culture environment, supporting the large-scale expansion and maintenance of canine MSCs for clinical applications.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.2
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• pp.201-208
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• 2010

The $SO_2$ and $NO_x$ removal with an activated coke catalyst was conducted by a two-stage reaction which first $SO_2$ was oxidized to $H_2SO_4$ and then $NO_x$ was reduced to $N_2$. But if unreacted sulfur dioxide entered in the second stage, the $NO_x$ reduction was hindered by the reaction with ammonia. In this study, experimental investigations by using lab-scale column apparatus on the product and the reactivity of $SO_2$ with ammonia over coke catalyst which was activated with sulfuric acid was carried out through ultimate analysis DTA, TGA and SEM of catalyst before and after the reaction. Also, the effect of reaction emperature on the reactivity of $SO_2$ with ammonia was determined by means of breakthrough curves with time. The obtained results from this study were summarized as following; Activated cokes were decreased carbon component and increased oxygen and sulfur components in comparison with original cokes. The products over coke catalyst were faced fine crystal of $(NH_4)_2SO_4$, which results in the pressure loss of reacting system. The order of general reactivity in terms of the reaction temperature after breakthrough for $SO_2$ was found to be $150^{\circ}C$ > $200^{\circ}C$ > $100^{\circ}C$. This was related to adsorption amounts of ammonia on the activated cokes.

• Journal of the Korean Society of Marine Environment & Safety
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• v.29 no.6
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• pp.697-703
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• 2023

As a part of the global industrial efforts to reduce environmental pollution owing to air pollution, regulations have been established by the International Maritime Organization (IMO). The IMO has implemented various regulations such as EEXI, EEDI, and CII to reduce air pollution emissions from ships. They are also promoting measures to decrease the power consumption in ships, aiming to conserve energy. Most of the power used in ships is consumed by electric motors. Among the motors installed on ships, the engine room blower that takes up a significant load, operates at a constant irrespective of demand. Therefore, energy savings can be expected through frequency control. In this study, we demonstrated the efficacy of energy savings by controlling the frequency of the electric motor of the generator blower that supplies combustion air to the generator's turbocharger. The system was modeled based on the output data of the turboharger outlet temperature in response to the blower frequency inpu. A PI control system was established to control the frequency with the target being the turbocharger outlet temperature. By maintaining the turbocharger design standard outlet temperature and controlling the blower frequency, we achieved an annual energy saving of 15,552kW in power consumption. The effectiveness of energy savings through frequency control of blower fans was verified during the summer (April to September) and winter (March to October) periods. Based on this, we achieved annual fuel cost savings of 6,091 thousand won and reduction of 8.5 tons of carbon dioxide, 2.4 kg of SOx, and 7.8 kg of NOx air pollutants on the training ship.

• Economic and Environmental Geology
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• v.52 no.2
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• pp.119-128
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• 2019

About 300,000 tones of oyster shell are annually produced in Korea and, thus, a massive recycling plan is required. Many desulfurizing studies using oyster shells with chemical composition of $CaCO_3$ have been performed so far; however, most of them have focused on dry desulfurization. This study investigates the possibility of using oyster shells for wet desulfurization after calcination. For this, a simulated wet desulfurization facility of spray type was devised and compared the SOx-stripping characteristics of calcined oyster shell with those of limestone. The calcined oyster shell slurry indicate a better desulfurizability than the slurries of raw shell or limestone because the oyster shell transformed to a more reactive phase ($Ca(OH)_2$) by the calcination and hydration. Because of this reason, when the calcined oyster shell slurries were used, the reaction residue showed the higher gypsum ($CaSO_4{\cdot}2H_2O$) contents than any other cases. In the continuous desulfurization experiments, calcined oyster shell slurry showed a wider pH variation than limestone or raw oyster shell slurries, another clear indication of high reactivity of calcined oyster shells for $SO_2$ absorption. Our study also shows that the efficiency of wet desulfurization can be improved by the use of calcined oyster shells.

• Journal of The Korean Society of Agricultural Engineers
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• v.64 no.3
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• pp.25-32
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• 2022

To understand the distribution characteristics of PM_2.5 concentration in the Saemangeum Reclamation Area and nearby areas, three points of the background area, the occurrence area, and the affected area were selected and samples were collected for each season. The chemical composition was determined. As a result of analyzing the chemical composition contained in PM_2.5, NO₃^- (7.2 ㎍/m³), SO₄^2- (4.3 ㎍/m³), NH₄+ (4.3 ㎍/m³), OC (2.5 ㎍/m³), Si (1.3 ㎍/m³) m³) and EC (0.5 ㎍/m³) seemed to be the main components, and NO₃-, SO₄^2-, NH₄⁺, which are components that form secondary particles, occupied a large proportion. The composition ratio of PM_2.5 was investigated in the order of ion component (56.8%) > Unknown (27.4%) > carbon component (11.8%) > heavy metal component (4.0%). During the PM_2.5 high concentration case days, the ionic component accounted for 90.7% during atmospheric stagnation cases, whereas the chemical composition ratio was in the order of ionic component (51.7%) > heavy metal component (41.5%) > carbon component (6.8%) during yellow dust cases. It was found that the characteristic of PM_2.5 in the Saemangeum reclaimed land and surrounding areas is mainly influenced by outside (domestic and overseas) throughout the year. Ion components accounted for the largest portion of PM_2.5 components in this area, but there were few sources of SOx and NOx emission in the Seamangeum area, which are precursors for secondary particle formation. Therefore, it is judged that most of these are generated and influenced as a secondary reaction in the atmosphere from the outside.

• 한국신재생에너지학회:학술대회논문집
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• 2009.11a
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• pp.513-513
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• 2009

최근 바이오디젤의 원료로 미세조류가 많은 관심을 끌고 있다. 미세조류는 물, 이산화탄소와 태양광을 이용해 광합성 성장이 가능하며, 지질(오일) 성분이 풍부하여 바이오디젤의 원료로 이용할 수 있다. 미세조류는 단위 면적당 오일 생산량이 곡물류의 50-100배 이상이며, 이산화탄소를 기질로 이용하므로 온실가스 직접 저감이 가능하다. 또한 배양시 비경작지나 황무지를 사용할 수 있으므로 기존 식용작물과 경쟁하지 않으며, 하수, 해수, 폐수 등 다양한 물자원을 이용할 수 있다. 본 연구에서는 고농도 $CO_2$에 내성을 지닌 Chlorella sp. KR-1을 대상으로 1 L 규모 기포탑 광생물반응기에서 균체 성장 및 지질(바이오오일) 합성에 대한 $CO_2$ 농도, 가스 공급속도, 질산염 농도 등 환경 및 영향 조건의 영향을 조사하였다. 가스 공급속도 0.4 L/min에서 $CO_2$ 농도를 0.03-20% 범위에서 조사하였을 때 최대 균체성장은 $CO_2$ 10%에서 관찰되었다. 균체내 지방산 함량은 $CO_2$ 농도 0.03%에서 가장 낮았고, 5-20% 범위에서는 $CO_2$ 농도 증가에 따라 감소하는 경향이 관찰되었다. 가스 공급속도를 0.2 L/min에서 0.8 L/min으로 증가시켰을 때 최대 균체농도는 0.6-0.8 L/min의 범위에서 관찰되었고, 생체내 최대 지방산 함량은 0.4 L/min에서 관찰되었다. Nitrate 농도 1-20 mM 범위에서 최종 균체농도는 nitrate 농도 증가에 따라 10mM까지 증가하였으나 그 이상에서는 증가하지 않았다. 반면 지방산 함량은 nitrate 농도 증가에 따라 감소하는 경향이 관찰되었다. 본 실험에서 얻은 최대 지방산 생산량은 1,100 mg/L이었으며, 주요 지방산은 C10:0(1.7%), C16:0(28.5%), C18:0(11%), C18:1n9c(25.9%), C18:2n6c(26.3%), C18:3n3(6.6%)이었다. 현재 100 L 규모 광생물반응기에서 석탄발전소 배가스($CO_2$ 12-15%, SOx 50ppm, NOx 100ppm)를 이용한 바이오오일 생산 실험이 진행 중이다.