• Journal of Plant Biotechnology
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• v.48 no.1
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• pp.34-43
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• 2021

Targeted genome editing using the CRISPR/Cas9 system is a ground-breaking technology that is being widely used to produce plants with useful traits. However, for woody plants, only a few successful attempts have been reported. These successes have used Agrobacterium-mediated transformation, which has been reported to be very efficient at producing genetically modified trees. Nonetheless, there are unresolved problems with plasmid sequences that remain in the plant genome. In this study, we demonstrated a DNA-free genome editing technique in which purified CRISPR/Cas9 ribonucleoproteins (RNPs) are delivered directly to the protoplasts of a hybrid poplar (Populus alba × Populus glandulosa). We designed three single-guide RNAs (sgRNAs) to target the stress-associated protein 1 gene (PagSAP1) in the hybrid poplar. Deep sequencing results showed that pre-assembled RNPs had a more efficient target mutagenesis insertion and deletion (indel) frequency than did non-assembled RNPs. Moreover, the RNP of sgRNA3 had a significantly higher editing efficacy than those of sgRNA1 and sgRNA2. Our results suggest that the CRISPR/Cas9 ribonucleoprotein-mediated transfection approach is useful for the production of transgene-free genome-edited tree plants.

• Korean Journal of Food Science and Technology
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• v.54 no.3
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• pp.288-296
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• 2022

This study aimed to verify the effect of Citrus unshiu peel water extract (CUP) on a mouse model of acute gastritis (AG) induced by HCl/ethanol. Several studies have found that CUP has anti-inflammatory effects. The AG model was induced by oral administration of 150 mM HCl/60% ethanol (550 µL) to all groups except the control group. Also, for drug treatment, sucralfate (10 mg/kg) and CUP (100 or 200 mg/kg) were orally administered for 90 minutes before induction. The effect of CUP treatment was confirmed by gross gastric mucosal damage measurement, and the levels of Glutamic Oxaloacetic Transaminase (GOT), Glutamic Pyruvic Transaminase (GPT), and myeloperoxidase were reduced as well as the levels of oxidative stress biomarkers and their related proteins. In addition, the levels of inflammatory proteins, mediators, and cytokines were significantly downregulated byPI3K/Akt signaling. Taken together, these results show that CUP treatment alleviates AG by regulating PI3K/Akt signaling.

• Nutrition Research and Practice
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• v.16 no.1
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• pp.14-32
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• 2022

BACKGROUND/OBJECTIVES: Peroxisome proliferator-activated receptor-gamma co-activator-1α (PGC-1α) has a central role in regulating muscle differentiation and mitochondrial metabolism. PGC-1α stimulates muscle growth and muscle fiber remodeling, concomitantly regulating lactate and lipid metabolism and promoting oxidative metabolism. Gynostemma pentaphyllum (Thumb.) has been widely employed as a traditional herbal medicine and possesses antioxidant, anti-obesity, anti-inflammatory, hypolipemic, hypoglycemic, and anticancer properties. We investigated whether G. pentaphyllum extract (GPE) and its active compound, gypenoside L (GL), affect muscle differentiation and mitochondrial metabolism via activation of the PGC-1α pathway in murine C2C12 myoblast cells. MATERIALS/METHODS: C2C12 cells were treated with GPE and GL, and quantitative reverse transcription polymerase chain reaction and western blot were used to analyze the mRNA and protein expression levels. Myh1 was determined using immunocytochemistry. Mitochondrial reactive oxygen species generation was measured using the 2'7'-dichlorofluorescein diacetate assay. RESULTS: GPE and GL promoted the differentiation of myoblasts into myotubes and elevated mRNA and protein expression levels of Myh1 (type IIx). GPE and GL also significantly increased the mRNA expression levels of the PGC-1α gene (Ppargc1a), lactate metabolism-regulatory genes (Esrra and Mct1), adipocyte-browning gene fibronectin type III domain-containing 5 gene (Fndc5), glycogen synthase gene (Gys), and lipid metabolism gene carnitine palmitoyltransferase 1b gene (Cpt1b). Moreover, GPE and GL induced the phosphorylation of AMP-activated protein kinase, p38, sirtuin1, and deacetylated PGC-1α. We also observed that treatment with GPE and GL significantly stimulated the expression of genes associated with the anti-oxidative stress response, such as Ucp2, Ucp3, Nrf2, and Sod2. CONCLUSIONS: The results indicated that GPE and GL enhance exercise performance by promoting myotube differentiation and mitochondrial metabolism through the upregulation of PGC-1α in C2C12 skeletal muscle.

• Tunnel and Underground Space
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• v.31 no.6
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• pp.561-577
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• 2021

The compacted bentonite buffer in a geological repository for high-level radioactive waste disposal is saturated due to groundwater inflow. Saturation of the bentonite buffer results in bentonite swelling and bentonite penetration into the rock discontinuities present around the disposal hole. The penetrated bentonite is exposed to groundwater flow and can be eroded out of the repository, resulting in bentonite mass loss which can affect the physical integrity of the engineered barrier system. Hence, the evaluation of buffer-rock interactions and coupled behavior due to groundwater inflow and bentonite penetration is necessary to ensure long-term disposal safety. In this study, the effects of the bentonite penetration and swelling on the physical properties of jointed rock mass were evaluated using the quasi-static resonant column test. Jointed rock specimens with bentonite penetration were manufactured using Gyeongju bentonite and hollow cylindrical granite rock discs obtained from the KAERI underground research tunnel. The effects of vertical stress and saturation were assessed using the P-wave and S-wave velocities for intact rock, jointed rock and jointed rock with bentonite penetration specimens. The joint normal and joint shear stiffnesses of each joint condition were inferred from the wave velocity results assuming an equivalent continuum. The joint normal and joint shear stiffnesses obtained from this study can be used as input factors for future numerical analysis on the performance evaluation of geological waste disposal considering rock discontinuities.

• Journal of the Korea institute for structural maintenance and inspection
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• v.26 no.4
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• pp.73-80
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• 2022

This study evaluated changes in fluidity and rheological properties over time for 3D printed composite materials, and evaluated compressive strength and splitting tensile strength properties for laminated and molded specimens. The composite material for 3D printing starts to change rapidly after 30 minutes of extrusion, and the viscosity of the material tends to be maintained up to 90 minutes, but it was confirmed that construction within 60 minutes after mixing is effective. The compressive strength of the laminated test specimen showed equivalent or better performance at all ages compared to the molded test specimen. In the stress-strain curve of the laminated specimen, the initial slope was similar to that of the molded specimen, but the descending slope was on average 1.9 times higher than that of the molded specimen, indicating relatively brittle behavior. The splitting tensile strength of the P-V laminated specimen was about 6% lower than that of the molded specimen. It is judged that this is because the interfacial adhesion force against the vertical load is affected by the pattern direction of the laminated test specimen.

• Korean Journal of Remote Sensing
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• v.37 no.2
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• pp.211-220
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• 2021

Land Surface Temperature (LST) is one of the useful parameters to diagnose the growth and development of crop and to detect crop stress. Unmanned Aerial Vehicle (UAV)-based LST (LSTUAV) can be estimated in the regional spatial scale due to miniaturization of thermal infrared camera and development of UAV. Given that meteorological variable, type of instrument, and surface condition can affect the LSTUAV, the evaluation for accuracy of LSTUAV is required. The purpose of this study is to evaluate the accuracy of LSTUAV using LST measured at ground (LSTGround) under various meteorological conditions and growth phases of garlic crop. To evaluate the accuracy of LSTUAV, Relative humidity (RH), absolute humidity (AH), gust, and vegetation index were considered. Root mean square error (RMSE) after minimizing the bias between LSTUAV and LSTGround was 2.565℃ under above 60% of RH, and it was higher than that of 1.82℃ under the below 60% of RH. Therefore, LSTUAV measurement should be conducted under the below 60% of RH. The error depending on the gust and surface conditions was not statistically significant (p-value < 0.05). LSTUAV had reliable accuracy under the wind speed conditions that allow flight and reflected the crop condition. These results help to comprehend the accuracy of LSTUAV and to utilize it in the agriculture field.

• Journal of the Computational Structural Engineering Institute of Korea
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• v.35 no.4
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• pp.243-248
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• 2022

Weight optimization was performed for a rotorcraft shaft system using one-dimensional Euler-Bernoulli beam elements. Torsion, shaft support stiffness such as bearings, flange mass are all considered. To guarantee structural dynamic stability, eigenvalue analysis was performed to avoid critical speed and tooth mesh excitation form the gearbox. The weight optimization was performed by adjusting the thickness and radius while the length of the shaft was fixed, and the optimization process was divided into two stages. In the first, the weight is optimized with the torsional strength constraint. In the second, the difference between the primary mode of shaft and the critical speed is maximized so that the primary mode of the shaft can avoid the critical speed while the constraint on the torsional strength of the shaft is satisfied according to the standard for shaft system stability (AMC P 706-201, 1974). The proposed method was verified by comparing the results of the optimal design using the given one-dimensional beam elements with the stress results of the 3D finite element and the actual manufactured shaft.

• Biomolecules & Therapeutics
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• v.31 no.1
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• pp.97-107
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• 2023

Aristolochic acid (AA), extracted from Aristolochiaceae plants, plays an essential role in traditional herbal medicines and is used for different diseases. However, AA has been found to be nephrotoxic and is known to cause aristolochic acid nephropathy (AAN). AA-induced acute kidney injury (AKI) is a syndrome in AAN with a high morbidity that manifests mitochondrial damage as a key part of its pathological progression. Melatonin primarily serves as a mitochondria-targeted antioxidant. However, its mitochondrial protective role in AA-induced AKI is barely reported. In this study, mice were administrated 2.5 mg/kg AA to induce AKI. Melatonin reduced the increase in Upro and Scr and attenuated the necrosis and atrophy of renal proximal tubules in mice exposed to AA. Melatonin suppressed ROS generation, MDA levels and iNOS expression and increased SOD activities in vivo and in vitro. Intriguingly, the in vivo study revealed that melatonin decreased mitochondrial fragmentation in renal proximal tubular cells and increased ATP levels in kidney tissues in response to AA. In vitro, melatonin restored the mitochondrial membrane potential (MMP) in NRK-52E and HK-2 cells and led to an elevation in ATP levels. Confocal immunofluorescence data showed that puncta containing Mito-tracker and GFP-LC3A/B were reduced, thereby impeding the mitophagy of tubular epithelial cells. Furthermore, melatonin decreased LC3A/B-II expression and increased p62 expression. The apoptosis of tubular epithelial cells induced by AA was decreased. Therefore, our findings revealed that melatonin could prevent AA-induced AKI by attenuating mitochondrial damage, which may provide a potential therapeutic method for renal AA toxicity.

• Journal of Veterinary Science
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• v.24 no.2
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• pp.24.1-24.13
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• 2023

Background: Ergothioneine (EGT) is a natural amino acid derivative in various animal organs and is a bioactive compound recognized as a food and medicine. Objectives: This study examined the effects of EGT supplementation during the in vitro maturation (IVM) period on porcine oocyte maturation and subsequent embryonic development competence after in vitro fertilization (IVF). Methods: Each EGT concentration (0, 10, 50, and 100 μM) was supplemented in the maturation medium during IVM. After IVM, nuclear maturation, intracellular glutathione (GSH), and reactive oxygen species (ROS) levels of oocytes were investigated. In addition, the genes related to cumulus function and antioxidant pathways in oocytes or cumulus cells were investigated. Finally, this study examined whether EGT could affect embryonic development after IVF. Results: After IVM, the EGT supplementation group showed significantly higher intracellular GSH levels and significantly lower intracellular ROS levels than the control group. Moreover, the expression levels of hyaluronan synthase 2 and Connexin 43 were significantly higher in the 10 μM EGT group than in the control group. The expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and NAD(P)H quinone dehydrogenase 1 (NQO1) were significantly higher in the oocytes of the 10 μM EGT group than in the control group. In the assessment of subsequent embryonic development after IVF, the 10 μM EGT treatment group improved the cleavage and blastocyst rate significantly than the control group. Conclusions: Supplementation of EGT improved oocyte maturation and embryonic development by reducing oxidative stress in IVM oocytes.

• Molecules and Cells
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• v.45 no.3
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• pp.134-147
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• 2022

The anti-oxidant enzyme heme oxygenase-1 (HO-1) is known to exert anti-inflammatory effects. From a library of pyrazolo[3,4-d]pyrimidines, we identified a novel compound KKC080096 that upregulated HO-1 at the mRNA and protein levels in microglial BV-2 cells. KKC080096 exhibited anti-inflammatory effects via suppressing nitric oxide, interleukin1β (IL-1β), and iNOS production in lipopolysaccharide (LPS)-challenged cells. It inhibited the phosphorylation of IKK and MAP kinases (p38, JNK, ERK), which trigger inflammatory signaling, and whose activities are inhibited by HO-1. Further, KKC080096 upregulated anti-inflammatory marker (Arg1, YM1, CD206, IL-10, transforming growth factor-β [TGF-β]) expression. In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridinetreated mice, KKC080096 lowered microglial activation, protected the nigral dopaminergic neurons, and nigral damage-associated motor deficits. Next, we elucidated the mechanisms by which KKC080096 upregulated HO-1. KKC080096 induced the phosphorylation of AMPK and its known upstream kinases LKB1 and CaMKKbeta, and pharmacological inhibition of AMPK activity reduced the effects of KKC080096 on HO-1 expression and LPS-induced NO generation, suggesting that KKC080096-induced HO-1 upregulation involves LKB1/AMPK and CaMKKbeta/AMPK pathway activation. Further, KKC080096 caused an increase in cellular Nrf2 level, bound to Keap1 (Nrf2 inhibitor protein) with high affinity, and blocked Keap1-Nrf2 interaction. This Nrf2 activation resulted in concurrent induction of HO-1 and other Nrf2-targeted antioxidant enzymes in BV-2 and in dopaminergic CATH.a cells. These results indicate that KKC080096 is a potential therapeutic for oxidative stress-and inflammation-related neurodegenerative disorders such as Parkinson's disease.