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Positive Regulator, a Rice C3H2C3-type RING Finger Protein H2-3(OsRFPH2-3), in Response to Salt Stress

  • Min Seok Choi;Cheol Seong Jang
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2023.04a
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    • pp.156-156
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    • 2023
  • Salinity is a major abiotic stress that limits rice productivity in many regions of the world. In order to develop salt stress tolerant rice plants, genetic engineering is a promising approach. We characterized the molecular function of rice C3H2C3 as a really interesting new gene (RING). Oryza sativa RING finger protein H2-3 (OsRFPH2-3) was highly expressed in 100 mM NaCl. To identify the localization of OsRFPH2-3, we fused vectors that include C-terminal GFP protein (35S;;OsRFPH2-3-GFP). OsRFPH2-3 was expressed in the nucleus in rice protoplasts. An in vitro ubiquitin assay demonstrated that OsRFPH2-3 possessed E3-ubiquitin ligase activity. However, the mutated OsRFPH2-3 were not possessed any E3-ubiquitin ligase activity. Under salinity conditions, OsRFPH2-3-overexpressing plants exhibited higher chlorophyll, proline, SOD, POD, CAT, and soluble sugar contents and lower H2O2 accumulation than wild-type plants, supporting transgenic plants with enhanced salinity tolerance phenotypes. OsRFPH2-3-overexpressing plants exhibited low Na+ accumulation and Na+/K+ ratios in their roots. Theses results suggest that overexpression of OsRFPH2-3 can make plant insensitivity about salinity conditions.

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Comparative Analysis on Antioxidant Activity in Various Human Skin Fibroblasts and Mesenchymal Stem Cells (사람의 피부 섬유아세포 및 중간엽 줄기세포에서 항산화 활성의 반응에 대한 비교 분석)

  • Kong, Ji-Weon;Park, Ryeok;Park, Joon-Woo;Lee, Joo-Yeong;Choi, Yeon-Joo;Moon, Sun-Ha;Kim, Hyeon-Ji;Jeon, Byeong-Gyun
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.2
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    • pp.394-406
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    • 2019
  • The cellular senescence may be due to damage by the reactive oxygen species (ROS). This study has compared the antioxidant activity in the human cell lines of various origins, including 10S and 50S-derived normal skin fibroblasts, and 10S bone marrow, dental tissue and adipose-derived adult stem cells. After being exposed to $H_2O_2$, half inhibitory concentration ($IC_{50}$) values by cytotoxicity assay was significantly (P<0.05) lower in 50S-derived skin fibroblasts, than in 10S-derived skin fibroblasts and various adult stem cell lines. The cell population doubling time (PDT) and the cell frequency with high senescence associated-${\beta}$-galactose activity were remarkably increased in 50S-derived fibroblasts exposed to 50 ppm $H_2O_2$ for 7 days, than those of 10S-derived fibroblasts and various adult stem cell lines. Further, the expression level of antioxidant-related genes, glutathione peroxidase (GPX) and catalase (CAT), was investigated in 10S and 50S-derived skin fibroblasts, and 10S-derived various adult stem cells by reverse transcription polymerase chain reaction (RT-PCR). The expression level of GPX was higher in most of cell lines, compared to CAT, and a significantly (P<0.05) higher expression level of GPX was observed in 10S-derived skin fibroblasts and adult stem cell lines, compared to 50S-derived skin fibroblasts. We concluded that old-aged skin fibroblasts seemed to be less resistant against ROS than young-aged skin fibroblasts and adult stem cells.

In vitro Comparision of Biological Activities of Solvent Fraction Extracts from Orostachys japonicus (와송(Orostachys japonicus) 용매별 분획 추출물의 항산화, 항균 및 암세포 독성 비교)

  • Kim, Seung Mi;Park, Jeong Hun;Boo, Hee Ock;Song, Sang Gi;Park, Hyeon Yong
    • Korean Journal of Plant Resources
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    • v.30 no.2
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    • pp.133-143
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    • 2017
  • This study was conducted to evaluate the contents of total polyphenol and flavonoid, and the effect of antioxidant, antimicrobial activities and cytotoxicity in vitro by different solvent fractions from Orostachys japonicus. The ethylacetate fraction extract for O. japonicus contained $634.48{\mu}g/g$ polyphenol and $205.20{\mu}g/g$ flavonoid. The ABTS radical scavenging ability of ethylacetate fraction extract at 1 mg/ml was higher than 95% which is comparable to ascorbic acid of 97%. The APX enzymatic activity and CAT activity were $1125.89{\mu}mol$ ascorbate oxidized/min/mg protein and 119.87 H2O2 decomposed/min/mg protein, respectively. In disc agar plate diffusion assay, the extract gave rise to a larger inhibition circle with Listeria monocytogenes, Staphylococcus epidermidis, Staphylococcus aureus and Malassezia furfur strains compared with antibiotics kanamycin suggestive of high antibiotic activity. The cytotoxicity of extracts of O. japonicus was significant differences between solvent fractions. That is, the cytotoxic effect against human cancer cell was higher in ethylacetate fraction extract than other fraction extracts. These results suggest that fraction extract of O. japonicus might be very effective and economical in developing natural antioxidant and antimicrobial.

Extracts from Gracilaria vermiculophylla Prevent Cellular Senescence and Improve Differentiation Potential in Replicatively Senescent Human Bone Marrow Mesenchymal Stem Cells (홍조류인 Gracilaria vermiculophylla 추출물에 의한 노화 골수유래 중간엽줄기세포의 항노화 및 분화능력 개선 효과)

  • Jeong, Sin-Gu;Cho, Tae Oh;Cho, Goang-Won
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1042-1047
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    • 2018
  • The red algae Gracilaria vermiculophylla is widespread on seashores worldwide and has been used as food in Asian countries. Previous studies have reported that extracts of Gracilaria red algae have beneficial anti-oxidant and anti-inflammatory effects. The present study examined the anti-senescence effects of Gracilaria vermiculophylla extracts (GV-Ex) in replicatively senescent human bone marrow mesenchymal stem cells (hBM-MSCs). GV-Ex pretreatment improved the cellular viability of hBM-MSCs that had been injured by oxidative stress. These effects of GV-Ex were confirmed by MTT assay and immunoblot analysis using the apoptotic proteins p53 and cleaved caspase-3. The reactive oxygen species (ROS) levels were examined in long-term cultured Passages 17 (P-17) mesenchymal stem cells (MSC) and compared to P-7 MSC. The ROS accumulation was greater in the P-17 than in the P-7. However, these increased ROS levels in the P-17 were decreased significantly after treatment with GV-Ex, and restoration of the levels of the anti-oxidant enzymes SOD1, SOD2, and CAT was also observed under these conditions. In addition, P-17 hBM-MSC treated with GV-Ex had decreased levels of the senescence proteins p53, p21, and p16. The results show that the ability of P-17 hBM-MSC to differentiate into osteocytes and adipocytes was improved by GV-Ex treatment, suggesting that GV-Ex ameliorates the functional decline of senescent stem cells.

Antioxidative effects of fermented sesame sauce against hydrogen peroxide-induced oxidative damage in LLC-PK1 porcine renal tubule cells

  • Song, Jia-Le;Choi, Jung-Ho;Seo, Jae-Hoon;Kil, Jeung-Ha;Park, Kun-Young
    • Nutrition Research and Practice
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    • v.8 no.2
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    • pp.138-145
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    • 2014
  • BACKGROUND/OBJECTIVES: This study was performed to investigate the in vitro antioxidant and cytoprotective effects of fermented sesame sauce (FSeS) against hydrogen peroxide ($H_2O_2$)-induced oxidative damage in renal proximal tubule LLC-PK1 cells. MATERIALS/METHODS: 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical ($^{\bullet}OH$), and $H_2O_2$ scavenging assay was used to evaluate the in vitro antioxidant activity of FSeS. To investigate the cytoprotective effect of FSeS against $H_2O_2$-induced oxidative damage in LLC-PK1 cells, the cellular levels of reactive oxygen species (ROS), lipid peroxidation, and endogenous antioxidant enzymes including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-px) were measured. RESULTS: The ability of FSeS to scavenge DPPH, $^{\bullet}OH$ and $H_2O_2$ was greater than that of FSS and AHSS. FSeS also significantly inhibited $H_2O_2$-induced ($500{\mu}M$) oxidative damage in the LLC-PK1 cells compared to FSS and AHSS (P < 0.05). Following treatment with $100{\mu}g/mL$ of FSeS and FSS to prevent $H_2O_2$-induced oxidation, cell viability increased from 56.7% (control) to 83.7% and 75.6%, respectively. However, AHSS was not able to reduce $H_2O_2$-induced cell damage (viability of the AHSS-treated cells was 54.6%). FSeS more effectively suppressed $H_2O_2$-induced ROS generation and lipid peroxidation compared to FSS and AHSS (P < 0.05). Compared to the other sauces, FSeS also significantly increased cellular CAT, SOD, and GSH-px activities and mRNA expression (P < 0.05). CONCULUSIONS: These results from the present study suggest that FSeS is an effective radical scavenger and protects against $H_2O_2$-induced oxidative damage in LLC-PK1 cells by reducing ROS levels, inhibiting lipid peroxidation, and stimulating antioxidant enzyme activity.

Cytoprotective Effect by Antioxidant Activity of Codonopsis lanceolata and Platycodon grandiflorum Ethyl Acetate Fraction in Human HepG2 Cells (인간 HepG2 세포에서 더덕 및 도라지 에틸아세테이트 분획물의 항산화 효과에 의한 세포보호 효과)

  • Kim, Soo-Hyun;Choi, Hyun-Jin;Oh, Hyun-Taek;Chung, Mi-Ja;Cui, Cheng-Bi;Ham, Seung-Shi
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.696-701
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    • 2008
  • The objective of this study was to determine whether Codonopsis lanceolata or Platycodon grandiflorum ethyl acetate fraction (CLEA or PGEA) protect cells against sodium nitroprusside (SNP)-induced oxidative stress via the expression of various antioxidant systems. The HepG2 cells exposed for 24 hr to 0.5 mM SNP showed a reduction in the cell viability by an MTT assay. Pretreatment with CLEA and PGEA resulted in an inhibition of SNP-induced cell death. In addition, the effects of CLEA and PGEA on the expression of antioxidant systems via RT-PCR analyses was assessed. The levels of catalase (CAT), glucose-6-phosphate dehydrogenase (G6PD) and metallothionein (MT)-1A mRNA were increased after 24 hr of CLEA exposure. The levels of Mn superoxide dismutase CAT, G6PD, MT-1A, and MT-2A mRNA were increased after PGEA treatment. In conclusion, CLEA and PGEA exert indirect antioxidant effects, perhaps via the induction of a variety of antioxidant systems which, may protect cells against oxidative stress.

Anti-oxidative and Anti-hyperglycemia Effects of Dung Beetle Extracts on the High Fat Diet SD Rats (고지방식이로 유발한 흰쥐에서 쇠똥구리 추출물의 항산화 효과 및 혈당강하에 미치는 영향)

  • Kim, Ha Jeong;Kim, Ban Ji;Ahn, Mi Young
    • Journal of Life Science
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    • v.26 no.7
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    • pp.772-781
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    • 2016
  • Dung beetle (Catharsius molossus, CA) is a well-known group of insects thanks to their exploitation of animal feces, a behavioral trait with a global impact on earth′s ecosystems. This study was conducted to investigate the effect of CA extract on a high-fat diet in SD rats. Male rats were divided into 5 groups. Animals were fed on a high-fat diet for seven weeks before and dung beetle extract for a month during the administration. Weight gain was decreased in ethanol extract from CA group. Administration of CA extract reduced the organ weight of testis and kidney, and adipose tissue weight. Lipid oxidative stress was evaluated measuring malondialdehyde level in liver. There were no significant differences in groups. Protein oxidative stress was evaluated measuring protein carbonyl content in blood. The protein carbonyl in blood was significantly decreased in ethanol and acetone extracted dung beetle groups (p<0.05). Meanwhile, the protein carbonyl in hepatocyte was not significant among the groups. Fibronectin and laminin by using D-HUVEC cell in vitro were measured by ELISA assay. There was significance in CA extract. The level of IL-10, IL-1β, VEGF, eNOS was evaluated by ELISA. There was significance in IL-10 compared to control (p<0.05). SOD and GPx tended to increase by CA extract. Furthermore, CAT was increased significantly by CA extract (p<0.05). After administration of CA extracts the composition of saturated fatty acid in adipose tissue tend to decrease, while unsaturated fatty acid increases. In conclusion, dung beetle had anti-hyperglycemia effects of oxidative stress and antioxidant activity.

Anti-inflammatory Efficacy of HK Shiitake Mushroom Mycelium in LPS-treated RAW 264.7 Cells Through Down-regulation of NF-κB Activation (LPS로 활성화한 RAW 264.7 세포에서 HK표고버섯균사체의 NF-κB 활성 억제를 통한 항염증 효과)

  • Song, Chae Yeong;Oh, Tae Woo;Kim, Hoon Hwan;Lee, Yu Bin;Kim, Jeong Ok;Kim, Gon Sup;Ha, Yeong Lae
    • Journal of Life Science
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    • v.32 no.7
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    • pp.491-500
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    • 2022
  • HK shiitake mushroom mycelium (HKSMM), containing 14% β-glucan, is a health functional food ingredient individually approved by the Korea Ministry of Food and Drug Safety for liver health. The anti-inflammatory effect of a 50% aqueous ethanol extract of HKSMM (designated HKSMM50) was studied in RAW 264.7 macrophage cells treated with lipopolysaccharide (LPS). An active hexose correlated compound (AHCC) was used as a positive control. LPS-activated RAW 264.7 cells were treated with HKSMM50 and AHCC (0, 20, 100, 500 ㎍/ml) and cultured for 24 hr. Inflammation-related elements in the supernatant were measured using enzyme-linked immunosorbent assay (ELISA) kits, and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins in the cells was analyzed by Western blotting. The HKSMM50 lowered iNOS and COX-2 protein expressions, and nuclear factor-kappa B (NF-κB), nitric oxide (NO) and prostaglandin E2 (PGE2) contents in a concentration-dependent manner as compared to LPS treatment. Similarly, the HKSMM50 lowered the content of pro-inflammatory cytokines interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and interleukin-6 (IL-6) contents and increased the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). The efficacy of the AHCC treatment was similar to that of the HKSSM50 treatments. These results indicate that HKSMM50 showed an anti-inflammatory effect in LPS-treated RAW 264.7 cells by down-regulation of NF-κB signaling and suggest that HKSMM could be used as a health functional food ingredient to help improve immune function.

Induction of systemic resistance in Panax ginseng against Phytophthora cactorum by native Bacillus amyloliquefaciens HK34

  • Lee, Byung Dae;Dutta, Swarnalee;Ryu, Hojin;Yoo, Sung-Je;Suh, Dong-Sang;Park, Kyungseok
    • Journal of Ginseng Research
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    • v.39 no.3
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    • pp.213-220
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    • 2015
  • Background: Korean ginseng (Panax ginseng Meyer) is a perennial herb prone to various root diseases, with Phytophthora cactorum being considered one of the most dreaded pathogens. P. cactorum causes foliar blight and root rot. Although chemical pesticides are available for disease control, attention has been shifted to viable, eco-friendly, and cost-effective biological means such as plant growth-promoting rhizobacteria (PGPR) for control of diseases. Methods: Native Bacillus amyloliquefaciens strain HK34 was isolated from wild ginseng and assessed as a biological control agent for ginseng. Leaves from plants treated with HK34 were analyzed for induced systemic resistance (ISR) against P. cactorum in square plate assay. Treated plants were verified for differential expression of defense-related marker genes using quantitative reverse transcription polymerase chain reaction. Results: A total of 78 native rhizosphere bacilli from wild P. ginseng were isolated. One of the root-associated bacteria identified as B. amyloliquefaciens strain HK34 effectively induced resistance against P. cactorum when applied as soil drench once (99.1% disease control) and as a priming treatment two times in the early stages (83.9% disease control). A similar result was observed in the leaf samples of plants under field conditions, where the percentage of disease control was 85.6%. Significant upregulation of the genes PgPR10, PgPR5, and PgCAT in the leaves of plants treated with HK34 was observed against P. cactorum compared with untreated controls and only pathogen-treated plants. Conclusion: The results of this study indicate HK34 as a potential biocontrol agent eliciting ISR in ginseng against P. cactorum.

Prevalence of Toxoplasma gondii infection from domestic pigs in Gyeongnam province (경남지역의 돼지톡소플라즈마병 감염실태 조사)

  • Kim, Eun-Gyeong;Park, Ho-Jung;Son, Byeong-Guk;Jung, Myeong-Ho;Heo, Jung-Ho;Hwang, Bo-Won
    • Korean Journal of Veterinary Service
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    • v.33 no.4
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    • pp.345-351
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    • 2010
  • Toxoplasma gondii is a species of parasitic protozoa in the genus Toxoplasma. The definitive host of T. gondii is the cat, but the parasite can be carried by the vast majority of warm-blooded animals, including humans. It is often found in the tissues of food animals including pigs and sheep. To determine the regional prevalence of infection with T. gondii, bloods (n=300) from domestic pigs and tissues (n=200) from slaughter pigs in Gyeongnam province were tested using an enzyme-linked immunosorbent assay (ELISA) and a polymerase chain reaction (PCR) for detection of antibody and antigen. A total of 115 sero-positive pigs were identified for a prevalence rate of 38.3%. Of the 50 herds from domestic pigs tested, 34 had at least one sero-positive pig for a herd prevalence rate of 68.0%. Sero-positive rates of pigs in fattening farm were higher than that of pigs in breeding company. Sero-positive rates of sows were higher than that of growing pigs. Seasonally, sero-positive rates of pigs were highest in winter (80.0%) and lowest in spring (23.8%). According to farm size, sero-positive rates of pigs were higher in small size farms (${\leq}$2,000) than that of big size farms (>2,000). However, none of the bloods (n=300) from domestic pigs and tissues (n=200) from slaughter pigs were positive for T. gondii specific DNA by PCR.