• Title/Summary/Keyword: DNA protection

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Development of novel microsatellite markers to analyze the genetic structure of dog populations in Taiwan

  • Lai, Fang-Yu;Lin, Yu-Chen;Ding, Shih-Torng;Chang, Chi-Sheng;Chao, Wi-Lin;Wang, Pei-Hwa
    • Animal Bioscience
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    • v.35 no.9
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    • pp.1314-1326
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    • 2022
  • Objective: Alongside the rise of animal-protection awareness in Taiwan, the public has been paying more attention to dog genetic deficiencies due to inbreeding in the pet market. The goal of this study was to isolate novel microsatellite markers for monitoring the genetic structure of domestic dog populations in Taiwan. Methods: A total of 113 DNA samples from three dog breeds-beagles (BEs), bichons (BIs), and schnauzers (SCs)-were used in subsequent polymorphic tests applying the 14 novel microsatellite markers that were isolated in this study. Results: The results showed that the high level of genetic diversity observed in these novel microsatellite markers provided strong discriminatory power. The estimated probability of identity (P(ID)) and the probability of identity among sibs (P(ID)sib) for the 14 novel microsatellite markers were 1.7×10-12 and 1.6×10-5, respectively. Furthermore, the power of exclusion for the 14 novel microsatellite markers was 99.98%. The neighbor-joining trees constructed among the three breeds indicated that the 14 sets of novel microsatellite markers were sufficient to correctly cluster the BEs, BIs, and SCs. The principal coordinate analysis plot showed that the dogs could be accurately separated by these 14 loci based on different breeds; moreover, the Beagles from different sources were also distinguished. The first, the second, and the third principal coordinates could be used to explain 44.15%, 26.35%, and 19.97% of the genetic variation. Conclusion: The results of this study could enable powerful monitoring of the genetic structure of domestic dog populations in Taiwan.

Construction of an Agroinfectious Clone of a Korean Isolate of Sweet Potato Symptomless Virus 1 and Comparison of Its Infectivity According to Agrobacterium tumefaciens Strains in Nicotiana benthamiana

  • Phuong T. Ho;Hee-Seong Byun;Thuy T. B. Vo;Aamir Lal;Sukchan Lee;Eui-Joon Kil
    • The Plant Pathology Journal
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    • v.39 no.3
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    • pp.255-264
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    • 2023
  • Sweet potato symptomless virus 1 (SPSMV-1) is a single-stranded circular DNA virus, belonging to the genus Mastrevirus (family Geminiviridae) that was first identified on sweet potato plants in South Korea in 2012. Although SPSMV-1 does not induce distinct symptoms in sweet potato plants, its co-infection with different sweet potato viruses is highly prevalent, and thus threatens sweet potato production in South Korea. In this study, the complete genome sequence of a Korean isolate of SPSMV-1 was obtained by Sanger sequencing of polymerase chain reaction (PCR) amplicons from sweet potato plants collected in the field (Suwon). An infectious clone of SPSMV-1 (1.1-mer) was constructed, cloned into the plant expression vector pCAMBIA1303, and agro-inoculated into Nicotiana benthamiana using three Agrobacterium tumefaciens strains (GV3101, LBA4404, and EHA105). Although no visual differences were observed between the mock and infected groups, SPSMV-1 accumulation was detected in the roots, stems, and newly produced leaves through PCR. The A. tumefaciens strain LBA4404 was the most effective at transferring the SPSMV-1 genome to N. benthamiana. We confirmed the viral replication in N. benthamiana samples through strand-specific amplification using virion-sense- and complementary-sense-specific primer sets.

Anti-Obesity and Inhibitory Effect of Lipid Accumulation of The Cone of Pinus rigida × Pinus taeda in 3T3-L1 Cells

  • Da-Yoon Lee;Tae-Won Jang;So-Yeon Han;Seo-Yoon Park;Woo-Jin Oh;Jae-Ho Park
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2023.04a
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    • pp.55-55
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    • 2023
  • With the COVID-19 pandemic, there is increasing interest in anti-obesity strategies. According to the National Statistical Office, the obesity rate in Korea was 38.3% in 2020 and 37.1% in 2021. Obesity is a risk factor for several severe diseases, including stroke, heart disease, type 2 diabetes, and certain types of cancer. Pinus rigida × Pinus taeda is a hybrid of Pinus rigida Mill and Pinus taeda Linn, and its cones are considered a by-product. Although previous studies have investigated their pharmacological effects on antioxidant activity and protection against oxidative DNA damage, few researchers have explored their potential as functional natural materials. Therefore, we evaluated the anti-obesity effects of the cone of ethyl acetate fraction of P. rigida × P. taeda (ERT), specifically its ability to inhibit lipid accumulation. Our analysis showed that ERT contains phytochemicals (catechin and caffeic acid) which are known to improve immune function and inhibit cell damage. ERT inhibited lipid droplet accumulation at the cellular levels through Oil Red O staining. Furthermore, ERT suppressed the expression of adipogenic transcription factors (PPARγ and CEBP/α) as well as downstream lipogenic target genes (FAS and SREBP-1) thereby inhibiting adipogenesis. ERT also down-regulated key adipogenic markers, including aP2α, while inducing the phosphorylation of AMPK. It has been reported that PPARγ and CEBP/α are expressed in the early stages of adipose differentiation, while SREBP-1 is expressed in the late stage. Therefore, our findings suggest that ERT activates AMPK signaling pathways, which inhibits adipogenic transcription factors (PPARγ, C/EBPα, and SREBP1) and lipogenic genes (FAS and aP2α), thereby blocking lipid accumulation and preventing obesity and related disorders. ERT showed potential as a new resource for developing a functional material for anti-obesity agents.

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Re-identification of Colletotrichum gloeosporioides Species Complex Isolates in Korea and Their Host Plants

  • Le Dinh Thao;Hyorim Choi;Yunhee, Choi;Anbazhagan Mageswari;Daseul Lee;Dong-Hyun Kim;Hyeon-Dong Shin;Hyowon Choi;Ho-Jong Ju;Seung-Beom Hong
    • The Plant Pathology Journal
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    • v.40 no.1
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    • pp.16-29
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    • 2024
  • The Colletotrichum gloeosporioides species complex includes many phytopathogenic species, causing anthracnose disease on a wide range of host plants and appearing to be globally distributed. Seventy-one Colletotrichum isolates in the complex from different plants and geographic regions in Korea were preserved in the Korean Agricultural Culture Collection (KACC). Most of them had been identified based on hosts and morphological features, this could lead to inaccurate species names. Therefore, the KACC isolates were re-identified using DNA sequence analyses of six loci, comprising internal transcribed spacer, gapdh, chs-1, his3, act, and tub2 in this study. Based on the combined phylogenetic analysis, KACC strains were assigned to 12 known species and three new species candidates. The detected species are C. siamense (n = 20), C. fructicola (n = 19), C. gloeosporioides (n = 9), C. aenigma (n = 5), C. camelliae (n = 3), C. temperatum (n = 3), C. musae (n = 2), C. theobromicola (n = 2), C. viniferum (n = 2), C. alatae (n = 1), C. jiangxiense (n = 1), and C. yulongense (n = 1). Of these, C. jiangxiense, C. temperatum, C. theobromicola and C. yulongense are unrecorded species in Korea. Host plant comparisons showed that 27 fungus-host associations are newly reported in the country. However, plant-fungus interactions need to be investigated by pathogenicity tests.

Combination of red ginseng and velvet antler extracts prevents skin damage by enhancing the antioxidant defense system and inhibiting MAPK/AP-1/NF-κB and caspase signaling pathways in UVB-irradiated HaCaT keratinocytes and SKH-1 hairless mice

  • Van-Long Truong;Yeon-Ji Bae;Ji-Hong Bang;Woo-Sik Jeong
    • Journal of Ginseng Research
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    • v.48 no.3
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    • pp.323-332
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    • 2024
  • Background: Studies have reported that the combination of two or more therapeutic compounds at certain ratios has more noticeable pharmaceutical properties than single compounds and requires reduced dosage of each agent. Red ginseng and velvet antler have been extensively used in boosting immunity and physical strength and preventing diseases. Thus, this study was conducted to elucidate the skin-protective potentials of red ginseng extract (RGE) and velvet antler extract (VAE) alone or in combination on ultraviolet (UVB)-irradiated human keratinocytes and SKH-1 hairless mice. Methods: HaCaT cells were preincubated with RGE/VAE alone or in combination for 2 h before UVB (30 mJ/cm2) irradiation. SKH-1 mice were orally given RGE/VAE alone or in combination for 15 days before exposure to single dose of UVB (600 mJ/cm2). Treated cells and treated skin tissues were collected and subjected to subsequent experiments. Results: RGE/VAE pretreatment alone or in combination significantly prevented UVB-induced cell death, apoptosis, reactive oxygen species production, and DNA damage in keratinocytes and SKH-1 mouse skins by downregulating mitogen-activated protein kinases/activator protein 1/nuclear factor kappa B and caspase signaling pathways. These extracts also strengthened the antioxidant defense systems and skin barriers in UVB-irradiated HaCaT cells and SKH-1 mouse skins. Furthermore, RGE/VAE co-administration appeared to be more effective in preventing UVB-caused skin injury than these extracts used alone. Conclusion: Overall, these findings suggest that the consumption of RGE/VAE, especially in combination, offers a protective ability against UVB-caused skin injury by preventing inflammation and apoptosis and enhancing antioxidant capacity.

INHIBITORY EFFECT OF ALPHA-LIPOIC ACID ON MITOCHONDRIAL DYSFUNCTION AND INTERLEUKIN-8 EXPRESSION IN INTERLEUKIN-1BETA-STIMULATED ATAXIA TELANGIECTASIA FIBROBLASTS

  • J. YOON;H. LEE;J.W. LIM;H. KIM
    • The Korean Journal of Physiology and Pharmacology
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    • v.71 no.1
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    • pp.155-165
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    • 2020
  • Ataxia telangiectasia (A-T) is an inherited neurodegenerative disease caused by mutation in the ataxia telangiectasia mutated (ATM) gene, leading to loss of function in the encoded protein ATM. Because ATM functions to reduce oxidative stress by up-regulating antioxidant enzymes, oxidative stress is a prevalent A-T phenotype and a mediator of the inflammation that drives A-T pathology. Reactive oxygen species (ROS) levels and the expression of pro-inflammatory cytokine interleukin-8 (IL-8) were higher in A-T cells than in normal cells. ROS are related to mitochondrial dysfunction and activation of nuclear factor kappa B (NF-κB) to induce IL-8 expression. Alpha-lipoic acid (α-LA), a naturally occurring thiol compound, shows an antioxidant effect in various cells. This study is aimed to determine if α-LA confers protection against NF-κB activation, IL-8 expression, and mitochondrial dysfunction in A-T cells which are exposed to the inflammatory cytokine IL-1β. A-T fibroblasts were treated with or without α-LA. The levels of intracellular and mitochondrial ROS, mRNA and protein levels of IL-8, mitochondrial membrane potential (MMP), ATP levels, and DNA binding activity of NF-κB were determined. As a result, IL-1β increased NF-κB activation, IL-8 expression, intracellular and mitochondrial ROS levels, but decreased MMP and ATP level in A-T cells. Pretreatment of A-T cells with α-LA inhibited IL-1β-induced activation of NF-κB, IL-8 expression, and mitochondrial dysfunction by reducing ROS levels. In conclusion, supplementation with α-LA may be beneficial for reducing the oxidative stress-induced mitochondrial dysfunction and IL-8 production associated with A-T.

Assaying Mitochondrial COI Sequences and Their Molecular Studies in Hexapoda, PART I: From 2000 to 2009 (육각강에서 보고된 미토콘드리아 COI 염기서열과 이들을 이용한 분자 연구 논문 분석, 파트 I: 2000년~2009년)

  • Lee, Wonhoon;Park, Jongsun;Akimoto, Shin-Ichi;Kim, Sora;Kim, Yang-Su;Lee, Yerim;Kim, Kwang-Ho;Lee, Si Hyeock;Lee, Yong-Hwan;Lee, Seunghwan
    • Korean journal of applied entomology
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    • v.52 no.4
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    • pp.395-402
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    • 2013
  • Since 2000, a large number of molecular studies have been conducted in Hexapoda with generating large amount of mitochondrial sequences. In this study, to review mitochondrial COI sequences and their molecular studies reported in Hexapoda from 2000 to 2009, 488 molecular studies conducted based on 58,323 COI sequences were categorized according to 26 orders and the positions of COI sequences (5', 3', and entire regions). The numbers of molecular studies in which the three regions utilized varied largely among the 26 orders; however, seven orders showed preferred positions of COI sequences in the researches: Diptera and Orthoptera revealed the largest number of studies in the 5' region; while, Coleoptera, Phthiraptera, Odonata, Phasmatodea, and Psocoptera, showed the largest number of studies in the 3' region. From comparing 84 molecular studies published before 2000, we observed the possibilities that molecular studies in Coleoptera, Diptera, Phthiraptera, and Phasmatodea from 2000 to 2009 had been followed classical studies using the positions of COI sequences well-known until 1999. This study provides useful information to understand the overall trends in COI sequence usages as well as molecular studies conducted from 2000 to 2009 in Hexapoda.

Interleukin-2 production and alteration of T cell subsets in mice infected with Naegleria fowleri (Naegleria fowleri 감염 마우스에 있어서 interleukin-2 생성 및 T 림프구 아형변동)

  • Yu, Cheol-Ju;Sin, Ju-Ok;Im, Gyeong-Il
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.249-258
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    • 1993
  • Naegleria fowleri is the cause of primary amoebic meningoencephalitis in man, IL-2 levels after stimulation of T lymphocytes by PHA or N.fowleri lysates. the amounts of T lymphocyte subsets and the blastogenic responses of T lymphocytes in mice after Infected with pathogenic N. fowleri were studied comparing between two study groups, one $1{\;}{\times}{\;}10^4$ trophozoites inoculated mice and the other $1{\;}{\times}{\;}10^5$ trophozoites inoculated mice. All experimental samples were obtained on the day 7, 14 and 24 after inoculation. The mice inoculated with $1{\;}{\times}{\;}10^4$ trophozoites showed a 14.3% mortality rate, and 72.2% in the mice inoculated with $1{\;}{\times}{\;}10^5$ trophozoites. The IL-2 levels on day 14 of two experimental groups were significantly decreased as compared with the control group. Thy 1.2+T cells in the total spleen Iymphocytes of $1{\;}{\times}{\;}10^5$ trophozoites inoculated group on day 7 were significantly increased compared with the control group. There was no significant difference between $1{\;}{\times}{\;}10^4$ trophozoites inoculated group and the control group. $L3T4^{+}{\;}T$ cells and $Ly2^{+}$ T cells in the total spleen Iymphocytes of $1{\;}{\times}{\;}10^5$ trophozoites inoculated group on day 7 were sigrlificantly increased compared with the control group. The DNA S fraction of T cells in the spleen of $1{\;}{\times}{\;}10^5$ trophozoites inoculated group was significantly increased on day 7. The amount of S fractions of DNA were sequentially decreased on day 14 and 24 but they were also signiacantly increased compared with the control group. The results obtained in the experiments indicats that cell mediated immunity after N.fowleri infection acts on very important host's protection immunity around the 7th day after infection. IL-2 level was much suppressed on day 14 which resulted from the exhaustion of host immune response. It was observed that the level of IL-2 production ability and the amounts of T lymphocytes subsets and the blastogenic responses of T lymphocytes were not well correlated during the observation period.

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Antidiabetic Effect of Korean Red Ginseng by Puffing Process in Streptozotocin-Induced Diabetic Rats (Streptozotocin 유발 당뇨쥐에서 팽화가공 처리한 홍삼의 항당뇨 효과)

  • Kim, Shin-Hee;Kang, Ju-Seop;Lee, Sang-Jun;Chung, Young-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.6
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    • pp.701-707
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    • 2008
  • Antidiabetic effect of Korean red ginseng (RG) processed by puffing in streptozotocin (STZ)-induced diabetic (DM) rats was investigated. Five week-old SD rats were divided into four groups; normal control (NC) group, DM group, red ginseng (RG) group and puffed red ginseng (PG) group. The RG and PG groups were orally provided with RG or PG dissolved in water (500 mg/kg) respectively for seven weeks after single injection of STZ (50 mg/kg, i.v.) followed by identification of DM. NC group received saline vehicle instead of STZ. At the end of feeding of RG or PG, the changes of fasting blood glucose, serum insulin and amylase level and serum lipid profiles were evaluated. Also, oral glucose tolerance test (OGTT), comet assay and histopathological examination were performed. At 7th week, the fasting blood glucose levels of the RG and PG groups were reduced compared to the DM group by 11.54% and 20.22%, respectively. The result of OGTT did not show significant differences among DM and two red ginseng groups. While serum insulin and TG levels were predominantly improved in PG group (p<0.05), serum amylase level was increased in RG group. Alkaline comet assay for checking the oxidative damage of DNA showed that TL (tail length, ${\mu}m$) and TM (tail moment) in the blood lymphocyte of PG group significantly decreased in contrast with DM group. Histopathological results of pancreas showed that destruction of exocrine as well as endocrine might be cured by the administration of RG and PG. These results suggest that PG could exert more protection against STZ-induced toxicity than RG group.

Construction and In vitro Study of a Prx 6/Luc Vector System for Screening Antioxidant Compounds in the Transgenic Mice (항산화반응을 유발하는 물질의 검색에 적용할 수 있는 형질전환 마우스 생산을 위한 새로운 Prx 6/Luc 벡터시스템의 제조 및 폐암세포주에서 반응성 확인)

  • Lee, Young Ju;Nam, So Hee;Kim, Ji Eun;Hwang, In Sik;Lee, Hye Ryun;Choi, Sun Il;Kwak, Moon Hwa;Lee, Jae Ho;Jung, Young Jin;An, Beum Soo;Hwang, Dae Youn
    • Journal of Life Science
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    • v.23 no.2
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    • pp.167-174
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    • 2013
  • Peroxiredoxin 6 (Prx 6) is a member of the thiol-specific antioxidant protein family, which may play a role in protection against oxidative stress and in regulating phospholipid turnover. The aim of this study was to determine whether a human Prx 6/Luc vector was stably expressed and responded to antioxidants in a lung cell line (NCI-H460). To achieve this, the luciferase signal, hPrx 6 mRNA expression, and superoxide dismutase (SOD) activity were measured in transfectants with a hPrx 6/Luc plasmid after treatment with four antioxidant extracts, including Korea white ginseng (KWG), Korea red ginseng (KRG), Liriope platyphylla (LP), and red Liriope platyphylla (RLP). First, the hPrx 6/Luc plasmid was successfully constructed with DNA fragments of human Prx 6 promoter, amplified by PCR using genomic DNA isolated from NCI-H460 cells, and cloned into the pTransLucent reporter vector. The orientation and sequencing of the hPrx 6/Luc plasmid were identified with restriction enzyme and automatic sequencing. A luciferase assay revealed significant enhancement of luciferase activity in the four treatment groups compared with a vehicle-treated group, although the ratio of the increase was different within each group. The KRG- and LP-treated groups showed higher activity than the KWG- and RLP-treated groups. Furthermore, the luciferase activity against RLP occurred roughly in a dose-dependent manner. However, the level of endogenous hPrx 6 mRNA did not change in any group treated with the four extracts. The SOD activity was in agreement with the luciferase activity. Therefore, these results indicate that the hPrx 6/Luc vector system may successfully express and respond to antioxidant compounds in NCI-H460 cells. The data also suggest that the Prx 6/Luc vector system may be effectively applied in screening the response of hPrx 6 to antioxidant compounds in transgenic mice.