• Title/Summary/Keyword: cytochrome b

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Effects of Dandelion on Oxygen Free Radical Generating and Scavenging System of Brain in Streptozotocin-Induced Diabetic Rats (서양민들레가 Streptozotocin으로 유발한 당뇨 횐쥐의 뇌조직 중 유해 활성산소 생성 및 제거 효소계에 미치는 영향)

  • 김명주;조수열
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.3
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    • pp.500-505
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    • 2002
  • Many studies have shown that hyperglycemia leads to an increase of lipid peroxidation in diabetic patients and animals, reflecting a rise reactive oxygen species production. It is increasingly recognized that brain is another site of diabetic organ damage. Accordingly, this study was to investigate the effect of dandelion on oxygen free radical generating and scavenging system of brain in streptozotocin (STZ)-induced diabetic rats. Male Wistar rats were divided into diabetic (control) and diabetic-dandelion supplemented groups. Dandelion was supplemented for 4 weeks with dandelion leaf and root powder (DLP, DRP) or dandelion leaf and root water extract (DLW, DRW) based on 11.4 g of raw dandelion/kg diet. Diabetes was induced by single injection STZ (55 mg/kg B.W., i.p.)in a citrate buffer. Oxygen free radical generating enzymes, cytochrome P-450, amino-pyrine N-demethylase, aniline hydroxylase and xanthine oxidase, were lowered in dandelion supplemented-groups compared to the control group. Superoxide dismutase, catalase and gluthathione peroxidase activities of brain were also lower in dandelion leaf and root supplemented-group than in the control group, whereas glutathione S-transferase activity and gluthathione content were increased in dandelion supplemented-groups compared to the control group. With regard to the lipid peroxidation products, the malondialdehyde content of brain was lower in dandelion supplemented groups. Therefore, it could be suggested that powder and water extract of dandelion leaf or root are beneficial in preventing diabetic complication from lipid peroxidation and free radical in brain of diabetic rat brain.

PCR-RFLP for the Identification of Mammalian Livestock Animal Species

  • Han, Sang-Hyun;Park, Seon-Mi;Oh, Hong-Shik;Kang, Geunho;Park, Beom-Young;Ko, Moon-Suck;Cho, Sang-Rae;Kang, Yong-Jun;Kim, Sang-Geum;Cho, In-Cheol
    • Journal of Embryo Transfer
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    • v.28 no.4
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    • pp.355-360
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    • 2013
  • Precise, rapid and simple methods for species identification in animals are among the most important techniques in the livestock industry and research fields including meat classification. In this study, polymerase chain reaction (PCR) based molecular identification using inter species polymorphisms were examined by PCR-restriction fragment length polymorphism (RFLP) analysis for mitochondrial DNA (mtDNA) cytochrome b (CYTB) gene sequences among four mammalian livestock animals (cattle, horse, goat and pig). The results from PCR-RFLP analysis using the AluI restriction enzyme were also provided for the species-specific band patterns among CYTB gene sequences in these four species. The AluI-digestion for CYTB genes provided interesting migration patterns differentially displayed according to each species. Cattle and horse had one AluI-recognition site at different nucleotide positions and their AluI-digested fragments showed different band patterns on the gels. Pig had two AluI-recognition sites within the amplified CYTB sequences and produced three bands on the gels. Goat had no AluI-recognition site and was located at the same position as the uncut PCR product. The results showed the species-specific band patterns on a single gel among the four livestock animal species by AluI-RFLP. In addition, the results from blind tests for the meat samples collected from providers without any records showed the identical information on the species recorded by observing their phenotypes before slaughter. The application of this PCR-RFLP method can be useful and provide rapid, simple, and clear information regarding species identification for various tissue samples originating from tested livestock species.

Occurrence of a Natural Intergeneric Hybrid between a Female Tanakia lanceolata and a Male Rhodeus pseudosericeus (Cypriniformes: Cyprinidae) in Daecheoncheon Stream Flowing into the Yellow Sea in the Republic of Korea (서해안 독립 하천 대천천에서 납자루 Tanakia lanceolata (♀)와 한강납줄개 Rhodeus pseudosericeus(♂)의 자연 속간잡종 출현)

  • Kim, Yong Hwi;Sung, Mu Sung;Yun, Bong Han;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.33 no.2
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    • pp.45-56
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    • 2021
  • A male, presumed to be an intergeneric hybrid between Tanakia lanceolata and Rhodeus pseudosericeus, was collected in the Boryeong Daecheoncheon Stream flowing into the Yellow Sea in the Republic of Korea. Morphological and molecular phylogenetic analyses were performed to discriminate the definite origin of the estimated natural hybrid. As a result of the morphological analysis, the color of the dorsal and anal fin rays edges of the natural hybrid individual, the upper and lower body colors followed the morphological characteristics of T. lanceolata, and that blue longitudinal stripe in the center of the caudal peduncle, the incomplete lateral line, and the barbels absent followed the morphological characteristics of R. pseudosericeus. In addition, as a result of the cytochrome b (cytb) gene analysis of mitochondrial DNA (mtDNA), the natural hybrid showed a nucleotide sequence similarity of 99.82 to 100% with T. lanceolata, and the maternal species was identified as T. lanceolata. As a result of the recombination activating gene 1 (rag1) gene analysis of nuclear DNA (nDNA), the natural hybrid showed double peaks pattern reflecting both the single nucleotide polymorphism sites (38 bp) between T. lanceolata and R. pseudosericeus, and the paternal species was identified as R. pseudosericeus. Therefore, a natural hybrid estimated male of Acheilognathinae analyzed in this study was found to be an intergeneric hybrid between a female T. lanceolata and a male R. pseudosericeus.

Phylogenetic Analysis of Carassius auratus and C. cuvieri in Lake Yedang Based on Variations of Mitochondrial CYTB Gene Sequences (예당호 붕어와 떡붕어의 CYTB 유전자를 이용한 유연관계 분석)

  • Kim, Gye-Woong;Joe, Sung-Duck;Kim, Hack-Youn;Park, Hee-Bok
    • Journal of Life Science
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    • v.30 no.12
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    • pp.1063-1069
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    • 2020
  • Two crucian carp species (Carassius auratus and C. cuvieri) inhabit Lake Yedang in South Korea, and C. auratus is known to be native to Korea. Classification of these two freshwater fish species is often confused because of their morphological similarity. To distinguish the two species, we conducted phylogenetic and population genetic analyses of C. auratus and C. cuvieri based on their mitochondrial DNA sequences of the cytochrome b gene (CYTB). We also compared our partial CYTB sequence (<1,056 bp) with 10 Chinese, nine Japanese, and two Russian crucian carp fishes. The results of our phylogenetic analysis showed that C. auratus and C. cuvieri were clearly divided into two phylogroups. The nucleotide diversity (π) of C. auratus from Korea, China, and Japan showed a range of 0.146%~0.421%, while the range of π of C. cuvieri from Korea and Japan was lower than those of C. auratus (0.0%~0.054%). Moreover, the comparison of CYTB divergence among crucian carp fishes in China, Japan, and Korea indicated that Korean Carassius fishes were distantly related to those from China and Japan, with two exceptions: the pairwise Fst value between Korean C. auratus and northern Chinese C. auratus was not significantly different. In addition, no significant genetic divergence between Korean and Japanese C. cuvieri was detected. We conclude that, despite the morphological similarities, C. auratus and C. cuvieri should be considered as separate freshwater fish resources in conservation efforts for genetic diversity.

Licochalcone C Inhibits the Growth of Human Colorectal Cancer HCT116 Cells Resistant to Oxaliplatin

  • Seung-On Lee;Sang Hoon Joo;Jin-Young Lee;Ah-Won Kwak;Ki-Taek Kim;Seung-Sik Cho;Goo Yoon;Yung Hyun Choi;Jin Woo Park;Jung-Hyun Shim
    • Biomolecules & Therapeutics
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    • v.32 no.1
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    • pp.104-114
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    • 2024
  • Licochalcone C (LCC; PubChem CID:9840805), a chalcone compound originating from the root of Glycyrrhiza inflata, has shown anticancer activity against skin cancer, esophageal squamous cell carcinoma, and oral squamous cell carcinoma. However, the therapeutic potential of LCC in treating colorectal cancer (CRC) and its underlying molecular mechanisms remain unclear. Chemotherapy for CRC is challenging because of the development of drug resistance. In this study, we examined the antiproliferative activity of LCC in human colorectal carcinoma HCT116 cells, oxaliplatin (Ox) sensitive and Ox-resistant HCT116 cells (HCT116-OxR). LCC significantly and selectively inhibited the growth of HCT116 and HCT116-OxR cells. An in vitro kinase assay showed that LCC inhibited the kinase activities of EGFR and AKT. Molecular docking simulations using AutoDock Vina indicated that LCC could be in ATP-binding pockets. Decreased phosphorylation of EGFR and AKT was observed in the LCC-treated cells. In addition, LCC induced cell cycle arrest by modulating the expression of cell cycle regulators p21, p27, cyclin B1, and cdc2. LCC treatment induced ROS generation in CRC cells, and the ROS induction was accompanied by the phosphorylation of JNK and p38 kinases. Moreover, LCC dysregulated mitochondrial membrane potential (MMP), and the disruption of MMP resulted in the release of cytochrome c into the cytoplasm and activation of caspases to execute apoptosis. Overall, LCC showed anticancer activity against both Ox-sensitive and Ox-resistant CRC cells by targeting EGFR and AKT, inducing ROS generation and disrupting MMP. Thus, LCC may be potential therapeutic agents for the treatment of Ox-resistant CRC cells.

Development of a Kit for Diagnosing AtCYP78A7 Protein in Abiotic-tolerant Transgenic Rice Overexpressing AtCYP78A7 (AtCYP78A7 과발현 환경스트레스 내성 형질전환 벼의 단백질 진단 키트 개발)

  • Nam, Kyong-Hee;Park, Jung-Ho;Pack, In-Soon;Kim, Ho Bang;Kim, Chang-Gi
    • Journal of Life Science
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    • v.28 no.7
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    • pp.835-840
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    • 2018
  • Quantitative determination of the protein expression levels is one of the most important parts in assessment of the safety of foods derived from genetically modified (GM) crops. Overexpression of AtCYP78A7, a gene encoding cytochrome P450 protein, has been reported to improve tolerance to abiotic stress, such as drought and salt stress, in transgenic rice (Oryza sativa L.). In the present study, an enzyme-linked immunosorbent assay (ELISA) kit for diagnosing AtCYP78A7 protein including AtCYP78A7-specific monoclonal antibody was developed. GST-AtCYP78A7 recombinant protein was induced and purified by affinity column. Four monoclonal antibodies (mAb 6A7, mAb 4C2, mAb 11H6, and mAb 7E8) against recombinant protein were also produced and biotinylated with avidin-HRP. After pairing test using GST-AtCYP78A7 protein and lysate of rice samples, mAb 4C2 and mAb 7E8 were selected as a capture antibody and a detecting antibody, respectively, for ELISA kit. Product test using rice samples indicated that percentages of detected protein in total protein were greater than 0.1% in AtCYP78A7-overexpressing transgenic rice (Line 10B-5 and 18A-4), whereas those in negative control non-transgenic rice (Ilpum and Hwayoung) were less than 0.1%. The ELISA kit developed in this study can be useful for the rapid detection and safety assessment of transgenic rice overexpressing AtCYP78A7.

Chemical Composition and Antitumor Apoptogenic Activity of Methylene Chloride Extracts from the Leaves of Zanthoxylum schinifolium (Zanthoxylum schinifolium잎의 methylene chloride 추출물의 화학적 조성 및 암세포에 대한 세포자살 유도활성과 그 작용기전)

  • Kim Jun-Seok;Jun Do-Youn;Woo Mi-Hee;Rhee In-Koo;Kim Young-Ho
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.546-554
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    • 2006
  • To understand antitumor activity of Zanthoxylum schinfolium, which has been used as an aromatic and medicinal plant in Korea, the cytotoxic effect of various organic solvent extracts of its leaves on human tumor cells were investigated. Among these extracts such as methanol extract (SL-13), methylene chloride extract (SL-14), ethyl acetate extract (SL-15), n-butanol extract (SL-16), and residual fraction (SL-17), SL-14 appeared to contain the most cytotoxic activity against leukemia and breast cancer cells tested. The methylene chloride extra.1 (SL-14) possessed an apoptogenic activity causing apoptotic DNA fragmentation of human acute leukemia Jurkat T cells via mitochondrial cytochrome c release into cytoplasm, subsequent activation of caspase-9 and caspase-3, and cleavage of PARP, which could be negatively regulated by antiapoptotic protein Bcl-xL. The GC-MS analysis of SL-14 revealed that the twenty-two ingredients of SL-14 were 9,19-cyclolanost-24-en-3-ol (15.1%), 2-a-methyl-17, b-hop-21-ene (15.1%), 15-methyl-2,3-dihydro-1H benzazepin (11.95%), phytol (10.38%), lupeol (9.92%), 12-methylbenzofuran (8.23%), hexadecanoic acid (5.96%), cis,cis,cis-9,12,15-octadecatrienoic acid-methyl-ester (5.49%), 9,12,15-octadecatrienoic acid-methylester (3.59%), 15-methyl-4-(1-methylethylidene)-2-(4-nitrophenyl) (3.36%), hexadecanoic acid methyl ester (1.93%), vitamine E (1.88%), beta-amyrin (0.96%), and auraptene (0.89%). These results demonstrate that the cytotoxicity of the methylene chloride extract of the leaves of Z. schinifolium toward Jurkat T cells is mainly attributable to apoptosis mediated by mitochondria-dependent caspase cascade regulated by Bcl-xL, and provide an insight into the mechanism underlying antitumor activity of the edible plant Z. schinifolium.

Mitigation Effects of Foliar-Applied Hydrogen Peroxide on Drought Stress in Sorghum bicolor (과산화수소 엽면 처리에 의한 수수에서 한발 스트레스 완화 효과)

  • Shim, Doo-Do;Lee, Seung-Ha;Chung, Jong-Il;Kim, Min Chul;Chung, Jung-Sung;Lee, Yeong-Hun;Jeon, Seung-Ho;Song, Gi-Eun;Shim, Sang-In
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.2
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    • pp.113-123
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    • 2020
  • Global climatic change and increasing climatic instability threaten crop productivity. Due to climatic change, drought stress is occurring more frequently in crop fields. In this study, we investigated the effect of treatment with hydrogen peroxide (H2O2) before leaf development on the growth and yield of sorghum for minimizing the damage of crops to drought. To assess the effect of H2O2 on the growth of sorghum plant, 10 mM H2O2 was used to treat sorghum leaves at the 3-leaf stage during growth in field conditions. Plant height, stem diameter, leaf length, and leaf width were increased by 7.6%, 9.6%, 8.3% and 11.5%, respectively. SPAD value, chlorophyll fluorescence (Fv/Fm), photosynthetic rate, stomatal conductance, and transpiration rate were increased by 3.0%, 4.9%, 26.0%, 23.4% and 12.7%, respectively. The amount of H2O2 in the leaf tissue of sorghum plant treated with 10 mM H2O2 was 0.7% of the applied amount after 1 hour. The level increased to approximately 1.0% after 6 hours. The highest antioxidant activity measured by the Oxygen Radical Absorbance Capacity assay was 847.3 µmol·g-1 at 6 hour after treatment. However, in the well-watered condition, the concentration of H2O2 in the plant treated by the foliar application of H2O2 was 227.8 µmol·g-1 higher than that of the untreated control. H2O2 treatment improved all the yield components and yield-related factors. Panicle length, plant dry weight, panicle weight, seed weight per plant, seed weight per unit area, and thousand seed weight were increased by 8.8%, 18.0%, 24.4%, 24.7%, 29.9% and 7.1%, respectively. Proteomic analysis showed that H2O2 treatment in sorghum increased the tolerance to drought stress and maintained growth and yield by ameliorating oxidative stress.

Induction of G2/M Arrest and Apoptosis by the Methanol Extract of Typha orientalis in Human Colon Adenocarcinoma HT29 Cells (포황 메탄올 추출물에 의한 인체 대장암 세포주 HT29의 G2/M Arrest 및 Apoptosis 유발)

  • Jin, Soojung;Yun, Seung-Geun;Oh, You Na;Lee, Ji-Young;Park, Hyun-Jin;Jin, Kyong-Suk;Kwon, Hyun Ju;Kim, Byung Woo
    • Microbiology and Biotechnology Letters
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    • v.41 no.4
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    • pp.425-432
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    • 2013
  • Typha orientalis, also known as bulrush or cattail, is a perennial herbaceous plant found in freshwater wetlands and has been widely used in constructed wetlands for wastewater treatment. Recent data has revealed that SH21B, a mixture composed of seven herbs including T. orientalis, exhibited an anti-adipogenic activity by the inhibition of the expression of adipogenic regulators. However, the anti-cancer effect of T. orientalis and its molecular mechanisms remain unclear. In this study, we evaluated the anti-cancer effect and its mechanism in the methanol extract of T. orientalis (METO) on human colon carcinoma HT29 cells. It was found that METO treatment showed cytotoxic activity in a dose-dependent manner, and induced G2/M cell cycle arrest and apoptosis in HT29 cells. The induction of G2/M arrest by METO was associated with the up-regulation of phospho-Cdc2 (Tyr15), an inactive form of Cdc2 and the down-regulation of Cdc25c phosphatase. METO also induced tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 (WAF1/CIP1) expression. In addition, METO-induced apoptosis was characterized by the proteolytic activation of caspase-3, degradation of poly ADP ribose polymerase (PARP), and up-regulation of death receptor FAS and pro-apoptotic Bax expression. Collectively, these results indicate that the cell cycle inhibition and apoptosis induction of METO in HT29 cells allows for the possibility of its use in anti-cancer therapies.

CYP450 1A1 and p53 expression and DNA adduct formation in the liver of rats treated with a single dose of aflatoxins (아플라톡신을 간회 투여한 랫드의 간에서 CYP450 1A1, p53의 발현과 DNA adduct의 형성)

  • Lee, Beom Jun;Lee, Sook Jin;Kim, Tae Myoung;Kim, Dae Joong;Nam, Sang Yoon;Hyun, Sang Hwan;Kang, Jong Koo;Hong, Jin Tae;Kim, Cheul Kyu;Yun, Young Won
    • Korean Journal of Veterinary Research
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    • v.44 no.4
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    • pp.507-513
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    • 2004
  • Aflatoxins are produced mainly by Aspergillus flavus and Aspergillus parasiticus that grow in improperly stored cereals. Aflatoxin B1 ($AFB_1$) is a potent hepatocarcinogen in a variety of experimental animals including human beings. In spite of a high attention to the hepatocarcinogenecity of $AFB_1$, the relative toxicity of aflatoxins ($AFB_2$ and $AFG_1$) is not fully clarified. Sprague-Dawley male rats were orally administered with $AFB_1$, $AFB_2$, and $AFG_1$ at the dose of 250 ${\mu}g/kg$ (additionally including a dose of $1250{\mu}g/kg $ for $AFB_1$) body weight. Animals were then killed at 12, 24 or 48 hrs following aflatoxin exposure. Subsequently the immunohistochemical examination of p53, cytochrome p450 1A1 (CYP450 1A1), and glutathione-S-transferase placental form (GST-P) were performed. The level of the 8-OxodG in the liver was determined. Expressions of CYP450 1A1 and p53 were high in the liver of rats through 48 hrs after treatment of $AFB_1$ at the single dose of $250{\mu}g/kg $. This pattern was more clear as increasing doses. The treatment of $AFB_2$ and $AFG_1$ did not affect the expression of CYP450 1A1 but it caused weak expression of p53. The activity of GST were not found in the liver of rats treated with aflatoxins. The formation of 8-OxodG by $AFB_1$ increased in a dose-dependent manner up to 24 hrs after a single treatment of $AFB_1$ thereafter decreased to the level of control. The treatment of $AFB_2$ and $AFG_1$ did not affect the levels of 8-OxodG in the liver of rats with increasing time. These results in the present study indicate that $AFB_1$ among aflatoxins with low comparable levels is the most toxic as determined by early biomarkers such as CYP450 1A1, p53, GST-P, and 8-OxodG.