• Title/Summary/Keyword: mitochondrial cytochrome b

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Identification of Deer Antler Species Using Sequence Analysis and PCR-RFLP of Mitochondrial DNA (사슴 미토콘드리아 DNA의 염기서열 및 PCR-RFLP분석에 의한 녹용의 종 감별)

  • Shin, Ki-Hyun;Shin, Sung-Chul;Chung, Ku-Young;Chung, Eui-Ryong
    • Food Science of Animal Resources
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    • v.28 no.3
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    • pp.276-282
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    • 2008
  • It is estimated that over 80% of deer antlers produced in the world are consumed in Korea. However, mislabeling or fraudulent replacement of costly antlers with cheaper ones is one of the most common problems in the domestic antler market. Therefore, there is a great need for the development of technology to identify species of antlers. This study was carried out to develop an accurate and reliable method for the identification and authentication of species or subspecies of antlers using DNA sequence analysis and comparison of mitochondrial cytochrome band D-loop region genes among antlers of five deer species, Cervus elaphus sibericus, Cervus elaphus canadensis, Cervus nippon, Cervus elaphus bactrianus and Rangifer tarandus. A variable region of cytochrome band D-loop genes was amplified using PCR with specifically designed primers and sequenced directly. The cytochrome band D-loop region genes showed different DNA sequences between the species of antlers and thus it is possible to differentiate between species on the basis of sequence variation. To distinguish between reindeer (Rangifer tarandus) antlers and other deer antlers, PCR amplicons of the cytochrome b gene were digested with the restriction enzymes NlaIV and TaqI, respectively, which generates a species-specific DNA profile of the reindeer. In addition, samples of 32 sliced antlers labeled Cervus elaphus sibericus from commercial markets were collected randomly and the mt DNA D-loop region of these antler samples was sequenced. Among the antler samples investigated, only 62.5% were from Cervus elaphus sibericus, and others were from Cervus elaphus bactrianus (25.0%), elk (Cervus elaphus canadensis) and reindeer (Rangifer tarandus). Our results suggest that DNA sequencing of mt DNA and PCR-RFLP methods using NlaIV and TaqI enzymes are useful for the identification and discrimination of deer antler species by routine analysis.

Pythium subutonaiense, A New Aquatic Oomycete from Southern China Based on Morphological and Molecular Characters

  • Chen, Jia-Jia;Zheng, Xiao-Bo
    • Mycobiology
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    • v.47 no.3
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    • pp.273-279
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    • 2019
  • A new species, Pythium subutonaiense, isolated from aquatic environments (lake) in China is being described based on morphological characters and molecular evidence. The isolates grew at temperatures between $5^{\circ}C$ and $38^{\circ}C$, and the optimum temperature was $30^{\circ}C$, with a radial growth rate of 17.6 mm at $25^{\circ}C$ per day. It is homothallic and characterized by globose to sub-globose shaped and mostly terminal or sometimes catenulate hyphal swellings, filamentous non-inflated sporangia, and smooth oogonia with hypogynous and monoclinous antheridia that contained one plerotic oospore. In phylogenetic analysis, inferred based on the internal transcribed spacer region of the ribosomal RNA gene and mitochondrial cytochrome c oxidase subunit 1 gene, the new species formed a distinct lineage in Pythium clade B. Differences between the new species and phylogenetically related and morphologically similar species are discussed.

Single Nucleotide Polymorphism Analysis of the COI Gene in Korean Native Chicken (한국재래닭 COI 유전자의 단일염기다형 분석)

  • Jin, S.D.;Seo, D.W.;Sim, J.M.;Baek, W.K.;Jung, K.C.;Jang, B.K.;Choi, K.D.;Lee, J.H.
    • Korean Journal of Poultry Science
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    • v.36 no.1
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    • pp.85-88
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    • 2009
  • One of the mitochondrial genes, called cytochrome c oxidase I (COI), has been widely used for the species identification (called bio-barcode) in birds. In this study, the bio-barcode has been applied to chicken breeds in Korea whether it also can be used as a molecular marker for breed identification. Data indicated that Korean native chicken has the mixed SNP (single nucleotide polymorphism) patterns between White Leghorn (Layer) and Cornish (Broiler) and ultimately, it can not be used as the marker for breed identification. However, this result indicates the mixed use of the Korean native chicken, since it has been used for dual purpose for producing meat and egg for a long time. In order to use as a marker for species identification, more reliable mitochondrial and/or nuclear DNA markers need to be developed.

Hyperglycemia Influences Apoptosis and Autophagy in Porcine Parthenotes Developing In Vitro

  • Xu, Yong-Nan;Li, Ying-Hua;Lee, Sung Hyun;Kwon, Jung-Woo;Lee, Seul Ki;Heo, Young-Tae;Cui, Xiang-Shun;Kim, Nam-Hyung
    • Reproductive and Developmental Biology
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    • v.37 no.2
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    • pp.65-73
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    • 2013
  • The objective of this study was to examine the effects of high concentrations of glucose on porcine parthenotes developing in vitro. Addition of 55 mM glucose to the culture medium of embryos at the four-cell-stage significantly inhibited blastocyst formation, resulting in fewer cells in blastocyst-stage embryos and increased levels of apoptosis and autophagy compared to control. Quantitative reverse transcriptase (RT) PCR analysis revealed that the expression of pro-apoptotic genes (Caspase 3, Bax and Bak) and autophagy genes (Atg6 and Atg8/Lc3) were increased significantly by the addition of 55 mM glucose to the culture medium compared to control. MitoTracker Green fluorescence revealed a decrease in the overall mitochondrial mass compared to control. However, the addition of 55 mM glucose had no effect on mRNA expression of the nuclear DNA-encoded mitochondrial-related genes, cytochrome oxidase (Cox) 5a, Cox5b and Cox6b1. These results suggest that hyperglycemia reduced the mitochondrial content of porcine embryos developing in vitro and that this may hinder embryonic development to the blastocyst stage and embryo quality by increasing apoptosis and autophagy in these embryos.

Induction of G1 Phase Cell Cycle Arrest and Apoptotic Cell Death by 5-Fluorouracil in Ewing′s Sarcoma CHP-100 Cells (CHP-100 Ewing′s 육종세포에서 5-fluorouracil에 의한 G1 arrest 유도 및 apoptosis 유발에 관한 연구)

  • Kim, Sung Ok;Choi, Yung Hyun
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1015-1021
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    • 2016
  • 5-fluorouracil (5-FU), a pyrimidine analog, is a widely used anticancer drug, which works through irreversible inhibition of thymidylate synthase. In the present study, it was investigated the anti-proliferative effects and molecular mechanisms of 5-FU using Ewing's Sarcoma CHP-100 Cells. The present data indicated that treatment of 5-FU to CHP-100 cells induced a G1 phase arrest of the cell cycle in a time-dependent manner. 5-FU-induced G1 arrest was correlated with the accumulation of the hypophosphorylated form of the retinoblastoma protein (pRB) and association of pRB with the transcription factors E2F-1 and E2F-4. Although 5-FU treatment did affect the levels of cyclin-dependent kinases, the levels of cyclin A and B were markedly down-regulated as compared with the untreated control group. In addition, 5-FU-induced G1 arrest of CHP-100 cells was also associated with the induction of apoptosis, as determined by apoptotic cell morphologies, degradation of poly(ADP-ribose) polymerase and Annexin V staining. Furthermore, 5-FU induced the loss of mitochondrial membrane potential with up-regulated pro-apoptotic Bax expression, down-regulated anti-apoptotic Bcl-2 expression and cytochrome c release from mitochondria to cytosol. Collectively, the data suggest that 5-FU is effective in inducing cell growth reduction and apoptosis, in part, by reducing phosphorylation of pRB and activating mitochondrial dysfunction in CHP-100 cells.

Enhancement of TRAIL-Mediated Apoptosis by Genistein in Human Hepatocellular Carcinoma Hep3B Cells: Roles of p38 MAPK Signaling Pathway (인체간암세포에서 genistein의 TRAIL에 의한 apoptosis 유도 상승효과에서 미치는 p38 MAPK signaling pathway의 영향)

  • Jin, Cheng-Yun;Park, Cheol;Park, Sang-Eun;Hong, Sang-Hoon;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1549-1557
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    • 2011
  • Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can selectively induce apoptosis in many types of transformed cells; however, some human hepatocellular carcinoma cells are particularly resistant to the effects of TRAIL. Although genistein, a natural isoflavonoid phytoestrogen, has been shown to have pro-apoptotic activity against human cancer cell lines, little is known about the mechanism of genistein in terms of TRAIL-induced apoptosis. In the present study, it was investigated whether or not combined treatment with genistein and TRAIL synergistically induced apoptosis in Hep3B hepatocarcinoma cells. Results indicate that treatment with TRAIL in combination with nontoxic concentrations of genistein sensitized TRAIL-resistant Hep3B cells to TRAIL-induced apoptosis, which was associated with mitochondrial dysfunction. Further, the inhibition of p38 mitogen-activated protein kinase (MAPK) activation markedly decreased genistein and TRAIL-induced cell viability and apoptosis by enhanced truncation of Bid, increase of pro-apoptotic Bax, decrease of anti-apoptotic Bcl-2, and release of cytochrome c from mitochondria to cytoplasm. Activation of caspases and degradation of poly (ADP-ribose) polymerase induced by the combined treatment was also markedly increased by the inhibition of p38 MAPK, through the mitochondrial amplification step. In conclusion, our data suggest that genistein sensitizes TRAIL-induced-apoptosis via p38 MAPK-dependent pathway.

Analysis of Mitochondrial Gene Sequence in Etoxazole Resistant Two-Spotted Spider Mite, Tetranychus urticae (Etoxazole 저항성 점박이응애의 미토콘드리아 유전자 서열 분석)

  • Park, Sang-Eun;Koo, Hyun-Na;Yoon, Chang-Mann;Choi, Jang-Jeon;Kim, Gil-Hah
    • The Korean Journal of Pesticide Science
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    • v.16 no.1
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    • pp.54-61
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    • 2012
  • The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most important pest species devastating many horticultural and ornamental crops and fruit trees. Difficulty in managing this mite is largely attributed to its ability to develop resistance to many important acaricides. Development of 3,700-folds resistance to etoxazole was found in the population of T. urticae collected from rose greenhouses in Buyeo, Chungnam Province in August 2000. This population has been selected for eleven years with etoxazole (over 500 times), and increased over 5,000,000-folds in resistance as compared with susceptible strain. Also, etoxazole-resistant strain was shown to be maternally inherited. The objective of this study was to determine whether resistance of T. urticae to etoxazole was linked with point mutations in the mitochondrial gene. DNA sequencing of cytochrome c oxidase subunit I (COX1), COX2, COX3, cytochrome b (CYTB), NADH dehydrogenase subunit 1 (ND1), ND2, ND3, ND4, ND5, and ND6 were analyzed by comparing two etoxazole-susceptible and etoxazole-resistant strains. As a result, differences were not detected between the nucleotide sequences of two strains within a mitochondrial gene.

Infection of Taenia asiatica in a Bai Person in Dali, China

  • Wang, Li;Luo, Xuenong;Hou, Junling;Guo, Aijiang;Zhang, Shaohua;Li, Hailong;Cai, Xuepeng
    • Parasites, Hosts and Diseases
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    • v.54 no.1
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    • pp.67-70
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    • 2016
  • We report here a human case of Taenia asiatica infection which was confirmed by genetic analyses in Dali, China. A patient was found to have symptoms of taeniasis with discharge of tapeworm proglottids. By sequencing of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene, we observed nucleotide sequence identity of 99% with T. asiatica and 96% with T. saginata. Using the cytochrome b (cytb) gene, 99% identity with T. asiatica and 96% identity with T. saginata were found. Our findings suggest that taeniasis of people in Dali, China may be mainly caused by T. asiatica.

Evolutionary Relationship of Liobagrus mediadiposalis (Teleostei: Amblycipitidae) Populations in Korea Inferred from Cytochrome b DNA Sequences (한국 고유종인 자가사리(Liobagrus mediadiposalis) 지역개체군의 분자진화적 유연관계)

  • Kim, Maeng Jin;Han, Song-Hun;Yang, Hye-Young;Jo, Mi-Ran;Chung, Sang-Chul;Song, Choon Bok
    • Korean Journal of Ichthyology
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    • v.18 no.4
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    • pp.329-338
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    • 2006
  • Phylogenetic relationships and DNA polymorphism among local populations of the Korean native catfish species, Liobagrus mediadiposalis, have been investigated based on mitochondrial cytochrome b DNA sequences. As a result, three genetically distinct groups of local populations were recognized based on the phylogenetic tree constructed. The first group was called "Nakdong-river group" that included the local populations from Geumho-river, Gyeongho-river, and Deokcheon-river; the second one was "Geum-river group"; the third one was represented as "Seomjin-river group" that included the samples from Seomjin-river, Dongjin-river and Geokum-do. The phylogeny also implied that the ancestral group of L. mediadiposalis have first evolved to Nakdong-river group, and later two local populations (Geum-river and Seomjinriver group) were diverged from the other lineage. DNA polymorphisms we observed were 4.4~4.7% between Geum-river group and Seomjin-river group, 5.1~5.5% between Seomjinriver group and Nakdong-river group, and 5.5~5.7% between Nakdong-river group and Geumriver group. These results indicated the long period of geographic isolation due to the river system in Korea caused such high degrees of DNA polymorphisms between local populations of L. mediadiposalis.

Hesperidin Attenuates Ultraviolet B-Induced Apoptosis by Mitigating Oxidative Stress in Human Keratinocytes

  • Hewage, Susara Ruwan Kumara Madduma;Piao, Mei Jing;Kang, Kyoung Ah;Ryu, Yea Seong;Han, Xia;Oh, Min Chang;Jung, Uhee;Kim, In Gyu;Hyun, Jin Won
    • Biomolecules & Therapeutics
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    • v.24 no.3
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    • pp.312-319
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    • 2016
  • Human skin cells undergo pathophysiological processes via generation of reactive oxygen species (ROS) upon excessive exposure to ultraviolet B (UVB) radiation. This study investigated the ability of hesperidin ($C_{28}H_{34}O_{15}$) to prevent apoptosis due to oxidative stress generated through UVB-induced ROS. Hesperidin significantly scavenged ROS generated by UVB radiation, attenuated the oxidation of cellular macromolecules, established mitochondrial membrane polarization, and prevented the release of cytochrome c into the cytosol. Hesperidin downregulated expression of caspase-9, caspase-3, and Bcl-2-associated X protein, and upregulated expression of B-cell lymphoma 2. Hesperidin absorbed wavelengths of light within the UVB range. In summary, hesperidin shielded human keratinocytes from UVB radiation-induced damage and apoptosis via its antioxidant and UVB absorption properties.