• Title/Summary/Keyword: Cytochrome C

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Comparative Study on the Inhibition Effect on Apoptosis in Neuro2A Cell on the Region of Zizania Latifolia(Radix, Rhizoma, Herba) (고장초의 부위별(뿌리, 줄기, 전초) Neuro2A 신경세포고사에 대한 억제 효과 비교 연구)

  • Cha, Yun-Yeop
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.4
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    • pp.936-941
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    • 2006
  • To prevent human body injury from oxidative stress, antioxidants are very important and many research about antioxidants are generally being conducted. Hydrogen peroxide$(H_20_2)$ that is one of vitality oxygen species has been seen that cause various diseases, DNA damage and gene change. We have already known that the inhibition effect of Zizania latifolia Radix, Rhizoma on apoptosis induced by $H_2O_2$ in Neuro2A cell. And the purpose of this study was that we made a comparative study on the inhibition effect of apoptosis in Neuro2A cell on the region of Zizania latifolia(Radix, Rhizoma, Herba). Neuro2A cells were cultivated in RPMI(GibcoBRL) with 5% FBS and treated with $H_2O_2$ and Zizania latifolia(Radix, Rhizoma, Herba). Separately we measured the cell viability and analyzed DNA fragmentation. Activity of PARP, Cytochrome C, caspase-9, caspase-3, p53, p21, Bax and Bcl-2 in the cell was examined by using western blot. The results obtained were as Follows: The cell viability in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment (60ug/m1<) decreased significantly compared with that of none treatment(p<0.001). Zizania latifolia Radix increased cell viability was most effective of three regions. But we had no significant difference among three regions. All of Zizania latifolia (Radix, Rhizoma, Herba) increased cell viability about twice as much as that being injury by $H_2O_2$,(Zizania Latifolia (Radix, nhizoma, Herba) 20ug/m1, $H_2O_2$ 200uM, p<0.001). DNA fragmentation developed by $H_2O_2$, but was not developed in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment. PARP, Cytochrome C, caspase-9 and caspase-3 activated all by $H_2O_2$ but were not activated in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment. P53, P2l and Bax activated by $H_2O_2$, and Bcl-2 got into inactivation. But the opposite results appeared in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment. In conclusion, these results suggest that all of Zizania latifolia (Radix, Rhizoma, Herba) inhibit the development of DNA fragmentation and apoptosis by $H_2O_2$and the antioridant action of all of Zizania latifolia (Radix, Rhizoma, Herba) is effective.

Bosentan and Rifampin Interactions Modulate Influx Transporter and Cytochrome P450 Expression and Activities in Primary Human Hepatocytes

  • Han, Kyoung-Moon;Ahn, Sun-Young;Seo, Hyewon;Yun, Jaesuk;Cha, Hye Jin;Shin, Ji-Soon;Kim, Young-Hoon;Kim, Hyungsoo;Park, Hye-kyung;Lee, Yong-Moon
    • Biomolecules & Therapeutics
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    • v.25 no.3
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    • pp.288-295
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    • 2017
  • The incidence of polypharmacy-which can result in drug-drug interactions-has increased in recent years. Drug-metabolizing enzymes and drug transporters are important polypharmacy modulators. In this study, the effects of bosentan and rifampin on the expression and activities of organic anion-transporting peptide (OATP) and cytochrome P450 (CYP450) 2C9 and CYP3A4 were investigated in vitro. HEK293 cells and primary human hepatocytes overexpressing the target genes were treated with bosentan and various concentrations of rifampin, which decreased the uptake activities of OATP transporters in a dose-dependent manner. In primary human hepatocytes, CYP2C9 and CYP3A4 gene expression and activities decreased upon treatment with $20{\mu}M$ $bosentan+200{\mu}M$ rifampin. Rifampin also reduced gene expression of OATP1B1, OATP1B3, and OATP2B1 transporter, and inhibited bosentan influx in human hepatocytes at increasing concentrations. These results confirm rifampin- and bosentan-induced interactions between OATP transporters and CYP450.

The Effects of Fucoidan on the Activation of Macrophage and Anticancer in Gastric Cancer Cell (Fucoidan의 면역세포 활성 및 위암 세포주에서의 항암효과)

  • An, In-Jung;Cho, Sung-Dae;Kwon, Jung-Ki;Kim, Hye-Ri;Yu, Hyun-Ju;Jung, Ji-Youn
    • Journal of Food Hygiene and Safety
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    • v.27 no.4
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    • pp.406-414
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    • 2012
  • This study was designed to investigate the effect of fucoidan on the activation of macrophage and on induction of apoptosis in AGS cell. To measure the activity of macrophages, NO and TNF-${\alpha}$ assays were performed in Raw 264.7 cell. Treatment with fucoidan significantly increased production of NO and TNF-${\alpha}$, indicating activation of macrophages. The result of MTT assay shows that cell viability was significantly decreased in a dose and time-dependent manner. Fucoidan increased to enhance mitochondrial membrane permeability, as well as the cytochrome c release from the mitochondria. Fucoidan decreased Bcl-2 and XIAP expression, whereas the expression of Bax was increased in a time-dependent manner compared to the control. In addition, the active forms of caspase-9 were increased, and the inactivation of Akt was decreased in a time-dependent manner. Caspase inhibitor, z-VAD-FMK, canceled the apoptosis of fucoidan, expression of Bax and caspase-9 were decrease. These results indicate that fucoidan induces activation of macrophage and apoptosis through activation of caspase on AGS cell.

Monitoring of Commercial Cephalopod Products Sold on the South Korea Market using DNA Barcode Information (DNA 바코드를 이용한 국내 유통 두족류 제품의 원재료 모니터링 연구)

  • Yu, Yeon-Cheol;Hong, Yewon;Kim, Jung Ju;Kim, Hyung Soo;Kang, Tae Sun
    • Journal of Food Hygiene and Safety
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    • v.34 no.5
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    • pp.502-507
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    • 2019
  • Cephalopods are one of the most important fishery resources in the world because of their desirable taste and nutritional value. In south Korea, one of the countries in which a large amount of seafood is consumed, cephalopods (e.g., octopus, squid, and cuttlefish) have an annual consumption rate of over 400,000 metric tons. In this study, octopus and squid products (n=28) sold on the market were monitored by analyzing sequences of DNA barcode markers (cytochrome c oxidase subunit I and 16S ribosomal RNA genes). For species identification, the NCBI BLAST database was screened with the sequences and analyzed as a query. In this BLAST search, twelve squid products showed 99-100% sequence identity to Dosidicus gigas (n=3) and Todarodes pacificus (n=9). In the case of the other 16 products that were declared using octopus as raw materials on the labels, six products were identified as Cistopus taiwanicus (n=1), Amphioctopus marginatus (n=1), Scaeurgus unicirrhus (n=1), and Dosidicus gigas (n=3). Monitoring results indicated that a significant percentage (37.5%) of mislabeling was present in octopus products sold on the South Korean market.

Increased Apoptotic Efficacy of Decitabine in Combination with an NF-kappaB Inhibitor in Human Gastric Cancer AGS Cells (핵산합성 억제제인 decitabine과 NF-κB 활성 저해제인 PDTC의 병용 처리에 의한 인체 위암세포사멸 효과 증진)

  • Choe, Won Kyung;Choi, Yung Hyun
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1268-1276
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    • 2018
  • The cytidine analog decitabine (DEC) acts as a nucleic acid synthesis inhibitor, whereas ammonium pyrrolidine dithiocarbamate (PDTC) is an inhibitor of nuclear factor-${\kappa}B$. The aim of this study was to investigate the possible synergistic inhibitory effect of these two inhibitors on proliferation of human gastric cancer AGS cells. The inhibitory effect of PDTC on AGS cell proliferation was significantly increased by DEC in a concentration-dependent manner, and this inhibition was associated with cell cycle arrest at the G2/M phase and the induction of apoptosis. This induction of apoptosis by the co-treatment with PDTC and DEC was related to the induction of DNA damage, as assessed by H2AX phosphorylation. Further studies demonstrated that co-treatment with PDTC and DEC induced the disruption of mitochondrial membrane potential, increased the generation of intracellular reactive oxygen species (ROS) and the expression of pro-apoptotic Bax, and down-regulated the expression of anti-apoptotic Bcl-2, ultimately resulting in the release of cytochrome c from the mitochondria into the cytoplasm. Co-treatment with PDTC and DEC also activated caspase-8 and caspase-9, which are representative caspases of the extrinsic and intrinsic apoptosis pathways. Co-treatment also activated caspase-3, which was accompanied by proteolytic degradation of poly (ADP-ribose) polymerase. Taken together, these data clearly indicated that co-treatment with PDTC and DEC suppressed the proliferation of AGS cells by increasing DNA damage and activating the ROS-mediated extrinsic and intrinsic apoptosis pathways.

Development of TaqMan Quantitative PCR Assays for Duplex Detection of Dirofilaria immitis COI and Dog GAPDH from Infected Dog Blood (심장사상충에 감염된 개 혈액에서 Dirofilaria immitis의 COI와 개의 GAPDH를 이중 검출하기 위한 정량적 TaqMan PCR 분석법의 개발)

  • Oh, In Young;Kim, Kyung Tae;Gwon, Sun-Yeong;Sung, Ho Joong
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.1
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    • pp.64-70
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    • 2019
  • Dirofilaria immitis (D. immitis) is a filarial nematode that causes cardiopulmonary dirofilariasis in dogs. In the late stages of infection, infected dogs show one or more symptoms and advanced heart disorder with perivascular inflammation. To detect D. immitis specifically and efficiently in the early stages of infection, a duplex TaqMan qPCR assay was developed based on previous studies using primers and probes specialized to detect D. immitis cytochrome c oxidase subunit I (COI) and dog glyceraldehyde-3-phosphate dehydrogenase (GAPDH). As positive controls, plasmid DNAs were constructed from D. immitis COI or dog GAPDH and a TA-cloning vector. Simplex and duplex TaqMan qPCR assays were performed using the specific primers, probes, and genomic or plasmid DNA. The duplex reaction developed could detect D. immitis COI and dog GAPDH in the same sample simultaneously after optimization of the primer concentrations. The limit of detection was 25 copies for the simplex and duplex assays, and both showed good linearity, high sensitivity, and excellent PCR efficiency. The duplex assays for pathogen detection reduce the costs, labor, and time compared to simplex reactions. Therefore, the duplex TaqMan qPCR assay developed herein will allow efficient D. immitis detection and quantification from a large number of samples simultaneously.

Morphological and Genetic Species Identification in the Chironomidae Larvae Found in Tap Water Purification Plants in Jeju (제주 정수장에서 출현한 깔따구과 유충의 형태 및 유전학적 분석)

  • Kwak, Ihn-Sil;Park, Jae-Won;Kim, Won-Seok;Park, Kiyun
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.240-246
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    • 2021
  • The Chironomidae is a benthic macroinvertebrate commonly found in freshwater ecosystems, along with Ephemeroptera and Trichoptera, which can be used for environmental health assessments. There are approximately 15,000 species of Chironomidae worldwide, but there are limited studies on species identification of domestic Chironomidae larvae. In the present study, we carried out species classification of the Chironomidae larvae that found in Jeju's tap water purification plants using morphological characteristics and genetic identification based on cytochrome c oxidase subunit I (COI) gene of the mitochondrial DNA. Body shape, mentum, antenna, mandible in the head capsule, and claws were observed in the larvae for morphological classification. Analysis of 17 larvae collected from faucets and fire hydrants of domestic tap water purification plants revealed the presence of two species, including 14 Orthocladius tamarutilus and 3 Paratrichocladius tammaater. These results will aid the use of the criteria information about species classification of the Chironomidae for water quality management in water purification plants and diversity monitoring of freshwater environments.

Food Fraud Monitoring of Commercial Sciaenidae Seafood Product Using DNA Barcode Information (DNA barcode를 이용한 민어과 수산가공품 진위판별 모니터링)

  • Park, Eun-Ji;Jo, Ah-Hyeon;Kang, Ju-Yeong;Lee, Han-Cheol;Park, Min-Ji;Yang, Ji-Young;Shin, Ji-Young;Kim, Gun-Do;Kim, Jong-Oh;Seo, Yong-Bae;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.35 no.6
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    • pp.574-580
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    • 2020
  • In this study we sought to determine the food fraud by discriminating species of commercial seafood product such as Larimichthys polyactis, Larimichthys crocea, Pennahia argentatus, and Miichthys miiuy, which are difficult to morphologically discriminate. After amplifying the mitochondrial cytochrome c oxidase subunit I gene of the reference fish, the DNA sequences of the amplified PCR products were analyzed. As a result, a 655 bp sequence for species identification was selected for use as DNA barcodes. To confirm the DNA data and primer set, the DNA barcode sequence of each fish was compared to that in that in the NCBI. All of the DNA barcode data were matched with the gene sequence of each fish in the NCBI. A total of 32 processed seafood products (8 L. polyactis, 12 L. crocea, 3 Pennahia argentatus, and 9 Miichthys miiuy) were investigated. Homology of 97% or more in DNA sequences was judged as the same species. As a result of the monitoring, there were no discovered cases of forgery or alteration. However, the use of a raw material name having no matching standard name in the Korea Food Code may cause consumer confusion. Therefore, it is suggested that the standard name or scientific name be co-labeled with the raw material name on seafood products to prevent consumer confusion.

Food Fraud Monitoring of Raw Materials for Commercial Seafood Products Using DNA Barcode Information (DNA Barcode를 이용한 수산가공품 원재료 진위판별)

  • Park, Eun-Ji;Kang, Ju-Yeong;Lee, Han-Cheol;Park, Min-Ji;Yang, Ji-Young;Shin, Ji-Young;Kim, Gun-Do;Kim, Jong-Oh;Seo, Yong-Bae;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.36 no.4
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    • pp.331-341
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    • 2021
  • DNA barcode sequences of commercial seafood products, which are difficult to morphologically discriminate, were analyzed to determine cases of food fraud. The gene sequences were analyzed by amplifying the COX I (cytochrome C oxidase subunit I) gene region of mitochondrial DNA, which is mainly used for species identification. The DNA barcode sequences were compared with the gene sequence of each fish registered in the US National Center for Biotechnology. A total of 46 processed seafood products (12 Pagrus majo, 4 Oplegnathus fasciatus, 7 Dentex tumifrons, 2 Acanthopagrus schlegelii, 7 Oreochromis niloticus, 6 Branchiostegus japonicus, 8 Branchiostegus albus) were investigated. Having DNA sequence identity of more than 97% was judged as the same species. As a result of this study, no cases of forgery and alteration were detected. However, some disparities in the commercial names used in local markets and the standard names given in the Korea Food Code were found, which may cause confusion for consumers. It is therefore suggested that the standard name or scientific name be displayed on seafood product labels.

Anti-cancer effects of kelp extract in mouse melanoma B16-F0 cell line through apoptosis (마우스 흑색종 세포주 B16-F0에서 다시마 추출물의 세포사멸을 통한 항암 효과)

  • Lee, Seong-Uk;Kim, Yoon Hee
    • Korean Journal of Food Science and Technology
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    • v.54 no.2
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    • pp.134-140
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    • 2022
  • Kelp belongs to the brown algae family and has been reported to exert anti-cancer effects on some cancer types, however studies have not been reported on the anti-cancer effects of kelp extracts on melanoma. In this study, the anti-cancer effects of kelp extract in B16-F0 cells were investigated, and the underlying molecular mechanisms were assessed. Kelp extract was found to inhibit the proliferation of B16-F0 cells, induce cytotoxicity, inhibit cell colony formation, and induce DNA fragmentation and apoptosis. The molecular mechanism was found to involve kelp extract increasing the expression of cytochrome-c and activated caspase-9 in the intrinsic apoptotic pathway. In addition, kelp extract upregulated the expression of Fas-associated protein with death domain and activated caspase-8 in the extrinsic apoptosis pathway. Activation of caspase-9 and caspase-8 by kelp extract induced activation of caspase-3 and cleaved poly adenosine diphosphate-ribose polymerase, consequently inducing apoptosis. These data suggest that kelp extract represents a potential therapeutic agent for melanoma.