• Title/Summary/Keyword: Cytochrome b gene

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Development of Duplex PCR Method for Simultaneous Detection of Rabbit (Oryctolagus cuniculus) and Cat (Felis catus) Meats (Duplex PCR을 이용한 토끼(Oryctolagus cuniculus)와 고양이(Felis catus) 육류의 동시 검출법 개발)

  • Hong, Yeun;Kim, Mi-Ju;Yang, Seung-Min;Yoo, In-Suk;Kim, Hae-Yeong
    • Journal of Applied Biological Chemistry
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    • v.58 no.4
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    • pp.383-387
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    • 2015
  • A duplex polymerase chain reaction (PCR) detection method was developed to authenticate the use of cat and rabbit in food and to prevent unlawful distribution of illegally butchered meat in both domestic and imported food market. Species-specific primers were designed targeting mitochondrial cytochrome b gene. The sizes of PCR products were 191 bp for cat and 101 bp for rabbit, which were relatively small for better application of the detection method on processed foods. Specificities of primers were verified using 21 animal species including cat and rabbit. Limit of detection was examined by serial dilution of the sample DNA and confirmed as 0.005 ng for rabbit and 0.0005 ng for cat using Bioanalyzer. The developed duplex PCR method showed specificity and sensitivity in the identification of two target species.

HY253, a Novel Decahydrofluorene Analog, Induces Apoptosis via Intrinsic Pathway and Cell Cycle Arrest in Liver Cancer HepG2 Cells

  • Choi, Ko-woon;Suh, Hyewon;Jang, Seunghun;Kim, Dongsik;Lee, Chul-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.413-417
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    • 2015
  • Recently, we isolated HY253, a novel decahydrofluorene analog with a molecular structure of 7,8a-divinyl-2,4a,4b,5,6,7,8,8a,9,9a-decahydro-1H-fluorene-2,4a,4b,9a-tetraol from the roots of Aralia continentalis, which is known as Dokwhal (獨活), a traditional medicinal herb. Moreover, we previously reported its cytotoxic activity on cancer cell proliferation in human lung cancer A549 and cervical cancer HeLa cells. The current study aimed to evaluate its detailed molecular mechanisms in cell cycle arrest and apoptotic induction in human hepatocellular carcinoma HepG2 cells. Flow cytometric analysis of HepG2 cells treated with $60{\mu}M$ HY253 revealed appreciable cell cycle arrest at the G1 phase via inhibition of Rb phosphorylation and down-regulation of cyclin D1. Furthermore, using western blots, we found that up-regulation of cyclin-dependent kinase inhibitors, such as p21CIP1 and p27KIP1, was associated with this G1 phase arrest. Moreover, TUNEL assay and immunoblottings revealed apoptotic induction in HepG2 cells treated with $60{\mu}M$ HY253 for 24 h, which is associated with cytochrome c release from mitochondria, via down-regulation of anti-apoptotic Bcl-2 protein, which in turn resulted in activation of caspase-9 and -3, and proteolytic cleavage of poly(ADP-ribose) polymerase (PARP). Accordingly, we suggest that HY253 may be a potent chemotherapeutic hit compound for treating human liver cancer cells via up-regulation and activation of the p53 gene.

Evaluation of the preservation state of human skeletal remains using real-time PCR (출토 인골 DNA의 real-time PCR 정량에 의한 보존상태 평가 연구 - 부여 오수리 출토 인골을 중심으로 -)

  • Kwon, Eun-Sil;Cho, Eun-Min;Kim, Sue-Hoon;Kang, Soyeong
    • 보존과학연구
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    • s.32
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    • pp.171-183
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    • 2011
  • In this study molecular genetic analysis was carried out on 4 human skeletal remains from Osuri, Buyeo. We showed that real-time PCR is the method of the choice to assess the initial number of genuine ancient DNA molecules. Human mitochondrial DNA quantification was accomplished by the real-time PCR for the cytochrome b gene of the mitochondria. Histological results proved to be a good potentiality for biochemical analysis using biomolecule. The level of specimen's preservation state was proved that level of quantitative result was BO-04, BO-01, BO-03, BO-02. Continually, we showed that biochemical and biomolecule results for the level of preservation state were similar. This study will be useful to important material for predicting biochemistry and biology analysis of the ancient bone.

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DNA barcoding for fish species identification and diversity assessment in the Mae Tam reservoir, Thailand

  • Dutrudi Panprommin;Kanyanat Soontornprasit;Siriluck Tuncharoen;Santiwat Pithakpol;Korntip Kannika;Konlawad Wongta
    • Fisheries and Aquatic Sciences
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    • v.26 no.9
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    • pp.548-557
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    • 2023
  • The purposes of this research were to identify fish species using DNA barcodes or partial sequences of cytochrome b (Cytb) and to assess the diversity of fish in the Mae Tam reservoir, Phayao province, Thailand. Fish samples were collected 3 times, during the winter, summer, and rainy seasons, from 2 sampling sites using gillnets with 3 mesh sizes (30, 50, and 70 mm). A total of 34 representative samples were classified into 12 species, 7 families and 6 orders by morphological- and DNA barcoding-based identifications. However, one cichlid species, Cichlasoma trimaculatum, could only be identified using DNA barcoding. Family Cyprinidae had the greatest diversity, 50.00%. The diversity, richness and evenness indices ranged from 0.43-0.65, 0.64-1.46, and 0.27-0.40, respectively, indicating that fish diversity at both sampling sites was relatively low. A comparison of the catch per unit effort (CPUE) with 3 different mesh sizes found that the 50 mm mesh size was the best (474.80 ± 171.56 g/100 m2/night), followed by the 70 mm (417.41 ± 176.24 g/100 m2/night) and 30 mm mesh sizes (327.88 ± 115.60 g/100 m2/night). These results indicate that DNA barcoding is a powerful tool for species identification. Our data can be used for planning the sustainable management of fisheries resources in the Mae Tam reservoir.

Molecular identification of fruit bats, natural host of Nipah virus in Bangladesh, based on DNA barcode

  • Md. Maharub Hossain Fahim;Walid Hassan;Afia Afsin;Md. Mahfuzur Rahman;Md. Tanvir Rahman;Sang Jin Lim;Yeonsu Oh;Yung Chul Park;Hossain Md. Faruquee;Md. Mafizur Rahman
    • Journal of Ecology and Environment
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    • v.48 no.2
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    • pp.163-172
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    • 2024
  • Background: Fruit bats are natural carriers of Nipah virus (NiV). The primary objective of this study is to identify potential reservoir species in a selected geographic regions. It is necessary to determine an accurate species identification of the associated reservoir bat species distributed in a specific region. Results: In this study, we collected 20 different bat specimens from the NiV-prone area of the Kushtia district. Among these, 14 were tissue samples (BT-1-14) and six were fecal samples (BF-1-6). We used the mitochondrial gene cytochrome b, one of the most abundant and frequently used genetic markers, for polymerase chain reaction amplification and sequencing. Out of the 20 samples, 12 tissue samples and 2 fecal samples were successfully amplified and sequenced. However, two tissue samples and four fecal samples yielded chimeric sequences, rendering them unsuitable for annotation. The sequences of the successfully amplified samples were compared to those deposited in the National Center for Biotechnology Information database using basic local alignment search tool to identify the bat specimen collected. The study identified six different bat species using both morphological and genetic data, which may carriers of the NiV. Conclusions: Our results suggest that additional research should be conducted to gather more information on fruit bats from different localities across the country. The study contributes to the establishment of appropriate measures for NiV carrying disease control and management.

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.

Microarray analysis of hypoxia-induced changes in gene expression in BV-2 microglial cells (BV-2 microglia 세포주에서 저산소증의 유전자 발현에 대한 마이크로어레이 분석)

  • Kim, Bum-Shik;Seo, Jung-chul
    • Journal of Acupuncture Research
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    • v.20 no.4
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    • pp.85-92
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    • 2003
  • 목적 : 허혈시 발생되는 저산소중 상태에서는 세포독성을 유발한다고 알려져 있으나 정확한 기전은 아직 규명되지 않았다. 본 연구에서는 뇌허혈로 인한 세포독성의 기전을 유전자 발현을 통하여 살펴보고자 하였다. 방법 : 본 실험에서는 BV-2 microglia 세포주에 12시간 동안의 저산소 상태에서의 유전자 발현을 분석하기 위하여 마이크로에레이를 시행하였다. 결과 : 저산소 상태에서는 정상에 비하여 cathepsin F, growth factor independent 1, calcitonin/calcitonin-related poly, leucine-rich repeat LGI family membrane, dublecortin, cyclohydrolase 1, Ia-associated invariant chain, carbohydrate kinase-like과 erythrocyte protein band 4.1-like 3 등의 유전자 발현이 3배 이상 증가하였다. 한편 neuronal guanine nucleotide exchange factor, Bcl-2-related ovarian killer protein, chemokine (C-X-C motif) ligand 5, RNA binding motif protein 3, interleukin 2 receptor, alpha chain, crystallin zeta, cytochrome P450 subfamily IV B, asparagine synthetase과 moesin 등의 유전자 발현은 0.2배 이하로 감소하였다. 결론 : 이상의 결과는 저산소중에 관여하는 유전자 및 저산소중과 관련된 뇌경색 등의 질환의 기전을 밝히는데 기초적 자료로 이용될 수 있을 것이다.

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Study on Species Identification Error Caused by Comparing Feces Appearance of Felis catus and Prionailurus bengalensis in the Ecosystems

  • Lee, Yoon Jeong;Kim, Jong Hyun;Kim, Eui Kyeong
    • Journal of Forest and Environmental Science
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    • v.37 no.4
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    • pp.331-337
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    • 2021
  • This study is designed according to the fact that the feces presumed to be from a Prionailurus bengalensis was found in Ulleungdo Island, where Prionailurus bengalensis is not known to inhabit, and that visual observation of the feces may cause errors in species identification. The feces observed in Ulleungdo Island on October 21, 2019 and August 29, 2020, in Gyeongju on December 4, 2020, and in Jecheon on December 7, 2020 was found intactly on grass, not buried in the ground. Although it was difficult to distinguish and identify the feces of Felis catus and Prionailurus bengalensis with visual observation, the feces collected from Ulleungdo Island was closely related to the Felis catus according to the genetic analysis whereas the ones collected from Gyeongju and Jecheon was identified from Prionailurus bengalensis. Therefore through the gene analysis, this study proved that visual observation of feces with similar appearance, specifically the feces found in Ulleungdo Island, Gyeongju, and Jecheon, may cause errors in species identification. It is judged to be necessary to analyze fields signs and genes for the species identification when using the feces of Felis catus and Prionailurus bengalensis.

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.

CYP1A1 (Ile462Val), CYP1B1 (Ala119Ser and Val432Leu), GSTM1 (null), and GSTT1 (null) Polymorphisms and Bladder Cancer Risk in a Turkish Population

  • Berber, Ufuk;Yilmaz, Ismail;Yilmaz, Omer;Haholu, Aptullah;Kucukodaci, Zafer;Ates, Ferhat;Demirel, Dilaver
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3925-3929
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    • 2013
  • We aimed to investigate bladder cancer risk with reference to polymorphic variants of cytochrome p450 (CYP) 1A1, CYP1B1, glutathione S-transferase (GST) M1, and GSTT1 genes in a case control study. Polymorphisms were examined in 114 bladder cancer patients and 114 age and sex-matched cancer-free subjects. Genotypes were determined using allele specific PCR for CYP1A1 and CYP1B1 genes, and by multiplex PCR and melting curve analysis for GSTM1 and GSTT1 genes. Our results revealed a statistically significant increased bladder cancer risk for GSTT1 null genotype carriers with an odds ratio of 3.06 (95% confidence interval=1.39-6.74, p=0.006). Differences of CYP1A1, CYP1B1 and GSTM1 genotype frequencies were not statistically significant between patients and controls. However, the specific combination of GSTM1 null, GSTT1 null, and CYP1B1 codon 119 risk allele carriers and specific combination of GSTM1 present, GSTT1 null, and CYP1B1 432 risk allele carriers exhibited increased cancer risk in the combined analysis. We did not observe any association between different genotype groups and prognostic tumor characteristics of bladder cancer. Our results indicate that inherited absence of GSTT1 gene may be associated with bladder cancer susceptibility, and specific combinations of GSTM1, GSTT1 and CYP1B1 gene polymorphisms may modify bladder cancer risk in the Turkish population, without any association being observed for CYP1A1 gene polymorphism and bladder cancer risk.