• Title/Summary/Keyword: mitochondrial cytochrome b

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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.

Curcumin Conjugates Induce Apoptosis Via a Mitochondrion Dependent Pathway in MCF-7 and MDA-MB-231 Cell Lines

  • Singh, Durg Vijay;Agarwal, Shikha;Singh, Preeti;Godbole, Madan Madhav;Misra, Krishna
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.5797-5804
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    • 2013
  • In order to enhance the bioavailability of curcumin its conjugates with piperic acid and glycine were synthesized by esterifying the 4 and 4' phenolic hydroxyls, the sites of metabolic conjugation. Antiproliferative and apoptotic efficacy of synthesized conjugates was investigated in MCF-7 and MDA-MB-231 cell lines. $IC_{50}$ values of di-O-glycinoyl (CDG) and di-O-piperoyl (CDP) esters of curcumin were found to be comparable with that of curcumin. Both conjugates induced chromatin condensation fragmentation and apoptotic body formation. CDP exposure to MCF-7 cells induced apoptosis initiating loss of mitochondrial membrane potential (${\Delta}{\Psi}m$) followed by inhibition of translocation of transcription factor NF-${\kappa}B$ and release of Cytochrome-C. Reactive oxygen species (ROS) production was evaluated by fluorescent activated cell sorter. Change in ratio of Bcl2/Bclxl was observed, suggesting permeablization of mitochondrial membrane leading to the release of AIF, Smac and other apoptogenic molecules. DNA fragmentation as a hallmark for apoptosis was monitored by TUNEL as well as agrose gel electrophoresis. Thus, it was proven that conjugation does not affect the therapeutic potential of parent molecule in vitro, while these could work in vivo as prodrugs with enhanced pharmacokinetic profile. Pharmacokinetics of these molecules under in vivo conditions is a further scope of this study.

CM1 Ligation Induces Apoptosis via Fas-FasL Interaction in Ramos Cells, but via Down-regulation of Bcl-2 and Subsequent Decrease of Mitochondrial Membrane Potential in Raji Cells

  • Lee, Young-Sun;Kim, Yeong-Seok;Kim, Dae-Jin;Hur, Dae-Young;Kang, Jae-Seung;Kim, Young-In;Hahm, Eun-Sil;Cho, Dae-Ho;Hwang, Young-Il;Lee, Wang-Jae
    • IMMUNE NETWORK
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    • v.6 no.2
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    • pp.59-66
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    • 2006
  • Background: CM1 (Centrocyte/-blast Marker I) defined by a mAb developed against concanavalin-A activated PBMC, is expressed specifically on a subpopulation of centroblasts and centrocytes of human germinal center (GC) B cells. Burkitt lymphoma (BL) is a tumor consisting of tumor cells with the characteristics of GC B cell. Previously we reported that CM1 ligation with anti-CM1 mAb induced apoptosis in Ramos $(IgM^{high})$ and Raji $(IgM^{low})$ cells. Methods & Results: In the present study, we observed that CM1 ligation with anti-CM1 mAb induced Fas ligand and Fas expression in Ramos cells, but not in Raji cells. Furthermore, anti-Fas blocking antibody, ZB4, blocked CM1-mediated apoptosis effectively in Ramos cells, but not in Raji cells. Increased mitochondrial membrane permeabilization, which was measured by $DiOC_6$, was observed only in Raji cells. In contrast to no significant change of Bax known as pro-apoptotic protein, anti-apoptotic protein Bcl-2 was significantly decreased in Raji cells. In addition, we observed that CM1 ligation increased release of mitochondrial cytochrome c and upregulated caspase-9 activity in Raji cells. Conclusion: These results suggest that apoptosis induced by CM1-ligation is mediated by Fas-Fas ligand interaction in Ramos cells, whereas apoptosis is mediated by down-regulation of Bcl-2 and subsequent decrease of mitochondrial membrane potential in Raji cells.

Species and Sex Identification of the Korean Goral (Nemorhaedus caudatus) by Molecular Analysis of Non-invasive Samples

  • Kim, Baek Jun;Lee, Yun-Sun;An, Jung-hwa;Park, Han-Chan;Okumura, Hideo;Lee, Hang;Min, Mi-Sook
    • Molecules and Cells
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    • v.26 no.3
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    • pp.314-318
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    • 2008
  • Korean long-tailed goral (Nemorhaedus caudatus) is one of the most endangered species in South Korea. However, detailed species distribution and sex ratio data on the elusive goral are still lacking due to difficulty of identification of the species and sex in the field. The primary aim of this study was to develop an economical PCR-RFLP method to identify species using invasive or non-invasive samples from five Korean ungulates: goral (N. caudatus), roe deer (Capreolus pygargus), feral goat (Capra hircus), water deer (Hydropotes inermis) and musk deer (Moschus moschiferus). The secondary aim was to find more efficient molecular sexing techniques that may be applied to invasive or non-invasive samples of ungulate species. We successfully utilized PCR-RFLP of partial mitochondrial cytochrome b gene (376 bp) for species identification, and sex-specific amplification of ZFX/Y and AMELX/Y genes for sexing. Three species (goral, goat and water deer) showed distinctive band patterns by using three restriction enzymes (Xbal, Stul or Sspl). Three different sexing primer sets (LGL331/335 for ZFX/Y gene; SE47/48 or SE47/53 for AMELX/Y gene) produced sex-specific band patterns in goral, goat and roe deer. Our results suggest that the molecular analyses of non-invasive samples might provide us with potential tools for the further genetic and ecological study of Korean goral and related species.

Genetic Species Identification by Sequencing Analysis of Nuclear and Mitochondrial Genes for Albino Misgurnus Species from Korea (우리나라 미꾸리속(genus Misgurnus) 알비노 개체의 미토콘드리아 및 핵 유전자 염기서열 분석에 의한 유전적 동정)

  • Song, Ha-Youn;Moon, Shin-Joo;Kim, Keun-Sik;Bang, In-Chul
    • Korean Journal of Ichthyology
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    • v.29 no.2
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    • pp.139-145
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    • 2017
  • The spontaneous color mutant, albino individuals of genus Misgurnus, are rarely discovered in Korea and there are difficult to identify morphological species due to lack melanin pigmentation. In this study, we developed a genetic identification method for the species of albino Misgurnus individuals based on phylogenetic analysis by using recombination activating gene 1 (rag1) and cytochrome b (cytb) region of mitochondrial DNA. As a result of molecular phylogenetic analysis, three clades were identified as Misgurnus mizolepis, M. anguillicaudatus and M. mohoity. The homology of the cytb sequences of M. mohoity was best match to that of M. mohoity sequences in GenBank database. As a result of species identification of 25 albino Misgurnus individuals based on the phylogenetic tree, the red-eye type was identified as 16 M. anguillicaudatus and one M. mizolepis. The remaining three individuals were identified as one M. mizolepis ♀${\times}$M. anguillicaudatus ♂, and two M. mohoity ♀${\times}$M. anguillicaudatus ♂, respectively. In addition, the five black-eye type individuals were identified as one M. anguillicaudatus, three M. mizolepis and one M. mohoity. Therefore, this genetic identification method will be an useful techniques for species or hybrid identification in genus Misgurnus.

Molecular Identification and Morphological Characteristics of the Longjaw Bonefish, Albula argentea Collected in Korea (한국산 여울멸의 분자동정과 형태적 특징)

  • Kim, Maeng Jin;Kim, Byeong Yeob;Song, Choon Bok
    • Korean Journal of Ichthyology
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    • v.31 no.4
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    • pp.249-254
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    • 2019
  • One specimen of Albula argentea (296.3 mm in standard length), belonging to the family Albulidae, was collected in a set net in the coastal waters of Jejudo Island, Korea. As the adult A. argentea has not been caught from Korean waters, we described its morphological characters based on the collected specimen and compared them with those of the previous reports on A. argentea and A. koreaus. As a result, the present specimen was characterized by having 68~73 pored lateral line scales (vs. 77~78 for A. koreana), 72~73 vertebrae pored lateral line scales (vs. 77~80) and none streak on the check (vs. yellowish streak). Molecular analysis using the mitochondrial cytochrome b DNA sequences was also conducted to confirm the correctness of species identification and taxonomic status of the sample.

Role of HIV Vpr as a Regulator of Apoptosis and an Effector on Bystander Cells

  • Moon, Ho Suck;Yang, Joo-Sung
    • Molecules and Cells
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    • v.21 no.1
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    • pp.7-20
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    • 2006
  • The major event in human immunodeficiency virus type 1 (HIV-1) infection is the death of many cells related to host immune response. The demise of these cells is normally explained by cell suicide mechanism, apoptosis. Interestingly, the decrease in the number of immune cells, such as non-CD4+ cells as well as CD4+ T cells, in HIV infection usually occurs in uninfected bystander cells, not in directly infected cells. It has, therefore, been suggested that several soluble factors, including viral protein R (Vpr), are released from the infected cells and induce the death of bystander cells. Some studies show that Vpr interacts directly with adenine nucleotide translocator (ANT) to induce mitochondrial membrane permeabilization (MMP). The MMP results in release of some apoptogenic factors such as cytochrome-c (cyt-c) and apoptosis-inducing factor (AIF). Vpr also has indirect effect on mitochondria through enhancing the level of caspase-9 transcription and suppressing nuclear factor-kappa B (NF-${\kappa}B$). The involvement of p53 in Vpr-induced apoptosis remains to be studied. On the other hand, low level of Vpr expression has anti-apoptotic effect, whereas it's high level of expression induces apoptosis. Extracellular Vpr also exhibits cytotoxicity to uninfected bystander cells through apoptotic or necrotic mechanism. The facts that Vpr has cytotoxic effect on both infected cells and bystander cells, and that it exhibits both proand anti-apoptotic activity may explain its role in viral survival and disease progression.

Origin of the Korean Mandarin Fish, Siniperca scherzeri and Its Molecular Phylogenetic Relationships to Other Siniperca Fishes (한국산 쏘가리의 기원과 분자계통진화적 위치)

  • Kim, Maeng-Jin;Song, Choon-Bok
    • Korean Journal of Ichthyology
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    • v.23 no.2
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    • pp.95-105
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    • 2011
  • To explain the origin of the Korean mandarin fish (Siniperca scherzeri), phylogenetic relationships and DNA polymorphism among Siniperca fishes have been investigated based on mitochondrial cytochrome b DNA sequences. As a result, S. roulei were firstly differentiated early in the evolution of Siniperca fishes and the other six species (S. schezeri, S. undulata, S. fortis, S. obscura, S. knerii and S. chuatsi) were evolved slightly later. However, the order of species differentiation among six species was not clear because the nodes of their phylogeny were poorly resolved. The constructed molecular phylogeny revealed three genetically distinct groups of local populations of S. scherzeri. The first group (group 1) is the local populations of Korean peninsula and northern China including Lioaning and Henan. The second one (group 2) is the local populations of Anhui, Fujian and Guangxi. The third one (group 3) is the local population of Zhejiang. The number of nucleotide differences in base pairs were 31~43 between group 1 and 2; 37~44 between group 2 and 3; 27~29 between group 1 and 3; and 1~5 within group 1. Thus, the Korean mandarin fish was likely to be originated from the northern China local population which was isolated from the middle or southern China local populations during the Cenozoic Pliocene. Low level of sequence divergence between Korean mandarin fish populations and northern China population indicated a recent expansion of distribution ranges from northern China to Korean peninsula.

The Sanguinarine Apoptosis Induction of Hep3B Human Hepatocellular Carcinoma Cells is Dependent on the Activation of Caspase (Sanguinarine에 의한 Hep3B 인체 간암세포의 apoptosis 유도에 관한 연구)

  • Han, Min Ho;Choi, Sung Hyun;Hong, Su Hyun;Park, Dong Il;Choi, ung Hyun
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1340-1348
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    • 2017
  • Sanguinarine is a benzophenanthridine alkaloid derived from the roots of Sanguinaria canadensis L., which is used for the purpose of treating various diseases. Although studies of anticancer activities have been performed using various cancer cell lines, the phenomenon of inducing apoptosis in cancer cells by using sanguinarine requires more research. Therefore, this study investigated the anti-cancer activities and related mechanisms of sanguinarine used with Hep3B human hepatocellular carcinoma cells in terms of the regulation of apoptosis. Sanguinarine inhibited the proliferation of Hep3B cells in a concentration-dependent manner, which was associated with the induction of apoptosis. Sanguinarine also increased the activity of caspase-3, which is a typical effector caspase, and the activities of caspase-8 and caspase-9, which are key when initiating extrinsic and intrinsic apoptosis pathways, respectively. In addition, sanguinarine increased the expression of death receptor-related genes and pro-apoptotic BAX, which belongs to the Bcl-2 family, while suppressing the expression of anti-apoptotic Bcl-2. Sanguinarine promoted the truncation of Bid and enhanced the release of cytochrome c from the mitochondria to the cytoplasm due to a loss of mitochondrial membrane potential. Furthermore, the reduction of a survival rate that was induced by sanguinarine and the induction of apoptosis disappeared with the inhibition of artificial caspase activity. Therefore, the results of the study indicated that sanguinarine-induced apoptosis in Hep3B cells involves both extrinsic and intrinsic pathways; such apoptosis is a caspase-dependent phenomenon.

Discordance between Morphological and Molecular Variations of the Genus Macroramphosus (Macroramphosidae) from Korea (한국산 대주둥치속(대주둥치과) 어류의 형태와 분자 변이의 불일치)

  • Sohn, Min-Soo;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.32 no.4
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    • pp.199-209
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    • 2020
  • In order to clarify the taxonomic status of the Korean Macroramphosus species, which were previously confused, we investigated morphological and molecular variations of Macroramphosus (18 individuals) from Korea, and Macroramphosus (35 individuals) from Japan and Taiwan, and compared with those of M. scolopax from type locality (Mediterranean Sea). Although the Korean and Japanese specimens of Macroramphosus were clearly divided into two types in the first dorsal spine length (22.8~32.1% in A-type vs. 15.6~21.4% in B-type), distance between the first dorsal fin and second dorsal fin (6.4~9.7% vs. 8.6~13.3%), and body depth (20.0~28.0% vs. 17.3~22.6%), no genetic differences among all individuals of longspine snipefish between them were found at the specific level [d=0.0~3.3% in control region (CR); 0.0~1.3% in cytochrome b (cytb); 0.0~0.5% in cytochrome c oxidase subunit I (COI)]. Whereas, they were well distinguished in genetics (9.9~11.5% in CR; 3.8~4.6% in cytb; 1.2~3.6% in COI) from those of M. scolopax in Mediterranean Sea. It needs the scientific name of the longspine snipefish (M. scolopax) in Korea be changed as M. japonicus (and/or M. sagifue). However, our results could not find evidence of consistency between morphological and mitochondrial DNA variations which suggests that their differentiation event may occur fairly recently. Further studies using more sensitive markers such as microsatellite are needed to clarify the degree of gene flow between the two types.