• Title/Summary/Keyword: mtDNa

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Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea

  • Yu, Hak-Sun;Choi, Kyung-Hee;Kim, Hyo-Kyung;Kong, Hyun-Hee;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.161-170
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    • 2001
  • We conducted both the small subunit ribosomal DNA (SSU rDNA) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mitochondrial (mt) DNA RFLP analyses for a genetic characterization of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea. Twenty-three strains of Acanthamoeba from the American Type Culture Collection and twelve clinical isolates from Korean patients were used as reference strains. Thirty-nine isolates from contact lens storage cases were classified into seven types (KA/LS1 , KA/LS2, KA/LS4, KA/LS5, KA/LS7 KA/LS18, KA/LS31). Four types (KA/LS1 , KA/LS2, KA/LS5, KA/LS18) including 33 isolates were regarded as A. castellanii complex by riboprints. KA/LS1 type was the most predominant (51.3%) in the present survey area, followed by KA/LS2 (20.9%), and KA/LSS (7.7%) types. Amoebae of KA/LS1 type had the same mtDNA RFLP and riboprint patterns as KA/E2 and KA/E12 strains, clinical isolates from Korean keratitis patients. Amoebae of KA/LS2 type had the identical mtDNA RFLP patterns with A. castellanii Ma strain, a corneal isolate from an American patient as amoebae of KA/LS5 type, with KA/E3 and KA/E8 strains from other Korean keratitis patients. Amoebae of KA/LS 18 type had identical patterns with JAC/E1, an ocular isolate from a Japanese patient. Three types , which remain unidentified at species level, were not corresponded with any clinical isolate in their mtDNA RFLP and riboprint patterns. Out of 39 isolates analyzed in this study, mtDNA RFLP and riboprint patterns of 33 isolates (84.6%) were identical to already known clinical isolates, and therefore, they may be regarded as potentially keratopathogenic. These results suggest that contact lens wearers in Seoul should pay more attention to hygienic maintenance of contact lens storage cases for the prevention of Acanthamoeba keratitis.

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The Number of Nucleotide Substitutions per Sites of Mitochondrial DNA in the Four Pleuronectid Species (미토콘드리아 DNA에 의한 붕넙치과 어류 4종간의 염기치환수)

  • PARK Jung-Youn;KIM Yoon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.5
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    • pp.649-658
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    • 1995
  • In order to estimate the level of genetic differences among the pleuronectid species, mitochondrial DNAs were isolated from four species: brown sole, Limanda herensteini; marbled sole, Limanda Yokohamae; stone flounder, Kareius bicoloratus; starry flounder, Platichthys stellatus, and the number of nucleotide substitutions was calculated by the restriction fragment length polymorphisms (RFIPs) generated by f4 sin base recognition restriction endonucleases. Total lengths of the mitochondrial DNA were measured as about 17.6 kbp in all species. Ten different composite genotypes were observed in brown sole, four different genotypes in marbled sole, and two different genotypes in starry flounder. However, only one genotype was observed in stone flounder. The calculated haplotypic diversity value of brown sole was higher than that of marbled sole. The average number of nucleotide substitutions per sites in four species was estimated to be 0.0045 in the intraspecies, 0.0344 in the interspecies, and 0.0457 in the genera, respectively. From these results, we could estimate that the genetic differences among interspecies were not influenced by nucleotide substitutions but genetical discontinuous.

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Systematic Studies on Korean Rodents: VI. Analysis of Morphometric Characters, Chromosomal Karyotypes and Mitochondrial DNA in Two Species of Genus Rattus (한국에 서식하고 있는 설치류의 계통분류학적 연구: 6. 집쥐속 2종의 형태학적 형질, 염색체 핵형 및 미토콘디리아 DNA의 분석)

  • 고홍선
    • Animal Systematics, Evolution and Diversity
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    • v.8 no.2
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    • pp.231-242
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    • 1992
  • Samples of two species of genus Ratts(black rat, Rattus rasttus Linnaeus; common rat, Rattus norvegicus Berkenhaut) in Korea were trapped and their 31 morphometric charcters were analyzed statistically in order to determine the range of geographic variation within each species and the interspecific differences. In addition, chromosomal G-bands and C-bands were compared and the fragment patterns of mtDNA resulted from the digestion with restriction enzymes were also analyzed. Samples of black rats from six localities in Korea were similar with one another in their morphometric characters: in head and body length, length of tail vertebrae, conventional karyotype and C-bands, they are comparable to Rattus rattus tanezumi in Japan. Specimens of common rats from seven localities in Korea were similar with one another in their morphometric characters: in conventional karyotype, they are comparable to Rattus norvegicus caraco in eastern Asia. Common rats differ from black rats in their morphometric characteris, chromosomal karyotypes and mtDNA. It is confirmed that correct species name of black rat in Korea is Rattus rattus tanezumi Tempminck: species name of common rat in Korea is Rattus norvegicus caraco Pallas: the common rat is a species, which is distinct from the black rat.

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Effects of Pogonatherum paniceum (Lamk) Hack extract on anti-mitochondrial DNA mediated inflammation by attenuating Tlr9 expression in LPS-induced macrophages

  • Rungthip Thongboontho;Kanoktip Petcharat;Narongsuk Munkong;Chakkraphong Khonthun;Atirada Boondech;Kanokkarn Phromnoi;Arthid Thim-uam
    • Nutrition Research and Practice
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    • v.17 no.5
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    • pp.827-843
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    • 2023
  • BACKGROUND/OBJECTIVES: Mitochondrial DNA leakage leads to inflammatory responses via endosome activation. This study aims to evaluate whether the perennial grass water extract (Pogonatherum panicum) ameliorate mitochondrial DNA (mtDNA) leakage. MATERIALS/METHODS: The major bioactive constituents of P. paniceum (PPW) were investigated by high-performance liquid chromatography, after which their antioxidant activities were assessed. In addition, RAW 264.7 macrophages were stimulated with lipopolysaccharide, resulting in mitochondrial damage. Quantitative polymerase chain reaction and enzyme-linked immunosorbent assay were used to examine the gene expression and cytokines. RESULTS: Our results showed that PPW extract-treated activated cells significantly decrease reactive oxygen species and nitric oxide levels by reducing the p2phox and iNOS expression and lowering cytokine-encoding genes, including IL-6, TNF-α, IL-1β, PG-E2 and IFN-γ relative to the lipopolysaccharide (LPS)-activated macrophages. Furthermore, we observed that LPS enhanced the mtDNA leaked into the cytoplasm, increasing the transcription of Tlr9 and signaling both MyD88/Irf7-dependent interferon and MyD88/NF-κb p65-dependent inflammatory cytokine mRNA expression but which was alleviated in the presence of PPW extract. CONCLUSIONS: Our data show that PPW extract has antioxidant and anti-inflammatory activities by facilitating mtDNA leakage and lowering the Tlr9 expression and signaling activation.

Zinc Status Assessment by Analysis of Mononuclear Cell Metallothionein mRNA Using Competitive-Reverse Transcriptase-Polymerase Chain Reaction

  • Lee, Soo-Lim;Yoon, Jin-Sook;Kwon, Chong-Suk;Beattie, John H.;Kwun, In-Sook
    • Preventive Nutrition and Food Science
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    • v.9 no.3
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    • pp.276-282
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    • 2004
  • Marginal Zn deficiency is prevalent through the world and yet human zinc status has not been properly assessed due to the lack of a reliable diagnostic indicator. One potential possibility for zinc status assessment using Zn-binding protein, metallothionein (MT)-mRNA, has been proposed. The purpose of the present study was aimed to show whether measurement of mononuclear cell (MNC) MT mRNA, using a competitive-reverse transcriptase-polymerase chain reaction (competitive-RT-PCR) assay, could indicate zinc status in human subjects. In this study, MNC MT-mRNA expression was measured using a competitive-RT-PCR to compare before and after 14 days of zinc supplementation (50 mg Zn/das zinc gluconate). RT-PCR oligonucleotide primers which were designed to amplify both a 278 bp segment of the human MT-2A cDNA and a 198 bp mutant competitor cDNA template from MNCs, were prepared. MT-2A mRNA was normalized by reference to the housekeeping gene, $\beta$-actin, mRNA for which was also measured by competitive-RT-PCR. There was considerable inter-individual variation in MT-mRNA concentration and yet, the mean MT-2A mRNA level increased 4.7-fold after Zn supplementation, as compared to before Zn supplementation. This MT-2A mRNA level was shown as the same pattern and, even more sensitive assay, compared to the conventional plasma and red blood cells (RBCs) Zn assessment in which plasma and RBCs zinc levels increased 2.3- and 1.2-fold, respectively (p<0.05). We suggest that MT competitive-RT-PCR can be a useful assessment tool for evaluating human zinc status.

mtDNA Diversity and Phylogenetic State of Korean Cattle Breed, Chikso

  • Kim, Jae-Hwan;Byun, Mi Jeong;Kim, Myung-Jick;Suh, Sang Won;Ko, Yeoung-Gyu;Lee, Chang Woo;Jung, Kyoung-Sub;Kim, Eun Sung;Yu, Dae Jung;Kim, Woo Hyun;Choi, Seong-Bok
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.2
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    • pp.163-170
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    • 2013
  • In order to analyze the genetic diversity and phylogenetic status of the Korean Chikso breed, we determined sequences of mtDNA cytochrome b (cyt b) gene and performed phylogenetic analysis using 239 individuals from 5 Chikso populations. Five non-synonymous mutations of a total of 15 polymorphic sites were identified among 239 cyt b coding sequences. Thirteen haplotypes were defined, and haplotype diversity was 0.4709 ranging from 0.2577 to 0.6114. Thirty-five haplotypes (C1-C35) were classified among 9 Asia and 3 European breeds. C2 was a major haplotype that contained 206 sequences (64.6%) from all breeds used. C3-C13 haplotypes were Chikso-specific haplotypes. C1 and C2 haplotypes contained 80.5% of cyt b sequences of Hanwoo, Yanbian, Zaosheng and JB breeds. In phylogenetic analyses, the Chikso breed was contained into B. taurus lineage and was genetically more closely related to two Chinese breeds than to Korean brown cattle, Hanwoo. These results suggest that Chikso and Hanwoo have a genetic difference based on the mtDNA cyt b gene as well as their coat color, sufficient for classification as a separate breed.

Expression of Vitellogenin Gene by Androgens in Rasinbow Trout, Oncorhynchus mykiss (웅성호르몬에 의한 무지개송어의 vitellogenin 유전자 발현)

  • 권혁추;윤종만;이종영
    • Journal of Aquaculture
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    • v.13 no.1
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    • pp.79-85
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    • 2000
  • The effects of estrogen and androgens on Vg gene expression were examined in primary hepatocyte culture and livers of the immature male trout. Specific primers of Vg cDNA were designed with already reported Vg gene nucleotide sequences. PCR product was sequenced and verified with Vg cDNA of rainbow trout. Total RNA was extracted from the cultured hepatocytes and livers of steroid-treated rainbow trout and then it was analyzed by reverse transcriptase- polymerase chain reaction (RT-PCR) analysis. The Vg mRNA and Vg protein synthesis were increased in rainbow trout in vivo and in vitro with E$_2$ and methyltestosterone (MT) There were dose and time-related effects of E$_2$ and MT on vitellogesis. Androgens such as progesterone androsterone and testosterone also stimulated Vg mRNA expression in vitro. The results show that androgens as well as E$_2$ can induce expression of Vg mRNA in trout in vivo and in vitro.

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Taxonomic position of Pedicularis hallaisanensis Hurusawa, an endemic plant of Mt. Halla (한라산 고유 한라송이풀의 분류학적 위치)

  • Cho, Won-Bum;Choi, Byoung-Hee
    • Korean Journal of Plant Taxonomy
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    • v.41 no.2
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    • pp.130-137
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    • 2011
  • Pedicularis growing at Mt. Halla of Jeju Island is known as an endemic species of P. hallaisanensis Hurusawa. On the other hand, the plant is morphologically similar to P. amoena, P. spicata, and P. verticillata in gross morphology, so the taxonomic treatment of the taxon remains controversial. To clarify the taxonomic position of the plants, we examined external morphological characters and nuclear ribosomal DNA ITS sequences for P. hallaisanensis and its related species. The plants of Mt. Halla are clearly different from P. amoena and P. verticillata in the morphology of calyx lobes, the length of galea and lower lip, density of glandular hairs on plants, presences of the radical leaves after anthesis and molecular data. However, P. hallaisanensis is not clearly separated from P. spicata distributed in N. E. Asia on external morphological characters and DNA sequences of internal transcribed spacers. In this study, the morphological and molecular data suggested that P. hallaisanensis should be merged into the former species.