• Biomolecules & Therapeutics
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• 제27권6호
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• pp.530-539
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• 2019

Brain aging is an inevitable process characterized by structural and functional changes and is a major risk factor for neurodegenerative diseases. Most brain aging studies are focused on neurons and less on astrocytes which are the most abundant cells in the brain known to be in charge of various functions including the maintenance of brain physical formation, ion homeostasis, and secretion of various extracellular matrix proteins. Altered mitochondrial dynamics, defective mitophagy or mitochondrial damages are causative factors of mitochondrial dysfunction, which is linked to age-related disorders. Etoposide is an anti-cancer reagent which can induce DNA stress and cellular senescence of cancer cell lines. In this study, we investigated whether etoposide induces senescence and functional alterations in cultured rat astrocytes. Senescence-associated ${\beta}$-galactosidase (SA-${\beta}$-gal) activity was used as a cellular senescence marker. The results indicated that etoposide-treated astrocytes showed cellular senescence phenotypes including increased SA-${\beta}$-gal-positive cells number, increased nuclear size and increased senescence-associated secretory phenotypes (SASP) such as IL-6. We also observed a decreased expression of cell cycle markers, including PhosphoHistone H3/Histone H3 and CDK2, and dysregulation of cellular functions based on wound-healing, neuronal protection, and phagocytosis assays. Finally, mitochondrial dysfunction was noted through the determination of mitochondrial membrane potential using tetramethylrhodamine methyl ester (TMRM) and the measurement of mitochondrial oxygen consumption rate (OCR). These data suggest that etoposide can induce cellular senescence and mitochondrial dysfunction in astrocytes which may have implications in brain aging and neurodegenerative conditions.

• 한국축산식품학회지
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• 제25권2호
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• pp.218-225
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• 2005

본 연구는 mt DNA의 D-loop 영역을 probe로 이용한 Southern blot hybridization 분석 기법을 이용하여 한우에서 mt DNA의 RFLP를 분석하고 RFLP marker가 유생산에 미치는 영향을 분석하여 산유량 관련 DNA marker를 개발하고자 수행하였다. Mt DNA의 D-loop 영역내 404번부터 15061까지 1142 bp 크기의 염기 서열 부위를 특이적인 primer를 이용하여 PCR로 증폭하였다. Mt DNA를 HpaII, BamHI, XbaI, HinfI, EcoRI, HindII 및 RsaI 7종류의 제한효소를 이용하여 각각 절단한 후 DIG로 표지된 D-loop probe를 이용하여 검출한 결과 XbaI, RsaI, BamHI 및 HpaII 4종류의 제한효소에서 각각 RFLP 다형성이 검출되었고 EcoRI, HindIII 및 HinfI 3종류 제한효소는 변이가 존재하지 않았다. 다유 계통과 저유 계통 선발 집단간의 각 제한효소별 RFLP type의 출현빈도를 비교한 결과 BamHI 및 RsaI 제한효소에서 두 집단간의 RFLP type의 출현율에 각각 통계적 유의성(P<.05)이 인정되었다. 다유 및 저유성으로 극단의 육종가 값을 갖는 두 계통의 mt DNA D-loop영역의 염기 서열을 비교 분석한 결과 441번째 염기가 G/C, 469번째 염기는 T/C, 503번째 염기는 C/T, 569번째 염기는 G/A, 614번째 염기는 C/A그리고 644번째 염기는 C/T로 각각 염기가 치환되었고 특히, 다유 계통 개체의 677번째의 A염기가 저유 계통 개체에서는 결실되어 있다는 사실이 확인되었다. 한편, 한우 암소의 유생산에 영향을 미치는 효과를 규명하기 위하여 mt DNA RFLP 형과 송아지 이유시 체중, 생시체중 및 비유량을 측정하여 얻어진 육종가의 성적을 근거로 통계 분석한 결과 XbaI 제한효소의 RFLP type이 산유 능력 육종가와 유의적인 관련성이 확인되었다 (P<05). 즉, RFLP A type을 갖는 축군의 평균 육종가 추정치가 6.233으로 B type을 갖는 축군의 평균 육종가 추정치 0.757보다 월등히 높은 것으로 나타났다. 결론적으로 산유량과 관련성이 확인된 mt DNA RFLP type은 한우의 산유량 향상을 위한 DNA marker로 이용 가능할 것으로 기대된다.

• 한국어류학회지
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• 제32권4호
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• pp.239-244
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• 2020

마이크로네시아의 축 주변해에서 채집한 치어(체장 7.8 mm)의 형태적 특징은 체장과 지느러미의 발달단계를 제외하면 Omosudis sp.와 매우 유사하였다. 이 표본을 12S rRNA 마커를 이용하여 홍메치목의 Omosudis lowii로 동정하였다. 분자마커로 동정된 Omosudis lowii 치어의 형태적 특징은 이 종의 자치어 동정에 매우 유용할 것이다.

• Nutrition Research and Practice
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• 제16권2호
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• pp.147-160
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• 2022

BACKGROUND/OBJECTIVES: Patients with chronic kidney disease (CKD) have a high concentration of uremic toxins in their blood and often experience muscle atrophy. Indoxyl sulfate (IS) is a uremic toxin produced by tryptophan metabolism. Although an elevated IS level may induce muscle dysfunction, the effect of IS on physiological concentration has not been elucidated. Additionally, the effects of ursolic acid (UA) on muscle hypertrophy have been reported in healthy models; however, it is unclear whether UA ameliorates muscle dysfunction associated with chronic diseases, such as CKD. Thus, this study aimed to investigate whether UA can improve the IS-induced impairment of mitochondrial biogenesis. MATERIALS/METHODS: C2C12 cells were incubated with or without IS (0.1 mM) and UA (1 or 2 μM) to elucidate the physiological effect of UA on CKD-related mitochondrial dysfunction and its related mechanisms using real-time reverse transcription-polymerase chain reaction, western blotting and enzyme-linked immunosorbent assay. RESULTS: IS suppressed the expression of differentiation marker genes without decreasing cell viability. IS decreased the mitochondrial DNA copy number and ATP levels by downregulating the genes pertaining to mitochondrial biogenesis (Ppargc1a, Nrf1, Tfam, Sirt1, and Mef2c), fusion (Mfn1 and Mfn2), oxidative phosphorylation (Cycs and Atp5b), and fatty acid oxidation (Pdk4, Acadm, Cpt1b, and Cd36). Furthermore, IS increased the intracellular mRNA and secretory protein levels of interleukin (IL)-6. Finally, UA ameliorated the IS-induced impairment in C2C12 cells. CONCLUSIONS: Our results indicated that UA improves the IS-induced impairment of mitochondrial biogenesis by affecting differentiation, ATP levels, and IL-6 secretion in C2C12 cells. Therefore, UA could be a novel therapeutic agent for CKD-induced muscle dysfunction.

• Parasites, Hosts and Diseases
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• 제51권1호
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• pp.1-8
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• 2013

Taenia solium, T. saginata, and T. asiatica are taeniid tapeworms that cause taeniasis in humans and cysticercosis in intermediate host animals. Taeniases remain an important public health concerns in the world. Molecular diagnostic methods using PCR assays have been developed for rapid and accurate detection of human infecting taeniid tapeworms, including the use of sequence-specific DNA probes, PCR-RFLP, and multiplex PCR. More recently, DNA diagnosis using PCR based on histopathological specimens such as 10% formalin-fixed paraffin-embedded and stained sections mounted on slides has been applied to cestode infections. The mitochondrial gene sequence is believed to be a very useful molecular marker for not only studying evolutionary relationships among distantly related taxa, but also for investigating the phylo-biogeography of closely related species. The complete sequence of the human Taenia tapeworms mitochondrial genomes were determined, and its organization and structure were compared to other human-tropic Taenia tapeworms for which complete mitochondrial sequence data were available. The multiplex PCR assay with the Ta4978F, Ts5058F, Tso7421F, and Rev7915 primers will be useful for differential diagnosis, molecular characterization, and epidemiological surveys of human Taenia tapeworms.

• Journal of Ginseng Research
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• 제41권3호
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• pp.326-329
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• 2017

Background: Cultivated ginseng is often introduced as a substitute and adulterant of Russian wild ginseng due to its lower cost or misidentification caused by similarity in appearance with wild ginseng. The aim of this study is to develop a simple and reliable method to differentiate Russian wild ginseng from cultivated ginseng. Methods: The mitochondrial NADH dehydrogenase subunit 7 (nad7) intron 3 regions of Russian wild ginseng and Chinese cultivated ginseng were analyzed. Based on the multiple sequence alignment result, a specific primer for Russian wild ginseng was designed by introducing additional mismatch and allele-specific polymerase chain reaction (PCR) was performed for identification of wild ginseng. Real-time allele-specific PCR with endpoint analysis was used for validation of the developed Russian wild ginseng single nucleotide polymorphism (SNP) marker. Results: An SNP site specific to Russian wild ginseng was exploited by multiple alignments of mitochondrial nad7 intron 3 regions of different ginseng samples. With the SNP-based specific primer, Russian wild ginseng was successfully discriminated from Chinese and Korean cultivated ginseng samples by allele-specific PCR. The reliability and specificity of the SNP marker was validated by checking 20 individuals of Russian wild ginseng samples with real-time allele-specific PCR assay. Conclusion: An effective DNA method for molecular discrimination of Russian wild ginseng from Chinese and Korean cultivated ginseng was developed. The established real-time allele-specific PCR was simple and reliable, and the present method should be a crucial complement of chemical analysis for authentication of Russian wild ginseng.

• 생명과학회지
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• 제27권3호
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• pp.361-369
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• 2017

미토콘드리아의 항상성은 미토콘드리아 생합성과 마이토파지(자가포식에 의한 미토콘드리아 분해)로 불리는 2가지 주요 과정들에 의해 정교하게 조절되고 있다. 지구성 운동 훈련에 반응하여 골격근에서 미토콘드리아 생합성에 관한 기전들은 잘 정립되어 있는 반면 지구성 운동 훈련 후 골격근의 마이토파지 조절 기전과 마이토파지와 미토콘드리아 생합성의 협응을 조절하는 기전은 아직 명확히 밝혀져 있지 않다. 최근 연구들에 의하면 지구성 운동 훈련은 골격근에서 미토콘드리아 생합성, 미토콘드리아 역동성, 미토콘드리아 분해와 관련된 유전인자들의 발현을 증가시킨다고 하였다. 하지만 골격근에서 자가포식이 억제되었을 경우, 지구성 운동 훈련에 의한 미토콘드리아 생합성과 관련된 지표들인 Cox IV와 citrate synthase의 증가는 상쇄되었다. 따라서 자가포식과 마이토파지는 골격근의 미토콘드리아 생합성에 중요한 역할을 하며 정반대되는 이 두 과정(이화 또는 동화작용)의 협응 과정이 지구성 운동 훈련에 반응하여 대사적 기능과 지구력 운동 수행능력을 향상시키는 것과 같은 골격근의 적응에 중요한 듯하다. 지구성 운동은 미토콘드리아의 일정한 숫자를 유지시키기 위해 미토콘드리아 생합성, 미토콘드리아의 융합과 분열, 자가포식/마이토 파지들의 각각의 과정들을 조절하는 것으로 여겨진다. 지구성 운동 훈련은 골격근에서 마이토파지를 활성화시켜 미토콘드리아 양과 질을 조절하여 늙고 건강하지 않은 미토콘드리아를 젊고 건강한 미토콘드리아로 교체시킬 수 있다. 이 총론에서 미토콘드리아 생합성과 마이토파지의 분자학적 기전과 서로 상반되는 이 두 과정간의 협응이 골격근의 지구성 훈련에 대한 세포적 적응에 관련한다는 내용이 논의될 것이다.

• Journal of Plant Biotechnology
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• 제2권2호
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• pp.101-108
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• 2000

The liverwort Pellia borealis is a diploid, monoecious, allopolypliod species (n=18) that as it was postulated, originated after hybridization and duplication of chromosome sets of two cryptic species: Pellia epiphylta-species N (n=9) and Pellia epiphylla-species 5 (n=9). Our recent results have supported the allopolyploid origin of P.borealis. We have shown that the nuclear genome of P.borealis consists of two nuclear genomes: one derived from P.epiphylla-species N and the other from P.epiphylla-species 5. In this paper we show the origin of chloroplast and mitochondrial genomes in an allopolyploid species P.borealis. To our knowledge there is no information concerning the way of mitochondria and chloroplast inheritance in Brophyta. Using an allopolyploid species of p. borealis as a model species we have decided to look into chloroplast and mitochondrial genomes of P.borealis, P.epiphylla-species N and P.epiphylla-species S for nucleotide sequences that would allow us to differentiate between both cryptic species and to identify the origin of organelle genomes in the alloploid species. We have amplified and sequenced a chloroplast $tRNA^{Leu}$ gene (anticodon UAA) containing an intron that has shown to be highly variable in a nucleotide sequence and used for plant population genetics. Unfortunately these sequences were identical in all three liverwort species tested. The analysis of the nucleotide sequence of chloroplast, an intron containing $tRNA^{Gly}$ (anticodon UCC) genes, gave expected results: the intron nucleotide sequence was identical in the case of both P.borealis and P.epiphyllaspecies N, while the sequence obtained from P.epiphyllasperies S was different in several nucleotide positions. These results were confirmed by the nucleotide sequence of another chloroplast molecular marker the chloroplast, an intron-contaning $tRNA^{Lys}$ gene (anticodon UUU). We have also sequenced mitochondrial, an intron-containing $tRNA^{Ser}$ gene (anticodon GCU) in all three liverwort species. In this case we found that, as in the case of the chloroplast genome, P.borealis mitochondrial genome was inherited from P.epiphylla-species N. On the basis of our results we claim that both organelle genomes of P.borealis derived from P.epiphylla-species N.

• International Journal of Industrial Entomology and Biomaterials
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• 제35권1호
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• pp.51-57
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• 2017

The tiny dragonfly, Nannophya pygmaea (Odonata: Libellulidae), is an endangered insect in South Korea. Previously, a partial mitochondrial DNA sequence that corresponded to a DNA barcoding region has been used to infer genetic diversity and gene flow. In this study, we additionally sequenced the barcoding region from N. pygmaea that had been collected from three previously sampled populations (40 individuals) and these sequences were combined with the preexisting data. We also selected and sequenced an additional mitochondrial gene (ND5) to find further variable gene regions in the mitochondrial genome. DNA barcoding sequences of 108 individuals from five South Korean localities showed that genetic diversity was highest in Gangjin, Jeollanam-do Province. Muuido, which was previously occupied by a single haplotype, was also found to have an identical haplotype, which confirmed the low genetic diversity on this islet. Gene flow among populations is highly limited, and no clear distance- or region-based geographic partitioning was observed. Phylogenetic relationships among haplotypes showed that there were no discernable haplotypes in South Korea. ND5 provided slightly more haplotypes compared to the barcoding region in 40 individuals (14 vs. 10 haplotypes in the COI gene). It also had a slightly higher within-locality diversity estimate, which suggested that ND5 had potential as mitochondrial DNA-based marker for population genetic analysis.

• 한국가금학회지
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• 제36권1호
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• pp.85-88
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• 2009

조류에서 미토콘드리아내 유전자인 cytochrome c oxidase I(COI)는 종 구분을 위한 바이오마커로 많이 이용이 되고 있다. 본 연구는 이 유전자의 변이를 이용하여 닭의 품종 구분이 가능한지를 실험하기 위하여 실시하였다. 그 결과 한국 재래닭은 공시축으로 사용한 산란계인 White Leghorn과 육계인 Cornish가 가지고 있는 단일염기다형을 모두 가지고 있는 것으로 판명되어 품종구분을 위한 마커로 사용하기에는 적합하지 않은 것으로 확인되었다. 그러나 본 연구 결과를 통하여 한국재래계가 육계와 산란계의 특성을 모두 가지고 있다는 기존의 학설을 뒷받침하였으며, 품종 구분을 위하여 미토콘드리아와 genomic DNA 유래 단일염기다형 선발의 중요성을 확인할 수 있었다.