• Title/Summary/Keyword: Cell free DNA

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Protective effects of carnosine and homocarnosine on ferritin and hydrogen peroxide-mediated DNA damage

  • Kang, Jung-Hoon
    • BMB Reports
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    • v.43 no.10
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    • pp.683-687
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    • 2010
  • Previous studies have shown that one of the primary causes of increased iron content in the brain may be the release of excess iron from intracellular iron storage molecules such as ferritin. Free iron generates ROS that cause oxidative cell damage. Carnosine and related compounds such as endogenous histidine dipetides have antioxidant activities. We have investigated the protective effects of carnosine and homocarnosine against oxidative damage of DNA induced by reaction of ferritin with $H_2O_2$. The results show that carnosine and homocarnosine prevented ferritin/$H_2O_2$-mediated DNA strand breakage. These compounds effectively inhibited ferritin/$H_2O_2$-mediated hydroxyl radical generation and decreased the mutagenicity of DNA induced by the ferritin/$H_2O_2$ reaction. Our results suggest that carnosine and related compounds might have antioxidant effects on DNA under pathophysiological conditions leading to degenerative damage such as neurodegenerative disorders.

Protection of Paeoniae radix from H2O2-induced oxidative DNA damage

  • Lee, Seung-Cheol;Kwon, Yong-Soo;Heo, Moon-Young
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.282.1-282.1
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    • 2002
  • Paeoniae radix is commonly used for various woman's health problems in traditional korean medicine. In order to develop new antioxidant for woman use. the ethanolic extracts of paeoniae radix (PRE) were prepared and various biological activities were evaluated. PRE showed potent free radical scavenging activity and moderate antioxidative activity in vitro. and also showed the protective effect on H2O2-induced DNA damage in mammalian cell. (omitted)

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Construction and Transformation of an Endogenous Plasmid pBL1-free Brevibacterium lactofermentum (내재형 Plasmid pBL1이 제거된 Brevibacterium lactofermentum 개발과 형질전환)

  • 이규남;민본홍;윤기홍
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.164-169
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    • 1995
  • An endogenous cryptic plasmid, pBL1, which has been used to construct plasmid vectors for coryneform bacteria producing amino acids, was eliminated from Brevibacterium lactofermentum. The pBL1 was partially digested with Sau3AI and the resulting DNA fragments were subcloned into a suicide vector pEM1 which contains a kanamycin-resistant (km$^{r}$) gene. KM$^{r}$ B. lactofermentum transconjugants were obtained by conjugal transfer of the pEM1 derivatives containing pBL1 DNA fragments from Escherichia coli into B. lactofermentum. A km$^{r}$ transconjugant was analyzed to contain a plasmid pEB14, which occurred in vivo by homologous recombination between pBL1 and the conjugal-transferred plasmid. The pEB14 including the pEM1-derived km$^{r}$ gene was found to be lost concomitantly with km$^{r}$ phenotype, resulting in the construction of a pBL1-free strain of B lactofermentum. Based on transformation efficiencies and plasmid stability, the resultant pBL1- free strain is more useful than wild strain as a host cell for genetic manipulation. It could be concluded that foreign plasmid DNAs are efficiently isolated and analyzed from the pBL1-free strain because of the absence of endogenous pBL1 plasmid.

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The Effects of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin (TCDD) on Proliferation of MCF-7 and Hec-1B Cell Lines

  • Ryu, Y.H.;Seo, D.S.;Ko, Y.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.94-94
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    • 2003
  • Endocrine disrupters (EDs) are exogenous chemicals that interfere with the production, releasing, metabolism, excretion, binding of natural hormones, and whole endocrine systems. EDs are very dangerous since they are extremely stable, not easily degraded, and accumulated in fat and tissue. 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) is known as the most toxic EDs. Therefore, this study was conducted to investigate the effects of TCDD on proliferation of human breast cancer (MCF-7) and endometrial adenocarcinoma (Hec-1B) cells. 10, 100, and 1000 nM of TCDD were treated with steroid free condition. Viable cell counting, MTT, and BrdU assay was performed to investigate cell proliferation. Apoptosis was investigated using DNA laddering. Although, DNA fragmentation as the evidence of apoptosis was not detected, all of these cell lines showed restricted proliferation at 48 hrs after 100 and 1000 nM TCDD treatments. Recently, it has been reported that the expression of transforming growth factor $\beta$s (TGF-$\beta$s) are increased in TCDD treatment and also involved in regulation of cell cycle. Therefore, these results were considered that the decreased cell prolifcration by TCDD is related to the expression of TGF-$\beta$s.

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Naegleria fowleri Induces Jurkat T Cell Death via O-deGlcNAcylation

  • Lee, Young Ah;Kim, Kyeong Ah;Shin, Myeong Heon
    • Parasites, Hosts and Diseases
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    • v.59 no.5
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    • pp.501-505
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    • 2021
  • The pathogenic free-living amoeba Naegleria fowleri causes primary amoebic meningoencephalitis, a fatal infection, by penetrating the nasal mucosa and migrating to the brain via the olfactory nerves. N. fowleri can induce host cell death via lytic necrosis. Similar to phosphorylation, O-linked β-N-acetylglucosamine (O-GlcNAc) glycosylation (O-GlcNAcylation) is involved in various cell-signaling processes, including apoptosis and proliferation, with O-GlcNAc addition and removal regulated by O-GlcNAc transferase and O-GlcNAcase (OGA), respectively. However, the detailed mechanism of host cell death induced by N. fowleri is unknown. In this study, we investigated whether N. fowleri can induce the modulation of O-GlcNAcylated proteins during cell death in Jurkat T cells. Co-incubation with live N. fowleri trophozoites increased DNA fragmentation. In addition, incubation with N. fowleri induced a dramatic reduction in O-GlcNAcylated protein levels in 30 min. Moreover, pretreatment of Jurkat T cells with the OGA inhibitor PUGNAc prevented N. fowleri-induced O-deGlcNAcylation and DNA fragmentation. These results suggest that O-deGlcNAcylation is an important signaling process that occurs during Jurkat T cell death induced by N. fowleri.

Interferon-Stimulated Gene 15 in the Control of Cellular Responses to Genotoxic Stress

  • Jeon, Young Joo;Park, Jong Ho;Chung, Chin Ha
    • Molecules and Cells
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    • v.40 no.2
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    • pp.83-89
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    • 2017
  • Error-free replication and repair of DNA are pivotal to organisms for faithful transmission of their genetic information. Cells orchestrate complex signaling networks that sense and resolve DNA damage. Post-translational protein modifications by ubiquitin and ubiquitin-like proteins, including SUMO and NEDD8, are critically involved in DNA damage response (DDR) and DNA damage tolerance (DDT). The expression of interferon-stimulated gene 15 (ISG15), the first identified ubiquitin-like protein, has recently been shown to be induced under various DNA damage conditions, such as exposure to UV, camptothecin, and doxorubicin. Here we overview the recent findings on the role of ISG15 and its conjugation to target proteins (e.g., p53,$ {\Delta}Np63{\alpha}$, and PCNA) in the control of cellular responses to genotoxic stress, such as the inhibition of cell growth and tumorigenesis.

Evaluation of Digital PCR as a Technique for Monitoring Acute Rejection in Kidney Transplantation

  • Lee, Hyeseon;Park, Young-Mi;We, Yu-Mee;Han, Duck Jong;Seo, Jung-Woo;Moon, Haena;Lee, Yu-Ho;Kim, Yang-Gyun;Moon, Ju-Young;Lee, Sang-Ho;Lee, Jong-Keuk
    • Genomics & Informatics
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    • v.15 no.1
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    • pp.2-10
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    • 2017
  • Early detection and proper management of kidney rejection are crucial for the long-term health of a transplant recipient. Recipients are normally monitored by serum creatinine measurement and sometimes with graft biopsies. Donor-derived cell-free deoxyribonucleic acid (cfDNA) in the recipient's plasma and/or urine may be a better indicator of acute rejection. We evaluated digital PCR (dPCR) as a system for monitoring graft status using single nucleotide polymorphism (SNP)-based detection of donor DNA in plasma or urine. We compared the detection abilities of the QX200, RainDrop, and QuantStudio 3D dPCR systems. The QX200 was the most accurate and sensitive. Plasma and/or urine samples were isolated from 34 kidney recipients at multiple time points after transplantation, and analyzed by dPCR using the QX200. We found that donor DNA was almost undetectable in plasma DNA samples, whereas a high percentage of donor DNA was measured in urine DNA samples, indicating that urine is a good source of cfDNA for patient monitoring. We found that at least 24% of the highly polymorphic SNPs used to identify individuals could also identify donor cfDNA in transplant patient samples. Our results further showed that autosomal, sex-specific, and mitochondrial SNPs were suitable markers for identifying donor cfDNA. Finally, we found that donor-derived cfDNA measurement by dPCR was not sufficient to predict a patient's clinical condition. Our results indicate that donor-derived cfDNA is not an accurate predictor of kidney status in kidney transplant patients.

Mitochondrial genome mutations in mesenchymal stem cells derived from human dental induced pluripotent stem cells

  • Park, Jumi;Lee, Yeonmi;Shin, Joosung;Lee, Hyeon-Jeong;Son, Young-Bum;Park, Bong-Wook;Kim, Deokhoon;Rho, Gyu-Jin;Kang, Eunju
    • BMB Reports
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    • v.52 no.12
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    • pp.689-694
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    • 2019
  • Ethical and safety issues have rendered mesenchymal stem cells (MSCs) popular candidates in regenerative medicine, but their therapeutic capacity is lower than that of induced pluripotent stem cells (iPSCs). This study compared original, dental tissue-derived MSCs with re-differentiated MSCs from iPSCs (iPS-MSCs). CD marker expression in iPS-MSCs was similar to original MSCs. iPS-MSCs expressed higher in pluripotent genes, but lower levels in mesodermal genes than MSCs. In addition, iPS-MSCs did not form teratomas. All iPSCs carried mtDNA mutations; some shared with original MSCs and others not previously detected therein. Shared mutations were synonymous, while novel mutations were non-synonymous or located on RNA-encoding genes. iPS-MSCs also harbored mtDNA mutations transmitted from iPSCs. Selected iPS-MSCs displayed lower mitochondrial respiration than original MSCs. In conclusion, screening for mtDNA mutations in iPSC lines for iPS-MSCs can identify mutation-free cell lines for therapeutic applications.

Validation of fetus aneuploidy in 221 Korean clinical samples using noninvasive chromosome examination: Clinical laboratory improvement amendments-certified noninvasive prenatal test

  • Kim, Min-Jeong;Kwon, Chang Hyuk;Kim, Dong-In;Im, Hee Su;Park, Sungil;Kim, Ji Ho;Bae, Jin-Sik;Lee, Myunghee;Lee, Min Seob
    • Journal of Genetic Medicine
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    • v.12 no.2
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    • pp.79-84
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    • 2015
  • Purpose: We developed and validated a fetal trisomy detection method for use as a noninvasive prenatal test (NIPT) including a Clinical Laboratory Improvement Amendments (CLIA)-certified bioinformatics pipeline on a cloud-based computing system using both Illumina and Life Technology sequencing platforms for 221 Korean clinical samples. We determined the necessary proportions of the fetal fraction in the cell-free DNA (cfDNA) sample for NIPT of trisomies 13, 18, and 21 through a limit of quantification (LOQ) test. Materials and Methods: Next-generation sequencing libraries from 221 clinical samples and three positive controls were generated using Illumina and Life Technology chemistries. Sequencing results were uploaded to a cloud and mapped on the human reference genome (GRCh37/hg19) using bioinformatics tools. Based on Z-scores calculated by normalization of the mapped read counts, final aneuploidy reports were automatically generated for fetal aneuploidy determination. Results: We identified in total 29 aneuploid samples, and additional analytical methods performed to confirm the results showed that one of these was a false-positive. The LOQ test showed that the proportion of fetal fraction in the cfDNA sample would affect the interpretation of the aneuploidy results. Conclusion: Noninvasive chromosome examination (NICE), a CLIA-certified NIPT with a cloud-based bioinformatics platform, showed unambiguous success in fetus aneuploidy detection.

Ordered Fragmentation of pDNA induced by PEG-PLL block copolymer -Correlation between Condensation degree and Biological Activity by Cell-Free System-

  • Osada, Kensuke;Doi, Motoyoshi;Shiotani, Tomonori;Yamasaki, Yuichi;Kataoka, Kazunori
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.254-254
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    • 2006
  • The sensitivity of plasmid DNA (pDNA) to S1 nuclease, an enzyme to cleave a single-strand DNA, was dramatically modulated through a supramolecular assembly (polyion complex micelle) with a synthetic block copolymer, poly(ethylene glycol)-b-poly(L-lysine) (PEG-PLL). The pDNA condensed in stoichiometric charge ratio was cleaved into 7 fragments each being 10/12, 9/12, 8/12, 6/12, 4/12, 3/12, and 2/12 of the original DNA length, on the other hand, the pDNA condensed in higher charge ratios (>4), were digested into non-specific manner. Condensation of the pDNA was investigated from two viewpoints that how does the rigid DNA molecules fold and condense and how does the condensation influence their biological activity.

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