• Title/Summary/Keyword: Plasma DNA

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Relationships of Plasma Homocysteine Concentration and Oxidative Stress Markers in Korean Collage Students (일부 남녀 대학생에서 혈장 호모시스테인 농도와 산화 스트레스 지표와의 상관관계)

  • Kim, Jung-Shin;Park, Eun-Ju;Min, Hye-Sun;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.43 no.5
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    • pp.443-452
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    • 2010
  • Elevated plasma concentration of total homocysteine (ptHcy) is known as an independent risk factor of cardiovascular disease (CVD) and oxidative stress is also commonly implicated in CVD. An association between ptHcy and oxidative stress has recently been suggested. The study objective is to examine the relationship between ptHcy and oxidative stress markers in 103 healthy college students (62 males and 41 females). Plasma levels of ptHcy, oxidative stress markers (conjugated diene, erythrocyte catalase, TRAP, lymphocyte DNA damage), antioxidant vitamins ($\alpha$-tocopherol, $\gamma$-tocopherol, carotenoids), and lipid parameters (total cholesterol, triglyceride, HDL cholesterol) were determined. The results show that the concentration of ptHcy was significantly higher in male subjects ($22.17\;{\pm}\;2.14\;{\mu}mole/L$) than in female subjects ($12.28\;{\pm}\;0.45\;{\mu}mole/L$). There was a negative association between ptHcy and plasma ${\beta}$-carotene in male subjects (p $lt; 0.05), but no correlation between ptHcy and other plasma antioxidant vitamin levels in either gender. However, there were the negative correlations between ptHcy and plasma ${\alpha}$-carotene or ${\beta}$-carotene, and a positive correlation between ptHcy and lymphocyte DNA damage. A significantly low level of ${\alpha}$-carotene or ${\beta}$-carotene was found in male subjects with elevated ptHcy (${\geq}\;15\;{\mu}mol/L$), as compared to those with lower plasma homocysteine. These study results confirmed the views on the association between plasma homocysteine and oxidative stress markers in humans and support the hypothesis that homocysteine promotes the oxidative environment by counteracting the antioxidant defense mechanism.

Effects of Polycyclic Aromatic Hydrocarbons on DNA Damage and Plasma Protein Expression in Mouse

  • Oh, Sang-Nam;Oh, Eun-Ha;Im, Ho-Sub;Jo, Gyu-Chan;Sul, Dong-Geun;Kim, Young-Whan;Lee, Eun-Il
    • Molecular & Cellular Toxicology
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    • v.1 no.1
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    • pp.32-39
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    • 2005
  • Polycyclic aromatic hydrocarbons (PAHs) are an important class of environmentally prevalent xenobiotics that exert complex effects on the biological system and characterized as probably carcinogenic materials. Single cell gel electrophoresis assays were performed in order to evaluate DNA damage occurring in the T-and B lymphocytes, spleens (T/B-cell), bone marrow, and livers of mouse exposed to mixture of PAHs (Benzo(a)pyrene, Benzo(e)pyrene, Fluoranthene, Pyrene) at dose of 400, 800, or 1600 mg/kg body weight for 2 days. DNA damage of the cells purified from mice was increased in dose dependent manner. In the blood cells and organs, DNA damage was also discovered to vary directly with PAHs. Especially T-cells had been damaged more than B-cell. Plasma proteomes were separated by 2-dimensional electrophoresis with pH 4-7 ranges of IPG Dry strips and many proteins showed significant up-and -down expressions with the dose dependent manner. Of these, significant 4 spots were identified using matrix-assisted laser desorption/ionization-time of fight (MALDI-TOF) mass spectrometry. Identified proteins were related to energy metabolism and signal transduction.

Pyridoxine Deficiency on Neurotransmitters in the Developing Rat Brain - Catecholamine Metabolism- (Pyridoxine결핍이 뇌의 신경전달물질에 미치는 영향 - Catecholamine 대사 -)

  • Choi, Hay-Mie;Kang, Soon-Ah
    • Journal of Nutrition and Health
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    • v.17 no.3
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    • pp.199-209
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    • 1984
  • Pregnant rats were fed a pyridoxine deficient diet during the gestation and lactation. DEF I group received the deficient diet from delivery ; DEF II group, from the 15 th day of gestation. Body and brain weights, brain protein, DNA, RNA, plasma GOT and GPT, and catecholamines were measured. Effect of MAO inhibiting drug, pargyline, was determined. Brain protein, DNA, and RNA of offsprings of deficient groups were significantly lower than the control group, but RNA/ DNA, brain weight/DNA, and protein/DNA show that cell number were more affected than cell size by the pyridoxine deficiency during the 3rd week of gestation and lactation. Plasma GOT activities were more significantly different than plasma GPT between the control and deficient group. Brain norepinephrine of offsprings of deficient group were significantly lower than the control, but brain dopamine content was not significantly different from the control. At 2nd and 3rd week, norepinephrine was significantly depressed in deficient groups. Pargyline treatment affected a 1.2 fold increase in catecholamines in 3hr while the control had a 1.5 fold increase. Thus norepinephrine and dopamine synthesis was depressed in the deficient groups. Dopaminergic neurons may be less dependent on pyridoxine level than neurons from norepinephrine. Pyridoxine deficiency in maternal diet is not so critical to brain catecholamines of offspring except to the neonatal rats.

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Detection of KRAS mutations in plasma cell-free DNA of colorectal cancer patients and comparison with cancer panel data for tissue samples of the same cancers

  • Min, Suji;Shin, Sun;Chung, Yeun-Jun
    • Genomics & Informatics
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    • v.17 no.4
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    • pp.42.1-42.6
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    • 2019
  • Robust identification of genetic alterations is important for the diagnosis and subsequent treatment of tumors. Screening for genetic alterations using tumor tissue samples may lead to biased interpretations because of the heterogeneous nature of the tumor mass. Liquid biopsy has been suggested as an attractive tool for the non-invasive follow-up of cancer treatment outcomes. In this study, we aimed to verify whether the mutations identified in primary tumor tissue samples could be consistently detected in plasma cell-free DNA (cfDNA) by digital polymerase chain reaction (dPCR). We first examined the genetic alteration profiles of three colorectal cancer (CRC) tissue samples by targeted next-generation sequencing (NGS) and identified 11 non-silent amino acid changes across six cancer-related genes (APC, KRAS, TP53, TERT, ARIDIA, and BRCA1). All three samples had KRAS mutations (G12V, G12C, and G13D), which were well-known driver events. Therefore, we examined the KRAS mutations by dPCR. When we examined the three KRAS mutations by dPCR using tumor tissue samples, all of them were consistently detected and the variant allele frequencies (VAFs) of the mutations were almost identical between targeted NGS and dPCR. When we examined the KRAS mutations using the plasma cfDNA of the three CRC patients by dPCR, all three mutations were consistently identified. However, the VAFs were lower (range, 0.166% to 2.638%) than those obtained using the CRC tissue samples. In conclusion, we confirmed that the KRAS mutations identified from CRC tumor tissue samples were consistently detected in the plasma cfDNA of the three CRC patients by dPCR.

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.

A Study on the DNA Polymorphisms at $\beta$fibrinogen Loci and Plasma Fibrinogen Concentration

  • Lee, Mi-Hwa;Ohrr, Hee-Choul;Song, Kyung-Soon
    • Biomedical Science Letters
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    • v.7 no.3
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    • pp.151-159
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    • 2001
  • Many prospective studies have confirmed the predictive value of plasma fibrinogen levels for vascular diseases, including ischemic heart disease. Several polymorphisms of the $\beta$fibrinogen gene have been investigated in relation to plasma fibrinogen levels. The blood samples for DNA were collected from 109 healthy Koreans who have no relationship by blood (67 males and 42 females) in due consideration of some other factors such as gender, age, and smoking status. Four polymorphisms of the $\beta$fibrinogen gene that consist of HaeIII, AluI, MaII and BcII restriction fragment length polymorphisms (RFLPs) were investigated to examine the associations between RFLPs and plasma fibrinogen levels. In conclusion, the significant associations between HaeIII, AluI, MnII RFLPs(H$_1$H$_2$, M$_1$M$_2$, $A_1$A$_2$) and the concentration of plasma fibrinogen were shown by the smokers as well as by the old people more than 50, whereas the association between BcII and plasma fibrinogen were shown no connection with the status of age and smoking. The concentration of plasma fibrinogen was significantly shown higher by the old people ($\geqq$50) by the younger people ($\leqq$49) in male and also higher by the smokers than by the nonsmokers.

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Red beet(Beta vulgaris L.) leaf supplementation improves antioxidant status in C57BL/6J mice fed high fat high cholesterol diet

  • Lee, Jeung-Hee;Son, Chan-Wook;Kim, Mi-Yeon;Kim, Min-Hee;Kim, Hye-Ran;Kwak, Eun-Shil;Kim, Se-Na;Kim, Mee-Ree
    • Nutrition Research and Practice
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    • v.3 no.2
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    • pp.114-121
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    • 2009
  • The effect of diet supplemented with red beet (Beta vulgaris L.) leaf on antioxidant status of plasma and tissue was investigated in C57BL/6J mice. The mice were randomly divided into two groups after one-week acclimation, and fed a high fat (20%) and high cholesterol (1%) diet without (control group) or with 8% freeze-dried red beet leaf (RBL group) for 4 weeks. In RBL mice, lipid peroxidation determined as 2-thiobarbituric acid-reactive substances (TBARS value) was significantly reduced in the plasma and selected organs (liver, heart, and kidney). Levels of antioxidants (glutathione and $\beta$-carotene) and the activities of antioxidant enzyme (glutathione peroxidase) in plasma and liver were considerably increased, suggesting that antioxidant defenses were improved by RBL diet. Comet parameters such as tail DNA (%), tail extent moment, olive tail moment and tail length were significantly reduced by 25.1%, 49.4%, 35.4%, and 23.7%, respectively, in plasma lymphocyte DNA of RBL mice compared with control mice, and indicated the increased resistance of lymphocyte DNA to oxidative damage. In addition, the RBL diet controlled body weight together with a significant reduction of fat pad (retroperitoneal, epididymal, inguinal fat, and total fat). Therefore, the present study suggested that the supplementation of 8% red beet leaf in high fat high cholesterol diet could prevent lipid peroxidation and improve antioxidant defense system in the plasma and tissue of C57BL/6J mice.

A Study of Immunogenicity and Reactogenicity of Hepatitis B Vaccine Made by Recombinant DNA Techniques in Yeast (효모재조합 DNA B형 간염백신의 면역효과에 관한 연구)

  • Min, Chang-Hong;Kim, Kyo-Myung;Lee, Kyu-Man
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.2
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    • pp.243-249
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    • 1986
  • A study of the immunogenicity and reactogenicity of two doses of lot H(10, 20 mcg), two doses of lot L (20, 40 mcg) of the Smith Kline-RIT recombinant DNA yeast-derived hepatitis B vaccine and a 20-mcg dose of the Merck Sharp and Dohme plasma-derived hepatitis B vaccine was conducted in young adults under randomized, double-blind conditions. Immunization was carried out according to a 0-, 1-, and 6-month vaccination schedule. Results indicated that the yeast-derived hepatitis B vaccine was well tolerated and immunogenic. Reactogenicity to both yeast- and plasma-derived vaccines was mild in severity and low in incidence with no significant differences appearing between the study groups. One month after the third dose, the yeast-derived vaccines induced a high degree of soroconversion ranging between 95.0% and 100%. The response was not lot or dose-dependent. The administration of the plasma-derived vaccine resulted in anti-HBs geometric mean titres statistically signifirantly higher than those elicited by the different yeast-derived hepatitis B vaccines one month after the third dose of vaccine but the difference was not large enough to be of great clinical significance.

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Lymphocyte DNA damage and plasma antioxidant status in Korean subclinical hypertensive patients by glutathione S-transferase polymorphism

  • Han, Jeong-Hwa;Lee, Hye-Jin;Choi, Hee Jeong;Yun, Kyung Eun;Kang, Myung-Hee
    • Nutrition Research and Practice
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    • v.11 no.3
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    • pp.214-222
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    • 2017
  • BACKGROUND/OBJECTIVES: Glutathione S-transferase (GST) forms a multigene family of phase II detoxification enzymes which are involved in the detoxification of xenobiotics by conjugating substances with glutathione. The aim of this study is to assess the antioxidative status and the degree of DNA damage in the subclinical hypertensive patients in Korea using glutathione S-transferase polymorphisms. SUBJECTS/METHODS: We examined whether DNA damage and antioxidative status show a difference between GSTM1 or GSTT1 genotype in 227 newly diagnosed, untreated (systolic blood pressure $(BP){\geq}130mmHg$ or diastolic $BP{\geq}85mmHg$) subclinical hypertensive patients and 130 normotensive subjects (systolic BP < 120 mmHg and diastolic BP < 80 mmHg). From the blood of the subjects, the degree of the DNA damage in lymphocyte, the activities of erythrocyte superoxide dismutase, the catalase, and the glutathione peroxidase, the level of glutathione, plasma total radical-trapping antioxidant potential (TRAP), anti-oxidative vitamins, as well as plasma lipid profiles and conjugated diene (CD) were analyzed. RESULTS: Of the 227 subjects studied, 68.3% were GSTM1 null genotype and 66.5% were GSTT1 null genotype. GSTM1 null genotype had an increased risk of hypertension (OR: 2.104, CI: 1.38-3.35), but no significant association in GSTT1 null genotype (OR 0.982, CI: 0.62-1.55). No difference in erythrocyte activities of superoxide dismutase, catalase, or glutathione peroxidase, and plasma TRAP, CD, lipid profiles, and GSH levels were observed between GSTM1 or GSTT1 genotype. Plasma levels of ${\alpha}-tocopherol$ increased significantly in GSTT1 wild genotype (P < 0.05); however, plasma level of ${\beta}-carotene$ increased significantly in GSTT1 null genotype (P < 0.01). DNA damage assessed by the Comet assay was significantly higher in GSTM1 null genotype than wild genotype (P < 0.05). CONCLUSIONS: These results confirm the association between GSTM1 null genotype and risk of hypertension as they suggest that GSTM1 null genotype leads to an increased oxidative stress compared with wild genotype.