• Title/Summary/Keyword: Plasma DNA

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Increase of Plasma IL-12/p40 Ratio Induced by the Combined Therapy of DNA Vaccine and Lamivudine Correlates with Sustained Viremia Control in CHB Carriers

  • Im, Se-Jin;Yang, Se-Hwan;Yoon, Seung-Kew;Sung, Young-Chul
    • IMMUNE NETWORK
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    • v.9 no.1
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    • pp.20-26
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    • 2009
  • We previously reported that $IFN-{\gamma}$ producing T cell responses induced by the combined therapy of DNA vaccine and lamivudine for one year are important for the induction of sustained virological response (SVR). However, $IFN-{\gamma}$ production is not sufficient to predict sustained viremia control in chronic hepatitis B (CHB) carriers treated. Methods: Twelve CHB carriers were intramuscularly immunized 12 times at a 4-week interval with 8mg of HBV DNA vaccine during the standard lamivudine treatment (100mg/daily/1 year). The level of cytokines during and after the combined therapy in plasma of all 12 CHB carriers treated was determined by each ELISA kit. Six out of 12 CHB carriers revisited the clinic, and their HBV DNA levels were examined. Results: The combined therapy increased plasma IL-12 and IL-12/p40 ratio during the treatment (baseline vs. peak level: $41.8{\pm}8.3$ vs. $163.1{\pm}29.2\;pg/ml$; p<0.01 and $0.96{\pm}0.25$ vs. $3.58{\pm}0.86$; p<0.01, respectively), and the peak level of plasma IL-12 and IL-12/p40 ratio was evoked at 6 to 10 months during the combined therapy. In particular, CHB carriers with SVR had two and three-fold higher level of the peak plasma IL-12 and plasma IL-12/p40 ratio than non-virological responders (NVRs), respectively ($218.0{\pm}41.4$ vs. $108.1{\pm}28.6\;pg/ml$; p=0.09 and $5.35{\pm}1.38$ vs. $1.80{\pm}0.29$; p<0.05, respectively), while p40 level was consistent during the combined therapy. In addition, there was no significant temporal correlation between the peak IL-12/p40 ratio and the elevation of serum alanine amino-transferase (ALT) in this study, contrast to $IFN-{\alpha}$ therapy which induced peak IL-12 level following ALT flares. Conclusion: Our results indicate that the combined therapy induces the increase of plasma IL-12 and IL-12/p40 ratio, which are associated with long-term SVR in CHB carriers.

Action of atmospheric pressure non-thermal plasma on the biomolecules and bio-organism

  • Attri, Pankaj;Park, Ji Hoon;Kumar, Naresh;Ali, Anser;Kim, In Tae;Lee, Weontae;Choi, Eun Ha
    • Proceedings of the Korean Vacuum Society Conference
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    • 2015.08a
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    • pp.66.1-66.1
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    • 2015
  • Plasma medicine is an upcoming research area that has attracted the scientists to explore more deeply the utility of plasma. So, apart from the treating biomaterials and tissues with plasma, we have studied the effect of plasma with different feeding gases on modification of biomolecules. Additionally, we have checked the action of nanosecond pulsed plasma on the biomolecules. We have checked the plasma action on proteins ((Hemoglobin (Hb) Myoglobin (Mb) and lysoenzyme), calf thymus DNA and amino acids. The structural changes or structural modification of proteins and DNA have been studied using circular dichroism (CD), dynamic light scattering (DLS), gel electrophoresis, protein oxidation test, UV-vis spectroscopy and 1D NMR, while Liquid Chromatograph/Capillary Electrophoresis-Mass Spectrometer(LC/CE-MS) based qualitative bio-analysis have been used to study the modification of amino acids. We have also shown the effect of NaCl and ionic liquid on the formation of OH radicals using electron spin resonance and fluorescence techinques.

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Intragenic DNA Methylation Concomitant with Repression of ATP4B and ATP4A Gene Expression in Gastric Cancer is a Potential Serum Biomarker

  • Raja, Uthandaraman Mahalinga;Gopal, Gopisetty;Rajkumar, Thangarajan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5563-5568
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    • 2012
  • Based on our previous report on gastric cancer which documented ATP4A and ATP4B mRNA down-regulation in gastric tumors relative to normal gastric tissues, we hypothesized that epigenetic mechanisms could be responsible. ATP4A and ATP4B mRNA expression in gastric cancer cell lines AGS, SNU638 and NUGC-3 was examined using reverse transcriptase PCR (RT-PCR). AGS cells were treated with TSA or 5'-AzaDC and methylation specific PCR (MSP) and bisulfite sequencing PCR (BSP) analysis were performed. MSP analysis was on DNA from paraffin embedded tissues sections and plasma. Expression analysis revealed downregulation of ATP4A and ATP4B genes in gastric cancer cell lines relative to normal gastric tissue, while treatment with 5'-AzaDC re-activated expression of both. Search for CpG islands in their putative promoter regions did not indicate CpG islands (CGI) but only further downstream in the bodies of the genes. Methylation specific PCR (MSP) in the exon1 of the ATP4B gene and exon7 in ATP4A indicated methylation in all the gastric cancer cell lines tested. MSP analysis in tumor tissue samples revealed methylation in the majority of tumor samples, 15/19, for ATP4B and 8/8 for ATP4A. There was concordance between ATP4B and ATP4A down-regulation and methylation status in the tumour samples tested. ATP4B methylation was detectable in cell free DNA from gastric cancer patient's plasma samples. Thus ATP4A and ATP4B down-regulation involves DNA methylation and methylated ATP4B DNA in plasma is a potential biomarker for gastric cancer.

Cellular and Molecular Responses of a Filamentous Fungus Neurospora Crassa to Non-thermal Plasma at Atmospheric Pressure

  • Park, Gyung-Soon;Ryu, Young-Hyo;Hong, Young-June;Uhm, Han-Sup;Choi, Eun-H.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.476-476
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    • 2012
  • Although plasma is an efficient means of microbial sterilization, mechanism of plasma effect on microorganisms still needs to be clarified. In addition, a limited number of studies are available on eukaryotic microorganisms such as yeast and fungi in relation to plasma application. Thus, we investigated cellular and molecular aspects of plasma effects on a filamentous fungus, Neurospora crassa by making use of argon plasma jet at atmospheric pressure. The viability and cell morphology of N. crassa spores exposed to plasma were both significantly reduced depending on the exposure time when treated in water. The intracellular genomic DNA content was dramatically reduced in fungal tissues after a plasma treatment and the transcription factor tah-3 was found to be required for fungal tolerance to a harsh plasma environment.

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Plasma Real Time-Quantitative Polymerase Chain Reaction of Epstein-Barr Virus in Immunocompetent Patients with Hepatitis

  • Hong, Ji-Hye;Bae, Yon-Jung;Sohn, Joon-Hyung;Ye, Byung-Il;Chun, Jin-Kyong;Kim, Hwang-Min
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.15 no.1
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    • pp.38-43
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    • 2012
  • Purpose: Epstein-Barr virus (EBV) hepatitis is a usually asymptomatic and self-limiting disease in immunocompetent patients. However, the range of severity is wide, and the serological diagnosis is typically difficult until the convalescent phase. Thus, we examined the value of plasma EBV DNA real-time quantitative polymerase chain reaction (RT-qPCR) in EBV hepatitis for the timely diagnosis and the relationship between EBV viral load and clinical severity. Methods: Sixty samples were confirmed as having EBV infection by RT-qPCR with the EBV BALF5 gene sequence. We examined the clinical characteristics of EBV hepatitis by reviewing medical records. Results: The median total duration of fever was 8 days (range: 0-13 days). The mean peak value of aspartate aminotransferase (AST) was $241{\pm}214$ U/L, and the mean peak value of alanine aminotransferase (ALT) was $298{\pm}312$ U/L. There was no correlation between the serum levels of liver enzyme and plasma EBV DNA titer ($p$=0.1) or between median total duration of fever and EBV DNA titer ($p$=0.056). The median age of the EBV VCA IgM-negative group was lower compared with the EBV VCA IgM-positive group in EBV hepatitis (2 years vs. 6 years, $p$=0.0009). Conclusion: The severity of EBV hepatitis does not correlate with circulating EBV DNA load according to our data. Furthermore, we suggest that plasma EBV PCR may be valuable in young infants in whom the results of serology test for EBV infection commonly are negative.

Characteristics of Gene Structure of Bovine Ghrelin and Influence of Aging on Plasma Ghrelin

  • Kita, K.;Harada, K.;Nagao, K.;Yokota, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.723-727
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    • 2005
  • Ghrelin is a novel growth-hormone-releasing acylated peptide, which has been purified and identified in rat stomach. In the present study, the full-length sequence of bovine ghrelin cDNA was cloned by RT-PCR. The bovine ghrelin cDNA sequence derived in the present study included a 348 bp open reading frame and a 137 bp 3'UTR. The putative amino acid sequence of bovine prepro-ghrelin consisted of 116 amino acids, which contained the 27-amino acid ghrelin. The sequence analysis of the bovine ghrelin gene revealed that an intron existed between Gln$^{13}$ and Arg$^{14}$ of ghrelin. This exon-intron boundary matched the GT-AG rule of the splicing mechanism. Compared with rats, which have two tandem CAG sequences in the 3'end of intron, bovine ghrelin genome has only one CAG sequence. Therefore, although rats can produce 28 amino acid-ghrelin and 27 amino acid-des-Gln$^{14}$-ghrelin by alternative splicing, ruminant species, including bovines, might be able to produce only one type of ghrelin peptide, des-Gln$^{14}$-ghrelin. The influence of aging on plasma ghrelin concentration was also examined. Plasma ghrelin concentration increased after birth to approximately 600 days of age, and then remained constant.

Plasma Amino Acid and Urine Organic Acid Analyses in Leigh Syndrome (리증후군에서의 혈장 아미노산 및 소변 유기산 분석)

  • Na, Ji-Hoon;Lee, Hyunjoo;Lee, Hae-in;Huh, Euira;Lee, Young-Mock
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.22 no.1
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    • pp.28-36
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    • 2022
  • Purpose: Detection of abnormal metabolites in plasma amino acid (PAA) and urine organic acid (UOA) analyses has been used to diagnose clinical mitochondrial diseases, such as Leigh syndrome. In this study, the diagnostic values and effectiveness of PAA and UOA analyses were reviewed. Methods: This was a retrospective study of patients with Leigh syndrome who were diagnosed between 2003 and 2018 in a single tertiary care center. Through a whole mitochondrial sequencing and nuclear DNA associated mitochondrial gene panel analysis, 19 patients were found to be positive for mitochondrial DNA (mtDNA) mutation-associated Leigh syndrome, and 57 patients were negative. Their PAA and UOA analyses results were then compared. Results: In the comparison of the PAA and UOA analyses results between the two groups, no abnormal metabolites showed obvious differences between the mtDNA mutation-positive Leigh syndrome and mtDNA mutation-negative Leigh syndrome groups. Conclusion: PAA and UOA analyses are inappropriate test methods for diagnosing Leigh syndrome or screening of mtDNA mutation-associated Leigh syndrome. However, UOA analysis might still be a suitable screening test for Leigh syndrome.

The Effect of Alpha-tocopherol Supplementation on the Improvement of Antioxidant Status and Lymphocyte DNA Damage in Postmenopausal Women (비타민 E 보충섭취가 폐경기 여성의 혈장 항산화 영양상태 및 DNA 손상 개선에 미치는 영향)

  • Kim, Chang-Suk;Kang, Hae-Jin;Lee, Soon-Hee;Park, Yoo-Kyoung;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.40 no.8
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    • pp.708-718
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    • 2007
  • The purpose of this project was to evaluate whether vitamin E supplementation could improve the antioxidant status and lymphocyte DNA damage in Korean postmenopausal women. This was double blinded, placebo-controlled trial. Thirty-five subject were randomized to receive either placebo 400 mg/capsule or natural $\alpha$-tocopherol 400 IU/capsule, 2 times a day for 6 weeks. We measured plasma vitamin C, $\alpha$-tocopherol, $\gamma$-tocopherol, $\alpha$-carotenoid, $\beta$-carotenoid, lycopene concentration and tail length, %DNA in tail, tail moment in lymphocyte DNA damage index. Vitamin E supplementation group had significantly increased plasma vitamin C(p<0.05), $\alpha$-tocopherol(p<0.000), whereas $\gamma$-tocopherol(p<0.000) and tail length(p<0.05) were significantly decreased. However, placebo supplementation group also had significantly increased plasma vitamin C(p<0.05). In conclusion, our study shows that vitamin E supplementation to Korean postmenopausal women may partially improve antioxidant status and lymphocyte DNA damage.

THE EFFECT OF GENISTEIN CONCENTRATED POLYSACCHARIDE (GCP) SUPPLEMENTATION ON OXIDATIVE DNA DAMAGE AND PLASMA TOTAL ANTIOXIDANT POTENTIAL IN OLD FEMALE SPRAGUE-DAWLEY RATS.

  • Park, Eunju;Shin, Jang-In;Park, Ok-Jin;Kang, Myung-Hee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.10a
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    • pp.118-119
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    • 2001
  • The anti-cardiovascular effect of estrogen replacement therapy in postmenopausal women is known to be associated with its role as an antioxidant, its ability to protect cells from DNA damage. Genistein concentrated polysaccharide (GCP) is a functional food produced by fermentation of soybean isoflavone extracts with Basidiomycetes, containing rich content of genistein aglycones. The aim of this study was to investigate the effect of GCP on oxidative DNA damage and plasma total antioxidant potential, comparing to the effect of estrogen.(omitted)

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Effective Method for Extraction of Cell-Free DNA from Maternal Plasma for Non-Invasive First-Trimester Fetal Gender Determination: A Preliminary Study

  • Lim, Ji-Hyae;Park, So-Yeon;Kim, Shin-Young;Kim, Do-Jin;Kim, Mee-Jin;Yang, Jae-Hyug;Kim, Moon-Young;Kim, Min-Hyoung;Han, Ho-Won;Choi, Kyu-Hong;Ryu, Hyun-Mee
    • Journal of Genetic Medicine
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    • v.7 no.1
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    • pp.53-58
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    • 2010
  • Purpose: To find the most effective method for extraction of cell-free DNA (cf-DNA) from maternal plasma, we compared a blood DNA extraction system (blood kit) and a viral DNA extraction system (viral kit) for non-invasive first-trimester fetal gender determination. Materials and Methods: A prospective cohort study was conducted with maternal plasma collected from 44 women in the first-trimester of pregnancy. The cf-DNA was extracted from maternal plasma using a blood kit and a viral kit. Quantitative fluorescent-polymerase chain reaction (QF-PCR) was used to detect the SRY gene and AMEL gene. The diagnostic accuracy of the QF-PCR results was determined based on comparison with the final delivery records. Results: A total of 44 women were tested, but the final delivery record was only obtained in 36 cases which included 16 male-bearing and 20 female-bearing pregnancies. For the blood kit and viral kit, the diagnostic accuracies for fetal gender determination were 63.9% (23/36) and 97.2% (35/36), respectively. Conclusion: In non-invasive first-trimester fetal gender determination by QF-PCR, using a viral kit for extraction of cf-DNA may result in a higher diagnostic accuracy.