• Title/Summary/Keyword: Bisulfate

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FRET-Based Quantitative Discrimination of Bisulfite-Untreated DNA from Bisulfite-Treated DNA

  • Lee, Eun Jeong;Cho, Yea Seul;Song, Seongeun;Hwang, Sang-Hyun;Hah, Sang Soo
    • Bulletin of the Korean Chemical Society
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    • v.35 no.5
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    • pp.1455-1459
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    • 2014
  • We report a sensitive and reliable FRET-based nanotechnology assay for efficient detection and quantification of bisulfite-unmodified or modified DNA. Bisulfite-untreated DNA or bisulfite-treated DNA is subjected to PCR amplification with biotin-conjugated primers so that the amounts of bisulfite-untreated and treated DNA can be differentiated. Streptavidin-coated quantum dots (QDs) are used to capture biotinylated PCR products intercalated with SYBR Green, enabling FRET measurement. Key features of our method include its low intrinsic background noise, high resolution, and high sensitivity, enabling detection of as little as 1.75 ng of bisulfite-untreated DNA in the presence of an approximately 1,000-fold excess of bisulfite-untreated DNA compared to bisulfate-treated DNA, with the use of PCR reduced (as low as 15 cycles). SYBR Green as an intercalating dye as well as a FRET acceptor allows for a single-step preparation without the need for primers or probes to be chemically conjugated to an organic fluorophore. Multiple acceptors per FRET donor significantly enhance the signal-to-noise ratio as well. In consideration of the high relevance of bisulfite treatment to DNA methylation quantitation, our system for FRET measurement between QDs and intercalating dyes can be generally utilized to analyze DNA methylation and to potentially benefit the scientific and clinical community.

Cleaning of the Waste Reverse Osmosis Membrane Filters for the Household Water Purifier and Their Performance Enhancement Study (정수기용 역삼투 폐분리막 필터의 세정 및 성능 향상 연구)

  • Cho, Young Ju;Rhim, Ji Won
    • Membrane Journal
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    • v.27 no.3
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    • pp.232-239
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    • 2017
  • In this study, the regeneration investigation for waste reverse osmosis membrane filters which were discarded after use for the household water purifiers has been carried out. Sodium hydroxide, sodium bisulfate, and ethylenediamine tetra acetic acid(EDTA). as the chemical cleaning agents were used. And they were in-situ cleaned with the micro-bubble generator as well. The best result was obtained when both 0.1% EDTA and micro-bubbles were used for 30 min cleaning. Thus, when the performance of the brand new RO membrane and restorated RO membrane were compared, the flux, 19.9%, the recovery ratio 45% were enhanced while the salt rejection was reduced for NaCl 100 mg/L solution, in other words, it has been recovered to the original brand new RO membrane filter. Also the removal of pollutants on membrane surface was confirmed in a naked eye through the scanning electron microscopy. Finally, this research has provided the possibility of the re-use of the waste RO membrane filters of household water purifier which were reclaimed or incinerated after use.

Oxidation of Diphenylmethane Using Polyethylene glycols as Phase Transfer Catalysts (폴리에틸렌글리콜 상이동 촉매를 이용한 디페닐메탄의 산화반응)

  • Lee, Hwa-Soo;Moon, Jeong-Yeol;Park, Dae-Won;Park, Sang-Wook;Shin, Jung-Ho
    • Applied Chemistry for Engineering
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    • v.4 no.4
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    • pp.715-720
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    • 1993
  • Diphenylmethane (pKa=33.4), which is difficult to be oxidized in normal oxidation conditions, was oxidized to produce benzophenone at ambient temperature and atmospheric pressure by using phase transfer catalysts and solid potassium tert-butoxide as base. Quaternary salt such as benzyltriethylammonium chloride, tetrabutyl ammonium bisulfate, tetrabutylphosponium chloride, are ineffective catalysts for this reaction, but 18-crown-6 and polyethylene glycols showed catalytic activity. The conversion of diphenylmethane was increased with increasing chain length of PEG molecules when they are used as phase transfer catalysts both in equal molar and equal weight basis. The conversion of diphenylmethane was increased with the agitation speed, and aprotic solvent like DMF showed higher reaction rate compared with benzene.

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Promoter Methylation and Relative mRNA Expression of the p16 Gene in Cervical Cancer in North Indians

  • Gupta, Amita;Ahmad, Mohammad Kaleem;Mahndi, Abbas Ali;Singh, Renu;Pradeep, Yashodhara
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.4149-4154
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    • 2016
  • Background: Cervical carcinoma is one of the main causes of mortality in women worldwide as well as in India. It occurs as a result of various molecular events that develop from the combined influences of an individual's genetic predisposition and external agents such as smoking and menstrual hygiene, for example. However, infection with human papillomavirus (HPV) is the established major risk factor. The aim of the current study was to investigate p16 CpG island methylation and establish any correlation with mRNA expression in north Indian population. Materials and Methods: We analyzed 196 woman volunteer out of which 98 were cases and 98 healthy controls. For the analysis of methylation pattern, DNA extracted from blood samples was modified with a bisulfate kit and used as template for methylation specific PCR (MSP). Quantitative real-time PCR (QRT-PCR) was performed to check mRNA expression. Results: Correlation between methylation status of p16 gene and poor menstrual hygiene was significant (p=0.006), high parity cases showed methylation of p16 gene (p=0.031) with increased risk up to 1.86 times for cervical cancer and smoking was a strong risk factor associated with cervical cancer. We analyzed methylation pattern and found 60.3% methylation in cases with low mRNA expression level (0.014) as compare to controls (1.24). It was also observed that promoter methylation of p16 gene was significantly greater in FIGO stage III. Conclusions: We conclude that p16 methylation plays an important role in cervical cancer in the north Indian population and its methylation decreases mRNA expression. It can be used as an important and consistent blood biomarker in cervical cancer patients.

Lack of Significance of the BRCA2 Promoter Methylation Status in Different Genotypes of the MTHFR a1298c Polymorphism in Ovarian Cancer Cases in Iran

  • Darehdori, Ahmad Shabanizadeh;Dastjerdi, Mehdi Nikbakht;Dahim, Hajar;Slahshoor, Mohammadreza;Babazadeh, Zahra;Taghavi, Mohammad Mohsen;Taghipour, Zahra;Gaafarineveh, Hamidreza
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.1833-1836
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    • 2012
  • Objective: Promoter methylation, which can be regulated by MTHFR activity, is associated with silencing of genes. In this study we evaluated the methylation status (type) of the BRCA2 promoter in ovarian cancer patients carrying different genotypes of the MTHFR gene (A or C polymorphisms at position 1298). Methods: The methylation type of the BRCA2 promoter was evaluated using bisulfate-modified DNA in methylation-specific PCR and the MTHFRa1278c polymorphism was assessed by PCR-RFLP. Results: Analysis of the BRCA2 promoter methylation type of cases showed that 7 out of 60 cases (11.7%) were methylated while the remaining 53 (88.3%) were unmethylated. In methylated cases, one out of the 7 cases had a CC genotype and the remaining 6 methylated cases had an AC genotype. The AA genotype was absent. In unmethylated cases, 34, 18, and one out of these had AC, AA and CC genotype, respectively. Conclusion: There was no significant relationship between the methylation types of the BRCA2 promoter in different genotypes of MTHFRa1298c polymorphism in ovarian cancer; p=0.255. There was no significant relation between the methylation types of the BRCA2 promoter in different genotypes of the MTHFRa1298c polymorphism in ovarian cancer.

Aberrant DNA Methylation of P16, MGMT, and hMLH1 Genes in Combination with MTHFR C677T Genetic Polymorphism and Folate Intake in Esophageal Squamous Cell Carcinoma

  • Chen, Jing;Huang, Zhi-Jie;Duan, Yu-Qin;Xiao, Xin-Rong;Jiang, Jian-Qing;Zhang, Ru
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5303-5306
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    • 2012
  • Aim: The present case-control study was conducted to explore the association of MTHFR gene polymorphism and relations of P16, MGMT and HMLH1 to MTHFR and folate intake. Methods: A total of 257 cases of esophageal squamous cell carcinoma confirmed by histopathological examination were collected. Genotyping of P16, MGMT and HMLH1 was accomplished by methylation-specific polymerase chain reaction (PCR) after sodium bisulfate modification of DNA and the MTHFR C677T genetic polymorphism was detected by PCR-restriction fragment-length polymorphism (PCR-RFLP). Results: The proportions of DNA hypermethylation in P16, MGMT and hMLH1 in cancer tissues were significantly higher than in paracancerous normal tissue. The proportion of hypermethylation in at least one gene was 88.5% in cancer tissue, and was also significantly higher than that in paracancerous normal tissue. Our finding showed individuals with homozygotes (TT) of MTHFR C677T had significant risk of DNA hypermethylation of MGMT in cancer tissues, with an OR (95% CI) of 3.15 (1.12-6.87). Similarly, patients with high intake of folate also showed a slight high risk of DNA methylation of MGMT, with OR (95% CI) of 2.03 (1.05-4.57). Conclusion: Our study found the P16, MGMT and hMLH1 demonstrate a high proportion of hypermethylation in esophageal squamous cell cancer cancer tissues, which might be used as biomarkers for cancer detection.

Selective Catalytic Reduction (SCR) Technology Trend for the Removal of Nitrogen Oxide from Ship Flue Gas (선박 배가스 내 질소산화물 제거를 위한 선택적촉매환원법(SCR) 기술동향)

  • Won, Jong Min;Hong, Sung Chang
    • Prospectives of Industrial Chemistry
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    • v.22 no.5
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    • pp.25-40
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    • 2019
  • 전 세계적으로 환경문제를 해결하기 위한 방안으로 환경규제를 강화시키며 특히 다양한 대기오염 물질 중 최근 큰 이슈인 초미세먼지 저감을 위해 전구물질로 알려진 질소산화물을 제어하기 위한 다양한 기술개발이 가속화되고 있다. 특히, 다양한 처리기술 중에 기술적·경제적인 이점을 갖춘 선택적 촉매환원법(selective catalytic reduction, SCR) 기술을 통하여 질소산화물 제거를 위해 암모니아를 환원제로 반응에 참여시켜 인체에 무해한 H2O, N2로 전환하는 기술이 대표적이다. 최근 전 세계적으로 다양한 산업군에서 질소산화물이 배출되고 있으며, 점오염원뿐만이 아니라 비점오염원(mobile sources)에 대한 규제가 강화되고 있다. 디젤엔진이 장착된 선박 배가스 처리장치 내 SCR 기술이 주목을 받고 있으며, NH3-SCR에 사용되는 촉매는 주로 VOx/TiO2, VOx/W/TiO2 촉매가 대표적이다. 한편 선박 디젤엔진에 사용되는 연료에 따라 연소배가스 특성이 다르다. 이러한 연료가 연소됨에 따라 SO2, SO3가 발생되고 환원제인 NH3와 결합하여 황산암모늄염((NH4)2SO4), ABS (ammonium bisulfate, NH4HSO4)과 같은 염을 형성시켜 탈질촉매의 비활성화 문제가 발생된다. 이러한 비활성화 물질이 침적된 탈질촉매를 재활성화 시키기 위하여 열 산화를 통해 재생시키고 있다. 이처럼 선박용 SCR 촉매는 강화되는 배출규제 및 엔진기술의 발달로 저감되는 운전 온도에 대비하여 저온 활성 재생이 가능한 고활성, 고내구성 촉매기술 개발이 필요하다.

Ultrafine Particle Events in the Ambient Atmosphere in Korea

  • Maskey, Shila;Kim, Jae-Seok;Cho, Hee-Joo;Park, Kihong
    • Asian Journal of Atmospheric Environment
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    • v.6 no.4
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    • pp.288-303
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    • 2012
  • In this study, real time measurements of particle number size distribution in urban Gwangju, coastal Taean, and industrial Yeosu in Korea were conducted in 2008 to understand the occurrence of ultrafine particle (UFP) (<100 nm) events, the variation of its concentration among different sampling sites, and UFP formation pathways. Also, to investigate seasonal and long-term variation of the UFP number concentration, data were collected for the period of 5 years (2007, 2008, 2010, 2011, and 2012) in urban Gwangju. Photochemical and combustion events were found to be responsible for the formation of UFP in the urban Gwangju site, whereas only photochemical event led to the formation of UFP in the coastal Taean site. The highest UFP concentration was found in industrial Yeosu (the average UFP number fractions were 79, 59 and 58% in Yeosu, Gwangju, and Taean, respectively), suggesting that high amount of gas pollutants (e.g., $NO_2$, $SO_2$, and volatile organic carbon (VOC)) emitted from industries and their photochemical reaction contributed for the elevated UFP concentration in the industrial Yeosu site. The UFP fraction also showed a seasonal variation with the peak value in spring (61.5, 54.5, 50.5, and 40.7% in spring, fall, summer, and winter, respectively) at urban Gwangju. Annual average UFP number concentrations in urban Gwangju were $5.53{\times}10^3\;cm^{-3}$, $4.68{\times}10^3\;cm^{-3}$, $5.32{\times}10^3\;cm^{-3}$, $3.99{\times}10^3\;cm^{-3}$, and $2.16{\times}10^3\;cm^{-3}$ in the year 2007, 2008, 2010, 2011, and 2012, respectively. Comparison of the annual average UFP number concentration with urban sites in other countries showed that the UFP concentrations of the Korean sites were lower than those in other urban cities, probably due to lower source strength in the current site. TEM/EDS analysis for the size-selected UFPs showed that the UFPs were classified into various types having different chemical species. Carbonaceous particles were observed in both combustion (soot and organics) and photochemical events (sulfate and organics). In the photochemical event, an internal mixture of organic species and ammonium sulfate/bisulfate was identified. Also, internal mixtures of aged Na-rich and organic species, aged Ca-rich particles, and doughnut shaped K-containing particles with elemental composition of a strong C with minor O, S, and K-likely to be originated from biomass burning nearby agricultural area, were observed. In addition, fly ash particles were also observed in the combustion event, not in the photochemical event.

Genetic Diagnosis of Beckwith Wiedemann Syndrome using Methylation Specific PCR-RFLP Method (Methylation Specific PCR-RFLP 방법을 이용한 Beckwith Wiedemann Syndrome의 진단)

  • Kim, Gu-Hwan;Lee, Jin-Joo;Choi, Seung-Hoon;Lee, Joo-Yeon;Lee, Beom-Hee;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.7 no.2
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    • pp.133-137
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    • 2010
  • Purpose: Beckwith-Wiedemann syndrome (BWS) is an overgrowth malformation syndrome caused by a methylation abnormality at chromosome 11p15, consisting of two imprinting centers, BWSIC1 (IGF2, H19) and BWSIC2 (LIT1, KvDMR). This study evaluated the applicability of a methylation-specific (MS) PCR RFLP method for the genetic diagnosis of BWS. Materials and Methods: A total of 12 patients were recruited based on clinical findings. Karyotyping was performed using peripheral blood leukocytes, and genomic DNA was treated with bisulfate and amplified using methylation-specific primers. RFLP was conducted with restriction enzymes in differentially methylated regions of LIT1, H19, and IGF2. Results: The 12 BWS patients had normal karyotypes. Abnormal methylation patterns in the BWSIC2 (LIT1) region were identified in seven patients (58.3%) using the MS-PCR RFLP method. Conclusions: The MS-PCR RFLP method is a simple, economical genetic test. It detected genetic abnormalities in 50-60% of BWS patients, suggesting that it can be used as a screening test. A more precise method is required, however, to enhance the detection rate of genetic abnormalities, especially in BWSIC1 region.

Characterization of fine particulate matter during summer at an urban site in Gwangju using chemical, optical, and spectroscopic methods (화학적·광학적·분광학적 방법을 이용한 광주 도심지역 여름철 초미세먼지의 특성)

  • Son, Se-Chang;Park, Tae-Eon;Park, Seungshik
    • Particle and aerosol research
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    • v.17 no.4
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    • pp.91-106
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    • 2021
  • Daily PM2.5 was collected during summer period in 2020 in Gwangju to investigate its chemical and light absorption properties. In addition, real-time light absorption coefficients were observed using a dual-spot 7-wavelength aethalometer. During the study period, SO42- was the most important contributor to PM2.5, accounting for on average 33% (10-64%) of PM2.5. The chemical form of SO42- was appeared to be combination of 70% (NH4)2SO4 and 30% NH4HSO4. Concentration-weighted trajectory (CWT) analysis indicated that SO42- particles were dominated by local pollution, rather than regional transport from China. A combination of aethalometer-based and water-extracted brown carbon (BrC) absorption indicated that light absorption of BrC due to aerosol particles was 1.6 times higher than that due to water-soluble BrC, but the opposite result was found in absorption Ångström exponent (AAE) values. Lower AAE value by aerosol BrC particles was due to the light absorption of aerosol BrC by both water-soluble and insoluble organic aerosols. The BrC light absorption was also influenced by both primary sources (e.g., traffic and biomass burning emissions) and secondary organic aerosol formation. Finally the ATR-FTIR analysis confirmed the presence of NH4+, C-H groups, SO42-, and HSO42-. The presence of HSO42- supports the result of the estimated composition ratio of inorganic sulfate ((NH4)2SO4) and bisulfate (NH4HSO4).