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Lee, Hyungbeen,Lee, Sang Won,Lee, Gyudo,Lee, Wonseok,Nam, Kihwan,Lee, Jeong Hoon,Hwang, Kyo Seon,Yang, Jaemoon,Lee, Hyeyoung,Kim, Sangsig,Lee, Sang Woo,Yoon, Dae Sung The Royal Society of Chemistry 2018 Nanoscale Vol.10 No.2
<P>Here, we demonstrate a powerful method to discriminate DNA mismatches at single-nucleotide resolution from 0 to 5 mismatches (<I>χ</I>0 to <I>χ</I>5) using Kelvin probe force microscopy (KPFM). Using our previously developed method, we quantified the surface potentials (SPs) of individual DNA-capped nanoparticles (DCNPs, ∼100 nm). On each DCNP, DNA hybridization occurs between ∼2200 immobilized probe DNA (<I>p</I>DNA) and target DNA with mismatches (<I>t</I>DNA, ∼80 nM). Thus, each DCNP used in the bioassay (each <I>p</I>DNA-<I>t</I>DNA interaction) corresponds to a single ensemble in which a large number of <I>p</I>DNA-<I>t</I>DNA interactions take place. Moreover, one KPFM image can scan at least dozens of ensembles, which allows statistical analysis (<I>i.e.</I>, an ensemble average) of many bioassay cases (ensembles) under the same conditions. We found that as the <I>χ</I>n increased from <I>χ</I>0 to <I>χ</I>5 in the tDNA, the average SP of dozens of ensembles (DCNPs) was attenuated owing to fewer hybridization events between the <I>p</I>DNA and the <I>t</I>DNA. Remarkably, the SP attenuation <I>vs.</I> the <I>χ</I>n showed an inverse-linear correlation, albeit the equilibrium constant for DNA hybridization exponentially decreased asymptotically as the <I>χ</I>n increased. In addition, we observed a cascade reaction at a 100-fold lower concentration of <I>t</I>DNA (∼0.8 nM); the average SP of DCNPs exhibited no significant decrease but rather split into two separate states (no-hybridization <I>vs.</I> full-hybridization). Compared to complementary <I>t</I>DNA (<I>i.e.</I>, <I>χ</I>0), the ratio of no-hybridization/full-hybridization within a given set of DCNPs became ∼1.6 times higher in the presence of tDNA with single mismatches (<I>i.e.</I>, <I>χ</I>1). The results imply that our method opens new avenues not only in the research on the DNA hybridization mechanism in the presence of DNA mismatches but also in the development of a robust technology for DNA mismatch detection.</P>
Development of an infant formula certified reference material for the analysis of organic nutrients
Lee, Joonhee,Kim, Byungjoo,Lee, Sun Young,Choi, Jongoh,Kang, Dukjin,Lee, Hwasim,Choi, KiHwan,Lee, Hyeyoung,Sim, Hee-Jung,Baek, Song-Yee,Lee, Honghee,Hyung, Seok-Won,Ahn, Seonghee,Seo, Dongwon,Hwang, J Applied Science Publishers 2019 Food chemistry Vol.298 No.-
<P><B>Abstract</B></P> <P>Infant formula certified reference material (CRM, KRISS CRM 108-02-003) were developed for the analysis of organic nutrients. The CRM is a milk-based infant formula powder, packaged at 14 g per unit. Ten thousand units were prepared and stored at −70 °C. For the certification of each nutrient, ten units were analyzed for simultaneous value-assignment and homogeneity test. Analytical methods used were isotope dilution mass spectrometry (IDMS) based on liquid chromatography mass spectrometer (LC/MS) or gas chromatography mass spectrometer (GC/MS) as higher-order reference methods.13 vitamins, 3 fatty acids, and total cholesterol were certified. The between-unit relative standard deviation of measurement results for each nutrient ranged 0.2% to 2.5%, showing very good homogeneity. The expanded relative uncertainties of the certified values ranged from 1% to 8%, indicating that they have higher-order metrological quality. The values of proximates (proteins, lipids, carbohydrates, water, and ash) were assigned through inter-laboratory comparisons.</P> <P><B>Highlights</B></P> <P> <UL> <LI> An infant formula CRMs for the analysis of organic nutrients was developed. </LI> <LI> Organic nutrients were certified by IDMS approaches as higher-order reference methods. </LI> <LI> Homogeneities and stability of the CRM were evaluated by IDMS approaches. </LI> <LI> Metrological qualities of the certified values were proved by their small uncertainties. </LI> <LI> Five proximates were value-assigned by interlaboratory comparison. </LI> </UL> </P>
Seok, Seung-Hyeok,Koo, Hye Cheong,Kasuga, Asako,Kim, Yeun,Lee, Eun Gae,Lee, Hyeyoung,Park, Jong-Hwan,Baek, Min-Won,Lee, Hui-Young,Kim, Dong-Jae,Lee, Byeung-Hee,Lee, Yong-Soon,Cho, Sang-Nae,Park, Jae-H Elsevier 2006 Veterinary microbiology Vol.114 No.3
<P><B>Abstract</B></P><P>Skin ulcers, scoliosis, and dropsy-like scale edema were observed in laboratory-maintained zebrafish. Affected fish had multifocal granulomas not only in internal organs such as the liver, intestine, genital organs, kidney, muscle, and spleen but also in the fin, epithelium, gills, and sclera of the eyes. Large numbers of acid-fast-rod-shaped bacteria were observed within the necrotic centers of well-demarcated, multifocal granulomas with Gram's stain and Ziehl–Neelson's stain. The size of the <I>Mycobacterium</I> spp. was 1–2μm×2–3μm with a double-layered cell wall, based upon electron-microscopical features. Definitive diagnosis of these outbreaks was obtained by culture on selective media followed by PCR-restriction fragment length polymorphism analysis (PRA) of the <I>rpoB</I> gene for species identification. The amplified 360-bp products of the <I>rpoB</I> gene of mycobacteria isolated from zebrafish were digested with <I>Msp</I>I restriction enzyme, which revealed unique band patterns matching those of <I>Mycobacterium abscessus</I> and <I>Mycobacterium chelonae</I> which are responsible for skin and soft tissue infection caused by rapidly growing mycobacteria in humans. This is the first documentation of the precise identification of zoonotic non-tuberculous mycobacteria isolated from laboratory-maintained zebrafish by the PRA of the <I>rpoB</I> gene; this study thus provides a great deal of useful epidemiological information and reduces the likelihood that epizootics will occur.</P>
Estimation of CO<sub>2</sub> emissions from waste incinerators: Comparison of three methods
Lee, Hyeyoung,Yi, Seung-Muk,Holsen, Thomas M.,Seo, Yong-Seok,Choi, Eunhwa Elsevier 2018 waste management Vol.73 No.-
<P><B>Abstract</B></P> <P>Climate-relevant CO<SUB>2</SUB> emissions from waste incineration were compared using three methods: making use of CO<SUB>2</SUB> concentration data, converting O<SUB>2</SUB> concentration and waste characteristic data, and using a mass balance method following Intergovernmental Panel on Climate Change (IPCC) guidelines. For the first two methods, CO<SUB>2</SUB> and O<SUB>2</SUB> concentrations were measured continuously from 24 to 86 days. The O<SUB>2</SUB> conversion method in comparison to the direct CO<SUB>2</SUB> measurement method had a 4.8% mean difference in daily CO<SUB>2</SUB> emissions for four incinerators where analyzed waste composition data were available. However, the IPCC method had a higher difference of 13% relative to the direct CO<SUB>2</SUB> measurement method. For three incinerators using designed values for waste composition, the O<SUB>2</SUB> conversion and IPCC methods in comparison to the direct CO<SUB>2</SUB> measurement method had mean differences of 7.5% and 89%, respectively. Therefore, the use of O<SUB>2</SUB> concentration data measured for monitoring air pollutant emissions is an effective method for estimating CO<SUB>2</SUB> emissions resulting from waste incineration.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Fossil CO<SUB>2</SUB> emissions from waste incineration were compared using three methods. </LI> <LI> Direct CO<SUB>2</SUB> measurements, converting O<SUB>2</SUB> concentrations, and mass balance were used. </LI> <LI> The first and the second methods agreed well. </LI> <LI> The first and the third methods had a larger difference. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>
Lee, Hyang-Ah,Park, Joon Ha,Kim, Dae Won,Lee, Choong-Hyun,Hwang, In Koo,Kim, Hyeyoung,Shin, Myoung Cheol,Cho, Jun Hwi,Lee, Jae-Chul,Noh, Yoohun,Kim, Sung-Su,Won, Moo-Ho,Ahn, Ji Hyeon SPANDIDOS PUBLICATIONS 2018 MOLECULAR MEDICINE REPORTS Vol.17 No.3
<P>Oligodendrocytes are myelin-forming cells in the central nervous system. Research into the effects of aging on oligodendrocyte protein expression remains limited. The present study aimed to determine the alterations in oligodendrocyte-specific protein (OSP) expression in the gerbil hippocampus at 1, 2, 3, 4, 6 and 24 months of age with western blot and immunohistochemistry analyses. OSP expression levels in the hippocampus were highest at 6 months of age. OSP immunoreactivity was identified in numerous cell bodies at 1 month, although the number of OSP immunoreactive cells was different according to hippocampal subregion. The number of OSP immunoreactive cells significantly decreased at 2 months and, thereafter, numbers decreased gradually. The detection of OSP immunoreactive fibers was negligible in all layers in the hippocampal subregions until 4 months. OSP immunoreactive fibers were abundant at 6 and 24 months, although the fiber distribution patterns in the CA1-3 areas and dentate gyrus were different. The results demonstrated that OSP expression in the gerbil hippocampus was age-dependent. The detection of OSP immunoreactive cell bodies and fibers was significantly different according to the layers of hippocampal subregions, indicating that myelination may be continuously altered in the hippocampus during normal aging.</P>
이혜영,--,--,--,--,--,-- THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 2003 Journal of biomedical laboratory sciences Vol.9 No.3
Since water borne infection causes acute diseases and results in spread of diseases by secondary infection, the prevention is very important. Therefore, it is necessary to have a method that is rapid and effective to monitor pathogenic bacteria in drinking water. In this study, we employed a systematic method, Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis, to develop an effective monitoring system for possible bacterial contaminants in drinking water. For this purpose, PCR primers were derived from 992 bp region of the 16s rRNA gene that is highly conserved through the different species of prokaryotes. To test whether the PCR primers designed are indeed useful for detecting all the possible microbial contaminants in the water, the primers were used to amplify 16s rRNA regions of different microbial water-borne pathogens such as E. coli, Salmonella, Yersinia, Listeria, and Staphylococcus. As expected, all of tested microorganisms amplified expected size of PCR products indicating designed PCR primers for 16s rRNA indeed can be useful to amplify all different microbial water-borne pathogens in the water. Furthermore, to test whether these 16s rRNA based PCR primers can detect bacterial populations present in the water, water samples taken from diverse sources, such as river, tap, and sewage, were used for amplification. PCR products were for then subjected for cloning into a T-vector to generate a library containing 16s rRNA sequences from various bacteria. With cloned PCR products, RFLP analysis was done using PCR products digested with restriction enzyme such as Hae Ⅲ to obtain species-specific RFLP profiles. After PCR-RFLP, the bacterial clones which showed the same RFLP profiles were regarded as the same ones, and the clones which showed distinctive RFLP profiles were subsequently subjected for sequence analysis for species identification. By this PCR-RFLP analysis, we were able to reveal diverse populations of bacteria living in water. In brief, in unsterilized natural river water, over 60 different species of bacteria were found. On the other hand, no PCR products were detected in drinking tap-water. The results from this study clearly indicate that the PCR-RFLP-sequence analysis can be a useful method for monitoring diverse, perhaps pathogenic bacteria contaminated in water in a rapid fashion.