• Title/Summary/Keyword: Chemical marker

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Isolation of human mesenchymal stem cells from the skin and their neurogenic differentiation in vitro

  • Byun, Jun-Ho;Kang, Eun-Ju;Park, Seong-Cheol;Kang, Dong-Ho;Choi, Mun-Jeong;Rho, Gyu-Jin;Park, Bong-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.38 no.6
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    • pp.343-353
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    • 2012
  • Objectives: This aim of this study was to effectively isolate mesenchymal stem cells (hSMSCs) from human submandibular skin tissues (termed hSMSCs) and evaluate their characteristics. These hSMSCs were then chemically induced to the neuronal lineage and analyzed for their neurogenic characteristics in vitro. Materials and Methods: Submandibular skin tissues were harvested from four adult patients and cultured in stem cell media. Isolated hSMSCs were evaluated for their multipotency and other stem cell characteristics. These cells were differentiated into neuronal cells with a chemical induction protocol. During the neuronal induction of hSMSCs, morphological changes and the expression of neuron-specific proteins (by fluorescence-activated cell sorting [FACS]) were evaluated. Results: The hSMSCs showed plate-adherence, fibroblast-like growth, expression of the stem-cell transcription factors Oct 4 and Nanog, and positive staining for mesenchymal stem cell (MSC) marker proteins (CD29, CD44, CD90, CD105, and vimentin) and a neural precursor marker (nestin). Moreover, the hSMSCs in this study were successfully differentiated into multiple mesenchymal lineages, including osteocytes, adipocytes, and chondrocytes. Neuron-like cell morphology and various neural markers were highly visible six hours after the neuronal induction of hSMSCs, but their neuron-like characteristics disappeared over time (24-48 hrs). Interestingly, when the chemical induction medium was changed to Dulbecco's Modified Eagle Medium (DMEM) supplemented with fetal bovine serum (FBS), the differentiated cells returned to their hSMSC morphology, and their cell number increased. These results indicate that chemically induced neuron-like cells should not be considered true nerve cells. Conclusion: Isolated hSMSCs have MSC characteristics and express a neural precursor marker, suggesting that human skin is a source of stem cells. However, the in vitro chemical neuronal induction of hSMSC does not produce long-lasting nerve cells and more studies are required before their use in nerve-tissue transplants.

Trace level analysis of Pb in plasma by inductively coupled plasma/mass spectrometry (유도결합플라즈마 질량분석법을 이용한 혈장 중 극미량 납 분석)

  • Lee, Sung-Bae;Yang, Jeong-Sun;Choi, Sung-Bong;Shin, Ho-Sang
    • Analytical Science and Technology
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    • v.25 no.3
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    • pp.190-196
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    • 2012
  • The human exposure of lead has usually detected the amount of lead in the whole blood, however, this method has a shortcoming to give the information on the short-term exposure to lead. In that sense, it is desirable to estimates the level of lead in plasma to draw the chronic bio-marker of lead exposure even though it is difficult to measure lead of several ng/L. An inductively coupled plasma-mass spectrometry (ICP-MS) method was developed for determining lead in plasma as the chronic bio-marker of lead of workers. To minimize the contamination of lead from the environment, we constructed class 1,000 clean room and compared the amount of floating dust before and after the operation of the clean room. The limit of detection (LOD) and the limit of quantification (LOQ) of lead in fetal bovine serum were 4.3 ng/L and 12.2 ng/L by NIOSH method (statistical calculation method) and 7.0 ng/L and 22.1 ng/L by signal/noise ratio, respectively. The accuracy was in a range of 92.3-101.3%, and the precision of the assay was less than 4% in the samples spiked in the concentration of 20 ng/L and 2,000 ng/L. The method was simple, reproducible and sensitive enough to permit reliable analysis of lead to the ng/L level in plasma and/or serum. The method was also useful for the biological monitoring of chronic exposure to lead.

BEAD BASED CHEMICAL REACTION SYSTEM USING TEMPERATURE AND FLUID CONTROL FOR CANCER DETECTION (유체와 온도 조절을 이용한 생화학 물질 반응용 마이크로칩의 개발)

  • Kim, Min-Su;Lee, Bo-Rahm;Yoon, Hyo-Jin;Kim, Byung-Gee;Lee, Yoon-Sik;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 2008.07a
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    • pp.1466-1467
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    • 2008
  • We describe here a novel micro total analysis system for the purification and identification of the affinity-captured proteins. Also we demonstrated the mass analysis of the Carcinoembrionic antigen (CEA) and Alpha femtoprotein which were chosen as the target cancer marker. For MALDI-TOF analyses, the proteins should to be separated from a protein mixture and be concentrated when needed. This procedure usually takes a long time even before protease-digested samples are to be obtained from them. Here, we describe integrated and efficient micro chip for protein purification and digestion for MALDI-TOF analyses. At first, disease protein is purified by passing the micro chamber from a protein mixture or human whole serum and released from the micro affinity beads by thermal heating. Purified protein is then transfer to the hole for trypsin digestion. The final sample is analyzed by MALDI-TOF. All the processes could be finished successfully within one hour, which renders MALDI-TOF analyses of a target protein quite simple.

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Study on the Applicability Analysis of HPLC for Fuel Marker (Unimark 1494DB) in Petroleum Products (석유제품의 식별제(Unimark 1494DB) 분석을 위한 HPLC 적용가능성 분석 연구)

  • Hwang, In-ha;Youn, Ju-min;Doe, Jin-woo;Park, Tae-seong;Kang, Hyung-kyu;Ha, Jong-han;Na, Byung-gi
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.4
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    • pp.1076-1084
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    • 2017
  • For analyzing the qualitative and quantitative analysis of fuel marker in petroleum products, the applicability of HPLC was studied. For the qualitative analysis of fuel marker in kerosene and automotive diesel, optimal analytical conditions(ratio of mobile phase solvent, flow rate, etc) in HPLC were selected and calibration curve for quantitative analysis of fuel marker was prepared based on the result of qualitative analysis. In particular, the correlation coefficient of calibration curve in kerosene and automotive diesel was shown to be 0.999 in a certain concentration range and it could be applied to the quantitative analysis. The results of analysis using the UV/Vis spectrometer, which is the current analysis method of fuel marker, were compared with the analysis results using the HPLC. The kerosene showed a low deviation of about 7 % and the automotive diesel showed a somewhat large deviation of about 20 %.

Emodin stimulates the osteoblast differentiation via activating bone morphogenetic protein-2 gene expression at low concentration

  • Cheon, Myeong-Sook;Lee, Su-Ui;Kim, Ho-Kyoung;Kim, Young-Sup;Min, Yong-Ki;Kim, Seong-Hwan
    • Korean Journal of Oriental Medicine
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    • v.13 no.1 s.19
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    • pp.139-145
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    • 2007
  • Emodin is one of the main active components of a traditional Korean medicine isolated from the root and rhizomes of Rheum palmatum L. In this study, of 222 natural compounds to evaluate the anabolic activities, emodin activated bone morphogenetic protein (BMP)-2 promoter in the differentiation process of mouse osteoblastic MC3T3-E1 cells. Emodin was shown to significantly stimulate the activity and expression of alkaline phosphatase, an earlyphase marker of osteoblastic differentiation, on the differentiation day 7, and induce the osteopontin mRNA expression from the differentiation day 14. In addition, low concentration (up to 5 M) of emodin dramatically promoted the induction of mineralization in MC3T3-E1 subclone 4 cells. The stimulatory effect of emodin on the osteoblast differentiation/mineralization could be associated with its potential to stimulate the BMP-2 gene expression. Although further studies are needed to determine the precise mechanism, this study suggests that the use of herbal medicine containing natural compounds with anabolic activity such as emodin could have a beneficial effect on bone health.

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Identification of quantitative trait loci for physical and chemical properties of rice grain

  • Cho, Yong-Gu;Kang, Hyeon-Jung;Lee, Young-Tae;Jong, Seung-Keun;Eun, Moo-Young;McCouch, Susan R.
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.61-73
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    • 2010
  • Quantitative trait loci (QTL) associated with six physical traits of cooked rice and seven chemical properties of rice grain were identified using a recombinant inbred (RI) population of rice evaluated over 3 years at the National Honam Agricultural Research Institute in Korea. The RI population consisted of 164 lines derived from a cross between Milyang23 and Gihobyeo, and the genetic map consisted of 414 molecular markers. A total of 49 QTL were identified for the 13 physico-chemical properties using composite interval mapping. Of these, 13 QTL were identified for 2 or more years, while 36 were detected in only 1 year. Five QTL were identified over all 3 years and will be useful for marker-assisted improvement of rice grain quality in Korea. The two QTL with the highest LOD scores, adhesiveness1.2 and potassium content7.1, provide a valuable starting point for positional cloning of genes underlying these QTL.

Isolation of RNA Aptamers Targeting HER-2-overexpressing Breast Cancer Cells Using Cell-SELEX

  • Kang, Hye-Suk;Huh, Yong-Min;Kim, So-Youn;Lee, Dong-ki
    • Bulletin of the Korean Chemical Society
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    • v.30 no.8
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    • pp.1827-1831
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    • 2009
  • Ligand molecules that can recognize and interact with cancer cell surface marker proteins with high affinity and specificity should greatly aid the development of novel cancer diagnostics and therapeutics. HER-2/ErbB2/Neu (HER-2), a member of the epidermal growth factor receptor family, is specifically overexpressed on the surface of breast cancer cells and serves as both a useful biomarker and a therapeutic target for breast cancer. In this study, we aimed to isolate RNA aptamers that specifically bind to a HER-2-overexpressing human breast cancer cell line, SK-BR-3, using Cell-SELEX strategy. The selected aptamers showed strong affinity to SK-BR-3, but not to MDAMB- 231, a HER-2-underexpressing breast cancer cell line. In addition, we confirmed the specific targeting of HER-2 receptor by aptamers using an unrelated mouse cell line overexpressing human HER-2 receptor. The HER-2-targeting RNA aptamers could become a useful reagent for the development of breast cancer diagnostics and therapeutics.

Heat or radiofrequency plasma glow discharge treatment of a titanium alloy stimulates osteoblast gene expression in the MC3T3 osteoprogenitor cell line

  • Rapuano, Bruce E.;Hackshaw, Kyle;Macdonald, Daniel E.
    • Journal of Periodontal and Implant Science
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    • v.42 no.3
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    • pp.95-104
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    • 2012
  • Purpose: The purpose of this study was to determine whether increasing the Ti6Al4V surface oxide negative charge through heat ($600^{\circ}C$) or radiofrequency plasma glow discharge (RFGD) pretreatment, with or without a subsequent coating with fibronectin, stimulated osteoblast gene marker expression in the MC3T3 osteoprogenitor cell line. Methods: Quantitative real-time polymerase chain reaction was used to measure changes over time in the mRNA levels for osteoblast gene markers, including alkaline phosphatase, bone sialoprotein, collagen type I (${\alpha}1$), osteocalcin, osteopontin and parathyroid hormone-related peptide (PTH-rP), and the osteoblast precursor genes Runx2 and osterix. Results: Osteoprogenitors began to differentiate earlier on disks that were pretreated with heat or RFGD. The pretreatments increased gene marker expression in the absence of a fibronectin coating. However, pretreatments increased osteoblast gene expression for fibronectin-coated disks more than uncoated disks, suggesting a surface oxide-mediated specific enhancement of fibronectin's bioactivity. Heat pretreatment had greater effects on the mRNA expression of genes for PTH-rP, alkaline phosphatase and osteocalcin while RFGD pretreatment had greater effects on osteopontin and bone sialoprotein gene expression. Conclusions: The results suggest that heat and RFGD pretreatments of the Ti6Al4V surface oxide stimulated osteoblast differentiation through an enhancement of (a) coated fibronectin's bioactivity and (b) the bioactivities of other serum or matrix proteins. The quantitative differences in the effects of the two pretreatments on osteoblast gene marker expression may have arisen from the unique physico-chemical characteristics of each resultant oxide surface. Therefore, engineering the Ti6Al4V surface oxide to become more negatively charged can be used to accelerate osteoblast differentiation through fibronectin-dependent and independent mechanisms.

Development of Fingerprints for Quality Control of Acorus species by Gas Chromatography/Mass Spectrometry

  • Yu, Se-Mi;Kim, Eun-Kyung;Lee, Je-Hyun;Lee, Kang-Ro;Hong, Jong-Ki
    • Bulletin of the Korean Chemical Society
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    • v.32 no.5
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    • pp.1547-1553
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    • 2011
  • An effective analytical method of gas chromatography/mass spectrometry (GC/MS) was developed for the rapid determination of essential oils in the crude extract of Acorus species (Acorus gramineus, Acorus tatarinowii, and Acorus calamus). Major phenypropanoids (${\beta}$,${\alpha}$-asarone isomers, euasarone, and methyleugenol) and ${\beta}$-caryophyllene in Acorus species were used as marker compounds and determined for the quality control of herbal medicines. To extract marker compounds, various extraction techniques such as solvent immersion, mechanical shaking, and sonication were compared, and the greatest efficiency was observed with sonication extraction using petroleum ether. The dynamic range of the GC/MS method depended on the specific analyte; acceptable quantification was obtained between 10 and 2000 ${\mu}g/mL$ for ${\beta}$-asarone, 10 and 500 ${\mu}g/mL$ for ${\alpha}$-asarone, 10 and 200 ${\mu}g/mL$ for methyleugenol, and between 5 and 100 ${\mu}g/mL$ for ${\beta}$-caryophyllene. The method was deemed satisfactory by inter- and intra-day validation and exhibited both high accuracy and precision, with a relative standard deviation < 10%. Overall limits of detection were approximately 0.34-0.83 ${\mu}g/mL$, with a standard deviation (${\sigma}$)-to-calibration slope (s) ratio (${\sigma}$/s) of 3. The limit of quantitation in our experiments was approximately 1.13-3.20 ${\mu}g/mL$ at a ${\sigma}$/s of 10. On the basement of method validation, 20 samples of Acorus species collected from markets in Korea were monitored for the quality control. In addition, principal component analysis (PCA) and hierarchical cluster analysis (HCA) were performed on the analytical data of 20 different Acorus species samples in order to classify samples that were collected from different regions.

Source Profile of Road Dust for Statistical Apportionment Modeling in Seoul (통계 수용모델을 위한 서울시 도로변 화학성분 원인 프로파일)

  • Park, Da-Jeong;Han, Young-Ji;Lee, Ji-Yi;Lee, Kwang-Yul;Cho, In-Hwan;Park, Eun Ha;Yi, Seung-Muk;Bae, Min-Suk
    • Journal of Korean Society for Atmospheric Environment
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    • v.31 no.2
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    • pp.105-117
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    • 2015
  • Sources related to road dust is one of the biggest sources, which is responsible for a large portion of emission. In particular, PM2.5 is a potential cause for respiratory diseases, thus it should be managed and a mitigation plan using results of statistical apportionment modeling such as chemical mass balance needs to be established. Recently, identifying sources of PM2.5 and analyzing the contribution of the road dust through a contribution assessment is required. Therefore, this study provides the chemical source profiles of PM2.5 using IC, GC/MS, OCEC, and XRF for both paved sidewalk and paved roadway collected at seven different sampling sites. As a result, for paved sidewalk, $NH{_4}^+$ (70%), $NO{_3}^-$ (12%), $PO{_4}^-$ (9%), and $SO{_4}^{2-}$ (9%) have been analyzed in PM2.5 mass. Major molecular marker such as Si has been indicated as $12.0{\pm}3.4%$ and $13.6{\pm}6.9%$ for paved sidewalk and roadway, respectively. PAHs such as Fluoranthene, Pyrene, Chrysene, and 1,3,5-Triphenylbenzene are suggested as molecular markers for road dust.