• Title/Summary/Keyword: DNA dyes

Search Result 31, Processing Time 0.029 seconds

DNA Damage in Lymphocytes after Hair Dyeing and Related Factors among Women Volunteers (일부 자원 여성에서 모발염색 후 림프구의 DNA손상과 관련 요인)

  • Cho, Jin-A;Oh, Eun-Ha;Sull, Dong-Geun;Lee, Eun-Il
    • Journal of Preventive Medicine and Public Health
    • /
    • v.35 no.4
    • /
    • pp.275-281
    • /
    • 2002
  • Objectives : To evaluate the DNA damage by hair dyeing in human lymphocytes Methods : Comet assays were carried out to evaluate the DNA damage in lymphocytes by hair dyeing. Twenty subjects were selected from women volunteers whose age ranged from 55 to 67 year old. All subjects had no smoking history. Blood samples were collected before and 6 hours after hair dyeing. DNA damage was evaluated by means of the tail moments, which were quantified by a KOMET 4.0 image analysis system. Results : The tail moments before hair dyeing showed no significant differences among subjects except for the high frequency group. The mean values of the tail moments in subjects with low and high frequencies of hair dyeing were 1.39 and 1.77, respectively (p<0.05). The tail moments after hair dyeing increased significantly, The mean values of tail moments in subjects before and after hair dyeing were 1.45 and 1.79, respectively (p<0.01). However, the difference levels of DNA damage in lymphocytes before and after hair dyeing were found to be slightly lower in both the dietary supplement taking group and high frequency group. Conclusions : The high frequency group appears to have a higher level of DNA damage than the low frequency group before hair dyeing. DNA damage in lymphocytes was found to be significantly higher in the volunteers after hair dyeing. In this study, the related factors such as high frequency and taking dietary supplements appeard to reduce DNA damage in lymphocytes after hair dyeing.

Identification of Clostridium perfringens AB&J and Its Uptake of Bromophenol Blue

  • Kim, Jeong-Dong;An, Hwa-Yong;Yoon, Jung-Hoon;Park, Yong-Ha;Fusako Kawai;Jung, Chang-Min;Kang, Kook_-Hee
    • Journal of Microbiology and Biotechnology
    • /
    • v.12 no.4
    • /
    • pp.544-552
    • /
    • 2002
  • Several microorganisms from rat and human feces and lumen fluid of cows were screened for their ability to decolorize the synthetic dyes. Consequently, a novel dye-degrading strain AB&J was isolated. Taxonomic identification including 165 rDNA sequencing and phylogenetic analysis indicated that the isolate had 99.9% homology in its 165 rDNA base sequence with Clostridium perfringens. After 27 h Incubation with the strain, brilliant blue R, bromophenol blue, crystal violet, malachite green, methyl green, and methyl orange were decolorized by about 69.3%, 97.7%, 96.3%, 97.9%, 75.1%, and 97.2%, respectively. The triphenlmethane dye, bromophenol blue, was decolorized extensively by growing Clostridium perfringens AB&J cells in liquid cultures under anaerobic condition, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly decolorized at a relatively lower concentration of below 50 $\mu g \;ml^{-1}$, however, the growth of the cells was mostly suppressed at a dye concentration of 100 $\mu g \;ml^{-1}$. The decolorization activity in cell-free extracts was much higher in cytoplasm than in periplasm and cytoplasmic membrane. Therefore, the enzyme related uptake of bromophenol blue seemed to be localized in cytoplasm. The optimal pH and temperature of bromophenol blue uptake fur decolorization activities were 7.0 and 4$0^{\circ}C$, respectively.

Identification of Stenotrophomonas maltophilia LK-24 and its Degradability of Crystal Violet

  • Kim, Jeong-Dong;Yoon, Jung-Hoon;Park, Yong-Ha;Fusako Kawai;Kim, Hyun-Tae;Lee, Dae-Weon;Kang, Kook-Hee
    • Journal of Microbiology and Biotechnology
    • /
    • v.12 no.3
    • /
    • pp.437-443
    • /
    • 2002
  • A number of soil and wastewater samples were collected from the vicinity of an effluent treatment plant for the chemical industry. Several microorganisms were screened fur their ability to decolorize the triphenylmethane group of dyes. As a result, a novel crystal violet dye-degrading strain LK-24 was isolated. Taxonomic identification including 16S rDNA sequencing and phylogenetic analysis indicated that the isolate had a $99.5\%$ homology in its 16S rDNA base sequence with Stenotrophomonas maltophilia. The triphenylmethane dye, crystal violet, was degraded extensively by growing cells of Stenotrophomonas maltophilia LK-24 in agitated liquid cultures, although their growth was strongly inhibited in the initial stage of incubation. This group of dyes is toxic, depending on the concentration used. The dye was significantly degraded at a relatively lower concentration, below $100{\mu}g\;ml^-1$, yet the growth of the cells was totally suppressed at a dye concentration of $250{\mu}g\;ml^-1$. The degradation products of crystal violet were identified as 4,4'-bis(dimethylamino)-benzophenone and ${\rho}$-dimethylaminophenol by Gas chromatography-Mass spectrometry. The 4,4'-bis(dimethylamino)-benzophenone was easily obtained in a reasonable yield, as it was not metabolized further by S. maltophilia LK-24; however, the ${\rho}$-dimethylaminophenol was not easily identifiable, as it was further metabolized.

Optimization of Anthraquinone Dyes Decolorization Conditions with Response Surface Methodology by Aspergillus

  • Ge, Yufeng;Wei, Bin;Wang, Siyu;Guo, Zhiguo;Xu, Xiaolin
    • Korean Chemical Engineering Research
    • /
    • v.53 no.3
    • /
    • pp.327-332
    • /
    • 2015
  • A large amount of dye wastewater poses a threat to environmental safety. Disperse blue, an anthraquinone dye that is widely used in textile dyes, is difficult to degrade in wastewater. In this work, one fungus was screened according to the decolorization rate of disperse blue. The fungus was identified and named Aspergillus XJ-2 on the basis of its morphological characteristics and 18s rDNA. Response surface method was used to optimize culture conditions for A. XJ-2. The optimum values of obtained responses were as follows: temperature, $35^{\circ}C$; pH, 5.2; carbon-to-nitrogen ratio, 30:5.5; and rotation ratio, $175r{\cdot}min^{-1}$. Under optimized conditions, the decolorization rate of A. XJ-2 was up to 94.8% in 48 h.

New Normalization Methods using Support Vector Machine Regression Approach in cDNA Microarray Analysis

  • Sohn, In-Suk;Kim, Su-Jong;Hwang, Chang-Ha;Lee, Jae-Won
    • Proceedings of the Korean Society for Bioinformatics Conference
    • /
    • 2005.09a
    • /
    • pp.51-56
    • /
    • 2005
  • There are many sources of systematic variations in cDNA microarray experiments which affect the measured gene expression levels like differences in labeling efficiency between the two fluorescent dyes. Print-tip lowess normalization is used in situations where dye biases can depend on spot overall intensity and/or spatial location within the array. However, print-tip lowess normalization performs poorly in situation where error variability for each gene is heterogeneous over intensity ranges. We proposed the new print-tip normalization methods based on support vector machine regression(SVMR) and support vector machine quantile regression(SVMQR). SVMQR was derived by employing the basic principle of support vector machine (SVM) for the estimation of the linear and nonlinear quantile regressions. We applied our proposed methods to previous cDNA micro array data of apolipoprotein-AI-knockout (apoAI-KO) mice, diet-induced obese mice, and genistein-fed obese mice. From our statistical analysis, we found that the proposed methods perform better than the existing print-tip lowess normalization method.

  • PDF

Simple Method to Correct Gene-Specific Dye Bias from Partial Dye Swap Information of a DNA Microarray Experiment

  • KIM BYUNG SOO;KANG SOO-JIN;LEE SAET-BYUL;HWANG WON;KIM KUN-SOO
    • Journal of Microbiology and Biotechnology
    • /
    • v.15 no.6
    • /
    • pp.1377-1383
    • /
    • 2005
  • In a cDNA microarray experiment using Cy3 and Cy5 as labeling agents, particularly for the direct design, cDNAs from some genes incorporate one dye more efficiently than the other, which is referred to as the gene-specific dye bias. Dye-swaps, in which two dyes are switched on replicate arrays, are commonly used to control the gene-specific dye bias. We developed a simple procedure to extract the gene-specific dye bias information from a partial dye swap experiment. We detected gene-specific dye bias by identifying outliers in an X-Y plane, where the X axis represents the average log-ratio from two sets of dye swap pairs and the Y axis exhibits the average log ratio of four forward labeled arrays. We used this information for detecting differentially expressed genes, of which the additionally detected genes were validated by real-time RT-PCR.

Depurination of Nucleosides and Calf Thymus DNA Induced by 2-Bromopropane at the Physiological Condition

  • Sherchan, Jyoti;Choi, Ho-Young;Lee, Eung-Seok
    • Bulletin of the Korean Chemical Society
    • /
    • v.30 no.10
    • /
    • pp.2309-2317
    • /
    • 2009
  • Depurination, the release of purine bases from nucleic acids by hydrolysis of the N-glycosidic bond, gives rise to alterations of the cell genome. Though cells have evolved mechanisms to repair these lesions, unrepaired apurinic sites have been shown to have two biological consequences: lethality and base substitution errors. 2-Bromopropane (2-BP) is used as an intermediate in the synthesis of pharmaceuticals, dyes, and other organics. In addition, 2-BP has been used as a replacement for chloroflurocarbons and 1,1,1-trichloroethane as a cleaning solvent in electronics industry. However, 2-BP was found to cause reproductive and hematopoietic disorders in local workers exposed to it. Owing to the toxicity of 2-BP, there has been a tendency to use 1-BP as an alternative cleaning solvent to 2-BP. However, 1-BP has also been reported to be neurotoxic in rats. Though $N^7$-guanine adduct of 2-BP has been reported previously, massive depurination of the nucleosides and calf thymus DNA was observed in this study. We incubated the nucleosides (ddG, dG, guanosine, ddA, dA and adenosine) with excess amount 2-BP at the physiological condition (pH 7.4, $37\;{^{\circ}C}$), which were analyzed by HPLC and LC-MS/MS. In addition, the time and dose response relationship of depurination in nucleosides induced by 2-bromopropane at the physiological condition was investigated. Similarly, incubation of calf-thymus DNA with the excess amount 2-BP at the physiological condition was also performed. In addition, the time and dose response relationship of depurination in calf-thymus DNA induced by 2-BP at the physiological condition was investigated. Those results suggest that the toxic effect of 2-BP could be both from the depurination of nucleosides and DNA adduct formation.

Effect of missing values in detecting differentially expressed genes in a cDNA microarray experiment

  • Kim, Byung-Soo;Rha, Sun-Young
    • Bioinformatics and Biosystems
    • /
    • v.1 no.1
    • /
    • pp.67-72
    • /
    • 2006
  • The aim of this paper is to discuss the effect of missing values in detecting differentially expressed genes in a cDNA microarray experiment in the context of a one sample problem. We conducted a cDNA micro array experiment to detect differentially expressed genes for the metastasis of colorectal cancer based on twenty patients who underwent liver resection due to liver metastasis from colorectal cancer. Total RNAs from metastatic liver tumor and adjacent normal liver tissue from a single patient were labeled with cy5 and cy3, respectively, and competitively hybridized to a cDNA microarray with 7775 human genes. We used $M=log_2(R/G)$ for the signal evaluation, where Rand G denoted the fluorescent intensities of Cy5 and Cy3 dyes, respectively. The statistical problem comprises a one sample test of testing E(M)=0 for each gene and involves multiple tests. The twenty cDNA microarray data would comprise a matrix of dimension 7775 by 20, if there were no missing values. However, missing values occur for various reasons. For each gene, the no missing proportion (NMP) was defined to be the proportion of non-missing values out of twenty. In detecting differentially expressed (DE) genes, we used the genes whose NMP is greater than or equal to 0.4 and then sequentially increased NMP by 0.1 for investigating its effect on the detection of DE genes. For each fixed NMP, we imputed the missing values with K-nearest neighbor method (K=10) and applied the nonparametric t-test of Dudoit et al. (2002), SAM by Tusher et al. (2001) and empirical Bayes procedure by $L\ddot{o}nnstedt$ and Speed (2002) to find out the effect of missing values in the final outcome. These three procedures yielded substantially agreeable result in detecting DE genes. Of these three procedures we used SAM for exploring the acceptable NMP level. The result showed that the optimum no missing proportion (NMP) found in this data set turned out to be 80%. It is more desirable to find the optimum level of NMP for each data set by applying the method described in this note, when the plot of (NMP, Number of overlapping genes) shows a turning point.

  • PDF

Lineage Tracing: Computational Reconstruction Goes Beyond the Limit of Imaging

  • Wu, Szu-Hsien (Sam);Lee, Ji-Hyun;Koo, Bon-Kyoung
    • Molecules and Cells
    • /
    • v.42 no.2
    • /
    • pp.104-112
    • /
    • 2019
  • Tracking the fate of individual cells and their progeny through lineage tracing has been widely used to investigate various biological processes including embryonic development, homeostatic tissue turnover, and stem cell function in regeneration and disease. Conventional lineage tracing involves the marking of cells either with dyes or nucleoside analogues or genetic marking with fluorescent and/or colorimetric protein reporters. Both are imaging-based approaches that have played a crucial role in the field of developmental biology as well as adult stem cell biology. However, imaging-based lineage tracing approaches are limited by their scalability and the lack of molecular information underlying fate transitions. Recently, computational biology approaches have been combined with diverse tracing methods to overcome these limitations and so provide high-order scalability and a wealth of molecular information. In this review, we will introduce such novel computational methods, starting from single-cell RNA sequencing-based lineage analysis to DNA barcoding or genetic scar analysis. These novel approaches are complementary to conventional imaging-based approaches and enable us to study the lineage relationships of numerous cell types during vertebrate, and in particular human, development and disease.

Effects of Boshimgeonbi-tang on Gene Expression in Hypothalamus of Immobilization-stressed Mouse (보심건비탕(補心健脾湯) 투여가 Stress 유발 Mouse의 Hypothalamus 유전자 발현에 미치는 영향)

  • Lee Seoung-Hee;Chang Gyu-Tae;Kim Jang-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
    • /
    • v.19 no.6
    • /
    • pp.1585-1593
    • /
    • 2005
  • The genetic effects of restraint stress challenge on HPA axis and the therapeutic effect of Boshimgeonbi-tang on the stress were studied with cDNA microarray analyses, RT-PCR on hypothalamus using an immobilization-stress mice as an animal model. Male CD-1 mice were restrained in a tightly fitted and ventilated vinyl holder for 2hrs once a day, and this challenge was repeated for seven· consecutive days. In the change of body weight it showed that the Boshimgeonbi-tang is effected recovery on weight loss caused by the immobilization-stress. Seven days later, total RNA was extracted from the organs of the mouse, body-labeled with $CyDye^{TM}$ fluorescence dyes and then hybridized to CDNA microarray chip. Scanning and analyzing the array slides were carried out using GenePix4000 series scanner and GenePix $Pro^{TM}$ analyzing program, respectively. The expression profiles of 109 genes out of 6000 genes on the chip were significantly modulated in hypothalamus by the immobilization stress. Energy metabolism-, lipid metabolism-, apoptosis-, stress protein, transcriptional factor, and signal transduction-related genes were transcriptionally activated whereas DNA repair-, protein biosysthesis-, and structure integrity-related genes were down-regulated in hypothalamus. The 58 genes were up-regulated by the mRNA expression folds of 1.5 to 7.9. and the 51 genes were down-regulated by 1.5 - 5.5 fold. The 11 genes among them were selected to confirm the expression profiles by RT-PCR. The mRNA expression levels of Tnfrsf1a (apoptosis), Calm2 (cell cycle), Bag3 (apoptosis), Ogg1 (DNA repair), Aatk (apoptosis), Dffa (apoptosis), Fkbp5 (protein folding) were restored to the normal one by the treatment of Boshimgeonbi-tang.