Lee Young-Keun;Chang Hwa-Hyoung;Jang Yu-Sin;Huh Jae-Ho;Hyung Seok-Won;Chung Hye-Young
Korean Journal of Environmental Biology
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v.22
no.3
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pp.472-477
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2004
To study the radiation related gene expression in mutants of Bacillus lentimorbus WJ5 induced by gamm radiation, the simultaneous gene expression was analyzed by DNA micro array. We constructed DNA chips including two thousand randomly digested genome spots of B. lentimorbus WJ5 and compared its quantitative aspect with seven mutants induced by gamma radiation $(^{60}/Co)$. From the cluster analysis of gene expression pattern, totally 408 genes were expressed and 27 genes were significantly upregulated by the gamma radiation in all mutants. Especially, genes involved in repair (mutL, mutM), energy metabolism (acsA, sdhB, pgk, yhjB, citB), protease (npr), and reduction response to oxidative stress (HMM) were simultaneously upregulated. It seems that the induction of the direct and/or indirect repair related genes in mutants induced by gamma radiation could be remarkably different from the adaptive responses against acute exposure to radiation.
The most important progress in diagnostic sciences is the increased sensitivity and specificity in diagnostic procedures due to the development of micromethodologies and increasing availability of immunological and molecular biological reagents. The technological advances led to consider the diagnostic use of saliva for an array of analytes and DNA source. The purpose of the present study was to compare DNA from saliva with those from blood and buccal swab, to evaluate diagnostic and forensic application of saliva, to investigate the changes of genomic DNA in saliva according to the storage temperature and period of saliva samples, and to evaluate the integrity of the DNA from saliva stored under various storage conditions by PCR analysis. Peripheral venous blood, unstimulated whole saliva, stimulated whole saliva, and buccal swab were obtained from healthy 10 subjects (mean age: $29.9{\pm}9.8$ years) and genomic DNA was extracted using commercial kit. For the study of effects of various storage conditions on genomic DNA from saliva, stimulated whole saliva were obtained from healthy 20 subjects (mean age: $32.3{\pm}6.6$ years). After making aliquots from fresh saliva, they were stored at room temperature, $4^{\circ}C$, $-20^{\circ}C$, and $-70^{\circ}C$. Saliva samples after lyophilization and dry-out procedure were stored at room temperature. After 1, 3, and 5 months, the same experiment was performed to investigate the changes in genomic DNA in saliva samples. In case of saliva aliquots stored at room temperature and dry-out samples, the results in 2 weeks were also included. Integrity of DNA from saliva stored under various storage conditions was also evaluated by PCR amplification analysis of $\beta$-globin gene fragments (989-bp). The results were as follows: 1. Concentration of genomic DNA extracted from saliva was lower than that from blood (p<0.05), but there were no significant differences among various types of saliva samples. Purities of genomic DNA extracted from stimulated whole saliva and lyophilized one were significantly higher than that from blood (p<0.05). Purity of genomic DNA extracted from buccal swab was lower than those from various types of saliva samples (p<0.05). 2. Concentration of genomic DNA from saliva stored at room temperature showed gradual reduction after 1 month, and decreased significantly in 3 and 5 months (p<0.05, p<0.01, respectively). Purities of DNA from saliva stored for 3 and 5 months showed significant differences with those of fresh saliva and stored saliva for 1 month (p<0.05). 3. In the case of saliva stored at $4^{\circ}C$ and $-20^{\circ}C$, there were no significant changes of concentration of genomic DNA in 3 months. Concentration of DNA decreased significantly in 5 months (p<0.05). 4. There were no significant differences of concentration of genomic DNA from saliva stored at $-70^{\circ}C$ and from lyophilized one according to storage period. Concentration of DNA showed decreasing tendency in 5 months. 5. Concentration of genomic DNA immediately extracted from saliva dried on Petri dish were 60% compared with that of fresh saliva. Concentration of DNA from saliva stored at room temperature after dry-out showed rapid reduction within 2 weeks (p<0.05). 6. Amplification of $\beta$-globin gene using PCR was successful in all lyophilized saliva stored for 5 months. At the time of 1 month, $\beta$-globin gene was successfully amplified in all saliva samples stored at $-20^{\circ}C$ and $-70^{\circ}C$, and in some saliva samples stored at $4^{\circ}C$. $\beta$-globin gene was failed to amplify in saliva stored at room temperature and dry-out saliva.
Proceedings of the Korean Society of Embryo Transfer Conference
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2002.11a
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pp.111-111
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2002
Stroke occurs when local thrombosis, embolic particle or the rupture of blood vessele interrupts the blood floe to the brain. $\beta$-estradiol 17-valerate has been reported to exert neuroprotective effects when administered before an ischemic insult. Recently, the pathophysiology of cerebral ischemia has been studied extensively in rat with various methods. In the present study, we investigates whether $\beta$-estrodiol 17-valerate can protect against brain injury. RNA sample were extracted from the hippocampus of female rat, reverse-transcription in the presence of [$\alpha$32p] dATP. Differential gene express-ion profiles were revealed (Bone morphogenetic protein type 1A receptor, Protein disulphide isomerase, Leukemia inhibitor factor receptor, cytochrome bc- 1 complex-x core P, thiol-specific antioxidant protein). RT-PCR was used to validate the relative expression pattern obtained by the cDNA array. The precise relationship between the early expression of recovery genes and stroke is a matter of luther investigation. This Study was supported by the Korea Science and Engineering Foundation(KOSEF) through the Biohealth Products Research Center(BPRC), Inje University, Korea.
Due to advances in omics technologies, numerous genome-wide studies on human samples have been published, and most of the omics data with the associated clinical information are available in public repositories, such as Gene Expression Omnibus and ArrayExpress. While analyzing several public datasets, we observed that errors in gender information occur quite often in public datasets. When we analyzed the gender description and the methylation patterns of gender-specific probes (glucose-6-phosphate dehydrogenase [G6PD], ephrin-B1 [EFNB1], and testis specific protein, Y-linked 2 [TSPY2]) in 5,611 samples produced using Infinium 450K HumanMethylation arrays, we found that 19 samples from 7 datasets were erroneously described. We also analyzed 1,819 samples produced using the Affymetrix U133Plus2 array using several gender-specific genes (X (inactive)-specific transcript [XIST], eukaryotic translation initiation factor 1A, Y-linked [EIF1AY], and DEAD [Asp-Glu-Ala-Asp] box polypeptide 3, Y-linked [DDDX3Y]) and found that 40 samples from 3 datasets were erroneously described. We suggest that the users of public datasets should not expect that the data are error-free and, whenever possible, that they should check the consistency of the data.
Proceedings of the Korean Society for Bioinformatics Conference
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2006.02a
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pp.30-36
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2006
Tissue microarray (TMA) is an array-based technology allowing the examination of hundreds of tissue samples on a single slide. To handle, exchange, and disseminate TMA data, we need standard representations of the methods used, of the data generated, and of the clinical and histopathological information related to TMA data analysis. This study aims to create a comprehensive data model with flexibility that supports diverse experimental designs and with expressivity and extensibility that enables an adequate and comprehensive description of new clinical and histopathological data elements. We designed a Tissue Microarray Object Model (TMA-OM). Both the Array Information and the Experimental Procedure models are created by referring to Microarray Gene Expression Object Model, Minimum Information Specification For In Situ Hybridization and Immunohistochemistry Experiments (MISFISHIE), and the TMA Data Exchange Specifications (TMA DES). The Clinical and Histopathological Information model is created by using CAP Cancer Protocols and National Cancer Institute Common Data Elements (NCI CDEs). MGED Ontology, UMLS and the terms extracted from CAP Cancer Protocols and NCI CDEs are used to create a controlled vocabulary for unambiguous annotation. We implemented a web-based application for TMA-OM, supporting data export in XML format conforming to the TMA DES or the DTD derived from TMA-OM. TMA-OM provides a comprehensive data model for storage, analysis and exchange of TMA data and facilitates model-level integration of other biological models.
Seo, Hee-Won;Choi, Jin-Won;Yun, Tae-Won;Lee, Hong-Jo;Kim, Hee-Seung;Song, Yong-Sang;Song, Gwon-Hwa;Han, Jae-Yong
Journal of Animal Science and Technology
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v.52
no.6
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pp.491-498
/
2010
Copy-number variation (CNV) in particular genomic segments owing to deletions or duplications can induce changes in cellular gene expression patterns and may increase susceptibility to diseases such as cancer. The aim of this study was to examine CNVs related to the incidence of epithelial ovarian cancer in chickens. Genomic DNA was extracted from blood cells and cancerous ovaries collected from four 120-week-old White Leghorn chickens and were used for array-based comparative genome hybridization (CGH) analysis. As a result, 25 amplified and 10 deleted CNV regions were detected in chicken ovarian cancer. Of these, 10 amplified and two deleted CNV regions contained genes associated with human ovarian cancer. Our study using a chicken model may provide a better understanding of human epithelial ovarian cancer.
Murine Neuro-2a (N2a) cells have been widely used for the investigation of neuronal differentiation, trophic interaction and neurotoxic effects of various compounds and their associated mechanisms. N2a cells have many genomic variations such as gains or losses in DNA copy number, similar to other neuroblastoma cells, and no systematic or high-resolution studies of their genome-wide chromosomal aberrations have been reported. Presently, we conducted a systematic genome-wide determination of chromosomal aberrations in N2a cells using a high-throughput, oligonucleotide array-based comparative genomic hybridization (oaCGH) technique. A hidden Markov Model was employed to assign each genomic oligonucleotide to a DNA copy number state: double loss, single loss, normal, gain, double gain and amplification. Unlike most neuroblastoma cells, Mycn amplification was not observed in N2a cells. In addition, these cells showed gain only in the neuron-derived neurotrophic factor (NF), while other neurotrophic factors such as glial line-derived NF and brain-derived NF presented normal copy numbers. Chromosomes 4, 8, 10, 11 and 15 displayed more than 1000 aberrational oligonucleotides, while chromosomes 3, 17, 18 and 19 displayed less than 20. The largest region of gain was located on chromosome 8 and its size was no less than 26.7 Mb (Chr8:8427841-35162415), while chromosome 4 had the longest region of single deletion, with a size of 15.1 Mb (Chr4:73265785-88374165).
The purpose of this study was to characterize functional distinction between human dental pulp cells(PC) and periodontal ligament cells(PDLC) using cDNA micro array assay and to confirm the results of the microarray assay using RT-PCR. 3 genes out of 51 genes which were found to be more expressed(>2 fold) in PC were selected, and 3 genes out of 19 genes which were found to be more expressed(>2 fold) in PDLC were selected for RT-PCR as well. According to this study, the results were as follows: 1. From the micro array assay, 51 genes were more expressed (2 fold) from PC than PDLC. 2. RT-PCR confirmed that ITGA4 and TGF ${\beta}2$ were more expressed in PC than in PDLC 3. From the micro array assay, 19 genes were more expressed (2 fold) from PDLC than PC. 4. RT-PCR confirmed that LUM, WISP1. and MMP1 were more expressed in PDLC than in PC. From the present study, different expression of the genes between the PC and PDLC were characterized to show the genes which play an important role in dentinogenesis were more expressed from PC than PDLC, while the genes which were related with collagen synthesis were more expressed from PDLC than PC.
Lin, Tzu-Kang;Yu, Li-Chen;Ku, Chang-Hung;Chang, Kuo-Chun;Kiremidjian, Anne
Smart Structures and Systems
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v.8
no.1
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pp.119-137
/
2011
A bio-inspired two-mode structural health monitoring (SHM) system based on the Na$\ddot{i}$ve Bayes (NB) classification method is discussed in this paper. To implement the molecular biology based Deoxyribonucleic acid (DNA) array concept in structural health monitoring, which has been demonstrated to be superior in disease detection, two types of array expression data have been proposed for the development of the SHM algorithm. For the micro-vibration mode, a two-tier auto-regression with exogenous (AR-ARX) process is used to extract the expression array from the recorded structural time history while an ARX process is applied for the analysis of the earthquake mode. The health condition of the structure is then determined using the NB classification method. In addition, the union concept in probability is used to improve the accuracy of the system. To verify the performance and reliability of the SHM algorithm, a downscaled eight-storey steel building located at the shaking table of the National Center for Research on Earthquake Engineering (NCREE) was used as the benchmark structure. The structural response from different damage levels and locations was collected and incorporated in the database to aid the structural health monitoring process. Preliminary verification has demonstrated that the structure health condition can be precisely detected by the proposed algorithm. To implement the developed SHM system in a practical application, a SHM prototype consisting of the input sensing module, the transmission module, and the SHM platform was developed. The vibration data were first measured by the deployed sensor, and subsequently the SHM mode corresponding to the desired excitation is chosen automatically to quickly evaluate the health condition of the structure. Test results from the ambient vibration and shaking table test showed that the condition and location of the benchmark structure damage can be successfully detected by the proposed SHM prototype system, and the information is instantaneously transmitted to a remote server to facilitate real-time monitoring. Implementing the bio-inspired two-mode SHM practically has been successfully demonstrated.
Bisphenol A (BPA) is commonly used in the production of pharmaceutical, industrial, and housing epoxy, as well as polycarbonate plastics. Owing to its extensive use, BPA can contaminate the environment either directly or through derivatives of these products. BPA has been classified as an endocrine disruptor chemicals (EDCs), and the primary toxicity of these EDCs in males involves the induction of reproductive system abnormality. First, in order to evaluate the direct effects on the Y chromosome associated with reproduction, we evaluated Y chromosome abnormalities using a Y chromosome microdeletion detection kit. However, we detected no Yq abnormality as the result of BPA exposure. Secondly, we performed high-density oligonucleotide array-based comparative genome hybridization (CGH) to assess genomic alteration as a component of our toxicity assessment. The results of our data analysis revealed some changes in copy number. Seven observed features were gains or losses in chromosomal DNA (P-value<1.0e-5, average log2 ratio>0.2). Interestingly, 21 probes of chr7:7312289-10272836 (qA1-qA2 in cytoband) were a commonly observed amplification (P-value 3.69e-10). Another region, chr14:4551029-10397399, was also commonly amplified (P-value 2.93e-12, average of log2 ratios in segment>0.3786). These regions include many genes associated with pheromone response, transcription, and signal transduction using ArrayToKegg software. These results help us to understand the molecular mechanisms underlying the reproductive effects induced by BPA.
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