• IMMUNE NETWORK
• /
• v.14 no.1
• /
• pp.54-65
• /
• 2014

Bone marrow-derived mesenchymal stem cells (MSCs) are multipotent, with the ability to differentiate into different cell types. Additionally, the immunomodulatory activity of MSCs can downregulate inflammatory responses. The use of MSCs to repair injured tissues and treat inflammation, including in neuroimmune diseases, has been extensively explored. Although MSCs have emerged as a promising resource for the treatment of neuroimmune diseases, attempts to define the molecular properties of MSCs have been limited by the heterogeneity of MSC populations. We recently developed a new method, the subfractionation culturing method, to isolate homogeneous human clonal MSCs (hcMSCs). The hcMSCs were able to differentiate into fat, cartilage, bone, neuroglia, and liver cell types. In this study, to better understand the properties of neurally differentiated MSCs, gene expression in highly homogeneous hcMSCs was analyzed. Neural differentiation of hcMSCs was induced for 14 days. Thereafter, RNA and genomic DNA was isolated and subjected to microarray analysis and DNA methylation array analysis, respectively. We correlated the transcriptome of hcMSCs during neural differentiation with the DNA methylation status. Here, we describe and discuss the gene expression profile of neurally differentiated hcMSCs. These findings will expand our understanding of the molecular properties of MSCs and contribute to the development of cell therapy for neuroimmune diseases.

• Journal of Korean Society on Water Environment
• /
• v.29 no.2
• /
• pp.232-238
• /
• 2013

Water temperature is key factor influencing growth and reproduction of fish and its increase give rise to various physiological changes including gene expression. Heat shock protein (Hsp), one of the molecular chaperones, is highly conserved throughout evolution and its expression is induced by various stressors such as temperature, oxidative, physical and chemical stresses. Here, we isolated partial cDNA clones encoding 70-kDa Hsp (Hsp70) and $\beta$-actin using reverse transcriptase-PCR (RT-PCR) from gut of Rhynchocypris kumgangensis, a Korean indigenous species and cold-water fish, and investigated expression profiles of Hsp70 under an increase of water temperature using $\beta$-actin as an internal control for RT-PCR. Cloned Hsp70 cDNA of R. kumgangensis showed homology to Ctenopharyngodon idella (96%), Hypophthalmichthys molitrix (96%), Danio rerio (93%) and Oncorhynchus mykiss (81%) Hsp70. Cloned $\beta$-actin cDNA of R. kumgangensis showed homology to D. rerio (98%), H. molitrix (97%), C. idella (97%) and O. mykiss (90%) $\beta$-actin. Both mRNA of Hsp70 and $\beta$-actin were expressed in gut, brain, and liver in R. kumgangensis. Futhermore, expression of Hsp70, in brain, was highly augmented by an increase of water temperature. These results suggest that Hsp70 mRNA expression level in brain can be used as a biological molecular marker to represent physiological stress against an increase of water temperature.

• Proceedings of the Korea Society of Environmental Toocicology Conference
• /
• 2003.05a
• /
• pp.187-187
• /
• 2003

Methylmercury (MeHg), one of the heavy metal compounds, can cause severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. In this study, two methods, cDNA Microarray and SSH, were performed to assess the expression profile against MeHg and to identify differentially expressed genes by MeHg in neuroblastoma cell line. TwinChip Human-8K (Digital Genomics) was used with total RNA from SH-SY5Y (human neuroblastoma cell line) treated with solvent (DMSO) and 6.25 uM (IC50) MeHg. And we performed forward and reverse SSH method on mRNA derived from SH-SY5Y treated with DMSO and MeHg (6.25 uM). Differentially expressed cDNA clones were sequenced and were screened by dot blot and ribonuclease protection assay to confirm that individual clones indeed represent differentially expressed genes. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences may provide an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as environmental pollutants.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.12
• /
• pp.1336-1344
• /
• 2011

This study details and examines a novel ligation-mediated polymerase chain reaction (LM-PCR) method. Named the LM-PCR/Shifter, it relies on the use of a Class IIS restriction enzyme giving restriction fragments with different 4-base, 5' overhangs, this being the Shifter, and the ligation of appropriate oligonucleotide adapters. A sequence of 4-base, 5' overhangs of the adapter and a 4-base sequence of the 3' end of the primer(s) determine a subset of the genomic restriction fragments, which are amplified by PCR. The method permits the differentiation of bacterial species strains on the basis of the different DNA band patterns obtained after electrophoresis in polyacrylamide gels stained with ethidium bromide and visualized in UV light. The usefulness of the LM-PCR/Shifter method for genotyping is analyzed by a comparison with the restriction endonuclease analysis of chromosomal DNA by the pulsed-field gel electrophoresis (REA-PFGE) and PCR melting profile (PCR MP) methods for isolates of clinical origin. The clustering of the LM-PCR/Shifter fingerprinting data matched those of the REA-PFGE and PCR MP methods. We found that the LM-PCR/Shifter is rapid, and offers good discriminatory power and excellent reproducibility, making it a method that may be effectively applied in epidemiological studies.

• Journal of Microbiology and Biotechnology
• /
• v.12 no.1
• /
• pp.71-76
• /
• 2002

Soil samples were screened for actinomycete strains capable of producing phospholipase D, and a strain, Streptomyces sp. YU100, showing a high transphosphatidylation activity was isolated. This strain secreted phospholipase D in a culture broth after 12 h of cultivation, and its productivity continued to increase for 36 h of fermentation. In addition, its transphosphatidylation rate of phosphatidylcholine to phosphatidylserine was almost $68\%$ within 1 h. The morphological and chemotaxonomical characteristics showed that this strain could be classified as a number of the Streptomycetaceae family, particularly due to the spiral form of its spore chain consisting of 60-70 smooth spores $(0.75{\times}1.0{\mu}m$) on an aerial mycelium, FA-2c type of fatty acid profile in the cell wall, and LL-DAP component in the cell wall peptidoglycan. A phylogenetic analysis of the 16S rDNA provided a clue that the strain YU100 was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyes sp. ASB27, S. peucetius JCM9920, and S. griseus ATCC10137. A dendrogram based on the 16S rDNA sequences also showed a phylogenetic relationship between the strain YU100 and these strains. However, the strain YU100 has not yet been assigned to a particular species, because of absence of any other classified species with a high matching score.

• Molecular & Cellular Toxicology
• /
• v.3 no.3
• /
• pp.165-171
• /
• 2007

Mitomycin C (MMC), an antitumor antibiotic isolated from Streptomyces caespitosus, is used in chemotherapy of gastric, bladder and colorectal cancer. MMC is activated in vivo to alkylate and crosslink DNA, via G-G interstrand bonds, thereby inhibiting DNA synthesis and transcription. This study investigates gene expression changes in response to MMC treatment in order to elucidate the mechanisms of MMC-induced toxicity. MMC was admistered with single dose (0.32 and 1.6 ${\mu}M$) to TK6 cells. Applied Biosystem's DNA chips were used for identifying the gene expression profile by MMC-induced toxicity. We identified up- or down-regulated 90 genes including cyclin M2, cyclin-dependent kinase inhibitor 1A (p21, cip1), programmed cell death 1, tumor necrosis factor (ligand) superfamily, member 9, et al. The regulated genes by MMC associated with the biological pathways apoptosis signaling pathway. Further characterization of these candidate markers related to the toxicity will be useful to understand the detailed mechanism of action of MMC.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.8
• /
• pp.3543-3549
• /
• 2015

Background: Bladder cancer is one of the most common cancers worldwide. Gene expression profiling using microarray technologies improves the understanding of cancer biology. The aim of this study was to determine the gene expression profile in Egyptian bladder cancer patients. Materials and Methods: Samples from 29 human bladder cancers and adjacent non-neoplastic tissues were analyzed by cDNA microarray, with hierarchical clustering and multidimensional analysis. Results: Five hundred and sixteen genes were differentially expressed of which SOS1, HDAC2, PLXNC1, GTSE1, ULK2, IRS2, ABCA12, TOP3A, HES1, and SRP68 genes were involved in 33 different pathways. The most frequently detected genes were: SOS1 in 20 different pathways; HDAC2 in 5 different pathways; IRS2 in 3 different pathways. There were 388 down-regulated genes. PLCB2 was involved in 11 different pathways, MDM2 in 9 pathways, FZD4 in 5 pathways, p15 and FGF12 in 4 pathways, POLE2 in 3 pathways, and MCM4 and POLR2E in 2 pathways. Thirty genes showed significant differences between transitional cell cancer (TCC) and squamous cell cancer (SCC) samples. Unsupervised cluster analysis of DNA microarray data revealed a clear distinction between low and high grade tumors. In addition 26 genes showed significant differences between low and high tumor stages, including fragile histidine triad, Ras and sialyltransferase 8 (alpha) and 16 showed significant differences between low and high tumor grades, like methionine adenosyl transferase II, beta. Conclusions: The present study identified some genes, that can be used as molecular biomarkers or target genes in Egyptian bladder cancer patients.

• Asian-Australasian Journal of Animal Sciences
• /
• v.18 no.8
• /
• pp.1080-1087
• /
• 2005

By screening specific genes related to the muscle growth of swine using cDNA microarray technology, a total of 5 novel genes (GF (growth factor) I, II, III, IV and V) were identified. Results of southern blotting to investigate the number of copies of these genes in the genome of swine indicated that GF I, GF III, and GF V existed as one copy and GF II, and GF IV existed as more than two copies. It was suggested that there are many isoforms of these genes in the genome of swine. Also, results of northern blotting to investigate whether these genes were expressed in grown muscle, using GF I, III, and V indicated that all the genes were much more expressed in the muscle of swine with body weight of 90 kg. Expression patterns of these genes in other organs, namely muscle and propagation and fat tissues, were investigated by extracting RNA from the tissues. These genes were not expressed in the propagation and fat tissues, but were expressed in the muscle tissue. To determine the mechanism of muscle growth, further studies should be preceded using the 3 specific genes related to muscle growth, that is GF I, III, and V.

• Proceedings of the Korean Society of Organic Agriculture Conference
• /
• 2009.12a
• /
• pp.302-302
• /
• 2009

유기농업에서 유기물은 양분의 공급, 토양의 이화학성 개선, 토양의 생물학적 건전성 유지 등 중요한 역할을 한다. 토양의 생물학적 건전성은 토양의 생태계적 기능을 지속적으로 유지시키는 토양미생물이 관여하고 있다. 따라서 본 연구는 유기물의 장기연용에 따른 밭토양 미생물의 다양성을 비교 분석하였다. 여러 가지 유기자원을 동일한 기준으로 매년 동일 장소에 처리하였다. 사용된 유기자원은 가축분퇴비, 채종유박인 유기질비료, 볏짚으로만 퇴비화한 볏짚퇴비와 겨울철 휴한기에 헤어리베치를 재배하여 이듬해 봄에 예취한 후 토양에 환원한 녹비처리구, NPK구, 가축분퇴비를 혼용처리한 NPK퇴비군, 양분을 전혀 시용하지 않은 무비구 등 총 7처리구였다. 각각의 처리구에서 토양(0-20 cm)을 채취하여 배양성 토양미생물은 희석평판법으로 해당 선백배지에 시료를 도말 하여 조사하였고 비배양성 미생물은 토양으로부터 genomic DNA를 추출하여 세균의 16S rDNA를 증폭시킨 후 denaturing gradient gel electrophoresis (DGGE)를 수행하여 분석하였다. 주요결과를 요약하면 밭토양에 서식하는 토양미생물의 균수는 처리별간의 차이를 보였으며 유기물처리구가 화학비료처리구보다 높았다. DGGE 분석을 통해 유기물 처리에 따른 군집의 다양성을 살펴본 결과 Fig. 1에서 보는바와 같이 Gel 상에서 다양한 위치의 밴드를 확인할 수 있었고 처리별로 특이 밴드가 있음을 확인할 수 있었다. Fig. 1에서 얻은 DGGE profile상의 밴드 강도와 수를 비교하여 Fig 2와 같은 dendrogram을 나타낼 수 있었다.

• The Korean Journal of Mycology
• /
• v.40 no.1
• /
• pp.11-18
• /
• 2012

This study was carried out to identify the genetic diversity of Penicillium isolates that were isolated from pears with postharvest decay in storage. URP-PCR was used to detect DNA diversity of 84 Penicillium isolates. Based on URP-PCR profiles, 18 Penicillium isolates were selected and their PCR polymorphic bands were produced by additional primers URP1F, URP2R, URP2F, and URP4R. UPGMA cluster analysis using the polymorphic bands showed four clustered groups and futhermore cultural and morphological features characterized the 18 Penicillium isolates. Group 1 was dominant, which occupies 70% in the four clustered groups and identified as P. expansum based on ITS sequence and morphological features.