• Korean Journal of Animal Reproduction
• /
• v.24 no.2
• /
• pp.125-142
• /
• 2000

13). Analysis of a purified preparation of eCG revealed that its $\beta$ -subunit consists of 149 amino acids, which was confirmed by the molecular cloning of its cDNA. There seem to be at least four to six, or even as many as 11, O-glycosylation sites on the extended C-tenninal region of the eCG $\beta$-subunit. Interestingly, eCG is a unique member of this family, as it appear to be a single molecule that possesses both LH- and FSH-like activities. Using the cDNA prepared from mRNA extracted from equine placental and pituitary tissues, we cloned the cDNA of eCG $\alpha$- and $\beta$ -subunits and eFSH $\beta$ -subunit. The mRNA expression of each subunit seems to be independently regulated, which may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. Thus, eCG is a distinct molecule from the view points of its biological function and glycoresidue structures. Recombinant eCGs including the mutants which lack oligosaccharides will be useful tools for analyzing the structure-function relationships of gonadotropins in the horse as well as other species. Similar experiments will also clarify the proposed structure and biological functions for the glycoprotein hormones. These experimental are now possible, and hopefully a resolution of the existing controversy will be forthcoming in the near future.

• Journal of Life Science
• /
• v.21 no.1
• /
• pp.44-55
• /
• 2011

This study aimed to investigate the effects of water extract of Rubus coreanus (RCE) on the expression and activity of endothelial nitric oxide synthase (eNOS), as well as its signal transduction pathways in human umbilical vein endothelial cells (HUVECs). The specific inhibitors of NOS show RCE treatment increases NO production in HUVECs due to the up-regulation of eNOS rather than iNOS. The real-time expression level of eNOS mRNA was also increased upon RCE treatment in HUVECs. While a PKC-specific inhibitor, RO-317549, did not alter RCE-induced NO production in HUVECs, tamoxifen (estrogen receptor-specific inhibitor), PD98059 (ERK-specific inhibitor) and LY-294002 (PI3K/Akt-specific inhibitor) did have suppressive effects. Increased NO production by RCE seems to result from a higher level of active eNOS (pSer1177). Specifically, inhibition of ERK not only decreased the level of active eNOS, but also increased the inactive form of the enzyme (pThr495) in HUVECs. This study suggests that RCE treatment increases NO production in HUVECs due to the increased expression and activity of eNOS. It is also shown that RCE-induced eNOS activation occurs partly through the binding of RCE to the estrogen receptor, along with ERK and PI3K/Akt-dependent signal transduction pathways. In addition, the regulatory binding proteins of eNOS including Hsp90 and caveolin-1 were related to these effects of RCE on eNOS activity in HUVECs.

• The Korean Journal of Mycology
• /
• v.47 no.4
• /
• pp.437-441
• /
• 2019

Verticillium wilt disease is caused by a fungal plant pathogen Verticillium dahliae, which attacks commercial crops such as chrysanthemum. The conventional methods so far used to identify this fungal pathogen require high expertise and are time-consuming. Therefore, in this study, we developed an assay for the rapid and specific detection of V. dahliae infection using loop-mediated isothermal amplification (LAMP) method. For this assay, four primers for LAMP were designed for targeting cellulose-growth-specific protein partial mRNA gene in Verticillium dahliae. Under standard condition, the optimum reaction temperature for amplification is around 60 ℃ within 60 minutes. This LAMP assay was designed to amplify only present in V. dahliae. When this LAMP assay applied to the DNAs for four other soil-borne fungi and host plants, no amplification was detected. Therefore, this LAMP assay we developed for V. dahliae is expected to do detection at the early stage of its infection. The fast and reliable detection method will allow us to develop effective management system to monitor and control infection of this pathogen in chrysanthemum plant.

• Development and Reproduction
• /
• v.8 no.1
• /
• pp.35-42
• /
• 2004

Oocyte maturation refers to the process that prophase I arrested germinal vesicle(GV) drives the progression of meiosis to metaphase II(MII) to have the capacity for fertilization and embryo development. To better understand the molecular mechanism(s) involved in oocyte maturation, we identified differentially expressed genes(DEGs) between GV and MII mouse oocytes using a new innovative annealing control primer (ACP) technology. Using 20 ACPs, we successfully cloned 32 DEGs between GV and Mll oocytes, and 26 out of these 32 DEGs were functionally known genes. Four genes including Pscd2 were GV-specific, 10 genes including PKD2 and CSN3 were highly expressed in GV oocytes(GV-selective), and 12 genes including Diva were highly expressed in MII oocytes (MII-selective). Ail of the genes identified in this study were first reported in the oocyte expression using ACP system and especially, we could characterize the existence of PKD-CSW signaling pathwayin the mouse oocytes. Results of the present study would provide insight for studying molecular mechanisms regulating oocyte maturation.

• Korean Journal of Ichthyology
• /
• v.18 no.2
• /
• pp.65-77
• /
• 2006

Gene transcripts potentially responsive to the heat stress were surveyed by cDNA microarray analysis in mud loach (Misgurnus mizolepis). Transcriptional profiles of hepatic tissue in the fish exposed to either $23^{\circ}C$ or $32^{\circ}C$ for 4 weeks were compared each other by 3 replicated hybridization assays using 1,124 unigene clones selected from mud loach liver expressed sequence tags (ESTs). A total of 93 clones showed the substantially increased mRNA levels (>2-fold) in $32^{\circ}C$-exposed group when compared in $23^{\circ}C$control group. It includes various enzymes and proteins involved in energy pathway, protease/protein metabolisms, immune/antioxidant functions, cytoskeleton/cell structure, transport and/or signal transduction. Maximum level of increase was up to 15-fold relative to $23^{\circ}C$ treatment. Heat exposure also resulted in the significant decrease (less than 50% relative to $23^{\circ}C$-exposed fish) of the transcriptional activities in 85 genes. Besides the above categories, yolk protein (vitellogenin) and ribosomal proteins were notably down regulated in the fish exposed to heat stress. A number of novel gene transcripts were also detected in both up-regulated and down-regulated groups.

• Korean Journal of Clinical Laboratory Science
• /
• v.49 no.4
• /
• pp.413-419
• /
• 2017

Four imipenem-resistant bacteria were isolated from the clinical specimens of a patient with pneumonia. To identify the isolates, we used the GN card of Vitek II system and performed a phylogenetic analysis based on 16S rRNA gene sequence. The isolates were identified as P. aeruginosa (2 strains), P. monteilii (1 strain), and P. putida (1 strain), and were tested for antibiotic resistance after determining the MIC of imipenem to be $${\geq_-}8{\mu}g/mL$$ using the AST-N225 card of Vitek II system. The imipenem-resistant genotypes were determined using PCR products amplified using specific ${\beta}-Lactamase$ gene primers. The MBL gene was identified in all four isolates. One strain of P. aeruginosa exhibited the VIM and SHV-1 type genes, while the other strain exhibited both VIM and OXA group II genes. According to the antimicrobial susceptibility test, the bacteria were more susceptible to amikacin than other antibiotics. DNA fingerprint analysis using ERIC-PCR to analyze the epidemiological relationship between strains estimated that both the P. aeruginosa isolates were similar, but exhibited different DNA band types. It is uncommon to find four strains of imipenem-resistant bacteria with different DNA band types in a single patient.

• Journal of Life Science
• /
• v.14 no.6 s.67
• /
• pp.1009-1017
• /
• 2004

Sialyltransferases cloned so far show the remarkable tissue-specific expression, which is correlated with the existence of cell type-specific sialylated sugar structure in glycoconjugates. In the previous studies, we found various mRNA isoforms of human sialyltransferases generated by alternative splicing and alternative promoter utilization. To understand the regulatory mechanisms for specific expression of human sialyltransferase genes and for production of their mRNA isoforms, in this study, we have isolated and characterized five kinds of human sialyltransferase genes: hST3Gal II, hST8Sia II, hST8Sia III, hST8Sia IV, and hST8Sia V. The hST3Gal II gene is composed of six exons, which span over 17kb, with exons ranging in size from 46 to over 1017 bp. The hST8Sia III gene comprises over 10 kb, and consists of only four exons, which is much smaller and simpler than other human sialyltransferase genes. In contrast, three genes (hST8Sia II, hST8Sia IV and hST8Sia V) span more than 70 kb, and comprise five or more exons. All exon-intron boundaries follow the GT-AG rule. In particular, the sialylmotif L, which is a highly conserved region in all cloned sialyltransferases, was found in one exon of hST8Sia III, whereas this motif is encoded by discrete exons in the other human sialyltransferases. Exon structures of these sialyltransferase genes show the structural diversity, as found in other human sialyltransferase genes reported so far. We determined the transcription start site of hST3Gal II gene by the 5'-RACE and cap site hunting experiments.

• Journal of Life Science
• /
• v.30 no.9
• /
• pp.772-782
• /
• 2020

Skeletal muscle differentiation or myogenesis is important to maintain muscle mass and metabolic homeostasis. Muscle-specific microRNAs (miRNAs) are known to play a critical role in skeletal myogenic differentiation. In this study, we examined the expression profiling of miRNAs during myogenic differentiation in rat L6 myoblasts using rat miRNA microarrays. We identified the upregulated expression of miR-128 as well as several well-known myogenic miRNAs, including miR-1, miR-133b, and miR-206. We additionally confirmed the increased expression of miR-128 observed on microarray through quantitative real-time PCR (qRT-PCR), which showed similarly upregulated expression of both primary miR-128 and mature miR-128, consistent with the microarray findings. Furthermore, transfection of miR-128 into rat L6 myoblasts induced gene expression of myogenic markers such as muscle creatine kinase (MCK), myogenin, and myosin heavy chain (MHC). Protein expression of MHC was increased as well. Inhibition of miR-128 by inhibitory peptide nucleic acids (PNAs) blocked the expression of those myogenic markers. In addition, the transfection of miR-128 into rat L6 myoblasts enhanced the phosphorylation of Erk and Akt proteins stimulated by insulin, while simultaneously reversing the inhibited phosphorylation of Erk and Akt due to insulin resistance. These findings suggest that miR-128 may play important roles in myogenic differentiation and insulin signaling.

• Annals of Clinical Microbiology
• /
• v.18 no.2
• /
• pp.37-43
• /
• 2015

Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.31 no.1 s.49
• /
• pp.103-109
• /
• 2005

UV irradiation leads to distinct changes in skin connective tissue, which is degradation of collagen. Many of these alterations in the extracellular matrix are mediated by matrix metalloproteinases. In this study, to develop a new anti-aging agent, we screened the antioxidant activity of solvent fractions from ethanolic extract of Melothria Heterophylla. Among the four solvent fractions tested, the EtOAc fraction exhibited the highest antioxidant activity. It was investigated the inhibitory effect of the EtOAc fraction on the expression and activity of MMP-1 in UVA-irradiated human dermal fibroblasts. The EtOAc fraction inhibited the activity of MMP-1 in a dose dependent manner with the $IC_{50}$ values of $9{\mu}g/mL$. Also, UVA-induced MMP-1 expression was reduced about $90\%$ by $100{\mu}g/mL$ of the EtOAc fraction but MMP-1 mRNA expression was not inhibited. Therefore, we conclude that the EtOAc fraction significantly inhibits MMP-1 expression at the protein level. From these results, we suggest that the EtOAc fraction from M. heterophylla could be used as a new anti-aging agent for the photo-damaged skin.