• Journal of Physiology & Pathology in Korean Medicine
• /
• v.23 no.3
• /
• pp.613-618
• /
• 2009

Although the effect of Eucommie umoides oliver in osteoporosis has been studied, direct action of Eucommis ulmoides Oliver on osteoclasts remains unknown. Here we examined whether Eucommiae cortex inhibits osteoclast differentiation and bone resorption, a process known to be involved in bone diseases such as osteoporosis. Water extract from Eucommiae cortex (WE-EC) inhibited differentiation of bone marrow macrophages (BMMs) into osteoclasts without causing cytotoxicity. WE-EC suppressed the phosphorylation of p38, ERK, and JNK in BMMs treated with RANKL. WE EC specifically suppressed the mRNA expression of NFATc1 induced by RANKL. However, WE-EC inhibited stability of c-Fos protein induced by RANKL. Furthermore, WE-EC inhibited osteoclast survival induced by RANKL and in turn suppressed bone resorption. Taken together, our results suggest that WE-EC may be better agents for therapeutic use in bone diseases.

• Korean Journal Plant Pathology
• /
• v.1 no.3
• /
• pp.199-206
• /
• 1985

Viroids are the smallest. well-characterized infectious agents presently known. and so far viroids have been found only in higher plants. The structures of viroid-molecules are single-stranded, covalently closed circular RNA molecules with a range of 240 to 380 nucleotides according to the various viroids. Viroids are remarkable not only as a new category of pathogen, which cause economically important diseases, but also as an excellent model system for biochemical and biophysical investigations because of their small size, relative stability and their self-replication. Four different patato spindle tuber viroid isolates, which express the different symptoms on the same host plant exchange only 2 to 6 nucleotides in the total number of 359 nucleotides, but now the mechanism of viroid pathogenicity is not explained fully. Viroid-melecules are replicated without any special viroid-associated proteins, and during the process of viroid replication oligomeric viroid-associated RNAs are detected at nuclei of viroid infected leaf tissue. The mechanism of viroid replication can now be illustrated according to a possible explanation of rolling-circle system. Although the rapid progress have been made in elucidation of the biochemical and biophysical properties of PSTV and other viroids, the mechanism of viroid replication and pathogenicity is less known and is still a matter of speculation. When these problems can be sufficiently explained, the viroid molecule could play an important role as an available vector in plant genetic engineering.

• Korean Journal of Microbiology
• /
• v.47 no.3
• /
• pp.255-262
• /
• 2011

Six protein-degrading bacteria were isolated from digestive organ of Harmonia axyridis. These isolates were categorized as Staphylococcus sciuri subsp. sciuri (3 strains), Bacillus subtilis (1 strain), and Bacillus thuringiensis (2 strains) by 16S rRNA gene sequence analysis. The Staphylococcus sp. CB2-3 was selected as a protease-producing bacterium which showed the highest protease activity of 58.5 U/ml at the pH 5.0 medium. The optimal pH and temperature of protease activity were pH 5.0 and $40^{\circ}C$, respectively. This acid protease had a relatively high stability of 80% between $30-50^{\circ}C$ at broad temperature range. The opimal medium compositions of carbon, nitrogen and mineral source for cell growth and protease activity were investigated. When sorbitol (0.5%) was used as carbon source, enzyme activity was increased about 2 times than that of the basal medium. When skim milk (0.5%) was used as nitrogen source, activity was increased about 2.5 times than that of the control. Cell growth and enzyme activity were increased by mineral source such as KCl, $K_2HPO_4$, $FeSO_4$, but was completely inhibited by divalent ions such as $Co^{2+}$, $Zn^{2+}$, $Mn^{2+}$, $Cu^{2+}$.

• The Journal of Korean Medicine
• /
• v.24 no.2
• /
• pp.166-178
• /
• 2003

Objectives : Nitric oxide (NO) plays an important role in normal and pathophysiological cells as a messenger molecule, neurotransmitter, microbiological agent, or dilator of blood vessels and arteriosclerosis, respectively. This study was undertaken to understand the mechanism of NO production and effect of Hyeolbuchukeo-tang (Xiefuzhuyu-tang) on NO production in cultured vascular smooth muscle cell (VSMC). Methods and Results : VSMC was isolated from aorta and cultured. Cultured primary cells were identified as VSMC with anti--smooth muscle actin antibody. A large amount of NO was produced in cultured VSMC treated with $IFN-{\gamma}$ plus TNF in a time- and dose-dependent manner. $TNF-{\alpha}$ was a more efficient stimulator than $IFN-{\gamma}$ in NO production of cultured VSMC. iNOS protein wasdetected within 3 hrs and it increased up to 12 hrs in a time-dependent manner. However, accumulated NO in cytokine-treated VSMC was not detected within 3 hrs. NO production in cytokine-treated VSMC showed the dose- and time-dependent manner, and increased up to 48 hrs. The activated VSMC produced a large amount of NO (about 60 uM). Hyeolbuchukeo-tang (Xiefuzhuyu-tang) alone did not induceNO production, but it potentiated the effect of $TNF-{\alpha}$ on NO production and increased NO production by about 20%. Hyeolbuchukeo-tang (Xiefuzhuyu-tang) did not affect the transcriptional activity of iNOS gene, but increased the accumulation of iNOS. These results indicate that Hyeolbuchukeo-tang (Xiefuzhuyu-tang) could modulate the translational level of iNOS. PKC did not modulate NO production, but calcium ionophore A23187 decreased NO production. However, Hyeolbuchukeo-tang (Xiefuzhuyu-tang) elevated the decreased NO production in A23187-treated VSMC by modulating the stability of iNOS transcripts. Half-life of the synthesized transcripts appeared to have about 6 hrs. PDTC, an $NF-{\kappa}B$ inhibitor, blocked the accumulation of iNOS mRNA, indicating that $NF-{\kappa}B$ served as an important modulator in the transcriptional regulation of iNOS. As Hyeolbuchukeo-tang (Xiefuzhuyu-tang) potentiated the effect of the $TNF-{\alpha}$ on NO production but had no additional effect on PDTC-modulated NO production, it is suggested that Hyeolbuchukeo-tang (Xiefuzhuyu-tang) enhances the $TNF-{\alpha}-mediated$ NO production of VSMC by modulating the iNOS activity and the stability of iNOS transcripts in activated VSMC having the elevated intracellular calcium ion. Conclusions : This study suggests that Hyeolbuchukeo-tang (Xiefuzhuyu-tang) has a potential capacity for preventing and treating diseases of the circulation system, including arteriosclerosis.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.18 no.12
• /
• pp.575-580
• /
• 2017

This study analyzed the effects of RGG box of hnRNP A1 on its subcellular localization and stabilization of hnRNP A1 over a three year period from October 2014. First, a 6R/K mutation in RGG box was generated, and pcDNA1-HA-hnRNP A1(6R/K) was constructed. The subcellular localization of hnRNP A1(6R/K) from the HeLa cells transfected with this plasmid DNA was analyzed by immunofluorescence microscopy. HA-hnRNP A1(6R/K) was found to exhibit nuclear and cytoplasmic fluorescence. The stability of hnRNP A1(6R/K) was checked by Western blot analysis using the expressed protein from the HeLa cells transfected with the pcDNA1-HA-hnRNP A1(6R/K). The results show that HA-hnRNP A1(6R/K) has a smaller size. These confirm that HA-hnRNP A1(6R/K) is localized both in the nuclear and cytoplasm, not because 6R/K mutation affects the nuclear localization of hnRNP A1, but because 6R/K mutation causes hnRNP A1(6R/K) to cleave at the mutation or near the mutation site. The cleaved protein fragment, which lacks the M9 domain (i.e. nuclear localization signal of hnRNP A1), did not exhibit nuclear fluorescence. This suggests that the arginines of RGG box in hnRNP A1 play an important role in stabilizing hnRNP A1. An analysis of the RNA-binding ability of hnRNP A1(6R/K) expressed and purified from bacteria will be a subsequent research project.

• Journal of Life Science
• /
• v.28 no.4
• /
• pp.389-397
• /
• 2018

The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

• Journal of Animal Science and Technology
• /
• v.58 no.4
• /
• pp.14.1-14.9
• /
• 2016

Background: Peroxisome proliferator-activated receptor gamma (PPARG), CCAAT/enhancer binding protein alpha (CEBPA) and retinoid X receptor alpha (RXRA) are nuclear transcription factors that play important roles in regulation of adipogenesis and fat deposition. The objectives of this study were to characterise the variability of these three candidate genes in a mixed sample panel composed of several cattle breeds with different meat quality, validate single nucleotide polymorphisms (SNPs) in a local crossbred population (Angus - Hereford - Limousin) and evaluate their effects on meat quality traits (backfat thickness, intramuscular fat content and fatty acid composition), supporting the association tests with bioinformatic predictive studies. Results: Globally, nine SNPs were detected in the PPARG and CEBPA genes within our mixed panel, including a novel SNP in the latter. Three of these nine, along with seven other SNPs selected from the Single Nucleotide Polymorphism database (SNPdb), including SNPs in the RXRA gene, were validated in the crossbred population (N = 260). After validation, five of these SNPs were evaluated for genotype effects on fatty acid content and composition. Significant effects were observed on backfat thickness and different fatty acid contents (P < 0.05). Some of these SNPs caused slight differences in mRNA structure stability and/or putative binding sites for proteins. Conclusions: PPARG and CEBPA showed low to moderate variability in our sample panel. Variations in these genes, along with RXRA, may explain part of the genetic variation in fat content and composition. Our results may contribute to knowledge about genetic variation in meat quality traits in cattle and should be evaluated in larger independent populations.

• Biomolecules & Therapeutics
• /
• v.21 no.3
• /
• pp.222-228
• /
• 2013

Although the role of ${\alpha}$-synuclein aggregation on Parkinson's disease is relatively well known, the physiological role and the regulatory mechanism governing the expression of ${\alpha}$-synuclein are unclear yet. We recently reported that ${\alpha}$-synuclein is expressed and secreted from cultured astrocytes. In this study, we investigated the effect of valproic acid (VPA), which has been suggested to provide neuroprotection by increasing ${\alpha}$-synuclein in neuron, on ${\alpha}$-synuclein expression in rat primary astrocytes. VPA concentration-dependently increased the protein expression level of ${\alpha}$-synuclein in cultured rat primary astrocytes with concomitant increase in mRNA expression level. Likewise, the level of secreted ${\alpha}$-synuclein was also increased by VPA. VPA increased the phosphorylation of Erk1/2 and JNK and pretreatment of a JNK inhibitor SP600125 prevented the VPA-induced increase in ${\alpha}$-synuclein. Whether the increased ${\alpha}$-synuclein in astrocytes is involved in the reported neuroprotective effects of VPA awaits further investigation.