• The Korean Journal of Physiology and Pharmacology
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• 제18권4호
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• pp.297-305
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• 2014

Flavonoids have an ability to suppress various ion channels. We determined whether one of flavonoids, cyanidin-3-glucoside, affects adenosine 5'-triphosphate (ATP)-induced calcium signaling using digital imaging methods for intracellular free $Ca^{2+}$ concentration ([$Ca^{2+}$]i), reactive oxygen species (ROS) and mitochondrial membrane potential in PC12 cells. Treatment with ATP ($100{\mu}M$) for 90 sec induced [$Ca^{2+}$]i increases in PC12 cells. Pretreatment with cyanidin-3-glucoside ($1{\mu}g/ml$ to $100{\mu}g/ml$) for 30 min inhibited the ATP-induced [$Ca^{2+}$]i increases in a concentration-dependent manner ($IC_{50}=15.3{\mu}g/ml$). Pretreatment with cyanidin-3-glucoside ($15{\mu}g/ml$) for 30 min significantly inhibited the ATP-induced [$Ca^{2+}$]i responses following removal of extracellular $Ca^{2+}$ or depletion of intracellular [$Ca^{2+}$]i stores. Cyanidin-3-glucoside also significantly inhibited the relatively specific P2X2 receptor agonist 2-MeSATP-induced [$Ca^{2+}$]i responses. Cyanidin-3-glucoside significantly inhibited the thapsigargin or ATP-induced store-operated calcium entry. Cyanidin-3-glucoside significantly inhibited the ATP-induced [$Ca^{2+}$]i responses in the presence of nimodipine and ${\omega}$-conotoxin. Cyanidin-3-glucoside also significantly inhibited KCl (50 mM)-induced [$Ca^{2+}$]i increases. Cyanidin-3-glucoside significantly inhibited ATP-induced mitochondrial depolarization. The intracellular $Ca^{2+}$ chelator BAPTA-AM or the mitochondrial $Ca^{2+}$ uniporter inhibitor RU360 blocked the ATP-induced mitochondrial depolarization in the presence of cyanidin-3-glucoside. Cyanidin-3-glucoside blocked ATP-induced formation of ROS. BAPTA-AM further decreased the formation of ROS in the presence of cyanidin-3-glucoside. All these results suggest that cyanidin-3-glucoside inhibits ATP-induced calcium signaling in PC12 cells by inhibiting multiple pathways which are the influx of extracellular $Ca^{2+}$ through the nimodipine and ${\omega}$-conotoxin-sensitive and -insensitive pathways and the release of $Ca^{2+}$ from intracellular stores. In addition, cyanidin-3-glucoside inhibits ATP-induced formation of ROS by inhibiting $Ca^{2+}$-induced mitochondrial depolarization.

• Molecules and Cells
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• 제20권1호
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• pp.74-82
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• 2005

Glutaredoxins (Grxs) are thioloxidoreductases which are required for maintaining thiol/disulfide equilibrium in living cells. The Grx3 gene, which encodes one of the three monothiol Grxs in the fission yeast Schizosaccharomyces pombe, was characterized, and its transcriptional regulation studied. Genomic DNA encoding Grx3 was isolated by PCR, and a plasmid pTT3 carrying this DNA was produced. The DNA sequence has 1,267 bp, which would encode a monothiol Grx of 166 amino acids with a molecular mass of 18.3 kDa. The putative protein has 27% homology with Grx5, and contains many hydrophobic amino acid residues in its N-terminal region. S. pombe cells harboring pTT3 had increased Grx activity and enhanced survival on minimal medium plates containing aluminum (5 mM), BSO (0.05 mM), menadione (0.01 mM) or cadmium (0.2 mM). The 568 bp upstream region of Grx3 was fused into the promoterless b-galactosidase gene of the shuttle vector YEp367R to generate fusion plasmid pMJS10. Potassium chloride (KCl) and metals including aluminum and cadmium enhanced the synthesis of ${\beta}$-galactosidase from the fusion gene. The synthesis of ${\beta}$-galactosidase was also enhanced, in a Pap1-dependent manner, by fermentable carbon sources such as glucose (at low concentrations) and sucrose, but not by non-fermentable carbon sources such as ethanol and acetate. Grx3 mRNA increased in response to treatment with BSO. These observations indicate that S. pombe Grx3 is involved in the response to stress, and is regulated by stress.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• 제27권2호
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• pp.63-81
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• 1997

This study was performed in order to identify changes of the plasma membrane proteins in rat submandibular gland tumors induced by 7,12-dimethylbenz[a]anthracene [DMBA] and X-irradiation. Two kinds of tumor associated membrane proteins (protein A and B) were isolated with 3 M KCl extraction from rat submandibular gland tumors induced by DMBA and X-irradiation. To identify their antigenicities, immunoelectrophoresis and double immunodiffusion was carried out with various proteins extracted from liver, heart, skin and pancreas of adult rats and from embryonic liver, heart and skin. The rabbit antisera against the protein A did not cross-react with any of the proteins extracted from the above mentioned tissues, suggesting that protein A might be tumor specific antigen. However, the rabbit antisera against protein B was precipitated with proteins extracted from the liver of adult and embryonic rats. Polyacrylamide gel electrophoresis of these two proteins (A and B) showed that protein A was a dimer with molecular weights of 69,000 and 35,000 dalton, whereas protein B was a monomer with molecular weight of 50,000 dalton.

• Korean Journal of Soil Science and Fertilizer
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• 제23권2호
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• pp.128-134
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• 1990

Croping pattern, fertilizer application, Soil chemical characteristics, plant nutritional condition and growth status were investigated in three major vinyl house farms near Yesan. Croping pattern changed from tomato to pumpkin due to tomato diseases such as wilting and blossom-end rot. Wilting seemed to be closely related with high EC, nitrogen and root cyst nematodes. Calcium deficiency seemed to be due to high potassium, EC in soil and high uptake of iron. Chinese cabbages in summer showed poor growth (80% inhibition) due to high EC(1.8mmho/cm) and easily got wenny root disease that might due to high phosphorus (1055ppm) in soil. Summer radish showed poor growth (50% inhibition) due to high EC(1.6mmho/cm), K and Mg resulting in base imbalance. Farmers used 5 kinds of compound fertilizer as basal application and one without P for top dressing. Urea and KCL were used for top dressing. Heavy application of livestock manure and chemical fertilizer for every crop made eutrophic even in subsoil.

• KOREAN JOURNAL OF CROP SCIENCE
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• 제51권3호
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• pp.191-198
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• 2006

Priming of seeds in 150 mM KCI at $10^{\circ}C$ for 6 days was most effective in increasing germination rate and shortening germination time. Germination rate of primed seeds was increased to 96% which is about 14% higher than that of non-primed seeds. Germination temperature for better priming effects from primed onion seeds was 10 to $15^{\circ}C$ which is relatively lower than that of normal germination temperature of $20^{\circ}C$. Drying primed seeds at $20^{\circ}C$ for 5 hours resulted in best priming effects.

• Journal of Korean Neurosurgical Society
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• 제57권1호
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• pp.1-5
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• 2015

Objective : Cerebral vessels, such as intracerebral perforating arterioles isolated from rat brain, have been widely used as an ex vivo model to study the cerebrovascular function associated with cerebrovascular disorders and the therapeutic effects of various pharmacological agents. These perforating arterioles, however, have demonstrated differences in the vascular architecture and reactivity compared with a larger leptomeningeal artery which has been commonly implicated in cerebrovascular disease. In this study, therefore, we developed the method for studying cerebrovascular function utilizing the olfactory artery isolated from the mouse brain. Methods : The olfactory artery (OA) was isolated from the C57/BL6 wild-type mouse brain. After removing connective tissues, one side of the isolated vessel segment (approximately $-500{\mu}m$ in length) was cannulated and the opposite end of the vessel was completely sealed while being viewed with an inverted microscope. After verifying the absence of pressure leakage, we examined the vascular reactivity to various vasoactive agents under the fixed intravascular pressure (60 mm Hg). Results : We found that the isolated mouse OAs were able to constrict in response to vasoconstrictors, including KCl, phenylephrine, endothelin-1, and prostaglandin $PGH_2$. Moreover, this isolated vessel demonstrated vasodilation in a dose-dependent manner when vasodilatory agents, acetylcholine and bradykinin, were applied. Conclusion : Our findings suggest that the isolated olfactory artery would provide as a useful ex vivo model to study the molecular and cellular mechanisms of vascular function underlying cerebrovascular disorders and the direct effects of such disease-modifying pathways on cerebrovascular function utilizing pharmacological agents and genetically modified mouse models.

• The Korean Journal of Mycology
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• 제26권2호통권85호
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• pp.275-280
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• 1998

Factors affecting protoplasts isolation and regeneration of Fomitella fraxinea were investigated. Lytic enzyme mixture of Novozym 234, Cellulase onozuka R-10 and ${\beta}-Glucuronidase$ was found to be the best for the protoplasts isolation. Osmotic stabilizer of 0.6 M sucrose was observed as the best for protoplasts isolation. The highest number of protoplasts was obtained from the F. fraxinea mycelium with lytic enzyme mixture and osmotic stabilizer that had been cultured for 3 hours. The highest regeneration rate of 0.02 % was achieved when the 0.6 M sorbitol was employed as osmotic stabilizer.

• KSBB Journal
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• 제14권6호
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• pp.661-664
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• 1999

Proteins were extracted from an aqueous phase with reversed micelles. The effect of pH, and salt concentration on the solubilization of lysozyme in AOT/isooctane solution was studied to explore the potential for employing this solvent system in the large-scale recovery and concentration of proteins using liquid extraction. For pH values below the isoelectric point, pl of the protein, solubilization was high, probably owing to strong electrostatic interactions between the positively charged proteins and the anionic surfactant heads forming the inner micelle wall. At low ionic strength complete solubilization of the protein was observed. A pH higher than the pl of lysozyme and a salt concentration lower than that of the water pool were required for the recovery aqueous phase to ensure the back extraction of lysozyme from the AOT reversed micelles.

• Particle and aerosol research
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• 제12권3호
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• pp.51-56
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• 2016

Due to recent development in nanotechnology and increasing usage and production of nanomaterials, numerous studies related to environment, sanitation and safety handling of nanoparticle are being conducted. Since nanoparticles can be easily absorbed into human bodies through breathing process, based on their toxic substances and their large specific surface, these particles can cause serious health damage. Therefore, to reduce nanoparticle emissions, nanofiltration technology is becoming a serious issue. Filtration is a separation process during which a fluid passes through a barrier by removing the particles from the stream. Barrier filters can be made of various materials and shapes. One of the most common type of barrier filter is the fibrous filter. Fibrous filters are divided in two types: nonwoven and woven fabrics. Polypropylene is a thermoplastic material, used as a base material for melt blown nonwoven fabric. In this study, we examined filtration property of KCl nanoparticles with a mean particle diameter of 75 nm using multilayer meltblown filter samples. These experiments verify that the penetration of nanoparticle in the filter correlate with pressure drop; the meltblown layer MB1 has the greatest effect on dust collection efficiency of the filter. Among all tested samples, dust collection efficiency of 2-layer filter was best. However, when considering the overall pressure drop and dust collection efficiency, the 4-layer filter has the highest quality factor for particles smaller than 70 nm.

• Korean Journal of Agricultural Science
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• 제13권2호
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• pp.299-310
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• 1986

The ovine squamous cell carcinoma (OSCC)-specific and immunosuppressive properties of OSCC extracts were investigated by using the techniques of lymphocyte blastogenicity, acid dissociation-ultrafiltration and gradient polyacrylamide gel electrophoresis. It was found that OSCC extracts contained two major and one minor protein peaks by Sephadex gel fractionation. Two major peaks bear substantial amount of immunoglobulins, antigen-antibody complex and OSCC-specific fractions, and the minor peak includes immunosuppressive materials. OSCC-specific components were detected at the molecular weights of 10,000 to 100,000 daltons in the major peaks and immunosuppressive materials at the fractions with the molecular weight of 10,000 to 100,000 and < 10,000 daltons in the minor peak. When the fractions were further separated by gradient polyacrylamide gel electrophoresis, the OSCC-specific antigens were found in the slice number 4 to 6 in fraction III, and immunosuppressive materials, in the slice numbers 9 to II in fraction V. The present results were considered to provide a basis for preparation and purification of OSCC-specific and immunosuppressive materials from the crude OSCC extracts.