• Proceedings of the Korean Biophysical Society Conference
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• 2003.06a
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• pp.54-54
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• 2003

Prostate gland contains neuroendocrine cells (PNECs) are playing important roles in physiological and pathophysiological processes of the prostate gland. Here, we investigated the role of purinoceptors in PNECs freshly isolated from rat ventral prostate (RPNECs) that show immunoreactivity to chromogranin A. Fura-2 ratiometry revealed that ATP evokes both fast Ca$\^$2+/ influx and store Ca$\^$2+/ release in RPNECs. A whole-cell patch clamp study demonstrated fast inactivating cationic current activated by ATP or by ${\alpha}$,${\beta}$-MeATP, which was blocked by ATP-TNP. The activation of P2X inward current was tightly associated with a sharp increase in [Ca$\^$2+/]$\sub$c/. The presence of P2X1/3 subtypes were proved by RT-PCR analysis. For the stored Ca$\^$2+/ release, ATP and UTP showed similar effects, suggesting the dominant role or P2Y2 subtypes, also confirmed by RT-PCR. Both P2X (${\alpha}$,${\beta}$-MeATP) and P2Y (UTP) stimulation induced changes in the cell morphology (initial shrinkage and blob formation on the surface) reversibly. Exocytotic membrane trafficking events were monitored with the membrane-bound fluorescent dye, FM1-43 using confocal microscopy. In spite of the similar Ca$\^$2+/ responses, UTP was far less effective in triggering exocytosis than ${\alpha}$,${\beta}$ -MeATP. Since serotonin is reportedly stored in the secretory granule of PNECs, we directly examined whether the aforementioned agonists elicit release of serotonin using carbon fiber electrode-amperometry. In accordance with the results of FM1 -43 experiments, ${\alpha}$,${\beta}$-MeATP efficiently evoke serotonin secretion while not with UTP. In summary, the P2X-mediated Ca$\^$2+/ influx plays crucial roles in the exocytosis of RPNECs. Although a global increase in [Ca$\^$2+]$\sub$c/ might be related with the morphological changes, a sharp rise of [Ca$\^$2+/]$\sub$c/ in the putative sub-plasmalemmal ‘microdomains’ might be a decisive factor for the exocytosis.

• Bulletin of the Korean Chemical Society
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• v.30 no.8
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• pp.1703-1710
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• 2009

The single-crystal structure of |$Ca_{35.5}$|[$Si_{121}Al_{71}O_{384}$]-FAU, $Ca_{35.5}Si_{121}Al_{71}O_{384}$ per unit cell, a = 24.9020(10) $\AA$, dehydrated at 673 K and 2 ${\times}\;10^{-6}$Torr, has been determined by single-crystal X-ray diffraction techniques in the cubic space group Fd$\overline{3}$m at 294 K. The large single crystals of zeolite Y (Si/Al = 1.70) were synthesized up to diameters of ${\mu}m\;and\;Ca^{2+}$-exchanged zeolite Y were prepared by ion exchange in a batch method of 0.05 M aqueous Ca($NO_3)_2$ for 4 hrs at 294 K. The structure was refined using all intensities to the final error indices (using only the 971 reflections for which $F_o\;>\;4{\sigma}(F_o))\;R_1$ = 0.038 (based on F) and $R_2$ = 0.172 (based on $F^2$). About 35.5 $Ca^{2+}$ ions per unit cell are found at an unusually large number of crystallographically distinct positions, four. Nearly filling site I (at the centers of the double 6-rings), 14.5 octahedrally coordinated $Ca^{2+}$ ions (Ca-O = 2.4194(24) $\AA$ and O-Ca-O = 87.00(8) and 93.00($8^o$) are found per unit cell. One $Ca^{2+}$ ion per unit cell is located at site II’ in the sodalite cavity and extends 0.50 $\AA$ into the sodalite cavity from its 3-oxygen plane (Ca-O = 2.324(13) $\AA$ and O-Ca-O = 115.5(10)o). The remaining twenty $Ca^{2+}$ ions are found at two nonequivalent sites II (in the supercages) with occupancies of 10 and 10 ions, respectively. Each of these $Ca^{2+}$ ions coordinates to three framework oxygens, either at 2.283(3) or 2.333(5) $\AA$, respectively, and extends either 0.24 or 0.54 $\AA$, respectively, into the supercage from the three oxygens to which it is bound. In this crystal, site I is the most populated; sites II’ and II are only sparsely occupied.$Ca^{2+}$+ appears to fit the octahedral site I best. No cations are found at sites III or III’, which are clearly less favorable for $Ca^{2+}$ ions in dehydrated zeolite Y.

• Journal of Mushroom
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• v.20 no.3
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• pp.178-182
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• 2022

Despite the long history of mushroom use, studies examining the genetic function of mushrooms and the development of new varieties via bio-molecular methods are significantly lacking compared to those examining other organisms. However, owing to recent developments, attempts have been made to use a novel gene-editing technique involving CRISPR/Cas9 technology and genetic scissors in mushroom studies. In particular, research is actively being conducted to utilize ribonucleoprotein particles (RNPs) that can be genetically edited with high efficiency without foreign gene insertion for ease of selection. However, RNPs are too large for Cas9 protein to pass through the cell membrane of the protoplasmic reticulum. Furthermore, guide RNA is unstable and can be easily decomposed, which remarkably affects gene editing efficiency. In this study, nanoparticles were used to mitigate the shortcomings of RNP-based gene editing techniques and to obtain transformants stably. We used Lentinula edodes (shiitake mushroom) Sanjo705-13 monokaryon strain, which has been successfully used in previous genome editing experiments. To identify a suitable osmotic buffer for the isolation of protoplast, 0.6 M and 1.2 M sucrose, mannitol, sorbitol, and KCl were treated, respectively. In addition, with various nanoparticle-forming materials, experiments were conducted to confirm genome editing efficiency via the formation of nanoparticles with calcium phosphate (CaP), which can be bound to Cas9 protein without any additional amino acid modification. RNPs/NP complex was successfully formed and protected nuclease activity with nucleotide sequence specificity.

• Proceedings of the KSAR Conference
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• 2000.10a
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• pp.10-12
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• 2000

Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

• Korean Journal of Animal Reproduction
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• v.24 no.4
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• pp.357-364
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• 2000

Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$-subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

• Journal of the Korea Concrete Institute
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• v.19 no.5
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• pp.613-621
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• 2007

Due to the increasing significance of durability, much researches on carbonation, one of the major deterioration phenomena are carried out. However, conventional researches based on fully hardened concrete are focused on prediction of carbonation depth and they sometimes cause errors. In contrast with steel members, behaviors in early-aged concrete such as porosity and hydrates (calcium hydroxide) are very important and may be changed under carbonation process. Because transportation of deteriorating factors is mainly dependent on porosity and saturation, it is desirable to consider these changes in behaviors in early-aged concrete under carbonation for reasonable analysis of durability in long term exposure or combined deterioration. As for porosity, unless the decrease in $CO_2$ diffusion due to change in porosity is considered, the results from the prediction is overestimated. The carbonation depth and characteristics of pore water are mainly determined by amount of calcium hydroxide, and bound chloride content in carbonated concrete is also affected. So Analysis based on test for hydration and porosity is recently carried out for evaluation of carbonation characteristics. In this study, changes in porosity and hydrate $(Ca(OH)_2)$ under carbonation process are performed through the tests. Mercury Intrusion Porosimetry (MIP) for changed porosity, Thermogravimetric Analysis (TGA) for amount of $(Ca(OH)_2)$ are carried out respectively and analysis technique for porosity and hydrates under carbonation is developed utilizing modeling for behavior in early-aged concrete such as multi component hydration heat model (MCHHM) and micro pore structure formation model (MPSFM). The results from developed technique is in reasonable agreement with experimental data, respectively and they are evaluated to be used for analysis of chloride behavior in carbonated concrete.

• Economic and Environmental Geology
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• v.28 no.3
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• pp.185-197
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• 1995

Tungsten skarns in the Chungju mine which consists mainly of strata-bound type iron ore deposits are found in the vicinity of the contact between the age-unknown Kyemeongsan Formation and granitic rock intrusions of Mesozoic age($134{\pm}2Ma$). Tungsten skarns were formed extensively from alumina and silica-rich schistose rocks by the introduction of calcium and iron from hydrothermal solution. The skarns comprise a metasomatic column and are subdivided into four facies; garnet facies, wollastonite facies, epidote facies and chlorite facies. The skarn process in time-evolutional trend can be divided broadly into the four facies in terms of the paragenetic sequence of calc-silicates and their chemical composition. Skarn and ore minerals were formed in the following sequence; (1) garnet facies, adjacent to biotite granite, containing mainly garnet(>Ad96) and magnetite, (2) wollastonite facies containing mainly wollastonite and garnet(Ad95~60), (3) epidote facies, containing mainly epidote(Ps35~31), quartz, andradite-grossular(Ad63~50), and scheelite, (4) chlorite facies, adjacent to and replacing schist, containing mainly chrolite, muscovite, quartz, calcite, epidote(Ps31~25), hematite and sulfides. The mineral assemblage and mineral compositions. suggest that the chemical potentials of Ca and Fe increased toward the granitic rock, and the component Al, Mg, K, and Si decreased from the host rock to granitic rock. The homogenization temperature and salinity of fluid inclusion in scheelite, quartz and epidote of epidote facies skarn is $300-400^{\circ}C$ and 3-8wt.% eqiv. NaCl, respectively. ${\delta}^{34}S$ values of pyrite and galena associated with chlorite facies skarn is $9.13{\sim}9.51%_{\circ}$ and $5.85{\sim}5.96%_{\circ}$, respectively. The temperature obtained from isotopic com· position of coexisting pyrite-galena is $283{\pm}20^{\circ}C$. Mineral assemblages and fluid inclusion data indicate that skarn formed at low $X_{CO_2}$, approximately 0.01. Temperature of the skarn mineralization are estimated to be in the range of $400^{\circ}C$ to $260^{\circ}C$ and pressure to be 0.5 kbar. The oxygen fugacity($fo_2$) of the skarn mineralization decreased with time. The early skarn facies would have formed at log $fo_2$ values of about -25 to -27, and late skarn facies would have formed at log $fo_2$ values of -28 to -30. The estimated physicochemical condition during skarn formation suggests that the principal causes of scheelite mineralization are reduction of the ore·forming fluid and a decrease in temperature.

• Journal of the Korean Applied Science and Technology
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• v.36 no.4
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• pp.1096-1107
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• 2019

This study on the physicochemical property of physiological activity substance in mung bean (Phaseolus aureus L.) was performed for the use as an functional food materialization. The proximate composition in the vacuum freeze dried mung bean was carbohydrate 57.20±0.29%, crude protein 26.40±0.69%, moisture 9.90±0.16%, crude ash 3.54±0.43%, and crude fat 2.96±0.26%, respectively. The vitamin content of mung bean was vitamin B₅ 0.62±0.013 mg/100 g, vitamin E 0.17±0.001 mg/100 g, vitamin B₁ 0.13±0.016 mg/100 g, and β-carotene 87.37±0.754 ㎍ RE/100 g, respectively. The mineral content of mung bean was potassium (K) 12,428.55±147.55 mg kg^-1, magnesium (Mg) 2,053.32±14.13 mg kg^-1, calcium (Ca) 1,966.40±14.53 mg kg^-1, sodium (Na) 1,063.99±7.75 mg kg^-1, iron (Fe) 63.77±0.98 mg kg^-1, and manganese (Mn) 14.67±0.22 mg kg^-1. The compositions of fatty acid were saturated fatty acid 29.23±0.03%, monoenes 20.30±0.04%, and polyenes 50.46±0.06%. Protein bound amino acid content of mung bean was 21.75±0.24 g%. And major amino acids were glutamic acid 3.93±0.03 g%, aspartic acid 2.68±0.03 g%, respectively. The composition of free amino acid of mung bean was 336.77±8.66 mg%, and major free amino acids were arginine, glutamic acid, asparagine, and aspartic acid. As a results of these experiment, Mung bean could be used a natural resouce and functional biohealth food substance.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.9
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• pp.840-850
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• 2010

The present study was attempted to find the effects of structural properties of phosphorus on the water quality of Gyehwa reservoir in Saemangeum. Relationship of phosphorus fractions between water and sediment properties was closely examined, and a few types of phosphorus were found from the sample sediment as : Saloid-P, Al-P, Fe-P, Ca-P, Red-P and Occd-P. Saloid-P (1.4%), Al-P (0.5%), Fe-P (39.8%), Ca-P (56.6%), Red-P (0.4%), Occd-P (1.3%) were extracted in a mass basis from the sediment of Gyehwa reservoir. Approximately more than 97% of phosphorus were calcium related phosphorus (Ca-P, 56%) and iron bound phosphorus (Fe-P, 39.8%). The Fe-P closely relates with water quality of T-N (r=0.761, p<0.05), $NO_3$-N (r=0.754, p<0.05), $NH_4$-N (r=0.728, p<0.05), T-P (r=0.774, p<0.05) and $PO_4$-P (r=0.767, p<0.05) while the Ca-P did not show any consistent dependency on the water quality. On the other hand, the correlation of Ca-P with $P_2O_5$ was high with r=0.783 (p<0.05) in the sediment. The Fe-P was affected significantly on the Ignition Loss (r=0.569, p<0.05), T-N (r=0.715, p<0.05) and T-P (r=0.983, p<0.01). In the research of correlation between phosphorus fraction and heavy metals in the sediment, Ca-P did not show any specific relationships with heavy metals. The Fe-P showed a significant correlation with As (r=0.817, p<0.01), Cu (r=0.793, p<0.05), Cd (r=0.786, p<0.05), Zn (r=0.738, p<0.05), so that it can be stated that the presence of Fe-P may implicate the volume of various metallic elements.