• Journal of Microbiology and Biotechnology
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• v.18 no.6
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• pp.1101-1108
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• 2008

Geldanamycin and its analogs are important anticancer agents that inhibit the newly targeted heat-shock protein (Hsp) 90, which is a chaperone protein in eukaryotic cells. To resolve which geldanamycin biosynthetic genes are responsible for particular post-polyketide synthase (PKS) processing steps and in which order the reactions occur, we individually inactivated candidate genes in Streptomyces hygroscopicus subsp. duamyceticus JCM4427 and isolated and elucidated the structures of intermediates from each mutant. The results indicated that gel7 governs at least one of the benzoquinone ring oxidation steps. The gel16 was found to be involved in double-bond formation between C-4 and C-5 of 4,5-dihydrogeldanamycin, which confirmed our previous findings that this double bond is reduced during the post-PKS modification of the polyketide assembly. In addition, pro-geldanamycin, which does not possess a double bond at C-4/5, was purified from the gel7 and gel8 double-gene-inactivated mutant.

• Archives of Pharmacal Research
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• v.29 no.8
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• pp.657-665
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• 2006

We recently reported that dimethylsphingosine (DMS), a metabolite of sphingolipids, increased intracellular pH and $Ca^{2+}$ concentration in U937 human monocytes. In the present study, we found that dimethylphytosphingosine (DMPH) induced the above responses more robustly than DMS. However, phytosphingosine, monomethylphytosphingosine or trimethylsphingosine showed little or no activity. Synthetic C3 deoxy analogues of sphingosine did show similar activities, with the C16 analogue more so than C18. The following structure-activity relationships were observed between DMS derivatives and the intracellular pH and $Ca^{2+}$ concentrations in U937 monocytes; 1) dimethyl modification is important for the DMS-induced increase of intracellular pH and $Ca^{2+}$, 2) the addition of an OH group on C4 enhances both activities, 3) the deletion of the OH group on C3 has a negligible effect on the activities, and 4) C16 appears to be more effective than C18. We also found that W-7, a calmodulin inhibitor, blocked the DMS-induced pH increase, whereas, KN-62, ML9, and MMPX, specific inhibitors for calmodulin-dependent kinase II, myosin light chain kinase, and $Ca^{2+}$-calmodulin-dependent phosphodiesterase, respectively, did not affect DMS-induced increases of pH in the U937 monocytes.

• Journal of Pharmaceutical Investigation
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• v.40 no.3
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• pp.175-180
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• 2010

Albumin-modification has been viewed as one of the most effective ways of extending the short in vivo lifetimes of peptide drugs by delaying glomerular filtration. In this study, we describe a new type 2 anti-diabetic exendin-4 (Ex4) peptide derivative with significant binding ability to human serum albumin (HSA). This exendin-4 derivative consists of a 4-branched polyethylene glycol $(PEG)_{5k}$ (Mw: 20 kDa) modified with three stearylamines ($C_{18}-NH_2$) and one exendin-4 on its branches. PEG and stearylamine were selected to provide functionality to increase molecular size and bind to albumin, respectively. This derivative ($3C_{18}-4PEG_{5k}$-Ex4) was shown to have larger molecular size (Ca. 152 kDa) than actual (25.0 kDa) when subjected to size-exclusion chromatography, and the fluorescein-tagged $3C_{18}-4PEG_{5k}$-Ex4 displayed significant binding to the HSA-immobilized Sepharose CL-4B resin using confocal laser scanning microscopy. Furthermore, $3C_{18}-4PEG_{5k}$-Ex4 was found to have acceptable anti-hyperglycemic efficacy via three consecutive oral glucose tolerance testings (OGTT) in fasted type 2 diabetic db/db mice. The $HD_{total}$ value ($57.6{\pm}12.3%$) of $3C_{18}-4PEG_{5k}$-Ex4 at a 50 nmol/kg dose was 2-fold greater than that ($31.0{\pm}8.7%$) of native exendin-4 in non-fasted db/db mice. Especially, the blood glucose levels in the mice group treated with $3C_{18}-4PEG_{5k}$-Ex4 did not rebound to ~150 mg/dL until 24 h after the injection, which obviously shows the extended hypoglycemia. We believe that this derivative has great pharmaceutical potential as a novel long-acting type 2 anti-diabetic injection treatment.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1098-1102
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• 2009

Gamma-linolenic acid (GLA, C18:3 ${\Delta}^{6,9,12}$) is synthesized by a delta-6 fatty acid desaturase using linoleic acid (LA, C18:2 ${\Delta}^{9,12}$) as a substrate. To enable the production of GLA in the conventional yeast Pichia pastoris, we have isolated a cDNA encoding the delta-6 fatty acid desaturase from Cunninghamella echinulata MIAN6 and confirmed its function by heterogeneous expression in P. pastoris. Sequence analysis indicated that this cDNA sequence has an open reading frame of 1,404 bp, which encodes a 52 kDa peptide of 468 amino acids. This sequence has 64% identity to the previously reported delta-6 fatty acid desaturase from Rhizopus oryzae. The polypeptide has a cytochrome b5 domain at the N-terminus including the HPGG motif in the heme-binding region, as reported for other delta-6 fatty acid desaturases. In addition, this enzyme differs from other desaturases by the presence of three possible N-linked glycosylation sites. Analysis of the fatty acid composition demonstrated the accumulation of GLA to the level of 3.1% of the total fatty acids. Notably, the amounts of ginkgolic acid (C17:1) and palmitic acid (C16:0) were increased from 1.3% to 29.6% and from 15% to 33%, respectively. These results reveal that the modification of the fatty acid biosynthetic pathway by genetic manipulation in order to produce specific polyunsaturated fatty acids in P. pastoris is a promising technique.

• The Korean Journal of Nuclear Medicine
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• v.33 no.6
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• pp.527-536
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• 1999

Purpose: Radiotracers that bind to the central benzodiazepine receptor are useful for the investigation of various neurological and psychiatric diseases. [C-11]Flumazenil, a benzodiazepine antagonist, is the most widely used radioligand for central benzodiazepine receptor imaging by PET. We synthesized 3-(2-[F-18]fluoro)flumazenil, a new fluorine-18 ($t_{1/2}$= 110 min) labeled analogue of benzodiazepine receptor imaging agent, and evaluated in vivo for biodistribution in mice. Materials and Methods: Flumazenil (Ro 15-1788) was synthesized by a modification of the reported method. Precursor of 3-(2-[F-18]fluoro)flumazenil, the tosylated flumazenil derivative was prepared by the tosylation of the ethyl ester by ditosylethane. [F-18] labeling of tosyl substitued flumazenil precursor was performed by adding F-18 ion at $85^{\circ}C$ in the hot ceil for 20 min. The reaction mixture was trapped by C18 cartridge, washed with 10% ethanol, and eluted by 40% ethanol. Bidistribution in mice was determined after intravenous injection. Results: The total chemical yield of tosylated flumazenil derivative was ${\sim}40%$. The efficiency of labeling 3-(2-[F-18]fluoro)flumazenil was 66% with a total synthesis time of 50 min. Brain uptakes of 3-(2-[F-18]fluoro)flumazenil at 10, 30, 60 min after injection, were $2.5{\pm}0.37,\;2.2{\pm}0.26,\;2.1{\pm}0.11$ and blood activities were $3.7{\pm}0.43,\;3.3{\pm}0.07,\;3.3{\pm}0.09%ID/g$, respectively. Conclusion: We synthesized a tosylated flumazenil derivative which was successfully labeled with no-carrier-added F-18 by nucleophilic substitution.

• Journal of Life Science
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• v.32 no.3
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• pp.202-209
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• 2022

The green alga Chlamydomonas reinhardtii, a unicellular haploid eukaryote, has long been used by researchers and industries as a cell factory to produce high value-added microalgae substances using genetic modification. Microalga K01, presumed to be Chlamydomonas, was isolated from 12 freshwater samples from the Chungcheong and Jeolla regions to replace C. reinhardtii, an introduced species currently used in most basic and industrial research. The isolated K01 strain was identified as C. reinhardtii through morphological and phylogenetic studies of the 18S rDNA gene sequence (NCBI accession number KC166137). The growth and lipid content of the isolated C. reinhardtii K01 were compared with three wild and four mutant strains in TAP medium, and it was found that the K01 strain could produce 1.74×10⁷ cells/ml by the third day of culture. The growth rate of C. reinhardtii K01 was 1.5 times faster than UTEX2244, which showed the highest number of cells (1.20×10⁷ cells/ml) among the compared strains. The lipid content of the isolated C. reinhardtii K01 (20.67%) was similar to those of the wild strains, although the fatty acid oleate C18:1 was not detected in the isolated strain but was identified in the seven others. The cell density of the isolated strain increased to 0.87 g/l during a six-day culture in BG11 medium, where nitrate (NaNO₃) was introduced as a nitrogen source, while the seven acquired strains showed almost no cell proliferation.

• Polymer(Korea)
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• v.24 no.4
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• pp.571-577
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• 2000

A new type of filler for epoxy-clay nanocomposites has been prepared by the reaction of octadecyltrimethylammonium bromide and layered sodium montmorillonite (MMT) via an ion-exchange reaction. The gallery space was further modified by grafting the aminopropyl groups via a reaction between a octadecyltrimethylammonium-MMT and 3-aminopropyltriethoxysilane (APS). The interlayer modification of MMT was confirmed by XRD, IR, and solid-state $^{29}$ Si CP/MAS NMR. Furthermore, clay-polymer nanocomposites have been synthesized by the polymerization of diglycidyl ether of bisphenol A(DGEBA) and $C_{18}$ H$_{37}$ N($CH_3$)$_3$-APS-MMT. The resulting hybrid nanocomposites were characterized by XRD, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The results proved that the organomontmorillonite could be exfoliated and uniformly dispersed in the epoxy matrix.

• Education of Primary School Mathematics
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• v.18 no.2
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• pp.61-75
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• 2015

This study analyzed the characteristics of mathematical tasks including the level of cognitive demands set up by pre-service elementary school teachers. 50 pre-service teachers in G university of education who participated in their 4 weeks teaching practicum were selected as subjects. They planned and implemented mathematics lesson with their lesson plans. Lesson plans, video of their lessons, transcript of video were gathered and analyzed the characteristics of mathematical tasks used in their lesson. Through the analysis, several conclusions were drawn as follow. First, 78% of the subjects modified tasks in mathematics textbooks. Since modification or construction of mathematical tasks gives good chance for constructing mathematical task knowledge for teaching, more chance should be given to pre-service teachers to construct new tasks or modify tasks in mathematics textbooks. Second, types of modification done by pre-service teachers were categorized as number change(15.6%), situation change(78.1%) and material change(6.3%). As Chapman(2013) emphasized the importance of MtKT, pre-service teachers must have more MtKT by understanding the characteristics of mathematical tasks. Third, the level of cognitive demands required by mathematical tasks were relatively low. 74% of mathematical tasks was lower cognitive demands and only 26% was higher cognitive demands. The level of cognitive demands of tasks in mathematics textbooks tended to be lowered by the directions given right after the tasks were given. In this respect, the structure of mathematics textbooks need to be changed.

• Journal of the Korean Wood Science and Technology
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• v.43 no.4
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• pp.438-445
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• 2015

Among several thermal wood modification methods German oil heating technology was applied to Korean pine specimens by heat treatment at $180^{\circ}C$ and $200^{\circ}C$. In addition the matched specimens were heat-treated by Thermowood method. The heat-treated specimens were planed and their colors were measured with a colorimeter at 1 mm and 4 mm below the surfaces. The average lightness index (L*) of Oil-180 specimens was the largest followed by Oil-200 and Tmo-200 specimens. The average redness indexes (a*) of all specimens were increased by heat treatment, while the average yellowness indexes (b*) were variable. The average color differences (${\Delta}E*$) of Oil-200 and Tmo-200 specimens were above 30.0, while that of Oil-180 specimens was only 18.4. It can be concluded that the colors of the heat-teated specimens were uniformly changed because their average color difference indexes (${\Delta}E*$) between the inner layers were below 6.0.

• Korean Journal of Food Science and Technology
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• v.40 no.3
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• pp.360-363
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• 2008

Two barley malt samples were selected at two different stages of germination, a well-modified malt germinated for 96 hr and a poorly-modified malt for 60 hr, and were analyzed for total, insoluble, and soluble ${\beta}$-glucan contents. The total ${\beta}$-glucan content in raw barley was 3.96%, and the content was reduced during malting. The total ${\beta}$-glucan contents of the poorly- and well-modified malts were 1.02% and 0.18%, respectively. After 4 days of germination, approximately 95% of the ${\beta}$-glucan present in the barley was degraded. A significantly higher proportion of water-soluble ${\beta}$-glucan was found in the well-modified malt, suggesting that ${\beta}$-glucan solubility was dependent on cell wall modifications in the malt (${\beta}$-glucan breakdown). The proportion of water-soluble ${\beta}$-glucan was also affected by the extraction temperature. The two differently modified malts were mashed isothermally at 45, 55, 65, and 75oC for 2 hr. An increasing mashing temperature resulted in increased viscosity for the wort and the resulting beer. The viscosity of the wort from the well-modified malt was significantly low, due to its low initial malt ${\beta}$-glucan with increased solubility as well as a presumably sufficient ${\beta}$-glucanase activity during mashing.