• Title/Summary/Keyword: Target Synthesis

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Systematic Target Screening Revealed That Tif302 Could Be an Off-Target of the Antifungal Terbinafine in Fission Yeast

  • Lee, Sol;Nam, Miyoung;Lee, Ah-Reum;Lee, Jaewoong;Woo, Jihye;Kang, Nam Sook;Balupuri, Anand;Lee, Minho;Kim, Seon-Young;Ro, Hyunju;Choi, Youn-Woong;Kim, Dong-Uk;Hoe, Kwang-Lae
    • Biomolecules & Therapeutics
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    • v.29 no.2
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    • pp.234-247
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    • 2021
  • We used a heterozygous gene deletion library of fission yeasts comprising all essential and non-essential genes for a microarray screening of target genes of the antifungal terbinafine, which inhibits ergosterol synthesis via the Erg1 enzyme. We identified 14 heterozygous strains corresponding to 10 non-essential [7 ribosomal-protein (RP) coding genes, spt7, spt20, and elp2] and 4 essential genes (tif302, rpl2501, rpl31, and erg1). Expectedly, their erg1 mRNA and protein levels had decreased compared to the control strain SP286. When we studied the action mechanism of the non-essential target genes using cognate haploid deletion strains, knockout of SAGA-subunit genes caused a down-regulation in erg1 transcription compared to the control strain ED668. However, knockout of RP genes conferred no susceptibility to ergosterol-targeting antifungals. Surprisingly, the RP genes participated in the erg1 transcription as components of repressor complexes as observed in a comparison analysis of the experimental ratio of erg1 mRNA. To understand the action mechanism of the interaction between the drug and the novel essential target genes, we performed isobologram assays with terbinafine and econazole (or cycloheximide). Terbinafine susceptibility of the tif302 heterozygous strain was attributed to both decreased erg1 mRNA levels and inhibition of translation. Moreover, Tif302 was required for efficacy of both terbinafine and cycloheximide. Based on a molecular modeling analysis, terbinafine could directly bind to Tif302 in yeasts, suggesting Tif302 as a potential off-target of terbinafine. In conclusion, this genome-wide screening system can be harnessed for the identification and characterization of target genes under any condition of interest.

The prominin-like Gene Expressed in a Subset of Dopaminergic Neurons Regulates Locomotion in Drosophila

  • Ryu, Tae Hoon;Subramanian, Manivannan;Yeom, Eunbyul;Yu, Kweon
    • Molecules and Cells
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    • v.45 no.9
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    • pp.640-648
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    • 2022
  • CD133, also known as prominin-1, was first identified as a biomarker of mammalian cancer and neural stem cells. Previous studies have shown that the prominin-like (promL) gene, an orthologue of mammalian CD133 in Drosophila, plays a role in glucose and lipid metabolism, body growth, and longevity. Because locomotion is required for food sourcing and ultimately the regulation of metabolism, we examined the function of promL in Drosophila locomotion. Both promL mutants and pan-neuronal promL inhibition flies displayed reduced spontaneous locomotor activity. As dopamine is known to modulate locomotion, we also examined the effects of promL inhibition on the dopamine concentration and mRNA expression levels of tyrosine hydroxylase (TH) and DOPA decarboxylase (Ddc), the enzymes responsible for dopamine biosynthesis, in the heads of flies. Compared with those in control flies, the levels of dopamine and the mRNAs encoding TH and Ddc were lower in promL mutant and pan-neuronal promL inhibition flies. In addition, an immunostaining analysis revealed that, compared with control flies, promL mutant and pan-neuronal promL inhibition flies had lower levels of the TH protein in protocerebral anterior medial (PAM) neurons, a subset of dopaminergic neurons. Inhibition of promL in these PAM neurons reduced the locomotor activity of the flies. Overall, these findings indicate that promL expressed in PAM dopaminergic neurons regulates locomotion by controlling dopamine synthesis in Drosophila.

Application of Bayer-Villiger Reaction to the Synthesis of Dibenzo-18-crown-6, Dibenzo-21-crown-7 and Dihydroxydibenzo-18-crown-6

  • Utekar, Druman R.;Saman, Shriniwas D.
    • Journal of the Korean Chemical Society
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    • v.58 no.2
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    • pp.193-197
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    • 2014
  • Dibenzo-18-crown-6, dibenzo-21-crown-7 and dihydroxy dibenzo-18-crown-6 were synthesized by Bayer-Villiger oxidation strategy. Dibenzo-18-crown-6 and dibenzo-21-crown-7 could be synthesized through a three-step protocol starting from salicylaldehyde. Salicylaldehyde was reacted with bis-(2-chloroethyl)ether using $K_2CO_3$ in acetonitrile to link the two phenolic groups with the oxyethylene bridge followed by conversion of the formyl group to the hydroxy group via a Baeyer-Villiger reaction and finally linking the two phenolic group with appropriate oxyethylene bridge. The two target crown ethers were obtained in overall yield, 24% and 30%, respectively. This method has a great potential for synthesis of symmetrical as well as unsymmetrical dibenzo crowns with varying oxyethylene bridges. Baeyer-Villiger oxidation could be used to prepare dihydroxy derivative of dibenzo-18-crown-6 through acetylation of dibenzo-18-crown-6 followed by Baeyer-Villiger oxidation. The Baeyer-Villiger oxidation could be substantially accelerated using trifluoroacetic acid.

(Ba1-xCax)(Ti1-yZry)O3 Powder Synthesis Via Hydrothermal Treatment

  • Park, Byung-Hyun;Choi, Kyoon;Choi, Eui-Seok;Kim, Jong-Hee
    • Journal of the Korean Ceramic Society
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    • v.39 no.11
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    • pp.1017-1022
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    • 2002
  • $(Ba_{1-x}Ca_x)(Ti_{1-y}Zr_y)O_3(BCTZ)$ powders for the Ni-electrode Multilayer Ceramic Capacitor(MLCC) were synthesized via hydrothermal treatment using mixed aqueous solutions of $BaCl_2{\cdot}2H_2O,\Ca(NO_3)2{\cdot}4H_2O,\ ZrOCl_2{\cdot}8H_2O$ and $TiCl_4$. Two component and three component systems were also extensively studied for basic data. BT, CT and BZ powders were crystalline but CZ was determined to be amorphous under the same synthetic condition. In BTZ system, Zr and Ti were completely soluble and Ca would be substituted for Ba up to ∼6 mol% in BCT. The submicron-sized $(Ba_{0.95}Ca_{0.05})(Ti_{0.80}Zr_{0.20})O_3$ powder of the target composition was successfully synthesized at $150{\circ}$ for 12h.

Functional Analysis of the Residues C770 and G771 of E. coli 16S rRNA Implicated in Forming the Intersubunit Bridge B2c of the Ribosome

  • Kim, Hong-Man;Yeom, Ji-Hyun;Ha, Hye-Jung;Kim, Jong-Myung;Lee, Kang-Seok
    • Journal of Microbiology and Biotechnology
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    • v.17 no.7
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    • pp.1204-1207
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    • 2007
  • Structural analyses have shown that nucleotides at the positions 770 and 771 of Escherichia coli 16S rRNA are implicated in forming one of highly conserved intersubunit bridges of the ribosome, B2c. To examine a functional role of these residues, base substitutions were introduced at these positions and mutant ribosomes were analyzed for their protein synthesis ability using a specialized ribosome system. The results showed requirement of a pyrimidine at the position 770 for ribosome function regardless of the nucleotide identity at the position 771. Sucrose gradient profiles of ribosomes revealed that the loss of protein-synthesis ability of mutant ribosome bearing a base substitution from C to G at the position 770 stems from its inability to form 70S ribosomes. These findings indicate involvement of nucleotide at the position 770, not 771, in ribosomal subunit association and provide a useful rRNA mutation that can be used as a target to investigate the physical interaction between 16S and 23S rRNA.

Spirodiclofen Analogues as Potential Lipid Biosynthesis Inhibitors: A Convenient Synthesis, Biological Evaluation, and Structure-Activity Relationship

  • Ke, Shaoyong;Sun, Tingting;Zhang, Zhigang;Zhang, Ya-Ni;Liang, Ying;Wang, Kaimei;Yang, Ziwen
    • Bulletin of the Korean Chemical Society
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    • v.31 no.8
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    • pp.2315-2321
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    • 2010
  • Twenty spirodiclofen analogues have been designed and conveniently synthesized via three steps including esterification, one-pot heterocyclization, and acylation reactions. The target molecules have been identified on the basis of analytical spectra ($^1H$ NMR, $^{13}C$ NMR and ESI-MS) data. All newly synthesized compounds have been screened for their potential insecticidal and herbicidal activity by standard method. The preliminary assays indicated that some of analogues displayed moderate to good insecticidal activity against Plutella xylostella compared with spirodiclofen, and some compounds showed obvious activity against Brassica chinensis. Structure-activity relationship (SAR) is also discussed based on the experimental data.

Inhibition of melanogenesis by tyrosinase siRNA in human melanocytes

  • An, Sang-Mi;Koh, Jae-Sook;Boo, Yong-Chool
    • BMB Reports
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    • v.42 no.3
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    • pp.178-183
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    • 2009
  • Tyrosinase (TYR) plays a critical role in cellular melanogenesis and, thus, has been the major target of pharmacological approaches for the control of skin pigmentation. This study examined an alternative molecular approach using TYR-small interfering RNA (siRNA) to control melanogenesis in the human melanocytes. Both the mRNA and protein levels of TYR were significantly lowered by TYR-siRNA treatment, whereas TYR-related protein 1 and TYR-related protein 2 displayed no such changes. TYR-siRNA treatment inhibited the cellular melanin synthesis from the externally supplied TYR substrate L-tyrosine. TYR-siRNA also suppressed melanin synthesis and decreased the viability of cells exposed to ultraviolet radiation, supporting a critical role of melanin in protection against ultraviolet radiation. These results suggest that molecular approaches using siRNA targeted to the enzymes of melanogenic pathway may provide a novel strategy for the control of cell pigmentation.

Effects of Ball Milling Condition on Sintering of Cu, Zn, Sn and Se Mixed Powders (Cu, Zn, Sn, Se 혼합 분말의 소결특성에 미치는 볼밀링 영향)

  • Ahn, Jong-Heon;Jung, Woon-Hwa;Jang, Yun-Jung;Lee, Seong-Heon;Kim, Kyoo-Ho
    • Journal of Powder Materials
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    • v.18 no.3
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    • pp.256-261
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    • 2011
  • In order to make a $Cu_2ZnSnSe_4$ (CZTSe) sputtering target sintered for solar cell application, synthesis of CZTSe compound by solid state reaction of Cu, Zn, Sn and Se mixed powders and effects of ball milling condition on sinterability such as ball size, combination of ball size, ball milling time and sintering temperature, was investigated. As a result of this research, sintering at $500^{\circ}C$ after ball milling using mixed balls of 1 mm and 3 mm for 72 hours was the optimum condition to synthesis near stoichiometric composition of $Cu_2ZnSnSe_4$ and to prepare sintered pellet with high density relatively.

A Study On Vehicle Interior Noise Reduction Applying FRF Based Substructuring (주파수 응답함수 합성법을 이용한 차량 실내 소음 저감에 관한 연구)

  • Oh, Sang-Hoon;Kang, Yeon-June;Sun, Jong-Cheon;Song, Moon-Sung;Kim, Seong-Goo
    • Proceedings of the Korean Society for Noise and Vibration Engineering Conference
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    • 2006.05a
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    • pp.122-125
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    • 2006
  • The Substructure Synthesis means the technology to predict the dynamic properties of an assembly from the properties of its components, or to predict the effect of a modification on a structure. The FRF Based Substructuring method is a kind of the Substructure Synthesis and very useful to predict the efficiency of the product in the early stage of development. Especially, the Hybrid FBS method is very useful to predict the vehicle NVH characteristics after modifying some components of the vehicle. Target components can be established on the basis of test models and FE models of the prototype constructed in the early stage of development. In this study, the Hybrid FBS method was applied to vehicle subframe and car-body in order to reduce vehicle interior noise induced by engine exciting force.

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Synthesis of Nanoporous Metal Oxide Films Using Anodic Oxidation and Their Gas Sensing Properties

  • Suh, Jun Min;Kim, Do Hong;Jang, Ho Won
    • Journal of Sensor Science and Technology
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    • v.27 no.1
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    • pp.13-20
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    • 2018
  • Gas sensors based on metal oxide semiconductors are used in numerous applications including monitoring indoor air quality and detecting harmful substances like volatile organic compounds. Nanostructures, for example, nanoparticles, nanotubes, nanodomes, and nanofibers have been widely utilized to improve gas sensing properties of metal oxide semiconductors, and this increases the effective surface area, resulting in participation of more target gas molecules in the surface reaction. In the recent times, 1-dimensional (1D) metal oxide nanostructures fabricated using anodic oxidation have attracted great attention due to their high surface-to-volume ratio with large-area uniformity, reproducibility, and capability of synthesis under ambient air and pressure, leading to cost-effectiveness. Here, we provide a brief overview of 1D metal oxide nanostructures fabricated by anodic oxidation and their gas sensing properties. In addition, recent progress on thin film-based anodic oxidation for application in gas sensors is introduced.