• Title/Summary/Keyword: RT synthesis

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Synthesis of Heterocycle-linked Thioureas and Their Inhibitory Activities of NO Production in LPS Activated Macrophages

  • Cheon, Ye-Jin;Gim, Hyo-Jin;Jang, Hee-Ryun;Ryu, Jae-Ha;Jeon, Raok
    • Bulletin of the Korean Chemical Society
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    • v.31 no.1
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    • pp.27-30
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    • 2010
  • A series of thioureas were synthesized as inhibitors of NO production in lipopolysaccharide-activated macrophages. We investigated the effect of lipophilic moiety and N-substituents of the thioureas on the activity. Phenoxazine and carbazole-containing derivatives revealed higher activity than indole-containing thioureas. The appropriate spacer between lipophilic tail and thiourea head and methyl substituent at N3 position of thiourea brought beneficial effect on the inhibition of NO production. Among prepared compounds, phenoxazine-containing derivative 2a was the most potent with $2.32 {\mu}M$ of $IC_{50}$ value. RT-PCR analysis suggested that the prepared thioureas inhibited NO production through the suppression of iNOS mRNA expression.

Antimicrobial Activity of Licochalcone E Against Staphylococcus aureus and Its Impact on the Production of Staphylococcal Alpha-Toxin

  • Zhou, Tiezhong;Deng, Xuming;Qiu, Jiazhang
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.800-805
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    • 2012
  • Licochalcone E was firstly isolated from licorice root in 2005, which belongs to the retrochalcone family. Studies on the biological activities of licochalcone E were in the initial stage. In the study, we demonstrated that licochalcone E has potent antimicrobial property against Staphylococcus aureus. Furthermore, via hemolysis, Western blot, and real-time RT-PCR assays, we have shown that subinhibitory concentrations of licochalcone E dose-dependently reduces the production of ${\alpha}$-toxin in both methicillin-sensitive S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA). The data suggest that licochalcone E may deserve further investigation as a potential therapeutic against S. aureus infections, or the structure of licochalcone E may be used as a basis for chemical synthesis of novel anti-S. aureus compounds.

Anti-inflammatory Effect and Inhibition of Melanin Biosynthesis of Clematis mandshurica (위령선 추출물의 항염활성 및 멜라닌 생성 억제효과)

  • Kim, Ye Rim;Hong, Yun Jung;Yang, Ki Sook
    • YAKHAK HOEJI
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    • v.58 no.1
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    • pp.47-52
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    • 2014
  • Clematis mandshurica (Ranunculaceae) has traditionally been used as a remedy for antidiuretic, antifungal, rheumatic conditions and alleviate pain. We carried out to evaluate the anti-oxidative effect, anti-inflammatory effect and anti-melanogenic effect of ethanol extract and solvent fractions of Clematis mandshurica. The ethanol extract and the dichloromethane fraction of Clematis mandshurica showed an anti-oxidative effect in DPPH assay, the inhibitory activity of nitric oxide (NO) production in lipopolysaccharide (LPS) activated RAW 264.7 cell, and melanin synthesis and tyrosinase activity of B16F10 melanoma cells. They reduced NO production and melanin content in a dose-dependent manner at concentrations of $2.5{\sim}10{\mu}g/ml$. They also suppressed iNOS and tyrosinase protein and m-RNA expressions dose dependently, assayed by western blot analysis and RT-PCR experiment.

Synthesis of N-Aryl Phenylglycine Ο-Alkyl Esters and Its Substitution of Ester Moiety (N-Aryl Phenylglycine Ο-Alkyl Esters의 간편한 합성 및 에스테르 치환반응)

  • 박명숙;박해선
    • YAKHAK HOEJI
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    • v.47 no.5
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    • pp.276-282
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    • 2003
  • For the development of new synthetic method for unnatural amino acid esters, N-aryl phenylglycine Ο-alkyl esters 4a∼i were synthesized through esterification, bromination, C-N bond formation from commercially available phenylacetic acids. An efficient and practical reaction condition for esters 2a∼c was that the starting materials 1a∼c were refluxed in absolute methanol for 3 hours with catalytic concentrated hydrosulfuric acid. In addition, bromines 3a∼c were formated for 3h in dichloromethane at rt with N-bromosuccinimide. Bromines 3a∼c were also converted to 4a∼i through substitution of arylamines during refluxing for 24 hours in ethanol with triethylamine. Interestingly, ethyl esters 5a∼c were formed via transesterification reaction when the p-sulfamylanilino group was used as a nucleophile in ethanol solvent.

Salicylamide Enhances Melanin Synthesis in B16F1 Melanoma Cells

  • Ito, Yusuke;Sato, Kazuomi
    • Biomolecules & Therapeutics
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    • v.29 no.4
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    • pp.445-451
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    • 2021
  • Salicylamide, a non-steroidal anti-inflammatory drug (NSAID), is used as an analgesic and antipyretic agent. We have previously shown that several NSAIDs have anti-melanogenic properties in B16F1 melanoma cells. In contrast, we have found that salicylamide enhances melanin contents in B16F1 melanoma cells; however, the underlying mechanism is not known. Therefore, we investigated the mechanism through which salicylamide stimulates melanogenesis. Interestingly, salicylamide enhanced diphenolase activity in a cell-free assay. Western blotting and real-time RT-PCR revealed that salicylamide increased tyrosinase expression via transcriptional activation of the Mitf gene. Together, our results indicate that salicylamide could be used as an anti-hypopigmentation agent for skin and/or hair.

Effects of Dokhwalkisaeng-tang on Melanin Synthesis Inhibition and Gene Expression in B16F10 Melanoma Cells (독활기생탕(獨活寄生湯)이 멜라닌 생성억제 및 유전자 발현에 미치는 영향)

  • Oh, Won-Kyo;Kim, Ki-Byoung;Lim, Jin-Young;Lee, Su-Kyung;Kwon, Young-Dal;Yeom, Seung-Ryong;Song, Yung-Sun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.1
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    • pp.63-75
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    • 2009
  • The aim of this study was to elucidate the antimelanogenic effect of Dokhwalkisaeng-tang(Duohujisheng-tang) in B16F10 melanoma cells. Dokhwalkisaeng-tang(DKT) was used to develop the effective prescription of inhibition of melanin production. We determined inhibitory effects of DKT on melanin-release, melanin production, and tyrosinase activity in B16F10 melanoma cells. And to explicate the action-mechanism of DKT, melanin-related gene expressions were determined using RT-PCR and real time RT PCR technique in B16F10 melanoma cells. DKT inhibited melanin-release, melanin production in B16F10 melanoma cells considerably. DKT inhibited tyrosinase activity in vitro and in B16F10 melanoma cells. DKT inhibited the expression of tyrosinase, TRP-1, TRP-2 in B16F10 melanoma cells. DKT inhibited the expression of PKA, PKC, MMP-2 and MITF in B16F10 melanoma cells. On the other hand, DKT increased the expression of ERK-1, ERK-2, AKT-1 in B16F10 melanoma cells. From these results, we propose that DKT may have effect on the antimelanogenesis.

Effect of immobilization stress on the expression of TH, BDH and CRH gene in rat brain (부동스트레스가 흰쥐 뇌 조직 내 TH, BDH와 CRH 유전자 발현에 미치는 영향)

  • Qian, Yong-Ri;Kim, Yoon-Sik
    • Journal of Genetic Medicine
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    • v.4 no.2
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    • pp.179-185
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    • 2007
  • Purpose : Catecholamines are the neuro-transmitters in the sympathetic nervous system (SNS) and are activated by stress stimulus. Tyrosine hydroxylase (TH) and Dopamine-${\beta}$-Hydroxylase (DBH) are very important enzymes in the catecholamine synthesis. Corticotropin releasing hormone (CRH) is released in the process of reacting to stresses. The aim of this study is to find out what effects immobilization stresses have on the expression of TH, BDH and CRH mRNA in a rat's brains. Methods : We compare expression levels in rat's brains of TH, DBH and CRH mRNA induced by immobilization stresses between the test group and controled group. The expression levels of TH, DBH and CRH mRNA are measured by RT-PCR and the Western Blotting Analysis (WBA). Results : In brains and adrenal glands of the immobilization stress group, the expression levels of TH and DBH mRNAs are significantly two to three times higher (P<0.01), and CRH mRNAs are approximately one and a half times higher (P<0.05) than those of controlled group. Conclusion : This study suggest that the expression levels of TH, DBH and CRH mRNAs are activated by stress stimulus in a rat's brains and adrenal glands.

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Occurrence of Apple scar viroid-Korean strain (ASSVd-K) in Apples Cultivated in Korea

  • Lee, Ju-Hee;Park, Jean-Kyung;Lee, Dong-Hyuk;Uhm, Jae-Youl;Ghim, Sa-Youl;Lee, Jai-Youl
    • The Plant Pathology Journal
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    • v.17 no.5
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    • pp.300-304
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    • 2001
  • Apple is the most economically important fruit in Korea. The suspected viroid disease of dapple apple was found in apple fruits cultivated in Kyungpook province. Symptoms begin in mid-July as small circular spots, which stand out against the background color on the young fruit. Dappling of the fruit becomes more intense and easier to detect as the fruit approaches maturity; the affected spots remain yellowish as the fruit matures. no leaf or bark syndromes have been associated with this disease. The infected fruits are downgraded considerably during quality grading. The low molecular weight RNA containing viroid RNA molecules were extracted from the peels of the apples with dapple symptoms. The RNA molecules were extracted from the apples using Qiagen column chromatography. The purified RNAs were used for the synthesis of cDNA with RT-PCR. The PCR products were then ligated into a pGEM-T Easy vector, cloned and sequenced. The sequence of the viroid RNA molecule shows 331 nucleotides with one base difference ("G" insertion between the position of 133 and 134) compared with that of the Apple scar skin viroid (ASSVd) reported by Hashimoto and Koganezawa in Japan. This is the first report on the occurrence of the ASSVd in apple trees cultivated in Korea, as well as the identification of a new Korean strain of the ASSVd.the ASSVd.

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High-Speed RNA Isolation Using Magnetic Oligo(dT) Beads and Lateral Magnetophoresis (올리고-dT 자성입자와 측면방향 자기영동을 이용한 초고속 RNA 추출 기술)

  • Lee, Hwan-Yong;Han, Song-I;Han, Ki-Ho
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.35 no.12
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    • pp.1309-1316
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    • 2011
  • This paper presents a high-speed RNA microextractor for the direct isolation of RNA from blood lysate using magnetic oligo(dT) beads. The extraction is performed through lateral magnetophoresis, which is induced by a ferromagnetic wire array inlaid. With this RNA microextractor, more than 80% of the magnetic beads could be separated at a flow rate up to 20 ml/h, and the overall extraction procedure was completed within 1 min. The absorbance ratio of RNA to protein(A260/A280) was greater than 1.7, indicating that the extraction technique yields pure RNA. The feasibility of using this technique in reverse transcription polymerase chain reaction procedures was investigated by cDNA synthesis and PCR processes. The results confirmed that the RNA microextractor is a practical device for easy, fast, and high-precision RT-PCR using minimal amounts of reagent.

RANKL expression is mediated by p38 MAPK in rat periodontal ligament cells (백서 치주인대세포의 RANKL 발현에 대한 p38 MAPK의 역할)

  • Kim, Chong-Cheol;Kim, Young-Joon;Chung, Hyun-Ju;Kim, Ok-Su
    • Journal of Periodontal and Implant Science
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    • v.34 no.3
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    • pp.489-498
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    • 2004
  • Recent studies have demonstrated that human periodontal ligament cells express receptor activation of nuclear factor ${\kappa}B$ ligand (RANKL) which enhances the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The purpose of this study is to determine the effects of p38 MAPK and JNK kinase upon regulating RANKL and OPG in response to $IL-1{\beta}$(l ng/ml) in rat periodontal ligament cells. Soluble RANKL was measured by immunoassay. The effects of p38 MAPK on RANKL and OPG expression was determined by RT-PCR. The results were as follows: 1. Periodontal ligament cells which stimulated by $IL-1{\beta}$ increased soluble RANKL synthesis by dose-dependent pattern. 2. p38 MAP kinase inhibitor (SB203580) showed regulation of soluble RANKL expression by dose-dependent manners. 3. p38 MAP kinase inhibitor (SB203580) regulated the expression of RANKL, but it dose regulate the expresseion of OPG. 4. JNK (c-jun $NH_2-terminal$ kinase) inhibitor (PD98059) did not regulate mRANKL and mOPG. These results suggested that p38 MAPK play a significant role in RANKL gene expression.