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Melanogenesis regulatory constituents from Premna serratifolia wood collected in Myanmar

  • WOO, SO-YEUN
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.21-22
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    • 2019
  • Melanin is a mixture of pigmented biopolymers synthesized by epidermal melanocytes that determine the skin, eye, and hair colors. Melanocytes produce two different kinds of melanin, eumelanin (dark brown/black insoluble pigments found in dark skin and dark hair and pheomelanin (lighter red/yellow). The biological role of melanin is to prevent skin damage by ultraviolet (UV) radiation. However, the overproduction or deficiency of melanin synthesis could lead to serious dermatological problems, which include melasma, melanoderma, lentigo, and vitiligo. Therefore, regulating melanin production is important to prevent the pigmentation disorders. Myanmar has a rich in natural resources. However, the chemical constituents of these natural resources in Myanmar have not been fully investigated. In the effort to search for compounds with anti-melanin deposition activity from Myanmar natural resources, five plants were collected in Myanmar. Extracts of these collected five plants were tested for anti-melanin deposition activity against a mouse melanoma cell line (B16-F10) induced with ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) and 3-isobutyl-1-methylxanthine (IBMX), and their anti-melanin deposition activities were compared with the positive control, arbutin. Among the tested extracts, the CHCl3 extracts of the Premna serratifolia (syn: P. integrifolia) wood showed anti-melanin deposition activities with IC50 values of $81.3{\mu}g/mL$. Hence, this study aims to identify secondary metabolites with anti-melanin deposition activity from P. serratifolia wood of Myanmar. P. serratifolia belongs to the Verbenaceae family and is widely distributed in near western sea coast from South Asia to South East Asia, which include India, Malaysia, Vietnam, Cambodia, and Sri Lanka. People in Tanintharyi region located in the southern part of Myanmar utilize the P. serratifolia, Sperethusa crenulata, Naringi crenulata, and Limonia acidissima as Thanaka, traditional cosmetics in Myanmar. Thanaka is applied in the form of paste onto skins to make it smooth and clear, as well as to prevent wrinkles, skin aging, excessive facial oil, pimples, blackheads, and whiteheads. However, the chemical constituents responsible for their cosmetic properties are yet to be identified. Moreover, the chemical constituents of P. serratifolia was almost uncharacterized. Investigation of the P. serratifolia chemical constituents is thus an attractive endeavor to discover new anti-melanin deposition active compounds. The investigation of the chemical constituents of the active CHCl3 extract of P. serratifolia led to isolation of four new lignoids, premnan A (1), premnan B (2), taungtangyiol C (3), and 7,9-dihydroxydolichanthin B (4), together with premnan C (5) (assumed to be an artifact), one natural newlignoid,(3R,4S)-4-(1,3-benzodioxol-5-ylcarbonyl)-3-[(R)-1-(1,3-benzo dioxol-5-yl)-1-hydroxy methyl]tetrahydro-2-furanone (6), and five known compounds (7-11)1,2). The structures of all isolated compounds were determined on the basis of their spectroscopic data and by comparison with the reported literatures. The absolute configurations of 1-3 and 5 were also determined by optical rotation and circular dichroism (CD) data analyses1). The anti-melanin deposition activities of all the isolated compounds were evaluated against B16-F10 cell line. 7,9-Dihydroxydolichanthin B (4) and ($2{\alpha},3{\alpha}$)-olean-12-en-28-oic acid (11) showed strong anti-melanin deposition activities with IC50 values of 18.4 and $11.2{\mu}M$, respectively, without cytotoxicity2). On the other hand, compounds 1-3, 5, and 7 showed melanogenesis enhancing activities1). To better understand their anti-melanin deposition mechanism, the effects of 4 and 11 on tyrosinase activities were investigated. The assay indicated that compounds 4 and 11 did not inhibit tyrosinase. Furthermore, we also examined the mRNA expression of microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosinase-related protein-1 (TRP-1), and tyrosinase-related protein-2 (TRP-2). Compounds 4 and 11 down-regulated the expression of Tyr and Mitf mRNAs, respectively. Although the P. serratifolia wood has been used as traditional cosmetics in Myanmar for centuries, there are no scientific evidences to support its effectiveness as cosmetics. Investigation of the anti-melanin deposition activity of the chemical constituents of P. serratifolia thus provided insight into the effectiveness of the P. serratifolia wood as a cosmetic agent.

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Molecular Characterization and Expression Analysis of Clathrin-Associated Adaptor Protein 3-δ Subunit 2 (AP3S2) in Chicken

  • Oh, Jae-Don;Bigirwa, Godfrey;Lee, Seokhyun;Song, Ki-Duk
    • Korean Journal of Poultry Science
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    • v.46 no.1
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    • pp.31-37
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    • 2019
  • A chicken clathrin-associated adaptor protein $3-{\delta}$ subunit 2 (AP3S2) is a subunit of AP3, which is involved in cargo protein trafficking to target membrane with clathrin-coated vesicles. AP3S2 may play a role in virus entry into host cells through clathrin-dependent endocytosis. AP3S2 is also known to participate in metabolic disease developments of progressions, such as liver fibrosis with hepatitis C virus infection and type 2 diabetes mellitus. Chicken AP3S2 (chAP3S2) gene was originally identified as one of the differentially expressed genes (DEGs) in chicken kidney which was fed with different calcium doses. This study aims to characterize the molecular characteristics, gene expression patterns, and transcriptional regulation of chAP3S2 in response to the stimulation of Toll-like receptor 3 (TLR3) to understand the involvement of chAP3S2 in metabolic disease in chicken. As a result, the structure prediction of chAP3S2 gene revealed that the gene is highly conserved among AP3S2 orthologs from other species. Evolutionarily, it was suggested that chAP3S2 is relatively closely related to zebrafish, and fairly far from mammal AP3S2. The transcriptional profile revealed that chAP3S2 gene was highly expressed in chicken lung and spleen tissues, and under the stimulation of poly (I:C), the chAP3S2 expression was down-regulated in DF-1 cells (P<0.05). However, the presence of the transcriptional inhibitors, BAY 11-7085 (Bay) as an inhibitor for nuclear factor ${\kappa}B$ ($NF{\kappa}B$) or Tanshinone IIA (Tan-II) as an inhibitor for activated protein 1 (AP-1), did not affect the expressional level of chAP3S2, suggesting that these transcription factors might be dispensable for TLR3 mediated repression. These results suggest that chAP3S2 gene may play a significant role against viral infection and be involved in TLR3 signaling pathway. Further study about the transcriptional regulation of chAP3S2 in TLR3 pathways and the mechanism of chAP3S2 upon virus entry shall be needed.

CLK3 is a Novel Negative Regulator of NF-κB Signaling (NF-κB 신호경로에서 CLK3의 새로운 음성 조절자로서의 기능)

  • Byeol-Eun, Jeon;Chan-Seong, Kwon;Ji-Eun, Lee;Ye-Lin, Woo;Sang-Woo, Kim
    • Journal of Life Science
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    • v.32 no.11
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    • pp.833-840
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    • 2022
  • Chronic inflammation has been shown to be closely associated with tumor development and progression. Nuclear factor kappa B (NF-κB) is composed of a family of five transcription factors. NF-κB signaling plays a crucial role in the inflammatory response and is often found to be dysregulated in various types of cancer, making it an attractive target in cancer therapeutics. In this study, CDC-like kinase 3 (CLK3) was identified as a novel kinase that regulates the NF-κB signaling pathway. Our data demonstrate that CLK3 inhibits the canonical and non-canonical NF-κB pathways. Luciferase assays following the transient or stable expression of CLK3 indicated that this kinase inhibited NF-κB activation mediated by Tumor necrosis factor-alpha (TNFα) and Phorbol 12-myristate 13-acetate (PMA), which are known to activate NF-κB signaling via the canonical pathway. Consistent with data on the ectopic expression of CLK3, CLK3 knockdown using shRNA constructs increased NF-κB activity 1.5-fold upon stimulation with TNFα in HEK293 cells compared with the control cells. Additionally, overexpression of CLK3 suppressed the activation of this signaling pathway induced by NF-κB-inducing kinase (NIK) or CD40, which are well-established activators of the non-canonical pathway. To further examine the negative impact of CLK3 on NF-κB signaling, we performed Western blotting following the TNFα treatment to directly identify the molecular components of the NF-κB pathway that are affected by this kinase. Our results revealed that CLK3 mitigated the phosphorylation/activation of transforming growth factor-α-activated kinase 1 (TAK1), inhibitor of NF-κB kinase alpha/beta (IKKα/α), NF-κB p65 (RelA), NF-κB inhibitor alpha (IκBα), and Extracellular signal-regulated kinase 1/2-Mitogen-activated protein kinase (ERK1/2-MAPK), suggesting that CLK3 inhibits both the NF-κB and MAPK signaling activated by TNFα exposure. Further studies are required to elucidate the mechanism by which CLK3 inhibits the canonical and non-canonical NF-κB pathways. Collectively, these findings reveal CLK3 as a novel negative regulator of NF-κB signaling.

Whitening activity of Ficus carica L. fruits extract through inhibition of tyrosinase and MITF expression (무화과(Ficus carica L.) 열매 추출물의 tyrosinase 및 MITF 발현 억제를 통한 미백 활성)

  • Min Ji Kim;Si Eun Park;Geun soo Lee;Jin Hwa Kim;Sunwoo Kwon;Hyung Seo Hwang
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.204-212
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    • 2023
  • Whitening is inhibitory activity of the melanin synthesis of melanocytes. Recently, whitening materials have been developed on natural materials because of its side effects on skin. Figs (Ficus Carica L.) is a fruit belonging to the Moraceae family and whitening activity was reported in focusing on the fig's stem and leaf components, but whitening activity of the figs fruit was not known. Thus, in this study, we tried to observe its anti-melanogenesis as well as antioxidant and anti-inflammation. The radical scavenging activity of figs fruits extract (FFE) was observed as the level of 34.52±1.98%/60.71±1.26% compared to the control in the its maximum concentration in the DPPH/ABTS assay. Cytotoxicity of FFE was observed at 10% concentration by CCK8 assay, so the maximum concentration was set at 5% and applied to all experiments. FFE concentration dependently decreased NO production associated with inducible nitric oxide synthase, cyclooxygenase-2, interleukin-6 and tumor necrosis factor-α gene expression, these strongly suggesting anti-inflammatory activity. In melanin contents assay, FFE significantly down-regulated melanin production in α-MSH-stimulated B16F10 cell as well as tyrosinase inhibition in vitro. In addition, FFE decreased the Microphthalmia-associated transcription factor (MITF) mRNA expression about 94.34% compared to the α-MSH treatment group in RT-PCR. Finally, FFE significantly reduced the MITF, cAMP response element-binding protein and tyrosinase protein expression in the α-MSH stimulated B16F10 cell. Through these results, we found that FFE can not only directly inhibit tyrosinase enzyme activity but also suppress melanogenesis through regulation of MITF gene expression in α-MSH signal transduction.

Characteristics of Spatio-temporal Variation of the Water Quality in the Lower Keum River (금강 하류역에서 수질의 시공간적 변화특성)

  • YANG Han-Soeb;KIM Seong-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.23 no.3
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    • pp.225-237
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    • 1990
  • Various chemical constituents were measured from April to August 1988 at the down-ward 20 stations of Keum River, which is located in the Midwest of Korea, to understand the characteristics of water quality with respect to spatio-temporal variations of each constituent. The 24-hrs continuous measurements with 2-hrs interval were made simultaneously at station 2 near the estuary weir and station 9(Ganggyeong) of 35 km upstream from the weir in April. By the results observed for one day in April at station 2, salinity has a range of $7.88\~22.14\%_{\circ}$ and its temporal variability is identical to the pattern of tidal cycle in the neigh-bouring Kunsan Harbor. However, turbidity shows relatively high values only at an interval of 4~5 hours after the lowest salinity time, though hourly fluctuation of pH is very small. Silicate and dissolved inorganic nitrogen have inversively linear correlationships with salinity, implying the concentration of the two nutrients strongly regulated by estuarine mixing of sea and river waters. In contrast, phosphate sustains roughly a constant level over a wide salinity range and distinctly lower values than those corresponding to nitrate in the oceans. Such distributions of phosphate have been observed in some estuaries, and interpreted as driven by removal of dissolved phosphate into bottom sediments and the bufforing of phosphate by particulate matter. COD values at station 2 are relatively high in day-time(particularly afternoon) and in high-salinity periods. At station 9, saltwater intrusion was never found but water level changed to the extent of 2.5 m for one day. Although each parameter at this station exhibits very slight variations in their abundance for 24 hours compared with station 2, the contents of COD, silicate and ammonia are significantly higher than at station 2. Concentration of suspended matter is relatively high in the brackish water region up to $\~20$ km above the river mouth, probably due to strong tidal stirring of the bottom de-posits. Also, relatively high pH, COD and $O_2$ saturation at the upward stations of $40\~50$ km from the weir are presumably attributable to active photosynthesis of plants in the region. In general, COD and nutrients except phosphate are higher values at the upper stations than in the estuary zone, and show the highest abundances in July nearly at all stations. Finally, in the estuarine region tidal mixing of sea-river waters seems to be an important factor controlling the distributions of turbidity, COD, silicate and nitrate as well as salinity. However, water quality in the upward fresh-water zone is remarkably variable according to months or seasons.

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The Melodic Structure of the Bulmosan Youngsanjae, Ongho-ge (불모산 영산재 범패 옹호게의 선율구조)

  • Choi, Heon
    • (The) Research of the performance art and culture
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    • no.34
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    • pp.383-421
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    • 2017
  • Because the Jitsori and the Hotsori of the Beompae(the Korean Budhhist chant) has no meter and no Jangdan(a Rhythmic cycle of the Korean Music), so it is hard to analyze the melody of the Beompae. Also the melody of the Beompae is different from that of the other Korean traditional music, so studying of the Beompae has been out of the limelight of many scholars, studying the Korean music. But the melody of Beompae had been handed down for thousands of years in Korea, it and other Korean trditional music, had exchanged the impacts each other for a longtime. So I thinks that the Korean Beomapae have shared the similarity of the musical features with the other Korean traditional music. Because the Beompae of the Bulmosan Yeongsanjae on the Geongsangnamdo province has also no meters and no Jangdan, it is difficult to understand, too. But because the Onghoge of Bulmosan Yeongsanjae have a well-regulated melodic structure in comparison with the Beompae of the Seoul province, so called Geongjae Beompae, it seem to be easy to analyze its melody. So I will analyze the melody of Bulmosan Yeongsanjae Onghoge. This analyze should be contribute to investigate the rule of the melodic progress method on the convoluted Beompae melody. Onghoge has been sung on the procedure for Siryeon, Samsiniun(Goebuliun), Jojeonjeoman, Sinjungjakbeop. And the monk for the ritual has sung the chant first to purify the ritual place and to protect the soul. They has called the song, Onghoge a Jitsori at the Bulmosan Yeongsanjae preservation society of the Gyeongnam province. Commonly, there were Jitsori and Hotsori in the Beompae melody, and the melody of Jitsori is longer than that of the Hotsori. So, the melody of Onghoge is lengthened. In other word, the melody of the Onghoge show the lengthened and curved melodic feture of the Beompae very well. Hahn Manyeong, who had studied on the Beompae, Budhhist chant, said that the Hotsori has five letters in a phrase, and there were 4 phrases in a song. And he had insisted that the form of the song, Hotsori, is ABAB. I analyze the melody of the Onghoge by the Hahn's method. I will extract the Wonjeom(a primary tone of a skeletal melodic structure) from the melody of Onghoge, and in the progress of the Wonjeom of Onghoge melodies, I will arrange the repeat of the Wonjeom melody. It is a structural melody of Onghoge. The first phrase of Bulmosan Yeongsanjae Onghoge, 'Pal bu geum gang ho do ryang(八部金剛護道場)' have 4 structural melodies, the second phrase 'Gong sin sog bu bo cheon wang(空神速赴報天王)', the third phrase 'Sam gye je cheon ham le jip(三界諸天咸來集)', the firth phrase 'Yeo geum bul chal bo jeong sang(如今佛刹補禎祥)' have 2 structural melodies each. The structural melodies of Onghoge are 10 in total. And the structural melody of the Onghoge is formed the shape of 'Mi - La - do - La - Mi'. All of the Onghoge melodies is repeated 10 times by the melodic shape. The form of the Onghoge is not ABAB by Hahn, but is 10 times repeat of the shape.