Melanosomes are specific melanin-containing intracellular organelles of epidermal melanocytes. In epidermal melanocytes, there are three kinds of key player proteins. Rab27a, melanophilin or Slac2-a and Myosin 5a form a tripartite complex connects the melanosome. Mature melanosomes make movements through the tripartite protein complex along actin filaments.In this study, we found that the haplamine (6-Methoxyflindersine) induced melanosome aggregation around the nucleus in epidermal melanocyte. In an attempt to elucidate the inhibitory effect of haplamine on melanosome transport, effect of haplamineon the expression level of Rab27a, melanophilin and Myosin 5a was measured. The results indicated that haplamine up to 5M effectively suppressed mRNA and protein expression level of melanophilin.To determine the upstream regulator of melanophilin regulated by haplamine, we checked the level of MITF, c-JUN and USF1. Those are possible transcription factor of melanophilin. Among them,treatment of USF1 siRNA decreased mRNA and protein expression level of USF1 as well as melanophilin. Also, treatment of haplamine decreased mRNA and protein expression level of melanophilin as well as USF1 in a dose-dependent manner. Consequently, we found the inhibitory effect of haplamine on melanosome transport in melan-a melanocyte. Treatment of haplamine reduced melanophilin expression level which is a key protein of melanosome transport. We identified that USF1 could be a major transcription factor of melanophilin regulated by haplamine.
Since the epigenetic alteration in tumor cells can be reversed by the dietary polyphenol quercetin (Q) or butyrate (B) with chemopreventive activity, suggesting that Q or B can be used for chemopreventive as well as therapeutic agent against tumors. In this study the polyphenol flavonoid quercetin (Q) or sodium butyrate (B) suppressed human esophageal 9706 cancer cell growth in dose dependent manner, and Q combined with B (Q+B) could further inhibit Eca9706 cell proliferation than that induced by Q or B alone, compared with untreated control group (C) in MTT assay. The reverse expressions of global DNMT1, $NF-{\kappa}Bp65$, HDAC1 and Cyclin D1 were down-regulated, while expressions of caspase-3 and $p16INK4{\alpha}$ were up-regulated, compared with the C group in immunoblotting; the down-regulated HDAC1-IR (-immunoreactivity) with nuclear translocation, and up-regulated E-cadherin-IR demonstrated in immunocytochemistry treated by Q or B, and Q+B also displayed further negatively and positively modulated effects compared with C group. The order of methylation specific (MS) PCR of $p16INK4{\alpha}$: C>B/Q>Q+B group, while the order of E-cadherin expression level was contrary, Q+B>Q/B>C group. Thus, Q/B, especially Q+B display reverse effect targeting both altered DNA methylation and histone acetylation, acting as histone deacetylase inhibitor mediated via epigenetic-$NF-{\kappa}B$ cascade signaling.
This study was designed to investigate the emission characteristics of unregulated pollutants (Aldehyde, VOCs, PAHs) as well as regulated pollutants (CO, HC, NOx and PM) from diesel medium-duty trucks. The emission characteristics of unregulated and regulated pollutants were assessed based on regulation standards (EURO 4 and EURO 5) and intake weight (2.5 ton and 5 ton). The results show that unregulated and regulated pollutants remained almost unchanged at higher speeds but decreased at below 23.5 km/h. Reduction in unregulated and regulated pollutants was noticeable in vehicles of recent regulation standards and light intake weight. The analysis of aldehyde using UPLC showed that formaldehyde and acetaldehyde of aldehyde were most dominant. The GC/MS analysis showed that benzene, toluene, ethylbenzene and xylene of VOCs was over 80% followed by toluene, xylene, ethylbenzene and benzene. In addition, the analysis of PAHs using GC/TOF-MS indicated that bi- and tricyclic aromatic ring of aromatic compounds was 73% and 53% at 2.5 ton and 5 ton vehicles, respectively. The results of this study will be contributed to establish HAPs inventory.
The Swedish mutation (K595N/M596L) of amyloid precursor protein (APP-swe) has been known to increase abnormal cleavage of cellular APP by Beta-secretase (BACE), which causes tau protein hyperphosphorylation and early-onset Alzheimer's disease (AD). Here, we analyzed the effect of APP-swe in global gene expression using deep transcriptome sequencing technique. We found 283 genes were down-regulated and 348 genes were up-regulated in APP-swe expressing H4-swe cells compared to H4 wild-type cells from a total of approximately 74 million reads of 38 base pairs from each transcriptome. Two independent mechanisms such as kinase and phosphatase signaling cascades leading hyperphosphorylation of tau protein were regulated by the expression of APP-swe. Expressions of catalytic subunit as well as several regulatory subunits of protein phosphatases 2A were decreased. In contrast, expressions of tau-phosphorylating glycogen synthase kinase $3\beta$(GSK-3$\beta$), cyclin dependent kinase 5 (CDK5), and cAMP-dependent protein kinase A (PKA) catalytic subunit were increased. Moreover, the expression of AD-related Aquaporin 1 and presenilin 2 expression was regulated by APP-swe. Taken together, we propose that the expression of APP-swe modulates global gene expression directed to AD pathogenesis.
Objectives : This study was aimed to investigate the therapeutic effect of Bogijetongtanggammi-bang (BJTG) on injury of the peripheral nerve tissues. Methods : Rats were divided into 2 groups. The rats of the first group were injected with Taxol (1.25 mg/kg) to their sciatic nerves, once each. The sciatic nerves of the rats of the second group were crushed by forcept for 30 seconds. Rats were administered with BJTG (400 mg/kg) or 0.9% saline for 5 days. Changes of DRG neurons, Schwann cells, Cdc2, caspase 3. phospho-p44/42 Erk1/2, phospho-vimentin and ${\beta}1$ integrin were observed by fluorescent microscope and analysed in western blot. Results : In Taxol-treated SD rat models, BJTG up-regulated neurite outgrowth, Schwann cells, Cdc2 and phospho-Erk1/2, and down-regulated caspase 3. In pressure-injured rat models, BJTG up-regulated axons of sciatic nerve, Schwann cells, Cdc2, phospho-vimentin, ${\beta}1$ integrin, and down-regulated caspase 3. Conclusions : Taken together, BJTG was promotive of nerve regeneration on SNI as well as Taxol-induced nerve injury. BJTG had a pharmaceutical property enhancing recovery of injured peripheral nerves and could be a candidate for drug development after further research.
Kwon, Soo Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag Hyun;Boo, Hee Ock;Song, Beom-Heon;Woo, Sun-Hee
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
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pp.132-132
/
2017
Plants, including Platycodon grandiflorum have been used globally across varied cultures as a safe natural source of medicines. From time immemorial, humans have relied on plants that could meet their basic necessities such as food, shelter, fuel and health. This study was executed to profile proteins from the hormone induced diploid and tetraploid roots using high throughput proteome approach. Two dimensional gels stained with CBB, a total of 64 differential expressed proteins were identified from the diploid root using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 20 differential expressed protein spots ( ${\geq}1.5-fold$) were analyzed using LTQ-FTICR MS whereas a total of 13 protein spots were up regulated and 7 protein spots were down-regulated. However, in the case of tetraploid root, a total of 78 differential expressed proteins were identified from tetraploid root of which a total of 28 differential expressed protein spots (${\geq}1.5-fold$) were analyzed by mass spectrometry whereas a total of 16 protein spots were up regulated and a total of 12 protein spots were down-regulated. However, proteins identified using iProClass databases revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding, oxidoreductase activity, transporter activity and isomers activity. The exclusive protein profile may provide insight clues for better understanding the characteristics of protein function and its metabolic activity that can help for the development of the nutritional and breeding aspects of this economically important medicinal plant.
Kim, In Hwang;Son, Jee-Soo;Wen, Yancheng;Jeong, Sang-Min;Min, Ga-Young;Park, Na-Young;Lee, Keun-Woo;Cho, Yong-Joon;Chun, Jongsik;Kim, Kun-Soo
Journal of Microbiology and Biotechnology
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v.23
no.12
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pp.1791-1801
/
2013
Diketopiperazine is produced by various organisms, including bacteria, fungi, and animals, and has been suggested as a novel signal molecule involved in the modulation of genes with various biological functions. Vibrio vulnificus, which causes septicemia in humans, produces cyclo($\small{L}$-phenylalanine-$\small{L}$-proline) (cFP). To understand the biological roles of cFP, the effect of the compound on the expression of the total mRNA in V. vulnificus was assessed by next-generation sequencing. Based on the transcriptomic analysis, we classified the cFP-regulated genes into functional categories and clustered them according to the expression patterns resulted from treatment with cFP. From a total of 4,673 genes, excepting the genes encoding tRNA in V. vulnificus, 356 genes were up-regulated and 602 genes were down-regulated with an RPKM (reads per kilobase per million) value above 3. The genes most highly induced by cFP comprised those associated with the transport and metabolism of inorganic molecules, particularly iron. The genes negatively regulated by cFP included those associated with energy production and conversion, as well as carbohydrate metabolism. Noticeably, numerous genes related with biofilm formation were modulated by cFP. We demonstrated that cFP interferes significantly with the biofilm formation of V. vulnificus.
This paper is aimed to analyze how to evaluate the choice of optimal management measures and level of control in fisheries management under the costly and imperfect management system by comparing with costless and perfect management system that is commonly assumed in the analysis of fisheries regulations. Fishermen would set the level of fishing efforts at the point where the marginal fishing profit for fishing effort is equal to the marginal level of fine under costly and imperfect management system. Therefore, under the case where the marginal fishing profit is higher than the marginal level of fine, the level of fishing efforts would be made at the point which is higher than the level of fishing efforts made under costless and perfect management system and is not a point where the economic profit is maximized in regulated fisheries. From this conclusion, the fishermens avoidance activities against regulations as well as the level of control in fisheries management substantially have an influence on the choice of fisheries management instruments. According to the analysis of optimal fisheries management policy, the economic profits in regulated fisheries are determined by the level of fisheries enforcement costs and total fishing profits, in which as enforcement costs increase the economic profits decrease. In addition, the economic profits vary in response to the level of control in avoidance activities. That is, as avoidance costs decrease, the economic profits increase. The determination of optimal level of control in fisheries management should be made at the point where the marginal regulation costs are equal to the marginal profits from regulated fisheries, in which marginal regulation costs are different according to the type of management measures. And the level of profits changes in response to different levels of avoidance activities. The management measure that can maximize the difference between the marginal regulation costs and marginal profits from regulated fisheries should be chosen as an optimal fisheries management instrument.
Dayarathne, Lakshi A.;Ranaweera, Sachithra S.;Natraj, Premkumar;Rajan, Priyanka;Lee, Young Jae;Han, Chang-Hoon
Journal of Veterinary Science
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v.22
no.4
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pp.55.1-55.17
/
2021
Background: Naringenin and its glycoside naringin are well known citrus flavonoids with several therapeutic benefits. Although the anti-adipogenic effects of naringenin and naringin have been reported previously, the detailed mechanism underlying their anti-adipogenesis effects is poorly understood. Objectives: This study examined the anti-adipogenic effects of naringenin and naringin by determining differential gene expression patterns in these flavonoids-treated 3T3-L1 adipocytes. Methods: Lipid accumulation and triglyceride (TG) content were determined by Oil red O staining and TG assay. Glucose uptake was measured using a 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-d-glucose fluorescent d-glucose analog. The phosphorylation levels of AMP-activated protein kinase (AMPK) and acetyl Co-A carboxylase (ACC) were observed via Western blot analysis. Differential gene expressions in 3T3-L1 adipocytes were evaluated via RNA sequencing analysis. Results: Naringenin and naringin inhibited both lipid accumulation and TG content, increased phosphorylation levels of both AMPK and ACC and decreased the expression level of 3-hydroxy-3-methylglutaryl CoA reductase (HMGCR) in 3T3-L1 adipocytes. RNA sequencing analysis revealed that 32 up-regulated (> 2-fold) and 17 down-regulated (< 0.6-fold) genes related to lipid metabolism, including Acaca, Fasn, Scd1, Mogat1, Dgat, Lipin1, Cpt1a, and Lepr, were normalized to the control level in naringenin-treated adipocytes. In addition, 25 up-regulated (> 2-fold) and 25 down-regulated (< 0.6-fold) genes related to lipid metabolism, including Acaca, Fasn, Fabp5, Scd1, Srebf1, Hmgcs1, Cpt1c, Lepr, and Lrp1, were normalized to the control level by naringin. Conclusions: The results indicate that naringenin and naringin have anti-adipogenic potentials that are achieved by normalizing the expression levels of lipid metabolism-related genes that were perturbed in differentiated 3T3-L1 cells.
Du, Jian;Yang, Si-Tong;Liu, Jia;Zhang, Ke-Xin;Leng, Ji-Yan
Molecules and Cells
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v.42
no.5
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pp.397-405
/
2019
The regulatory role of long noncoding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) in both cancerous and noncancerous cells have been widely reported. This study aimed to evaluate the role of lncRNA GAS5 in heart failure caused by myocardial infarction. We reported that silence of lncRNA GAS5 attenuated hypoxia-triggered cell death, as cell viability was increased and apoptosis rate was decreased. This phenomenon was coupled with the down-regulated expression of p53, Bax and cleaved caspase-3, as well as the up-regulated expression of CyclinD1, CDK4 and Bcl-2. At the meantime, the expression of four heart failure-related miR-NAs was altered when lncRNA GAS5 was silenced (miR-21 and miR-142-5p were up-regulated; miR-30b and miR-93 were down-regulated). RNA immunoprecipitation assay results showed that lncRNA GAS5 worked as a molecular sponge for miR-142-5p. More interestingly, the protective actions of lncRNA GAS5 silence on hypoxia-stimulated cells were attenuated by miR-142-5p suppression. Besides, TP53INP1 was a target gene for miR-142-5p. Silence of lncRNA GAS5 promoted the activation of PI3K/AKT and MEK/ERK signaling pathways in a miR-142-5p-dependent manner. Collectively, this study demonstrated that silence of lncRNA GAS5 protected H9c2 cells against hypoxia-induced injury possibly via sponging miR-142-5p, functionally releasing TP53INP1 mRNA transcripts that are normally targeted by miR-142-5p.
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