• Title/Summary/Keyword: fusion expression

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A Study on the Whitening Effect of Mangifera indica L. Peel Extracts through Inhibition of Melanin Synthesis Factor (Melanin 생성 인자 억제 효과를 통한 Mangifera indica L. Peel의 미백효과 연구)

  • Kim, Hyo-Min;Yoo, Dan-Hee;Lee, In-Chul
    • Microbiology and Biotechnology Letters
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    • v.50 no.1
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    • pp.31-39
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    • 2022
  • The purpose of this study was to investigate the whitening effects of hot water (AMPW) and ethanol (AMPE) extracts of Mangifera indica L. peel. To verify the whitening effects, tyrosinase inhibitory activity was measured. 9.51% inhibitory activity, and 35.98% inhibitory activity at 1,000 ㎍/ml. The effects of AMPW and AMPE on cell viability were measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in B16-F10 melanoma cells. Greater than 95% cell viability was observed at 100 ㎍/ml. Thus, subsequent experiments were performed at concentrations less than 100 ㎍/ml. The whitening effects were confirmed by measuring the protein and mRNA expression levels of microphthalmia-associated transcription factor, tyrosinase, tyrosinase-related protein 1 (TRP-1), and TRP-2, which are factors involved in melanin synthesis. Western blotting and reverse transcription-polymerase chain reaction results confirmed that 100 ㎍/ml AMPW and AMPE showed superior inhibitory effects than the control treatment (alpha-melanocyte stimulating hormone only). Therefore, Mangifera indica L. peel extract had a whitening effect, and thus, has potential as a natural material for use in cosmetics.

Tat-Thioredoxin-like protein 1 attenuates ischemic brain injury by regulation of MAPKs and apoptosis signaling

  • Hyun Ju Cha;Won Sik Eum;Gi Soo Youn;Jung Hwan Park;Hyeon Ji Yeo;Eun Ji Yeo;Hyun Jung Kwon;Lee Re Lee;Na Yeon Kim;Su Yeon Kwon;Yong-Jun Cho;Sung-Woo Cho;Oh-Shin Kwon;Eun Jeong Sohn;Dae Won Kim;Duk-Soo Kim;Yu Ran Lee;Min Jea Shin;Soo Young Choi
    • BMB Reports
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    • v.56 no.4
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    • pp.234-239
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    • 2023
  • Thioredoxin-like protein 1 (TXNL1), one of the thioredoxin superfamily known as redox-regulator, plays an essential in maintaining cell survival via various antioxidant and anti-apoptotic mechanisms. It is well known that relationship between ischemia and oxidative stress, however, the role of TXNL1 protein in ischemic damage has not been fully investigated. In the present study, we aimed to determine the protective role of TXNL1 against on ischemic injury in vitro and in vivo using cell permeable Tat-TXNL1 fusion protein. Transduced Tat-TXNL1 inhibited ROS production and cell death in H2O2-exposed hippocampal neuronal (HT-22) cells and modulated MAPKs and Akt activation, and pro-apoptotic protein expression levels in the cells. In an ischemia animal model, Tat-TXNL1 markedly decreased hippocampal neuronal cell death and the activation of astrocytes and microglia. These findings indicate that cell permeable Tat-TXNL1 protects against oxidative stress in vitro and in vivo ischemic animal model. Therefore, we suggest Tat-TXNL1 can be a potential therapeutic protein for ischemic injury.

Transgenic Mice Overexpressing Cocaine-Amphetamine Regulated Transcript in the Brain and Spinal Cord (뇌와 척수에서 Cocaine-Amphetamine Regulated Transcript를 과발현하는 형질전환 생쥐)

  • Choi, S.H.;Lee, J.W.;Park, H.D.;Jahng, J.W.;Chung, K.S.;Lee, H.T.
    • Korean Journal of Animal Reproduction
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    • v.25 no.4
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    • pp.389-397
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    • 2001
  • Cocaine-amphetamine regulated transcript (CART), a satiety factor regulated by leptin, is associated with food intake and motor behavior. In knock out studies, Leu34Phe mutation of human CART gene resulted in obese phenotype but mice carrying a targeted deletion of the CART gene exhibited no dramatic increase of body weight on normal fat diet. To establish a new transgenic mouse model for determining the function of CART on feeding behavior in vivo, we constructed the fusion gene, CART gene under the control of neurofilament light chain promoter, which regulates gene expression at the stage of neuronal differentiation. Transgenic mice were generated by microinjection method and screened by PCR and Southern blot analyses. In these transgenic mice, overexpression of CART was detected by in situ hybridization in spinal cords and brains at 13.5 days post-coitum embryos. At six weeks of age, RT-PCR analysis showed that exogenous CART mRNA was expressed strongly in brains and spinal cords, but not much in other tissues. Our results suggest that these transgenic mice provide a new model to investigate the function of CART gene in neuronal network associated with feeding behavior.

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Production of Transgenic Bovine Embryos Following Nuclear Transfer of Bovine Fetal Fibroblasts Transfected by Foreign Genes (외래유전자를 도입한 소 태아세포의 핵치환에 의한 형질전환 소 수정란 생산)

  • Kil, K.S.;Uhm, S.J.;Kim, E.H.;Chung, H.J.;Kim, T.;Park, H.;Lee, H.T.;Chung, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.429-437
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    • 2000
  • This study investigated the successful introduction of genes of erythropoietin (EPO) and enhanced green fluorescent protein (EGFP) in bovine embryos following nuclear transfer of bovine fetal fibroblasts (bFF), which were transfected by retrovirus vector system. Non-starved bFF were, transferred into perivitelline space of enucleated oocytes. The bFF-oocyte units were accomplished by cell to cell fusion and activated with calcium inophore and 6-dimethylaminopurine. Reconstructed embryos were co-cultured with bovine oviduct epithelial cells in CRlaa medium for 8 days. Out of 187 (EPO) and 210 (EGFP) bovine eggs reconstructed by nuclear transfer, 149 (EPO : 80.0%) and 158 (EGFP : 75.2%) embryos were cleaved, and among them 36 (EPO : 24.2%) and 35 (EGFP : 22.2%) embryos developed to the blastocyst stage. Of these blastocysts, 100% integration of EPO gene in 36 embryos was determined by PCR, and 100% expression of EGFP gene in 35 embryos was observed under the fluorescent microscope. This result indicates that bovine oocytes reconstructed by nuclear transfer of transfected bFF can successfully develop to the blastocyst stage. Furthermore, this novel procedure may be presumably an attractive method efficiently to produce the transgenic cattles.

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A Study on the Stage Costume of Pansori Ballet Theater 'Dokkaeba! Dokkaeba!' Based on Storytelling (스토리텔링에 의한 판소리 발레극 '도깨바! 도깨바!'의 무대의상 연구)

  • Ryu Jinyoung
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.3
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    • pp.399-406
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    • 2023
  • Due to the increasing influence of the Korean Wave, traditional themes are emerging to the forefront of performance content development, emphasizing the necessity of applying the art of storytelling to the creative process. The purpose of this research is to examine the differentiated costume design of the Pansori ballet "Dokkaeba! Dokkaeba!," developed through the implementation of storytelling in its performance creation process. Choreographers, writers, and costume designer collaborated throughout the storytelling process from planning to content creation. This resulted in a clarified depiction of the Dokkaebi as divine beings with transcendental powers who are simultaneously friendly and familiar entities, often appearing in the form of humans and objects. Accordingly, the costume design based on this collaborative storytelling deliberately avoided the appearance of Dokkaebi reminiscent of 'Kwimyeonwa' or Japanese 'Oni', typically expressed with horns or decorations, and instead implemented novel design elements, such as fusion Hanbok with a variety of dimensionally aesthetic accessories, meaningful color contrast in modern clothing, textural elements indirectly expressing character and transmogrification, and daily clothing attire to illustrate the humanized Dokkaebi. Therefore, this study seeks to contribute to the production of new performance art showcasing the Dokkaebi by identifying the diversity of expression and direction within costume design.

The Taoist Ideology of Psychoanalytic Psychology and the Future Development Trend of Art Therapy (정신분석 심리학의 도교사상과 예술치료의 미래 발전 트렌드)

  • Li Huisshu
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.2
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    • pp.437-444
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    • 2023
  • With the development of modern society, society provides people with a quick and convenient life, but it also causes anxiety, depression, other psychological and mental illnesses, so art can also be used as a way and path to treating psychological disorders. Art therapy is a creative method of delivery used in therapy, resulting from the fusion of art and psychotherapy. Art is a service and public art activity, mainly aimed at solving human psychological problems, providing nonverbal expression and communication opportunities, and treating psychological and psychological entanglement by improving emotion, stability, and indirectly inappropriate behavior through multi-disciplinary convergence. Art therapy, as an independent discipline, began in Europe in the 3rd and 40th centuries and was mainly influenced by two psychologists, Sigmund Freud and Carl (Gustav Jung). We Starting with the theoretical foundation of psychoanalysis, this paper explores the effectiveness of modern people's psychotherapy, analyzes the similarities between the two ideas, explores the application of art therapy and the development of science and technology, and provides the public with various ways of art therapy and the validity of science and technology.

Integrative analysis of microRNA-mediated mitochondrial dysfunction in hippocampal neural progenitor cell death in relation with Alzheimer's disease

  • A Reum Han;Tae Kwon Moon;Im Kyeung Kang;Dae Bong Yu;Yechan Kim;Cheolhwan Byon;Sujeong Park;Hae Lin Kim;Kyoung Jin Lee;Heuiran Lee;Ha-Na Woo;Seong Who Kim
    • BMB Reports
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    • v.57 no.6
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    • pp.281-286
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    • 2024
  • Adult hippocampal neurogenesis plays a pivotal role in maintaining cognitive brain function. However, this process diminishes with age, particularly in patients with neurodegenerative disorders. While small, non-coding microRNAs (miRNAs) are crucial for hippocampal neural stem (HCN) cell maintenance, their involvement in neurodegenerative disorders remains unclear. This study aimed to elucidate the mechanisms through which miRNAs regulate HCN cell death and their potential involvement in neurodegenerative disorders. We performed a comprehensive microarray-based analysis to investigate changes in miRNA expression in insulin-deprived HCN cells as an in vitro model for cognitive impairment. miR-150-3p, miR-323-5p, and miR-370-3p, which increased significantly over time following insulin withdrawal, induced pronounced mitochondrial fission and dysfunction, ultimately leading to HCN cell death. These miRNAs collectively targeted the mitochondrial fusion protein OPA1, with miR-150-3p also targeting MFN2. Data-driven analyses of the hippocampi and brains of human subjects revealed significant reductions in OPA1 and MFN2 in patients with Alzheimer's disease (AD). Our results indicate that miR-150-3p, miR-323-5p, and miR-370-3p contribute to deficits in hippocampal neurogenesis by modulating mitochondrial dynamics. Our findings provide novel insight into the intricate connections between miRNA and mitochondrial dynamics, shedding light on their potential involvement in conditions characterized by deficits in hippocampal neurogenesis, such as AD.

THE EFFECT OF BONE MORPHOGENETIC PROTEIN 2(BMP2) ON THE GROWTH OF CRANIAL BONE AND EARLY MORPHOGENESIS OF THE CRANIAL SUTURE (Bone Morphogenetic Protein 2 가 두개골 성장 및 두개봉합부의 초기형태발생에 미치는 영향)

  • Jung, Hae-Kyung;Park, Mi-Hyun;Ryoo, Hyun-Mo;Nam, Soon-Hyeun;Kim, Young-Jin;Kim, Hyun-Jung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.2
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    • pp.217-228
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    • 2003
  • Co-ordinate growth of the brain and skull is achieved through a series of tissue interactions between the developing brain, the growing bones of the skull and the sutures that unite the bones. Craniosynostosis, the premature fusion of cranial sutures, presumably involves disturbance of these interactions. Bmp2, one of bone morphogenetic proteins (Bmps), is involved in the regulation of the shapes of individual bones and the relative proportions of the skeleton. Mutations in the homeobox gene Msx2, known as a downstream gene of Bmp, cause Boston-type human craniosynostosis. The phenotype of Dlx5 homozygote mutant mouse presents craniofacial abnormalities including a delayed ossification of calvarial bone. These facts suggest important roles of Bmp2, Msx2 and Dlx5 genes in the cranial bone growth and suture morphogenesis. To elucidate the function of these molecules in the early morphogenesis of mouse cranial sutures, we first analyzed by in situ hybridization the expression of Bmp2(E15-18), Msx2 and Dlx5 genes in the developing sagittal suture of calvaria during the embryonic stage. Bmp2 mRNA was intensely expressed in the osteogenic fronts and also at the low level in the periosteum of parietal bones during embryonic stage, Msx2 mRNA was intensely expressed in the sutural mesenchyme and mildly expressed in the dura mater during the embryonic stage. Dlx5 mRNA was intensely expressed osteogenic fronts and parietal bones. To further examine the role of Bmp signaling in cranial suture, we did in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of Bmp2-soaked beads onto the osteogenic fronts after 48 hours organ culture resulted in the increase of the tissue thickness and cell number around Bmp2 beads, compared to BSA control beads. In addition Bmp2 induced etopic expressions of Msx2 and Dlx5 genes. On the other hand, overexpression of FGF2 did not induce the expression of Msx2 and Dlx5. Taken together, these data indicate that Bmp2 signaling molecule has a important role in regulating the cranial bone growth and early morphogenesis of cranial suture. We also suggest that Bmp signaling is involved in all the stages of osteogenesis of cranial bones and the maintenance of cranial suture by regulating Msx2 and Dlx5 genes, and that Msx2 and Dlx5 genes are specific transcription factors of Bmp signaling pathway.

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Cloning and Characterization of Phosphoinositide 3-Kinase γ cDNA from Flounder (Paralichthys olivaceus) (넙치에서 분리된 phosphoinositide 3-kinase γ 유전자의 클로닝 및 특성 연구)

  • Jeong, Tae Hyug;Youn, Joo Yeon;Ji, Keunho;Seo, Yong Bae;Kim, Young Tae
    • Journal of Life Science
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    • v.24 no.4
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    • pp.343-351
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    • 2014
  • Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.

Purification of Human HtrA1 Expressed in E. coli and Characterization of Its Serine Protease Activity (E. coli에서 발현된 human HtrA1 단백질의 정제와 HtrA1의 serine protease 활성 조건에 관한 연구)

  • Kim, Kyung-Hee;Kim, Sang-Soo;Kim, Goo-Young;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1133-1140
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    • 2006
  • Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.