• Title/Summary/Keyword: Trans-human

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Molecular Cloning, Tissue Distribution and Expression of Porcine y+L Amino Acid Transporter-1

  • Zhi, Ai-min;Zhou, Xiang-yan;Zuo, Jian-jun;Zou, Shi-geng;Huang, Zhi-yi;Wang, Xiao-lan;Tao, Lin;Feng, Ding-yuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.2
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    • pp.272-278
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    • 2010
  • In this study, we cloned, sequenced and characterized porcine y+L Amino Acid Transporter-1 (y+LAT1). By screening a translated EST database with the protein sequence of the human $y^{+}$LAT1 and by using rapid amplification of cDNA ends (RACE), the full-length cDNA encoding porcine $y^{+}$LAT1 was isolated from porcine intestine RNA. It was 2,111 bp long, encoding a 511 amino acid trans-membrane glycoprotein composed of 12 transmembrane domains. The predicted amino acid sequence was found to be 91%, 90%, 87% and 87% identical to those of cattle, human, mouse and rat $y^{+}$LAT1 respectively. Real-time RT-PCR results indicated that the small intestine had the highest $y^{+}$LAT1 mRNA abundance and the lung had the lowest $y^{+}$LAT1 mRNA abundance. Baby hamster kidney (BHK) cells transfected with green fluorescent protein (GFP) tagged porcine $y^{+}$LAT1 cDNA indicated that the cellular localization of the gene product in BHK was on the plasma membrane.

Role of Retinoic Acid in Spontaneous Apoptosis of Human Neutrophils

  • Yang, Eun-Ju;Lee, Ji-Sook;Kim, Dong-Hee;Min, Bok-Kee;Hyun, Sung-Hee;Kim, In-Sik
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.279-285
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    • 2007
  • Although retinoic acid has been known as either anti-inflammatory or pro-inflammatory molecule, depending on the cell type, its exact role in mature human neutrophils has not been fully explored. In this study, we investigate the effects of retinoic acid on neutrophil apoptosis and the associated mechanism and found that 9-cis retinoic acid (9CRA) significantly inhibits the spontaneous apoptosis of neutrophils. Its effect is increased by co-treatment with $TNF-\alpha$ (P<0.05). The 9CRA-induced inhibition is blocked by the following enzyme inhibitors: Ly 294002, phosphoinoside (PI)-3 kinase inhibitor, U73122, a phospholipase C (PLC) inhibitor, PP2, Src family protein inhibitor, SB202190, p38 MAPK inhibitor, and BAY-11-7085, NF-kB inhibitor. This study also demonstrates that all-trans retinoic acid suppresses spontaneous apoptosis, similar to the mechanism of inhibition exhibited by 9CRA. Phosphorylation of p38 MAPK decreases by 9CRA treatment. $Ik-B{\alpha}$ is degraded until 30 minutes after a time-dependent 9CRA treatment, but degradation can be inhibited by Ly 294002. These results indicate that 9CRA decreases p38 MAPK activation, induces NF-kB activation via PI-3 kinase, and also blocks cleavage of caspase 3. As these findings suggest, 9CRA has a molecular mechanism which may help pro-inflammatory response by blocking neutrophil apoptosis.

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The effect of L-carnitine in the expression of matrix metalloproteinases by human dermal fibroblasts

  • Yoon, Eun-Jeong;Lee, Kyoung-Eun;Sim, Kwan-Sup;Lee, Bum-Chun;Pyo, Hyeong-Bae;Choe, Tae-boo
    • Proceedings of the SCSK Conference
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    • 2003.09b
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    • pp.12-25
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    • 2003
  • L-camitine ($\beta$ -hydroxy-${\gamma}$ -trimethyl-ammoniumbutyric acid) is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-camitine, we investigated the effects of in vitro MMP inhibition and activity and expression of UVA-induced MMP 1 in human skin fibroblasts. Fluorometric assays of the proteolytic activities of MMP-l were performed using fluorescent collagen substrates. ELISA (enzyme linked immuno sorbent assay), gelatin-substrate zymography, and RT-PCR ELISA techniques were used for the effects of L-camitine on MMP expression and activity, MMP mRNA expression in UVA irradiated fibroblast. L-camitine inhibited the activities of MMP-l in a dose-dependent manner and the $IC_{50}$/ values calculated from semi-log plots were 2.45mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP expression was reduced 40% by treated with L-carnitine, and MMP-l mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibition the MMP activity, regulation of MMP expression in protein and mRNA level. All these results suggest that L-carnitine may be useful as new anti-aging cofactor for protection against UVA induced MMP expression and activity.

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Green Fluorescent Protein-reporter Mammalian One-hybrid System for Identifying Novel Transcriptional Modulators for Human $p14^{ARF}$ Tumor Suppressor Gene

  • Lee, Hye Jin;Yang, Dong Hwa;Yim, Tae Hee;Rhee, Byung Kirl;Kim, Jung-Wook;Lee, Jungwoon;Gim, Jin Bae;Kim, JungHo
    • Animal cells and systems
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    • v.6 no.4
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    • pp.317-322
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    • 2002
  • To improve conventional yeast one-hybrid screening, we have developed an efficient mammalian one-hybrid system that allows rapid isolation of com-plementary DNAs which are able to induce human p14$^{ARF}$. tumor suppressor gene. A 1.5 kb promoter region of p14$^{ARF}$ was fused to EGFP to generate ARF promoter-EGFP reporter vector. This reporter plasmid was stably trans-fected into NIH3T3 cells for generation of reporter cell line. When the reporter cell line was infected with E2F-1 together with excess amounts of empty vector, the cells that received the positive modulator were readily identifiable by green fluorescence using FACS. The GFP-positive cells were cloned directly from the cultured cells and expanded in bulk culture. The genomic DNAs from GFP-positive cells were prepared and the CDNA insert in integrated retroviral genome was recovered by PCR using primers annealing to the retroviral vector sequences flanking the insert-cloning site. This system should be useful for efficient screening of expression CDNA libraries in mammalian cells to identify novel upstream regulators for spe-cific genes by one-hybrid interaction.ion.

Differentiation Induction of Dendritic Cell Phenotypes from Human Leukemic Cell Lines

  • Lee, Dae-Heui;Park, Jae-Sun;Eo, Wan-Kyu;Kim, Woo-Mi;Kang, Koo-Il
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.1
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    • pp.79-86
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    • 2001
  • Recent clinical studies have shown that a high proportion of patients with acute promyelocytic leukemia (APL) achieve complete remission after treatment with all-trans retinoic acid (ATRA). However, most patients who receive continuous treatment with ATRA relapse and develop ATRA-resistant leukemia. Dendritic cells (DCs) are important antigen-presenting cells in the development of antileukemic T-cell responses. In this study, we investigated the strategies to overcome ATRA resistance of APL cells by inducing the differentiation of DCs from human leukemic cell lines for the developtment of adoptive immunotherapy. CD83 was used as a mature DC marker in this study and the expression of CD83 mRNA was determined by RT-PCR method. The promyelocytic leukemic cell line HL-60, B lymphoblast cell lines RPMI 7666 and NC-37 could be induced to dendritic cells in vitro. Treatment of HL-60 with phorbol 12-myristate 13-acetate (PMA) resulted in the expression of myeloid-related DC phenotypes, while treatment of RPMI 7666 with fms-like tyrosine kinase 3 ligand (Flt3-ligand, FL) and treatment of NC-37 with PMA and FL led to the expression of lymphoid-related DC phenotypes. In conclusion, myeloid-related DC phenotypes and lymphoid-related DC phenotypes could be generated from HL-60, NC-37 and RPMI 7666 cell lines, respectively. These DC phenotypes can potentially be used to generate antileukemic T cells in vitro for adoptive immunotherapy.

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A Study on the Analysis of the Characteristics of the Real-time Behavior Space Design - Focused on the Works of onl and NOX - (물리구축환경의 지능적 부활로서의 실시간 행태 공간의 특성 분석 - onl과 NOX의 작품을 중심으로 -)

  • Lee Hanna;Park Hyun-Ok
    • Korean Institute of Interior Design Journal
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    • v.14 no.4 s.51
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    • pp.19-26
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    • 2005
  • Digital technology continually makes a space evolves. The real-time behavior design communicates the data with the situation of circumference of the space(visitors moving, interior and exterior situations). The space form was changed because it interfaces in real time. The purpose of this study was finding out the characteristics of real-time behavior space design through the analysis of space formative languages, sensorium, S-R and material. This study will be the one of basic references for the digital space design. The boundary of this study set limits to the works of digital space designer who applies the real-time exchanging data to their design among the digital space design works from 1996 to 2004. But it excepted from the real-time behavior space in virtual realty. Therefore, the objects of this study were the works of onl and NOX(paraSITE, Trans-port 2001, Muscle, MotormeCCa, Handdrawspace, Saltwater Pavilion, Son-O-House, H2O Expo). The method was the contents analysis of space formative languages(Greg Lynn's ten space formative languages; bleb, blob, branch, flower, fold, lattice, teeth, shred, skins and strand), sensorium, S-R and material. The results of the study are as follows: 1) The organizational elements; Space formative languages(bleb, blob, fold, shred, skins, strand), stimulation(Human Participation, Human Moving, Weather Conditions), and response(Spatial Moving, Sound Pattern, Lighting Pattern, color Pattern, Activating Particles, Moving Picture, Virtual Friend) 2) The material Use; Sound, lights, and network have been used in the space. Immaterial matter will be used the main material of space design in 21"'century, 3)The spatial types; formal changing of space, projecting immaterial elements, and changing the sound.

Characteristics of Daesoon Thought in Korean Modern Times - Focused on Transnationalism, Modern and Post-modern Values - (한국 근대시기 대순사상의 특질 - 초민족주의와 근대 및 탈근대 가치를 중심으로 -)

  • Park, Jae-hyun
    • Journal of the Daesoon Academy of Sciences
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    • v.24_1
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    • pp.255-289
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    • 2014
  • This study's aim is to identify modern and post-modern values and transnationalism embodied in Daesoon Thought and to seek for the new value to overcome irrationality of modern values in this society we live in. Several previous studies discussed about these issues, but most of them studied them on the basis of Korean new religions or Jeungsangyo, or in sociological theory perspective. Therefore, this study focused on Daesoon Thought encompassing ideological perspective as well as historical perspective of Daesoonjinrihoe. As for nationalism, while Eastern learning(Dong-Hak) is prone to ethnocentricity, Daesoon Thought shows trans-ethnic perspective. As for historical perspectivel, Mugeuikdo, a precursor to Daesoonjinrihoe showed non-relationship with any politics as contrasted with other new korean religious movement at that time. As for aspects of modern values, 3 perspectives (political system, social system, abolition of premodern values) were discussed. As for political system perspective, while Eastern learning advocates democratic modernity but accepted monarchy, Jeungsan denied monarchy. And While western political philosophy advocated rationality-based absolute person, Daesoon Thought proposes ideal human who can have political power and do religious indoctrination all together. As for social system perspective, while western humanism is based on all of he people's equity in front of God, Eastern learning on humans are Heaven (人乃天), Daesoon Thought is based on Injon thought(人尊思想) which encompasses spiritual world, human world and all of the universe. Daesoon Thought also proposes abolition of discrimination by gender, social position. As for abolition of premodern values, Daesoon Thought critics pre-modern formalism and advocate acceptance of other nations' culture, pragmatism, and humanism. As presented above, Daesoon Thought has not only modern values but also aspects of post-modernity and transnationalism. In the future, further studies are needed which tackle these issues and search for new values of Daesoon Thought which can overcome limitation of modern values.

PDZ Peptide of the ZO-1 Protein Significantly Increases UTP-Induced MUC8 Anti-Inflammatory Mucin Overproduction in Human Airway Epithelial Cells

  • Han Seo;Hyun-Chae Lee;Ki Chul Lee;Doosik Kim;Jiwook Kim;Donghee Kang;Hyung-Joo Chung;Hee-Jae Cha;Jeongtae Kim;Kyoung Seob Song
    • Molecules and Cells
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    • v.46 no.11
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    • pp.700-709
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    • 2023
  • Mucus hyperproduction and hypersecretion are observed often in respiratory diseases. MUC8 is a glycoprotein synthesized by epithelial cells and generally expressed in the respiratory track. However, the physiological mechanism by which extracellular nucleotides induce MUC8 gene expression in human airway epithelial cells is unclear. Here, we show that UTP could induce MUC8 gene expression through P2Y2-PLCβ3-Ca2+ activation. Because the full-length cDNA sequence of MUC8 has not been identified, a specific siRNA-MUC8 was designed based on the partial cDNA sequence of MUC8. siRNA-MUC8 significantly increased TNF-α production and decreased IL-1Ra production, suggesting that MUC8 may downregulate UTP/P2Y2-induced airway inflammation. Interestingly, the PDZ peptide of ZO-1 protein strongly abolished UTP-induced TNF-α production and increased IL-1Ra production and MUC8 gene expression. In addition, the PDZ peptide dramatically increased the levels of UTP-induced ZO proteins and TEER (trans-epithelial electrical resistance). These results show that the anti-inflammatory mucin MUC8 may contribute to homeostasis, and the PDZ peptide can be a novel therapeutic candidate for UTP-induced airway inflammation.

Alkaloids from Beach Spider Lily (Hymenocallis littoralis) Induce Apoptosis of HepG-2 Cells by the Fas-signaling Pathway

  • Ji, Yu-Bin;Chen, Ning;Zhu, Hong-Wei;Ling, Na;Li, Wen-Lan;Song, Dong-Xue;Gao, Shi-Yong;Zhang, Wang-Cheng;Ma, Nan-Nan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.21
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    • pp.9319-9325
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    • 2014
  • Alkaloids are the most extensively featured compounds of natural anti-tumor herbs, which have attracted much attention in pharmaceutical research. In our previous studies, a mixture of major three alkaloid components (5, 6-dihydrobicolorine, 7-deoxy-trans-dihydronarciclasine, littoraline) from Hymenocallis littoralis were extracted, analyzed and designated as AHL. In this paper, AHL extracts were added to human liver hepatocellular cells HepG-2, human gastric cancer cell SGC-7901, human breast adenocarcinoma cell MCF-7 and human umbilical vein endothelial cell EVC-304, to screen one or more AHL-sensitive tumor cell. Among these cells, HepG-2 was the most sensitive to AHL treatment, a very low dose ($0.8{\mu}g/ml$) significantly inhibiting proliferation. The non-tumor cell EVC-304, however, was not apparently affected. Effect of AHL on HepG-2 cells was then explored. We found that the AHL could cause HepG-2 cycle arrest at G2/M checkpoint, induce apoptosis, and interrupt polymerization of microtubules. In addition, expression of two cell cycle-regulated proteins, CyclinB1 and CDK1, was up-regulated upon AHL treatment. Up-regulation of the Fas, Fas ligand, Caspase-8 and Caspase-3 was observed as well, which might imply roles for the Fas/FsaL signaling pathway in the AHL-induced apoptosis of HepG-2 cells.

The Effect of Conjugated Linoleic Acid and Its Isomers on the Proliferation of Prostate TSU-Prl Cancer Cells (Conjugated Linoleic Acid (CLA)와 그 이성체가 전립선 암세포의 증식에 미치는 영향)

  • 오윤신;김은지;김종우;김우경;이현숙;윤정한
    • Journal of Nutrition and Health
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    • v.35 no.2
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    • pp.192-200
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    • 2002
  • Conjugated linoleic acid (CLA) is a collective term for positional and geometric isomers of octadecadienoic acid in which the double bonds are conjugated. CLA has anticancer activity in a variety of animal cancer models, and cis-9, trans-11 (c9t11) and trans-10, cis-12(t10c12) CLA are the most predominant isomers present in the synthetic preparations utilized in these animal studies. To compare the ability of c9t11, t10c12 and an isomeric mixture of CLA to inhibit TSU-Prl cell growth, cells were incubated in a serum-free medium with various concentrations of these fatty acids. The isomeric mixture inhibited cell growth in a dose-dependent manner (1-3 $\mu$M) with a 41 $\pm$ 1% inhibition observed at 3 $\mu$M concentration after 48 hours. T10c12 also inhibited cell proliferation in a dote-dependent manner, However, the efficacy and potency of this isomer was much greater than that of the isomeric mixture with a 49 $\pm$ 2% inhibition observed at 0.3 $\mu$M concentration after 48 hours. By contrast, c9t11 slightly increased cell proliferation. To determine whether the growth-inhibiting effect of CLA is related to the changes in production of insulin-like growth factors (IGF) and IGF-binding proteins (IGFBP) by these cells, serum-free conditioned media were collected. Immunoblot analysis of conditioned media using a monoclonal anti-IGF-II antibody showed that both the isomeric mixture and t10c12 inhibited secretion of both mature 7,500 Mr and higher Mr forms of pro IGF-II, whereas c9t11 had no effect. Ligand blot analysis with 125I-IGF-II revealed the presence of two types of IGFBPs : 24,000 Mr IGFBP-4 and 30,000 Mr IGFBP-6. The production of IGFBP-4 slightly decreased at the highest concentrations of the isomeric mixture and t10c12. These results indicate that CLA inhibits human prostate cancer cell growth, an effect largely due to the action of t10c12. The growth inhibition may result, at least in part, from decreased production of IGF-II and IGFBP-4 by these cells.