• Biomolecules & Therapeutics
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• 제23권6호
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• pp.539-548
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• 2015

Prostaglandin $E_2$ ($PGE_2$), a major product of cyclooxygenase, binds to four different prostaglandin $E_2$ receptors (EP1, EP2, EP3, and EP4) which are G-protein coupled transmembrane receptors (GPCRs). Although GPCRs including EP receptors have been shown to be associated with their specific G proteins, recent evidences suggest that GPCRs can regulate MAPK signaling via non-G protein coupled pathways including Src. EP2 is differentially expressed in various tissues and the expression of EP2 is induced by extracellular stimuli. We hypothesized that an increased level of EP2 expression may affect MAPK signaling. The overexpression of EP2 in HEK 293 cells resulted in significant increase in intracellular cAMP levels response to treatment with butaprost, a specific EP2 agonist, while overexpression of EP2 alone did not increase intracellular cAMP levels. However, EP2 overexpression in the absence of $PGE_2$ induced an increase in the level of p38 phosphorylation as well as the kinase activity of p38, suggesting that up-regulation of EP2 may promote p38 activation via non-G protein coupled pathway. Inhibition of Src completely blocked EP2-induced p38 phosphorylation and overexpression of Src increased the level of p38 phosphorylation, indicating that Src is upstream kinase for EP2-induced p38 phosphorylation. EP2 overexpression also increased the Src activity and EP2 protein was co-immunoprecipitated with Src. Furthermore, sequential co-immunoprecipitation studies showed that EP2, Src, and ${\beta}$-arrestin can form a complex. Our study found a novel pathway in which EP2 is associated with Src, regulating p38 pathway.

• 약학회지
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• 제49권4호
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• pp.275-283
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• 2005

The purpose of the present study was to kinetically investigate the carrier-mediated uptake in the hepatic transport of organic anions, and to simulate the 'in vivo counter-transport' phenomena, using kinetic model which was developed in this study. The condition that the mobility of carrier-ligand complex is greater than that of free carrier is not essential for the occurrence of 'counter-transport' phenomenon. To examine the inhibitory effects on the initial uptake of organic anions by the liver, it is necessary to judge whether the true counter-transport mechanism (trans-stimulation) is working or not. Effects of bromophenol blue (BPB) or bromosulfophthalein (BSP) on the plasma disappearance curves of a 1-anilino-8-naphthalene sulfonate (ANS) were then kinetically analyzed based on a flow model, in which the ligand is eliminated only from the peripheral compartment (liver compartment). Moreover, 'in vivo counter-transport' phenomena were simulated based on the perfusion model which incorporated the carrier-mediated transport and the saturable intracellular binding. The 'in vivo counter-transport' phenomena in the hepatic transport of a organic anions were well demonstrated by incorporating the carrier-mediated process. However, the 'in vivo counter-transport' phenomena may be also explained by the enhancement of back diffusion due to the displacement of intracellular binding. In conclusion, one should be more cautious in interpreting data obtained from so-called 'in vivo counter-transport' experiments.

• BMB Reports
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• 제44권5호
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• pp.298-305
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• 2011

Most intracellular reactive oxygen species (ROS), especially superoxide anion ($O_2^{{\bullet}_-}$) that is converted from oxygen, are overproduced by excessive electron leakage from the mitochondrial respiratory chain. Intracellular oxidative stress that damages cellular components can contribute to lifestyle-related diseases such as diabetes and arteriosclerosis, and age-related diseases such as cancer and neuronal degenerative diseases. We have previously demonstrated that the excessive mitochondrial $O_2^{{\bullet}_-}$ production caused by SDHC mutations (G71E in C. elegans, I71E in Drosophila and V69E in mouse) results in premature death in C. elegans and Drosophila, cancer in mouse embryonic fibroblast cells and infertility in transgenic mice. SDHC is a subunit of mitochondrial complex II. In humans, it has been reported that mutations in SDHB, SDHC or SDHD often result in inherited head and neck paragangliomas (PGLs). Recently, we established Tet-mev-1 conditional transgenic mice using our uniquely developed Tet-On/Off system, which equilibrates transgene expression to endogenous levels. These mice experienced mitochondrial respiratory chain dysfunction that resulted in $O_2^{{\bullet}_-}$ overproduction. The mitochondrial oxidative stress caused excessive apoptosis leading to low birth weight and growth retardation in the neonatal developmental phase in Tet-mev-1 mice. Here, we briefly describe the relationships between mitochondrial $O_2^{{\bullet}_-}$ and aging phenomena in mev-1 animal models

• Journal of Microbiology and Biotechnology
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• 제22권4호
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• pp.571-573
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• 2012

Glyceraldehyde-3-phosphate (G-3-P), the substrate of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), is a key intermediate in several metabolic pathways. Recently, we reported that G-3-P directly inhibits caspase-3 activity in a reversible noncompetitive mode, suggesting the intracellular G-3-P level as a cell fate decision factor. It has been known that apoptotic stimuli induce the generation of NO, and NO S-nitrosylates GAPDH at the catalytic cysteine residue, which confers GAPDH the ability to bind to Siah-1, an E3 ubiquitin ligase. The GAPDH-Siah-1 complex is translocated into the nucleus and subsequently triggers the apoptotic process. Here, we clearly showed that intracellular G-3-P protects GAPDH from S-nitrosylation at above a certain level, and consequently maintains the cell survival. In case G-3-P drops below a certain level as a result of exposure to specific stimuli, G-3-P cannot inhibit S-nitrosylation of GAPDH anymore, and consequently GAPDH translocates with Siah-1 into the nucleus. Based on these results, we suggest that G-3-P functions as a molecule switch between cell survival and apoptosis by regulating S-nitrosylation of GAPDH.

• 한국산학기술학회:학술대회논문집
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• 한국산학기술학회 2004년도 춘계학술대회
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• pp.253-257
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• 2004

Platelet aggregation is a complex phenomenon that probably involves several intracellular biochemical pathways. When activated, platelets change shape, aggregate and release the contents of their intracellular granules. The interactions between platelets and blood vessel walls are important in the development of thrombosis and cardiovascular diseases. When blood vessels are damaged, platelet aggregation occurs rapidly to form haemostatic Plugs or arterial thrombi at the sites of vessel injury or in regions where blood flow is disturbed. These thrombi are the source of thromboembolic complications of atherosclerosis, heart attacks, stroke, and peripheral vascular disease. Therefore, the inhibition of platelet function represents a promising approach for the prevention of thrombosis. Plants constitute a rich source of bioactive chemicals such as phenolics, terpenoids and alkaloids. Plant extracts may be an alternative to currently used medicinal source because they constitute a rich source of bioactive chemicals. This study was performed to investigate the antiplatelet activity of extract of Tabebuia impetiginosa Martius ex DC (Taheebo) and find out which fractions to this activity in rabbit platelet. Taheebo was methanol extracted and solvent fractionated in to five fractions (hexane, chloroform, ethylacetate, butanol and water). And each fractions were investigated inhibitory effects on platelet aggregation induced by various agonists using washed rabbit platelets in vitro.

• Molecules and Cells
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• 제45권7호
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• pp.454-464
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• 2022

DJ-1 is one of the causative genes of early-onset familial Parkinson's disease (PD). As a result, DJ-1 influences the pathogenesis of sporadic PD. DJ-1 has various physiological functions that converge to control the levels of intracellular reactive oxygen species (ROS). Based on genetic analyses that sought to investigate novel antioxidant DJ-1 downstream genes, pyruvate dehydrogenase (PDH) kinase (PDK) was demonstrated to increase survival rates and decrease dopaminergic (DA) neuron loss in DJ-1 mutant flies under oxidative stress. PDK phosphorylates and inhibits the PDH complex (PDC), subsequently downregulating glucose metabolism in the mitochondria, which is a major source of intracellular ROS. A loss-of-function mutation in PDK was not found to have a significant effect on fly development and reproduction, but severely ameliorated oxidative stress resistance. Thus, PDK plays a critical role in the protection against oxidative stress. Loss of PDH phosphatase (PDP), which dephosphorylates and activates PDH, was also shown to protect DJ-1 mutants from oxidative stress, ultimately supporting our findings. Further genetic analyses suggested that DJ-1 controls PDK expression through hypoxia-inducible factor 1 (HIF-1), a transcriptional regulator of the adaptive response to hypoxia and oxidative stress. Furthermore, CPI-613, an inhibitor of PDH, protected DJ-1 null flies from oxidative stress, suggesting that the genetic and pharmacological inhibition of PDH may be a novel treatment strategy for PD associated with DJ-1 dysfunction.

• 대한임상검사과학회지
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• 제49권4호
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• pp.435-438
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• 2017

비결핵 항상균(Nontuberculous Mycobacteria, NTM) M. avium-intracellular complex (MAC), M. fortuitum, M. chelonae, M. abscessus, M. kansasii 등을 함유한다. 비결핵 항상균은 자연 환경속에 서식하며 다양한 가축등에 감염한다. 그리고 사람한테는 기회감염을 유발하여 사회적 경제적 문제를 야기한다. 본 연구는 면역결핍 사람한테 기회감염을 유발하는 비결핵 항상균이 사람들이 흔히 이용하는 다중이용시설내 존재 유무를 확인하기위해 40개 에어컨 먼지를 채취하여 직접 AFB 염색, 직접 PCR, 간접 AFB 염색, 간접 PCR 등을 실시했다. 그 결과 채취한 샘플의 직접 AFB 염색, 직접 PCR 에서는 인형결핵균(MTB), 비결핵 항상균 모두 발견되지 않았다. 배양된 집락을 이용한 간접 AFB 염색, 간접 PCR 결과 인형결핵균 (MTB)은 모두 음성반응을 나타냈으며, 비결핵 항상균은 40개 샘풀 중 2개(5%)에서 양성반응을 나타냈다. 본 실험결과는 자연 내 비결핵 항상균이 존재한다는 사실을 증명했으며 비결핵 항상균이 기회감염의 원인임을 감안했을 때 다중이용시설의 청결한 위생처리가 중요함을 암시한다. 다만 비결핵 항상균의 종(species) 감별과 기회감염 유발 유무는 좀 더 연구가 필요하다고 사료된다.

• Journal of Microbiology and Biotechnology
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• 제33권9호
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• pp.1206-1212
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• 2023

The Wnt/β-catenin pathway plays essential roles in regulating various cellular behaviors, including proliferation, survival, and differentiation [1-3]. The intracellular β-catenin level, which is regulated by a proteasomal degradation pathway, is critical to Wnt/β-catenin pathway control [4]. Normally, casein kinase 1 (CK1) and glycogen synthase kinase-3β (GSK-3β), which form a complex with the scaffolding protein Axin and the tumor suppressor protein adenomatous polyposis coli (APC), phosphorylate β-catenin at Ser45, Thr41, Ser37, and Ser33 [5, 6]. Phosphorylated β-catenin is ubiquitinated by the β-transducin repeat-containing protein (β-TrCP), an F-box E3 ubiquitin ligase complex, and ubiquitinated β-catenin is degraded via a proteasome pathway [7, 8]. Colorectal cancer is a significant cause of cancer-related deaths worldwide. Abnormal up-regulation of the Wnt/β-catenin pathway is a major pathological event in intestinal epithelial cells during human colorectal cancer oncogenesis [9]. Genetic mutations in the APC gene are observed in familial adenomatous polyposis coli (FAP) and sporadic colorectal cancers [10]. In addition, mutations in the N-terminal phosphorylation motif of the β-catenin gene were found in patients with colorectal cancer [11]. These mutations cause β-catenin to accumulate in the nucleus, where it forms complexes with transcription factors of the T-cell factor/lymphocyte enhancer factor (TCF/LEF) family to stimulate the expression of β-catenin responsive genes, such as c-Myc and cyclin D1, which leads to colorectal tumorigenesis [12-14]. Therefore, downregulating β-catenin response transcription (CRT) is a potential strategy for preventing and treating colorectal cancer. Plant cytokinins are N⁶-substituted purine derivatives; they promote cell division in plants and regulate developmental pathways. Natural cytokinins are classified as isoprenoid (isopentenyladenine, zeatin, and dihydrozeatin), aromatic (benzyladenine, topolin, and methoxytopolin), or furfural (kinetin and kinetin riboside), depending on their structure [15, 16]. Kinetin riboside was identified in coconut water and is a naturally produced cytokinin that induces apoptosis and exhibits antiproliferative activity in several human cancer cell lines [17]. However, little attention has been paid to kinetin riboside's mode of action. In this study, we show that kinetin riboside exerts its cytotoxic activity against colon cancer cells by suppressing the Wnt/β-catenin pathway and promoting intracellular β-catenin degradation.

• Applied Microscopy
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• 제10권1_2호
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• pp.27-32
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• 1980

중동지방에 다녀온 48세의 한국인 남자 피부에 발생한 leishmania증 1예에 대하여 광학 및 전자현미경적 관찰을 하여 문헌 고찰과 아울러 보고하였다. 광학현미경적으로는 조직구 침윤을 주로 하는 만성 육아종성염증반응을 보였고 Giemsa염색상 다수의 leishmania충체를 조직구내에서 관찰할 수 있었다. 전자현미경적으로는 난원형의 단세포 또는 분열중인 충체가 조직구내에 존재하였고 충체는 이중막으로 싸였으며 그 바로밑에 microtubule이 배열되고 세포질내에는 신장된 mitochondrion내에 kinetoplast가 존재하고 그 전방에 flagella가 위치하였으며 기타 다수의 ribosome과 드물게 Golgi complex등을 관찰할 수 있었다. 이와같은 특징은 Leishmania tropica의 promastigote stage와 일치하였다. Leishmania증은 원칙적으로 열대병이나 열대지방과의 접촉이 빈번함에 따라 앞으로 우리나라에서도 보다 많은 증예가 발생할 것으로 사료된다.

• BMB Reports
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• 제37권3호
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• pp.298-303
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• 2004

In general, a SYPRO Ruby dye is well known as a sensitive fluorescence-based method for detecting proteins by one-or two-dimensional SDS-PAGE (1-DE or 2-DE). Based on the SYPRO Ruby dye system, the combined two-dimensional fibrin zymography (2-D FZ) with SYPRO Ruby staining was newly developed to identify the Bacillus sp. proteases. Namely, complex protein mixtures from Bacillus sp. DJ-4, which were screened from Doen-Jang (Korean traditional fermented food), showed activity on the zymogram gel. The gel spots on the SYPRO Ruby gel, which corresponded to the active spots showing on the 2-D FZ gel, were analyzed by a matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometric analysis. Five intracellular fibrinolytic enzymes of Bacillus sp. DJ-4 were detected through 2-D FZ. The gel spots on the SYPRO Ruby dye stained 2-D gel corresponding to 2-D FZ were then analyzed by MALID TOF MS. Three of the five gel spots proved to be quite similar to the ATP-dependent protease, extracellular neutral metalloprotease, and protease of Bacillus subtilis. Also, the extracellular proteases of Bacillus sp. DJ-4 employing this combined system were identified on three gels (e.g., casein, fibrin, and gelatin) and the proteolytic maps were established. This combined system of 2-D zymography and SYPRO Ruby dye should be useful for searching the specific protease from complex protein mixtures of many other sources (e.g., yeast and cancer cell lines).