• Title/Summary/Keyword: Cellular Formation

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Formation of DNA-Protein Crosslink at Oxidized Abasic Site Mediated by Human DNA Polymerase Iota and Mitochondrial DNA Polymerase Gamma

  • Son, Mi-Young;Jun, Hyun-Ik;Goo, Sun-Young;Sung, Jung-Suk
    • Biomedical Science Letters
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    • v.15 no.1
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    • pp.1-8
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    • 2009
  • Human genomic DNA is continuously attacked by oxygen radicals originated from cellular metabolic processes and numerous environmental carcinogens. 2-deoxyribonolactone (dL) is a major type of oxidized abasic (AP) lesion implicated in DNA strand scission, mutagenesis, and formation of covalent DNA-protein crosslink (DPC) with DNA polymerase (Pol) ${\beta}$. We show here that human DNA polymerase (Pol)${\iota}$ and mitochondrial $Pol{\gamma}$ give rise to stable DNA-protein crosslink (DPC) formation that is specifically mediated by dL lesion. $Pol{\gamma}$ mediates DPC formation at the incised dL residue by its 5'-deoxyribose-5-phosphate (dRP) lyase activity, while $Pol{\gamma}$ cross links with dL thorough its intrinsic dRP lyase and AP lyase activities. Reactivity in forming dL-mediated DPC was significantly higher with $Pol{\gamma}$ than with $Pol{\iota}$. DPC formation by $Pol{\gamma}$, however, can be reduced by an accessory factor of $Pol{\gamma}$ holoenzyme that may attenuate deleterious effects of crosslink adducts on mitochondrial DNA. Comparative kinetic analysis of DPC formation showed that the rate of DPC formation with either $Pol{\iota}$ or $Pol{\gamma}$ was lower than that with $Pol{\beta}$. These results revealed that the activity of catalytic lyase in DNA polymerases determine the efficiency of DPC formation with dL damages. Irreversible crosslink formation of such DNA polymerases by dL lesions may result in a prolonged strand scission and a suicide of DNA repair proteins, both of which could pose a threat to the genetic and structural integrity of DNA.

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Effect of Eicosapentaenoic Acid on Cellular Lipid Accumulation and Transcription Factors Involving Glucose Utilization (에이코사펜타에노익산에 의한 세포 내에서의 지방 축적 억제 효과 및 포도당 대사에 관여하는 전사인자의 변화)

  • Bu, So-Young
    • The Korean Journal of Food And Nutrition
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    • v.24 no.4
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    • pp.501-508
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    • 2011
  • Previous studies suggest that polyunsaturated fatty acids with long carbon chains such as eicosapentaenoic acid(EPA) and docosahexaenoic acid(DHA) have several health benefits. However metabolic consequences of these fatty acids themselves and their regulation of transcriptional activity involving glucose utilization are not well established. Thus, the purpose of this study was to investigate how EPA influx affects cellular lipid accumulation and gene expressions involving $de$ $novo$ lipogenesis in hepatocyte cultures. Compared to oleic acid treatment, EPA treatment showed remarkably decreased cellular TG conversion and accumulation, along with phospholipids at a lower extent. As expected, EPA increased mRNA expression involving fatty acid influx and lipid droplet formation, but did not affect mRNA expression involving glucose utilization. EPA increased transcriptional activity of PPAR-${\alpha}$ and glucose responsive transcription factor when transcription factor binding protein was activated. Taken together, these data suggest that EPA decreases lipid accumulation through increases of the ${\beta}$-oxidation pathway without interruption of glucose utilization.

Comparison of cytotoxicities and wound healing effects of diquafosol tetrasodium and hyaluronic acid on human corneal epithelial cells

  • Lee, Jong Heon;Lee, Jong Soo;Kim, Sujin;Lee, Ji Eun
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.2
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    • pp.189-195
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    • 2017
  • This study aimed to compare the cellular toxicities of three clinically used dry eye treatments; 3% diquafosol tetrasodium and hyaluronic acid at 0.3 and 0.18%. A methyl thiazolyltetrazoiun (MTT)-based calorimetric assay was used to assess cellular proliferation and a lactate dehydrogenase (LDH) leakage assay to assess cytotoxicity, using Human corneal epithelial cells (HCECs) exposed to 3% diquafosol tetrasodium, 0.3% hyaluronic acid (HA), or 0.18% HA or 1, 6 or 24 h. Cellular morphology was evaluated by inverted phase-contrast light microscopy and electron microscopy, and wound widths were measured 24 h after confluent HCECs were scratched. Diquafosol had a significant, time-dependent, inhibitory effect on HCEC proliferation and cytotoxicity. HCECs treated with diquafosol detached more from the bottoms of dishes and damaged cells showed degenerative changes, such as, reduced numbers of microvilli, vacuole formation, and chromatin of the nuclear remnant condensed along the nuclear periphery. All significantly stimulated reepithelialization of HCECs scratched, which were less observed in diquafosol. Therefore, epithelial toxicity should be considered after long-term usage of diquafosol and in overdose cases, especially in dry eye patients with pre-existing punctated epithelial erosion.

In vitro biocompatibility of a cement compositecontaining poly ($\varepsilon$-caprolactonemicrosphere) (PCL)

  • Jyoti, Md. Anirban;Min, Young-Ki;Lee, Byong-Taek;Song, Ho-Yeon
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2009.11a
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    • pp.42.1-42.1
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    • 2009
  • In recent years, it has been tried to develop the efficacy and bioactivity of Calcium Phosphate cements(CPC) as injectable bone substitute (IBS) by reinforcing them through varying the amount in its compositions and relative concentrations or adding other additives. In this study, the biocompatibility of are inforced Calcium Phosphate-Calcium Sulfate injectable bone substitute (IBS)containing poly ($\varepsilon$-caprolactone)PCL microspheres was evaluated which consisted of solution chitosan and Na-citrate as liquid phase and tetra calcium phosphate (TTCP), dicalciumphosphate anhydrous (DCPA) powder as the solid phase. The in vitrobiocompatibility of the IBS was done using MTT assay and Cellular adhesion and spreading studies. The in vitro experiments with simulated body fluid (SBF) confirmed the formation of apatite on sample surface after 7 and 14 days of incubation in SBF. SEM images for one cell morphologies showed that the cellular attachment was good. MG-63 cells were found to maintain their phenotype on samples and SEM micrograph confirmed that cellular attachment was well. In vitro cytotoxicity tests by an extract dilution method showed that the IBS was cytocompatible for fibroblast L-929.

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Rapid Separation of Cellular Cyclosophoraoses Produced by Rhizobium Species

  • Seo, Dong-Hyuk;Lee, Sang-Hoo;Park, Hey-Lin;Kwon, Tae-Jong;Jung, Seun-Ho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.522-525
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    • 2002
  • A very rapid and efficient separation technique for cellular rhizobial cyclosophoraoses was developed based on fractional precipitation and partition chromatography. Cyclosophoraoses are known to function in the osmotic regulation and root nodule formation of legumes during the nitrogen fixation process. Cyclosophoraoses are produced as unbranched cyclic (1longrightarrow12)-${\beta}$-D-glucans in Agrobacterium or Rhizobium species. Recent research has shown that cyclosophoraoses can form inclusion complexation with various unstable or insoluble guest chemicals, thereby implying great potential for industrial application. Typical separation of pure cellular cyclosophoraoses has been so far carried out by several time-consuming steps, including size exclusion, anion exchange, and desalting liquid chromatographies, with a relatively poor recovery. However, the proposed method demonstrated that the successive application of fractional ethanol precipitation and one step of silica gel-based flash column chromatography was enough to simultaneously purify neutral or anionic forms of cyclosophoraoses. This novel technique is very rapid and provides a high recovery.

Effect of the hedgehog signaling pathway on hair formation-related cells

  • Park, Jaehyun;Park, Sangkyu;Seo, Jeongmin;Roh, Sangho
    • International Journal of Oral Biology
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    • v.44 no.4
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    • pp.144-151
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    • 2019
  • Alopecia has emerged as one of the biggest interests in modern society. Many studies have focused on the treatment of alopecia, such as transplantation of hair follicles or inhibition of the androgen pathway. Hair growth is achieved through proper proliferation of the components such as keratinocytes and dermal papilla cells (DPCs), movement, and interaction between the two cells. The present study examined the effect of the hedgehog (Hh) signaling pathway, which is an important and fundamental signal in the cell, on the morphology and the viability of human keratinocytes and DPCs. Upregulation of Hh signaling caused a morphological change and an increase in epithelium-mesenchymal transition-related gene expression but reduced the viability of keratinocytes, while the alteration of Hh signaling did not cause any change in DPCs. The results show the possibility that the regulation of Hh signaling can be applied for the treatment of alopecia.

Nuclear Localization of Chfr Is Crucial for Its Checkpoint Function

  • Kwon, Young Eun;Kim, Ye Seul;Oh, Young Mi;Seol, Jae Hong
    • Molecules and Cells
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    • v.27 no.3
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    • pp.359-363
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    • 2009
  • Chfr, a checkpoint with FHA and RING finger domains, plays an important role in cell cycle progression and tumor suppression. Chfr possesses the E3 ubiquitin ligase activity and stimulates the formation of polyubiquitin chains by Ub-conjugating enzymes, and induces the proteasome-dependent degradation of a number of cellular proteins, including Plk1 and Aurora A. While Chfr is a nuclear protein that functions within the cell nucleus, how Chfr is localized in the nucleus has not been clearly demonstrated. Here, we show that nuclear localization of Chfr is mediated by nuclear localization signal (NLS) sequences. To reveal the signal sequences responsible for nuclear localization, a short lysine-rich stretch (KKK) at amino acid residues 257-259 was replaced with alanine, which completely abolished nuclear localization. Moreover, we show that nuclear localization of Chfr is essential for its checkpoint function but not for its stability. Thus, our results suggest that NLS-mediated nuclear localization of Chfr leads to its accumulation within the nucleus, which may be important in the regulation of Chfr activation and Chfr-mediated cellular processes, including cell cycle progression and tumor suppression.

Resource Allocation for Cooperative Relay based Wireless D2D Networks with Selfish Users

  • Niu, Jinxin;Guo, Wei
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.9 no.6
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    • pp.1996-2013
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    • 2015
  • This paper considers a scenario that more D2D users exist in the cell, they compete for cellular resources to increase their own data rates, which may cause transmission interference to cellular users (CU) and the unfairness of resource allocation. We design a resource allocation scheme for selfish D2D users assisted by cooperative relay technique which is used to further enhance the users' transmission rates, meanwhile guarantee the QoS requirement of the CUs. Two transmission modes are considered for D2D users: direct transmission mode and cooperative relay transmission mode, both of which reuses the cellular uplink frequency resources. To ensure the fairness of resource distribution, Nash bargaining theory is used to determine the transmission mode and solve the bandwidth allocation problem for D2D users choosing cooperative relay transmission mode, and coalition formation game theory is used to solve the uplink frequency sharing problem between D2D users and CUs through a new defined "Selfish order". Through theoretical analysis, we obtain the closed Nash bargaining solution under CUs' rate constraints, and prove the stability of the formatted coalition. Simulation results show that the proposed resource allocation approach achieves better performance on resource allocation fairness, with only little sacrifice on the system sum rates.

A STUDY OF THE RADIATION EFFECTS ON THE BASAL CELL OF THE RAT TONGUE EPITHELIUM ACCORDING TO THE EXPOSURE TIMING (방사선 조사시기에 따른 백서 설상피의 기저세포에 미치는 영향에 관한 연구)

  • Na Chun-Hwa;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.25 no.2
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    • pp.343-362
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    • 1995
  • The purpose of this study was to investigate chronic radiation effects on the basal cell of the rat tongue epithelium according to different irradiation timing. Forty-two female rats were devided into 5 experimental groups according to different irradiation timing and were irradiated single dose of 396cGy by MK cell irradiator using Cs-137. Experimental rats were sacrificed at the 2nd week, 4th week and 6th week after birth. The specimens were examined with light microscope and transmission electron microscope. The following results were obtained. 1. The first changes after irraditation were vacuoles. The vacuoles were chiefly observed in the cytoplasm, perinuclei area, and nuclei. 2. The most severe degenerative changes in the basal cell layer were observed in all experimental groups. ; cellular disarrangement, vacuole formation, widening of intercellular space, enlarged mitochondria & rER, and chromatin clumping were seen. 3. The cellular degenerative changes were most severe at the 4th week after birth in all experimental group, and the basal cell hyperplasia was seen at the 6th week in the most of experimental groups 4. The experimental groups 3 and 4 show more severe and more prolonged cellular degeneration than experimental groups 1 and 2, which were irradiated in pregnancy, and experimental group 5, which was irradiated after tongue maturation.

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Effects of mTORC1 inhibition on proteasome activity and levels

  • Park, Seo Hyeong;Choi, Won Hoon;Lee, Min Jae
    • BMB Reports
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    • v.55 no.4
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    • pp.161-165
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    • 2022
  • The mechanistic target of rapamycin (mTOR) regulates numerous extracellular and intracellular signals involved in the maintenance of cellular homeostasis and cell growth. mTOR also functions as an endogenous inhibitor of autophagy. Under nutrient-rich conditions, mTOR complex 1 (mTORC1) phosphorylates the ULK1 complex, preventing its activation and subsequent autophagosome formation, while inhibition of mTORC1 using either rapamycin or nutrient deprivation induces autophagy. Autophagy and proteasomal proteolysis provide amino acids necessary for protein translation. Although the connection between mTORC1 and autophagy is well characterized, the association of mTORC1 inhibition with proteasome biogenesis and activity has not been fully elucidated yet. Proteasomes are long-lived cellular organelles. Their spatiotemporal rather than homeostatic regulation could be another adaptive cellular mechanism to respond to starvation. Here, we reviewed several published reports and the latest research from our group to examine the connection between mTORC1 and proteasome. We have also investigated and described the effect of mTORC1 inhibition on proteasome activity using purified proteasomes. Since mTORC1 inhibitors are currently evaluated as treatments for several human diseases, a better understanding of the link between mTORC1 activity and proteasome function is of utmost importance.