• Title/Summary/Keyword: Cellular Uptake

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Cyclin-Dependent Kinase Inhibitor 2A is a Key Regulator of Cell Cycle Arrest and Senescence in Endothelial Colony-Forming Cells in Moyamoya Disease

  • Seung Ah Choi;Youn Joo Moon;Eun Jung Koh;Ji Hoon Phi;Ji Yeoun Lee;Kyung Hyun Kim;Seung-Ki Kim
    • Journal of Korean Neurosurgical Society
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    • v.66 no.6
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    • pp.642-651
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    • 2023
  • Objective : Endothelial colony-forming cells (ECFCs) have been reported to play an important role in the pathogenesis of moyamoya disease (MMD). We have previously observed stagnant growth in MMD ECFCs with functional impairment of tubule formation. We aimed to verify the key regulators and related signaling pathways involved in the functional defects of MMD ECFCs. Methods : ECFCs were cultured from peripheral blood mononuclear cells of healthy volunteers (normal) and MMD patients. Low-density lipoproteins uptake, flow cytometry, high content screening, senescence-associated β-galactosidase, immunofluorescence, cell cycle, tubule formation, microarray, real-time quantitative polymerase chain reaction, small interfering RNA transfection, and western blot analyses were performed. Results : The acquisition of cells that can be cultured for a long time with the characteristics of late ECFCs was significantly lower in the MMD patients than the normal. Importantly, the MMD ECFCs showed decreased cellular proliferation with G1 cell cycle arrest and cellular senescence compared to the normal ECFCs. A pathway enrichment analysis demonstrated that the cell cycle pathway was the major enriched pathway, which is consistent with the results of the functional analysis of ECFCs. Among the genes associated with the cell cycle, cyclin-dependent kinase inhibitor 2A (CDKN2A) showed the highest expression in MMD ECFCs. Knockdown of CDKN2A in MMD ECFCs enhanced proliferation by reducing G1 cell cycle arrest and inhibiting senescence through the regulation of CDK4 and phospho retinoblastoma protein. Conclusion : Our study suggests that CDKN2A plays an important role in the growth retardation of MMD ECFCs by inducing cell cycle arrest and senescence.

Effect of Ouabain on the Interaction of Mitochondria with Calcium (Mitochondria의 Calcium Uptake에 미치는 Ouabain의 영향)

  • Hong, Sa-Ack;Park, Chan-Woong;Kim, Myung-Suk;Chung, Myung-Hee
    • The Korean Journal of Pharmacology
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    • v.8 no.1
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    • pp.67-75
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    • 1972
  • Many studies on the mechanism of the inotropic action of cardiac glycosides have shown the possible intimate relationship between the mobilization of intracellular calcium and inotropic effect. Evidence obtained from recent studies suggests that cardiac glycosides may increase the intracellular $Ca^{++}$ concentration through the release of this ion from cellular or intracellular membrane. It seemed imperative to study the effect of ouabain on the interaction between mitochondria and $Ca^{++}$, because mitochondria are known to have a rather powerful $Ca^{++}$ pump mechanism which may have an important role on the regulation of intracellular $Ca^{++}$ concentration. The present investigations was made into the effect of ouabain on $Ca^{++}$ untake of mitochondria in the presence of ATP and its dependence on $K^+$ and $Na^+$ in the medium. The results are summarized as follows: 1. The rate of rise in the turbidity of superprecipitation was solely influenced by ionic strength of the medium, not by the species of ion, i.e. $Na^+$ or $K^+$. The higher ionic strength suppressed and the lower enhanced the rate of superprecipitation respectively. 2. No effect of ouabain was found on the rate of superprecipitation. 3. Mitochondria depressed the rate of superpretipitation, and the depressed rate of superprecipitation by mitochondria was reversed by ouabain, and the degree of this reversal was almost identical in $Na^+$ and $K^+$ medium. 4. $Ca^{++}$ uptake of mitochondria was inhibited by ouabain in the presence of ATP and the degree of inhibition showed the dose-response manner in terms of concentration of ouabain. 5. In the absence of ATP, mitochondria took or the $Ca^{++}$ in initial period but released it later. Such uptake and release of $Ca^{++}$ was not influenced by ouabain. 6. It is suggested that intracellular calcium mobilization by ouabain through the action upon the mitochondria was due to inhibition on ATP-dependent $Ca^{++}$ uptake by this agent, not to the action upon so called binding.

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20S-dihydroprotopanaxatriol modulates functional activation of monocytes and macrophages

  • Kim, Mi-Yeon;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.37 no.3
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    • pp.300-307
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    • 2013
  • 20S-dihydroprotopanaxatriol (2H-PPT) is a derivative of protopanaxatrol from ginseng. Unlike other components from Panax ginseng, the pharmacological activity of this compound has not been fully elucidated. In this study, we investigated the modulatory activity of 2H-PPT on the cellular responses of monocytes and macrophages to understand its immunoregulatory actions. 2H-PPT strongly upregulated the release of radicals in sodium nitroprusside-treated RAW264.7 cells and the surface levels of costimulatory molecule CD86. More importantly, this compound remarkably suppressed nitric oxide production, morphological changes, phagocytic uptake, cell-cell aggregation, and cell-matrix adhesion in RAW264.7 and U937 cells in the presence or absence of lipopolysaccharide, anti-CD43 antibody, fibronectin, and phorbal 12-myristate 13-acetate. Therefore, our results suggest that 2H-PPT can be applied as a novel functional immunoregulator of macrophages and monocytes.

Synthesis of 3β [L-Lysinamide-carbamoyl] Cholesterol Derivatives by Solid-Phase Method and Characteristics of Complexes with Antisense Oligodeoxynucleotides

  • Lee, Eun-Jung;Lee, Min-hyung;Park, Jong-Sang;Choi, Joon-Sig
    • Bulletin of the Korean Chemical Society
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    • v.27 no.7
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    • pp.1020-1024
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    • 2006
  • In this report, we describe the synthesis of mono- and di-valent cationic $3\beta$ [L-Lysinamide-carbamoyl] cholesterol (K-Chol) derivatives by solid-phase peptide synthesis method and the characteristics of K-Chol/antisense oligodeoxynucleotide (ODN) complexes. K-Chol was able to interact with antisense ODNs electrostatically and constructed nanometer-sized complexes of 50-100 nm in diameter. The formation of K-Chol/antisense ODN complexes was demonstrated by non-denaturing polyacrylamide gel electrophoresis assay and atomic force microscopy. The cell-associated radioactivity was measured to monitor the cellular uptake of the complexes containing radioactive antisense ODNs using HL 60 cells.

COMBINED IN VITRO ASSAY FOR 3T3 NRU PT ASSAY AND PHOTOHEMOLYSIS AS PART OF PHOTOTOXICITY TEST

  • Chunja Nam;Kim, Baehwan;Lee, Byoungseok;Seongjoon Moon;Ihseop Chang
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.05a
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    • pp.117-117
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    • 2001
  • The aim of this study was to assess a possible alternative method as replacement for in vivo phototoxicity test. The 3T3 mouse fibroblast neutral red uptake phototoxicity assay (3T3 NRU PT assay) is a screening method for studying DNA or cellular damage. Photohemolysis assay is a mechanistic study for investigating oxygen-dependent membrane damage.(omitted)

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Influence of $C_5$-Precursors on $\delta$-Aminolevlinic Acid Biosynthesis in Rhodocyclus gelatinosus KUP-74 (Rhodocyclus gelatinosus KUP-74에 의한 $\delta$-Aminolevulinic acid 생합성의 $C_5$-전구물질의 영향)

  • 최경민;임왕진;황세영
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.527-533
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    • 1993
  • Aminolevulinic acid(ALA) was shown to be synthesized via active pathways of either C4 or C5 ALA biosynthesis in cells of a photosynthetic bacterium, Rhodocyclus gelatinosus KUP-74, where the C5 pathway was appeared to be preferntially expressed in the cells. It was strongly suggested that L-glutamine might be utilized more effectively than L-glutamate to synthesize ALA via C5 pathway in this bacterium from the fact of relationship between the cellular uptake rates of glutamate and its Gamma-derivaties and corresponded ALA productivities in vitro and in vivo.

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Obesity-associated Morbidity (비만관련 합병증의 이해)

  • Choi, Kwang Hae
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.11 no.sup1
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    • pp.149-152
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    • 2008
  • Obesity tracks from childhood into adulthood, and the persistence of obesity rises with age among obese children. Obesity are independent risk factors for increased morbidity and mortality throughout the lifecycle. Obese individuals develop resistance to the cellular actions of insulin, characterized by an impaired ability of insulin to inhibit glucose output from the liver and to promote glucose uptake in fat and muscle. Insulin resistance is a key etiological factor for type 2 diabetes mellitus, dyslipidemia, hypertension, nonalcoholic steatohepatitis, polycystic ovarian syndrome.

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Effect of Palmiwon on the Streptozotocin induced Prediabetic Model in Panceratic Bita Cells (췌장베타세포에서 스트렙토초토신으로 유도한 당뇨병 실험 모델에 대한 팔미원의 영향)

  • 이인순;이인자
    • Biomolecules & Therapeutics
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    • v.6 no.4
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    • pp.371-377
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    • 1998
  • The aim of the present study was to investigate the effect of Palmiwon on the type 1-prediabetic models induced by streptozotocin (STZ) in RINm5F cells and HIT-T15 cells. Palmiwon increased the cell proliferation and insulin release when pre- and post-treated for the STZ-exposed pancreatic beta cells. The cell proliferation and insulin release of these beta cells were measured by $^3$H-thymidine uptake and RIA, respectively. We also analyzed nutrients such as sugars, fatty acid and amino acids and minerals containing in Palmiwon using by gas chromatography, amino acid analyzer and AA spectrometer, respectively. Palmiwon seems to have protective and recovery properties on the prediabetic model in cellular level, which were ascribe to various nutrients and minerals containing in Palmiwon. From these results, it could be suggested that Palmiwon may be available as preventive and therapeutic prescription of type 1 diabetes mellitus.

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Polyvinyl butyral DMN-conjugates for the controlled release of singlet oxygen in medical and antimicrobial applications

  • Posavec, Damir;Muller, Rainer;Bogner, Udo;Bernhardt, Gunther;Knor, Gunther
    • Biomaterials and Biomechanics in Bioengineering
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    • v.1 no.2
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    • pp.73-79
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    • 2014
  • Covalent attachment of 1, 4-dimethylnaphthalene (DMN) based endoperoxide forming subunits to a polyvinyl butyral (PVB) backbone has been achieved. The functionalized polymer materials prepared and characterized here can serve as biocompatible carrier systems for studying cellular uptake, intermediate storage and delayed release of singlet oxygen, which opens up new doors for optimizing a variety of medical applications of photogenerated DMN-endoperoxides such as antiviral, antibacterial, antiplasmodial and antitumor activity.

Control of Pierce's Disease through Degradation of Xanthan Gum

  • Lee, Seung-Don;Donald A. Cooksey
    • The Plant Pathology Journal
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    • v.20 no.1
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    • pp.1-6
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    • 2004
  • The diseases caused by Xylella fastidiosa are associated with aggregation of the bacteria m xylem vessels, formation of a gummy matrix and subsequent blockage of water uptake. In the closely related pathogen, Xanthomonas campestris, xanthan gum is known to be an important virulence factor, probably contributing to bacterial adhesion, aggregation and plugging of xylem. Xanthan gum, produced by X. campestris, is an extra-cellular polysaccharide consisting of a cellulose backbone ($\bate$-1,4-linked D-glucose) with trisaccharide side chains composed of mannose, glucuronic acid and mannose attached to alternate glucose residues in the backbone. We had constructed a mutant of X. campestris lacking gumI gene that is responsible for adding the terminal mannose for producing modified xanthan gum which is similar to xanthan gum fromX. fastidiosa. The modified xanthan gum degrading endgphytic bacterium Acineto-bacter johnsonii GX123 isolated from the oleander infected with leaf scorch disease.