• Title/Summary/Keyword: cellular uptake

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Effects of Tea Constituents on Intracellular Level of the Major Tea Catechin, (-)-Epigallocatechin-3-gallate

  • Hong, Jun-Gil;Yang, Chung-S.
    • Food Science and Biotechnology
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    • v.16 no.1
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    • pp.167-170
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    • 2007
  • (-)-Epigallocatechin-3-gallate (EGCG), a mai or tea catechin has been shown to have many interesting biological activities. In the present study, we studied the effects of green tea catechins, EGCG metabolites, and black tea theaflavins on accumulation of EGCG in HT-29 human colon cells. Intracellular levels of [$^3H$]-EGCG were not changed significantly in the presence of other tea catechins including (-)-epicatechin, (-)-epigallocatechin, and (-)-epicatechin-3-gallate. EGCG methyl metabolites and EGCG 4"-glucuronide did not affect cellular levels of [$^3H$]-EGCG. Black tea theaflavins and theasinensin A (TsA), an EGCG oxidative dimer, however, significantly decreased cellular accumulation of EGCG in HT-29 cells by 31-56%. This decrease was more pronounced when cells were incubated in the presence of theaflavin-3',3"-digallate (TFdiG) or TsA. When EGCG was added separately from TFdiG or TsA, the accumulation of EGCG in HT-29 cells was also significantly decreased regardless of when TFdiG or TsA was added during the uptake study (p<0.01). The results suggest that theaflavins and TsA may interrupt EGCG absorption through the gastrointestinal epithelium.

Characterization of Putrescine Uptake in Hamster Amelanocytic Melanoma AMEL-3 Cells

  • Garcia-Fernandez, Antonio J.;Rodriguez, Rosa A.;Perez-Pertejo, Yolanda;Balana-Fouce, Rafael
    • Molecules and Cells
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    • v.20 no.1
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    • pp.127-135
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    • 2005
  • The uptake of putrescine, spermidine and spermine by Fortner's hamster amelanocytic melanoma AMEL-3 cells was observed in this study to be time-dependent, temperature-sensitive, pH-dependent and saturable. Metabolic poisons nullified polyamine uptake, an indication that this is an energy-requiring mechanism. The presence of $Na^+$ ions was found to be requisite to full activity. Valinomycin, gramicidin, monensin and the calcium ionophore calcimycin were also observed to inhibit the process substantially. The transporter active site would seem to contain sulfhydryl groups. Other diamines and polyamine analogues, as well as cationic diamidines, suppressed putrescine uptake. The presence of the ornithine decarboxylase inhibitor DFMO in the culture medium induced putrescine inflows. Putrescine, in turn, induced the negative expression of the carrier, thus suggesting that this influx mechanism is governed by up/down regulation. The cationic diamidine CGP 40215A and its analogue CGP039937A competitively inhibited putrescine transport, with Ki values of 1.9 and $15{\mu}M$, respectively. The role of polyamine uptake in these cultures is discussed.

Effects of the Particulate Matter2.5 (PM2.5) on Lipoprotein Metabolism, Uptake and Degradation, and Embryo Toxicity

  • Kim, Jae-Yong;Lee, Eun-Young;Choi, Inho;Kim, Jihoe;Cho, Kyung-Hyun
    • Molecules and Cells
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    • v.38 no.12
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    • pp.1096-1104
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    • 2015
  • Particulate $matter_{2.5}$ ($PM_{2.5}$) is notorious for its strong toxic effects on the cardiovascular, skin, nervous, and reproduction systems. However, the molecular mechanism by which $PM_{2.5}$ aggravates disease progression is poorly understood, especially in a water-soluble state. In the current study, we investigated the putative physiological effects of aqueous $PM_{2.5}$ solution on lipoprotein metabolism. Collected $PM_{2.5}$ from Seoul, Korea was dissolved in water, and the water extract (final 3 and 30 ppm) was treated to human serum lipoproteins, macrophages, and dermal cells. $PM_{2.5}$ extract resulted in degradation and aggregation of high-density lipoprotein (HDL) as well as low-density lipoprotein (LDL); apoA-I in HDL aggregated and apo-B in LDL disappeared. $PM_{2.5}$ treatment (final 30 ppm) also induced cellular uptake of oxidized LDL (oxLDL) into macrophages, especially in the presence of fructose (final 50 mM). Uptake of oxLDL along with production of reactive oxygen species was accelerated by $PM_{2.5}$ solution in a dose-dependent manner. Further, $PM_{2.5}$ solution caused cellular senescence in human dermal fibroblast cells. Microinjection of $PM_{2.5}$ solution into zebrafish embryos induced severe mortality accompanied by impairment of skeletal development. In conclusion, water extract of $PM_{2.5}$ induced oxidative stress as a precursor to cardiovascular toxicity, skin cell senescence, and embryonic toxicity via aggregation and proteolytic degradation of serum lipoproteins.

Cellular Uptake and Transfection Efficiency of Plasmid DNA Using Low Molecular Weight Polyethylenimine (저분자량 폴리에틸렌이민을 이용한 유전자 송달 및 발현 유효성 연구)

  • Jeong, Gil-Jae;Park, Kui-Lye;Shin, Ji-Young;Choi, Han-Gon;Oh, Yu-Kyoung
    • Journal of Pharmaceutical Investigation
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    • v.34 no.4
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    • pp.263-267
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    • 2004
  • Branched and linear polyethylenimines (PEIs) have been studied as efficient and versatile agents for gene delivery in vitro and in vivo. PEIs exist in a linear or branched topology and are available in a wide range of molecular weight (Mw). Most studies have been done using PEIs with Mw higher than 10Kd. This study was aimed to test the transfection efficiency and the cell viability following gene delivery using PEI of Mw 2Kd, a relatively lower Mw cationic polymer. We used murine interleukin-2(mIL-2) plasmid DNA complexed with branched PEI 2Kd or 25Kd, and transfected them into a myoblast muscle cell line, C2C12. The cellular uptake of mIL-2 plasmid DNA was determined using quantitative polymerase chain reaction. RNA transcript levels were studied in the myoblast cells. Our results show that PEI 2Kd was as effective as PEI 25Kd in celluar gene delivery and transfection efficiency in C2C12 cells. Moreover, MTT assay indicated that PEI 2Kd/DNA complexes did not significantly reduce the cell viability regardless of N/P ratios. These results suggest that PEI of Mw 2Kd might play a role as effective and low toxic nonviral vector systems for muscular cell lines.

Effects of Bisphosphonates on Glucose Transport in a Conditionally Immortalized Rat Retinal Capillary Endothelial Cell Line (TR-iBRB Cells)

  • Lee, Na-Young;Park, Hyun-Joo;Kang, Young-Sook
    • Biomolecules & Therapeutics
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    • v.24 no.1
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    • pp.94-98
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    • 2016
  • The objective of the present study was to elucidate the effect of bisphosphonates, anti-osteoporosis agents, on glucose uptake in retinal capillary endothelial cells under normal and high glucose conditions. The change of glucose uptake by pre-treatment of bisphosphonates at the inner blood-retinal barrier (iBRB) was determined by measuring cellular uptake of $[^3H]3$-O-methyl glucose (3-OMG) using a conditionally immortalized rat retinal capillary endothelial cell line (TR-iBRB cells) under normal and high glucose conditions. $[^3H]3$-OMG uptake was inhibited by simultaneous treatment of unlabeled D-glucose and 3-OMG as well as glucose transport inhibitor, cytochalasin B. On the other hand, simultaneous treatment of alendronate or pamidronate had no significant inhibitory effect on $[^3H]3$-OMG uptake by TR-iBRB cells. Under high glucose condition of TR-iBRB cells, $[^3H]3$-OMG uptake was increased at 48 h. However, $[^3H]3$-OMG uptake was decreased significantly by pre-treatment of alendronate or pamidronate compared with the values for normal and high glucose conditions. Moreover, geranylgeraniol (GGOH), a mevalonate pathway intermediate, increased the uptake of $[^3H]3$-OMG reduced by bisphosphonates pre-treatment. But, pre-treatment of histamine did not show significant inhibition of $[^3H]3$-OMG uptake. The glucose uptake may be down regulated by inhibiting the mevalonate pathway with pre-treatment of bisphosphonates in TR-iBRB cells at high glucose condition.

Evaluation of Optimal Combination of Commercially Available Superparamagnetic Iron Oxide Nanoparticles and Transfection Agents for Labelling of Human Mesenchymal Stem Cells (인체 중간엽 줄기세포의 표지를 위한 상용화 된 Superparamagnetic Iron Oxide Nanoparticle과 Tansfection Agent의 적절한 병용을 위한 연구)

  • Kim, Sung-Hun;Oh, Soon-Nam;Park, Youn-Hee;Kang, Won-Kyung;Ahn, Kook-Jin;Chung, Soo-Kyo
    • Investigative Magnetic Resonance Imaging
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    • v.16 no.1
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    • pp.31-39
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    • 2012
  • Purpose : To determine the optimal combination of commercially available superparamagnetic iron oxide (SPIO) nanoparticles with transfection agents (TA). Materials and Methods: Protamine sulfate (Pro) and poly-L-lysin (PLL) were incubated with ferumoxide and ferucarbotran in human mesenchymal stem cells at various concentrations, and cellular viability were evaluated. Cellular iron uptake was qualitatively and quantitatively evaluated. Cell visibility was assessed via MR imaging and the T2-relaxation time was calculated. Results: The cellular viabilities with ferucarbotran were more significantly decreased than those with ferumoxide (p < 0.05). Iron uptake with ferumoxide was significantly higher than that for those with with ferucarbotran. The T2-relaxation time was observed to be shorter with ferumoxide in comparison to those with ferucarbotran (p < 0.05). Ferumoxide at a concentration of 25 ${\mu}g$/ml in combination with either Pro or PLL at a concentration of 3.0 ${\mu}g$/ml did not adversely impact cell viability, maximized iron uptake, and exhibited a lower T2-relaxation time in comparison to other combinations. Conclusion: Stem cells with ferumoxide exhibited a higher cellular viability and iron uptake in comparison to ferucarbotran-treated stem cells. A 25 ${\mu}g$/ml of ferumoxide with a 3.0 ${\mu}g$/ml of TA is sufficient to label mesenchymal stem cells.

Cholesterol Uptake by Lactobacillus acidophilus: Its Fate and Factors Influencing the Uptake (Lactobacillus acidophilus에 의한 콜레스테롤의 흡착)

  • Noh, Dong-Ouk
    • Journal of Dairy Science and Biotechnology
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    • v.14 no.2
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    • pp.195-206
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    • 1996
  • Cholesterol assimilated by Lactobacillus acidophilus ATCC 43121 was not metabolically degraded in that most of it was recovered with the cells. Cells grown in the presence of cholesterol micelles and bile salts were more resistant to Iysis by sonication than those grown in their absence, suggesting a possible alteration of cellular membranes. Cholesterol assimilation occurred during growth at pH 6.0, the amount of which was more than that by cells grown without pH control. Cholesterol assimilated by cells was recovered in the membrane fractions of cells both grown at pH 6.0 and without pH control. The effect of unsaturated fatty acids on cholesterol assimilation was not clear, since there was no significant (P> 0.05) difference in the amount taken up from micelles prepared using L-${\alpha}$-phosphatidylcholine, dioleoyl or L-${\alpha}$-phosphatidylcholine, distearoyl. Without Tween 80, little, if any, cell growth or cholesterol uptake was observed. In the presence of 0.05% Tween 80, cholesterol uptake increased dramatically as did growth. However, as the amount of Tween 80 increased beyond 0.05%, cholesterol uptake decreased while the amount of growth remained the same.

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Aluminium increase Iron uptake into Glial cells

  • Cheong, Jae-Hoon;Lim, Sung-Sup;Lee, Choong-Jae
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.293.3-294
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    • 2002
  • In the brain, glial cells serve in the role to sequester metal from the neural microenvironment and therefore play an important role as a cellular deposition site. The central nervous system is highly vulnerable to oxidative stress, and free iron can stimulate oxidative stress by the Fenton reaction. Aluminum may upregulates the transferrin-independent iron uptake system and stimulate oxidative stress. Nramp2. also known as DMT 1. is a 12-transmembrane(TM) domain protein responsible for dietary iron uptake as well as metal ions such as iron. lead, mangamese. zinc. copper, and cobait. (omitted)

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Effect of Hypoxia on the Doxorubicin Sensitivity of Human MCF-7 Breast Cancer Cells

  • Lim, Soo-Jeong;Kang, He-Kyung
    • Journal of Pharmaceutical Investigation
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    • v.37 no.5
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    • pp.287-290
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    • 2007
  • Intrinsic or acquired resistance to chemotherapeutic drugs is one of the major obstacles to effective cancer treatment. Hypoxia is widespread in solid tumors as a consequence of decreased blood flow in the tumor-derived neovasculature. The recent finding of a link between hypoxia and chemoresistance prompted us to investigate whether hypoxia induces doxorubicin resistance in human MCF-7 breast cancer cells. Low oxygen concentration decreased the doxorubicin sensitivity in MCF-7 cells. The expression of p-glycoprotein, a major MDR-related transporter, and those of apoptosis-related proteins (anti-apoptotic Bcl-2, Bcl-XL and pro-apoptotic Bax) were not altered by hypoxia in MCF-7 cells. Intracellular uptake of doxorubicin was significantly decreased under hypoxic conditions. Decreased cellular uptake of doxorubicin under hypoxia may contribute to causing doxorubicin resistance in these cells. The use of agents that can modulate the doxorubicin uptake for adjuvant therapy may contribute to improving the therapeutic efficacy of doxorubicin in breast cancer patients.

Deciphering Macrophage Phenotypes upon Lipid Uptake and Atherosclerosis

  • Jihye Lee;Jae-Hoon Choi
    • IMMUNE NETWORK
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    • v.20 no.3
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    • pp.22.1-22.21
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    • 2020
  • In the progression of atherosclerosis, macrophages are the key immune cells for foam cell formation. During hyperlipidemic condition, phagocytic cells such as monocytes and macrophages uptake oxidized low-density lipoproteins (oxLDLs) accumulated in subintimal space, and lipid droplets are accumulated in their cytosols. In this review, we discussed the characteristics and phenotypic changes of macrophages in atherosclerosis and the effect of cytosolic lipid accumulation on macrophage phenotype. Due to macrophage plasticity, the inflammatory phenotypes triggered by oxLDL can be re-programmed by cytosolic lipid accumulation, showing downregulation of NF-κB activation followed by activation of anti-inflammatory genes, leading to tissue repair and homeostasis. We also discuss about various in vivo and in vitro models for atherosclerosis research and next generation sequencing technologies for foam cell gene expression profiling. Analysis of the phenotypic changes of macrophages during the progression of atherosclerosis with adequate approach may lead to exact understandings of the cellular mechanisms and hint therapeutic targets for the treatment of atherosclerosis.