• Title/Summary/Keyword: Cellular Uptake

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Proteome analysis of sorghum leaf and root in response to heavy metal stress

  • Roy, Swapan Kumar;Cho, Seong-Woo;Kwon, Soo Jeong;Kamal, Abu Hena Mostafa;Lee, Dong-Gi;Sarker, Kabita;Lee, Moon-Soon;Xin, Zhanguo;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.24-24
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    • 2017
  • Heavy metals at toxic levels have the capability to interact with several vital cellular biomolecules such as nuclear proteins and DNA, leading to oxidative stress in plants. The present study was performed to explore the metal tolerance mechanism in Sorghum seedling. Morpho-physiological and metal ions uptake changes were observed prominently in the seedlings when the plants were subjected to different concentrations of $CuSO_4$ and $CdCl_2$. The observed morphological changes revealed that the plants treated with Cu and Cd displayed dramatically altered shoot lengths, fresh weights, and relative water content. In addition, the concentration of Cu and Cd was markedly increased by treatment with Cu and Cd, and the amount of interacting ions taken up by the shoots and roots was significantly and directly correlated with the applied level of Cu and Cd. Using the 2-DE method, a total of 24 and 21 differentially expressed protein spots from sorghum leaves and roots respectively, 33 protein spots from sorghum leaves under Cd stress were analyzed using MALDI-TOF/TOF MS. However, the over-expression of GAPDH plays a significant role in assisting Sorghum bicolor to attenuate the adverse effects of oxidative stress caused by Cu, and the proteins involved in resistance to stress helped the sorghum plants to tolerate high levels of Cu. Significant changes were absorbed in the levels of proteins known to be involved in carbohydrate metabolism, transcriptional regulation, translation and stress responses. In addition, the up-regulation of glutathione S-transferase and cytochrome P450 may play a significant role in Cd-related toxicity and stress responses. The results obtained from the present study may provide insights into the tolerance mechanism of seedling leaves and roots in Sorghum under heavy metal stress.

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Effects of Iron and chelators on Primary production and Nitrogen New Production in the Equatorial Pacific Upwelling System (적도 태평양 용승계에서 철과 킬레이트 화합물이 일차생산과 질소 신생산에 미치는 영향)

  • YANG, SUNG RYULL
    • 한국해양학회지
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    • v.28 no.1
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    • pp.52-68
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    • 1993
  • Effects of iron and/or chelator addition on primary production in the equatorial Upwelling system were studied during the TOGA(Tropical oceans and Global Atmosphere) and EPOCS (Equatorial Pacific ocean Climate Studies) cruises in June and November-December of 1989. Changes in the phytoplankton biomass and the degree of iron stress were estimated using the changes in vivo fluorescence before and after the addition of DCMU, which is an inhibitor of photosynthetic electron transposer system. Nitrate uptake was measured using /SUP 45/N labeled KNO$_3$ to estimate the new production. When samples were taken from the Upwelling area where nitrate concentration was higher than 5 uM, there were significant differences between the control and cheated iron treatments in vivo fluorescence and in nitrate uptake capacity. However, CFC (Cellular fluorescence capacity) did not show any significant difference between the control and treatments until nutrient limitation becomes severse and cells become shifted-down. Outside of the Upwelling area where surface nitrate concentration was low (below 0.5 uM), there was no significant difference between the control and treatments in vivo fluorescence and CFC. It is evident that primary and new production in the equatorial Pacific Upwelling region are limited by the availability of iron. However, the physiology of phytoplankton indigenous to this region does not appear to be iron stressed judging from CFC values.

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Solids Loss with Water Uptake during Soaking of Soybeans (대두의 침지과정중 침출액의 성분변화)

  • Lee, Yung-Heon;Jung, Hae-Ok;Rhee, Chong-Ouk
    • Korean Journal of Food Science and Technology
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    • v.19 no.6
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    • pp.492-498
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    • 1987
  • Dried soybeans (varieties: Saeal, Kwanggyo, Tanyob) took up water rapidly for first 3hr followed by a slower rate of uptake. The beans took up an equal weight of water (100% hydration)after approximately 3.5hr at $50^{\circ}C$, 5hr at $30^{\circ}C$ and 7.5hr at $20^{\circ}C$ respectively. pH of the soaking solutions decreased during the soaking period. This was undoubtedly caused by the ionization of the cellular components resulting in increased levels of hydrogen ions in the liquor. Soluble solids were leached out of the beans at fairly steady rate throughout the hydration and the amount was greater with higher temperature. This amounted to 0.4-0.7g at $20^{\circ}C$ and 10.2-15.0g at $50^{\circ}C$ per 100g soybeans. Temperature was the most important factor in determining the rate of water absorption and of solid losses. Of the total solids lost, 12-25% was protein. The proportion of protein loss increased as the soaking time and temperature increase. Amount of protein loss was 80-200mg at $20^{\circ}C$ and 440-480mg at $50^{\circ}C$ after 24hr soaking per 100g soybeans. About 5% of soluble sugars, including fructose, sucrose, raffinose, and stachyose, was removed from the beans after 24hr soaking at $20^{\circ}C$.

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Insulin-like Growth Factor-I Regulates the FAT/CD36 Expression in C2C12 Skeletal Muscle Cells (C2C12 골격근 세포에서 FAT/CD36 발현 조절에 있어 Insulin-like growth factor-I이 미치는 영향)

  • Kim, Hye Jin;Yoon, Hae Min;Kim, Tae Young;Lee, Won Jun
    • Journal of Life Science
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    • v.26 no.7
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    • pp.758-763
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    • 2016
  • Fatty acid transporters are key mediators of skeletal muscle lipid metabolism. Several protein groups have been implicated in cellular long-chain fatty acid uptake or oxidation, including fatty acid transporter proteins (FATPs), the plasma membrane fatty acid-binding protein (FABPpm), and the fatty acid translocase (FAT/CD36). FAT/CD36 is highly expressed in skeletal muscle and known to be regulated by various factors such as exercise and hormones. Insulin-like growth factor-I (IGF-I) is a well-known regulator of skeletal muscle cells. However, it has not been studied whether there is any interaction between IGF-I and FAT/CD36 in skeletal muscle cells. In this study, the effects of IGF-I treatment on FAT/CD36 induction were examined. Differentiated C2C12 cells were treated with 20 ng/ml of IGF-I at different time points. Treatment of C2C12 cells with IGF-I resulted in increased FAT/CD36 mRNA and protein expression. After 24 and 48 hr of IGF-I treatment, FAT/CD36 mRNA increased 89% and 24% respectively. The increase of both proteins returned to the control level after 72 hr of IGF-I treatment, suggesting that the FAT/CD36 gene is regulated pretranslationally by IGF-I in skeletal muscle cells. These results suggest that IGF-I can regulate the expression of FAT/CD36 in skeletal muscle cells. In conclusion, IGF-I induces a rapid transcriptional modification of the FAT/CD36 gene in C2C12 skeletal muscle cells and has modulating effects on fatty acid uptake proteins as well as oxidative proteins.

Potential of Fucoidan Extracted from Seaweeds as an Adjuvant for Fish Vaccine (해조류 유래 Fucoidan의 어류용 백신 항원보조제로서의 가능성에 대한 고찰)

  • Min, Eun Young;Kim, Kwang Il;Cho, Mi Young;Jung, Sung-Hee;Han, Hyun-Ja
    • Journal of Marine Life Science
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    • v.4 no.1
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    • pp.1-13
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    • 2019
  • Fucoidan is a physiologically functional ingredient extracted from seaweed brown algae, which is a sulfated polysaccharide containing fucose as a main molecule backbone. Fucoidan has a variety of immune-modulating or -stimulating effects, including promoting antigen uptake and enhancing anti-bacterial, anti-viral and anti-tumor effects. In addition, recent studies have suggested the possibility of use of fucoidan as a vaccine adjuvant in the field of human vaccine. Use of fucoidan as supplementary feeds have already been studied, but the development of fucoidan as an adjuvant of fish vaccine is still premature. However, the intracellular uptake of fucoidan differs depending on the molecular weight of fucoidan, and there is a limit to the study on specific immune response including the production of antibodies to fish caused by an artificial infection of pathogen. Although the safety of fucoidan has been demonstrated in animal cells, there is a need to confirm the safety of fucoidan in fish. Therefore, active research in this field is needed to use fucoidan as a vaccine adjuvant. This study discussed the effects of fucoidan on immune stimulation, humoraland cellular- immunity including humans and animals. The prospect of fucoidan as a vaccine adjuvant in fisheries also reviewed.

Use of Peristeum as a Source of Endothelial-like Cells (혈관내피유사세포 채취의 원천으로 골막의 활용)

  • Park, Bong-Wook;Kim, Shin-Won;Kim, Uk-Kyu;Hah, Young-Sool;Kim, Jin-Hyun;Kim, Deok-Ryong;Sung, Iel-Young;Cho, Yeong-Cheol;Son, Jang-Ho;Kim, Jong-Ryoul;Byun, June-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.33 no.5
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    • pp.385-391
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    • 2011
  • Purpose: The periosteum is a well-known source of osteogenic precursor cells for tissue-engineered bone formation. However, cultured endothelial or endothelial-like cells derived from periosteum have not yet been investigated. This study focused on endothelial-like cell culture from the periosteum. Methods: Periosteal tissues were harvested from the mandible during surgical extraction of lower impacted third molars. The tissues were treated with 0.075% type I collagenase in phosphate-buffered saline (PBS) for 1 hr at $37^{\circ}C$ to release cellular fractions. The collagenase was inactivated with an equal volume of DMEM/10% fetal bovine serum (FBS) and the infranatant was centrifuged for 10 min at 2,400 rpm. The cellular pellet was filtered through a $100{\mu}m$ nylon cell strainer, and the filtered cells were centrifuged for 10 min at 2,400 rpm. The resuspended cells were plated into T25 flasks and cultured in endothelial cell basal medium (EBM)-2. Results: Among the hematopoietic markers, CD146 was more highly expressed than CD31 and CD34. The periosteal-derived cells also expressed CD90 and CD166, mesenchymal stem cell markers. Considering that the expression of CD146 was constant and that the expression of CD90 was lower at passage 5, respectively, the CD146 positive cells in passage 5 were isolated using the magnetic cell sorting (MACS) system. These CD146 sorted, periosteal-derived cells formed tube-like structures on Matrigel. The uptake of acetylated, low-density lipoprotein, labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI-Ac-LDL) was also examined in these cells. Conclusion: These results suggest that the CD146-sorted positive cells can be referred to as periosteal-derived CD146 positive endothelial-like cells. In particular, when a co-culture system with endothelial and osteoblastic cells in a three-dimensional scaffold is used, the use of periosteum as a single cell source would be strongly beneficial for bone tissue engineering.

Characteristics and Cellular Protective Effects against UVA of Cationic Liposome Loaded with Quercetin and Rutin (쿼세틴과 루틴을 담지한 양이온 리포좀의 특성조사 및 UVA에 대한 세포 보호 효과)

  • Gu, Hyun A;Kim, Moon Jin;Kim, Hae Soo;Ha, Ji Hoon;Yu, Eun Ryung;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.26 no.2
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    • pp.165-172
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    • 2015
  • Quercetin and its glycoside, rutin, are flavonoids, which are well known as natural antioxidants. In this study, cationic liposomes loaded with flavonoids (quercetin or rutin) were investigated for their effects on cell and skin permeability, and protective effects against UVA. The particle size of the empty cationic liposomes was in the range of 100~130 nm, and the zeta potential was + 33.05 mV. The entrapment efficiency of 0.5R/CL was higher than that of 0.5 Q/CL. The cellular uptake of the cationic liposomes was five-fold higher than that of liposomes. The skin permeability of quercetin and rutin was investigated using Franz diffusion cells. Compared to the initial loading dose, the amount of quercetin or rutin delivered to the skin by cationic liposomes was higher than that delivered by conventional liposomes or phosphate-buffered saline. From the protective effect of cationic liposomes against UVA ($25J/cm^2$), we found that the cell viability in cationic liposomes containing flavonoids was higher than that of using UVA irradiation only. These results indicate that cationic liposomes provide enhanced delivery of flavonoids (quercetin and rutin) into the skin and may be used for antiaging and antioxidant cosmetics.

Cytotoxicity Assessments of Portulaca oleracea and Petroselinum sativum Seed Extracts on Human Hepatocellular Carcinoma Cells (HepG2)

  • Farshori, Nida Nayyar;Al-Sheddi, Ebtesam Saad;Al-Oqail, Mai Mohammad;Musarrat, Javed;Al-Khedhairy, Abdulaziz Ali;Siddiqui, Maqsood Ahmed
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6633-6638
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    • 2014
  • The Pharmacological potential, such as antioxidant, anti-inflammatory, and antibacterial activities of Portulaca oleracea (PO) and Petroselinum sativum (PS) extracts are well known. However, the preventive properties against hepatocellular carcinoma cells have not been explored so far. Therefore, the present investigation was designed to study the anticancer activity of seed extracts of PO and PS on the human hepatocellular carcinoma cells (HepG2). The HepG2 cells were exposed with $5-500{\mu}g/ml$ of PO and PS for 24 h. After the exposure, cell viability by 3-(4,5-dimethylthiazol-2yl)-2,5-biphenyl tetrazolium bromide (MTT) assay, neutral red uptake (NRU) assay, and cellular morphology by phase contrast inverted microscope were studied. The results showed that PO and PS extracts significantly reduced the cell viability of HepG2 in a concentration dependent manner. The cell viability was recorded to be 67%, 31%, 21%, and 17% at 50, 100, 250, and $500{\mu}g/ml$ of PO, respectively by MTT assay and 91%, 62%, 27%, and 18% at 50, 100, 250, and $500{\mu}g/ml$ of PO, respectively by NRU assay. PS exposed HepG2 cells with $100{\mu}g/ml$ and higher concentrations were also found to be cytotoxic. The decrease in the cell viability at 100, 250, and $500{\mu}g/ml$ of PS was recorded as 70%, 33%, and 15% by MTT assay and 63%, 29%, and 17%, respectively by NRU assay. Results also showed that PO and PS exposed cells reduced the normal morphology and adhesion capacity of HepG2 cells. HepG2 cells exposed with $50{\mu}g/ml$ and higher concentrations of PO and PS lost their typical morphology, become smaller in size, and appeared in rounded bodies. Our results demonstrated preliminary screening of anticancer activity of Portulaca oleracea and Petroselinum sativum extracts against HepG2 cells, which can be further used for the development of a potential therapeutic anticancer agent.

Solute Carrier SLC41A1 'A MINI REVIEW'

  • Basnet Hom Bahadur
    • Environmental Mutagens and Carcinogens
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    • v.25 no.2
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    • pp.60-65
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    • 2005
  • The human solute carrier, SLC41Al, is a $Mg^{2}+$ transporter that is regulated by extracellular magnesium. Although intracellular magnesium plays a fundamental role in cellular metabolism, little is known about how $Mg^{2}+$ is taken up and controlled by cells. Magnesium plays a fundamental role in cellular metabolism so that its control within the body is critical. Magnesium homeostasis is principally a balance between intestinal absorption of dietary magnesium and renal excretion of urinary magnesium. The kidney, mainly the distal convoluted tubule, controls magnesium reabsorption. Although renal reabsorption is under the influence of many hormones, selective regulation of magnesium transport is due to intrinsic control involving transcriptional processes and synthesis of transport proteins. Using microarray analysis, identification of the genetic elements involved with this transcriptional control has been begun. SLC41A1(GenBank Accession No. AJ514402), comprises 10 putative transmembrane domains, two of which are highly homologous to the integral membrane part of the prokaryote transports $Mg^{2}+$ and other divalent cations $Sr^2+,\;Zn^2+,\;Cu^2+,\;Fe^2+,\;Co^2+,\;Ba^2+,\;and\;Cd^2+,\;but\;not\;Ca^2+,\;Mn^2+,\;and\;Ni^2+.$ Transport of $Mg^{2}+$ by SLC41Al is rheogenic, voltage dependent, and not coupled to Na or Cl. Expressed SLC41Al transports a range of other divalent cations: $Mg^{2+},\;Sr^{2+},\;Zn^{2+},\;Cu^{2+},\;Fe^{2+},\;Co^{2+},\;Ba^{2+},\;and\;Cd^{2+}$. The divalent cations $Ca^{2+},\;Mn^{2+},\;and\;Ni^{2+}$and the trivalent ion $Gd^{3+}$ did not induce currents nor did they inhibit $Mg^{2+}$ transport. The nonselective cation $La^{3+}$ abolishes $Mg^{2+}$ uptake. Computer analysis of the SLC41Al protein structure reveals that it belongs to MgtE protein family & suggested that the human solute carrier, SLC41Al, might be a eukaryotic $Mg^{2+}$ transporter closely related $(60-70\%)$ protein encoded by SLC41A2 is a $Mg^{2}+$ transporter that might be involved in magnesium homeostasis in epithelial cells also transports a range of other divalent cations: $Ba^2,\;Ni^2,\;CO^2,\;Fe^2,\;or\;Mn^2,\;but\;not\;Ca^2,\;Zn^2,\;or\;Cu^{2+}$ that may have related functional properties.

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Pharmacokinetic and Pharmacodynamic Interaction between Metformin and (-)-Epigallocatechin-3-gallate

  • Ko, Jeong-Hyeon;Jang, Eun-Hee;Park, Chang-Shin;Kim, Hyoung-Kwang;Cho, Soon-Gu;Shin, Dong-Wun;Yi, Hyeon-Gyu;Kang, Ju-Hee
    • Molecular & Cellular Toxicology
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    • v.5 no.4
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    • pp.298-303
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    • 2009
  • (-)-Epigallocatechin-3-gallate (EGCG), a major flavonoid in green tea has multiple health benefits including chemoprevention, anti-inflammatory, anti-diabetic, and anti-obesity effects. In connection with these effects, EGCG can be a candidate to help the treatment of metabolic diseases. Metformin is a widely used anti-diabetic drug regulating cellular energy homeostasis via AMP-activated protein kinase (AMPK) activation. Therefore, the combination of metformin with EGCG may have additive or synergistic effects on treatment of type 2 diabetes. Nevertheless, there is no report for the pharmacokinetic and/or pharmacodynamic interaction of EGCG with metformin. Here, we evaluated the pharmacokinetic and pharmacodynamic interaction between metformin and EGCG in rats. Pharmacokinetics parameters of metformin were measured after oral administration of metformin in rats pre-treated with EGCG (10 mg/kg) or saline for 7 days. The results showed that there is no significant difference in pharmacokinetic parameters between saline control and EGCG-treated group. In addition, the hepatic AMPK activation by metformin in EGCG-treated rats was also similar to the control. The lack of additive effects of EGCG on AMPK activation or intracellular uptake of metformin was also evaluated in cells in the presence or absence of EGCG. Treatment of HepG2 cells with EGCG inhibited the metformin-induced AMPK activation. Combined results suggested that EGCG has no effect on the pharmacokinetics of metformin but may contribute to metformin action.