• Title/Summary/Keyword: chelation

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Inhibition of Microsomal $H^+\;-\;ATPase$ Prepared from Tomato Roots by Various Anions (음이온에 의한 토마토 뿌리조직 마이크로솜 $H^+-ATPase$ 활성 저해)

  • Shin, Dae-Seop;Kim, Young-Kee
    • Applied Biological Chemistry
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    • v.44 no.2
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    • pp.67-72
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    • 2001
  • $H^+-ATPase$ located on plasma and vacuolar membranes play major roles in various cellular physiological processes. In order to investigate the physiological roles of $H^+-ATPase$, microsomes were prepared from tomato roots and the effects of various anions were measured on the activities of $H^+-ATPase$. $H^+-ATPase$ was inhibited by various anions. Citrate and phosphate were chosen to investigate detailed inhibitory mechanisms on $H^+-ATPase$ since they showed different levels of inhibition. Inhibitory effect of citrate was observed at the concentrations above 3 mM. When 20 mM citrate was added, the ATPase activity was decreased by 50-60%. However, the inhibitory effect of citrate was decreased by increasing the concentration of$Mg^{2+}$ The citrate-induced inhibited activity was recovered by the addition of $Mg^{2+}$ Addition of 7 mM $Mg^{2+}$ completely removed the inhibitory effect of citrate and the activity recovered to the level of the control experiment. These results imply that citrate chelates $Mg^{2+}$ and thus inhibits $H^+-ATPase$. Meanwhile, the inhibitory effect of phosphate was observed at the concentration above 3 mM and the activity was decreased by 50% in the presence of 30 mM phosphate. Further addition of $Mg^{2+}$ showed no recovery on the activity. These results imply that the inhibitory effect of phosphate is not dependent upon the concentration of $Mg^{2+}$.

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Antioxidant and Antihypertensive Activities of Solvent Extract from Styela clava Tunic, Fishery Waste (수산폐기물인 미더덕껍질 용매 추출물의 항산화 및 항고혈압 활성)

  • Lee, Sang-Mee;Lee, Ye-Ram;Cho, Kwang-Sik;Park, Soo-Yun;Jang, Eun-Young;Hwang, Dae-Youn;Jung, Young-Jin;Son, Hong-Joo
    • Journal of Environmental Science International
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    • v.24 no.7
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    • pp.917-925
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    • 2015
  • Styela clava tunic is generated in large amounts as a waste from S. clava processing plants and leads to environmental problems. It destroys the beach scenery and causes a bad smell and pollution by trashing on the seashore. Therefore, purpose of this study was to investigate antioxidant and antihypertensive activities of different solvent extracts from S. clava tunic for recycling of fishery waste. Antioxidant and antihypertensive activities of all extracts were concentration-dependent. Of extracts, hot water extract showed the highest DPPH radical scavenging activity with the lowest effective concentration ($EC_{50}$) value (0.733 mg/ml). Chloroform extract exhibited the highest metal chelation activity with the lowest $EC_{50}$ value (2.696 mg/ml). Autoclaved water extract showed the highest NO radical scavenging activity with the lowest $EC_{50}$ value (0.491 mg/ml) and n-hexane extract showed the highest reducing power ($A_{700}=1.897$ at 100 mg/ml). And n-butanol extract showed the highest SOD-like activity with the lowest $EC_{50}$ value (19.116 mg/ml) and ACE inhibition activity with the lowest inhibitory concentration($IC_{50}$) value (0.149 mg/ml). These results indicate that extracts obtained from S. clava tunic may potential candidate to reduce diseases caused by various oxidative stresses and hypertension.

ANTIBACTERIAL EFFECT OF POLYPHOSPHATES ON MUTANS STREPTOCOCCI (Mutans streptococci에 대한 polyphosphate의 항균효과)

  • Kang, Kye-Sook;Choi, Yeong-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.1
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    • pp.80-91
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    • 2003
  • Mutans streptococci, especially S. mutans and S. sobrinus strongly implicated in pathogenesis of dental caries, the major cause of tooth loss in children. Use of an antibacterial agent controlling dental caries has been rationalized. The present study was performed to observe the antibacterial effect of inorganic polyphosphates (polyP) on S. mutans and S. sobrinus. S. mutans GS5 and S. sobrinus 6715 were grown in brain-heart infusion broth with or without polyP. Minimal inhibitory concentration (MIC) of polyP for S. mutans GS5 was determined to be 0.08% and that for S. sobrius 6715 was 0.17%. PolyP 15 added to the growing culture of S. mutans GS5 and S. sobrinus 6715 at their exponential phase was as effective in inhibiting the growth of S. mutans GS5 and S. sobrinus 6715 as polyP added at the very beginning of the culture. More than 85% of the cells lost their viability determined by viable cell count when polyP 15 was added to the culture of growing S. mutans GS5 at MIC, suggesting that polyP 15 has bacterial effect on the bacterium. And more than 99.9% of the cells lost their viability determined by viable cell count when polyP 15 was added to the culture of growing S. sobrinus 6715 at MIC, suggesting that polyP 15 has bacterial effect on the bacterium. Intracellular nucleotide release from S. mutans CS5 and S. sobrinus 6715 was increased in the presence of polyP 15 for 5h but was not really reversed by the addition of divalent cations like $Ca^{++}\;and\;Mg^{++}$. The majority of the cells appeared to be atypical in their shape, demonstrating accumulation of highly electron-dense granules and ghost cells. The overall results suggest that polyP have a strong bactericidal activity against S. mutans and S. sobrinus in which lysis in relation to chelation may not play the major role but unknown mechanism that possibly affects the viability of the bacterium may be involved. PolyP may be used as an agent for prevention of dental caries.

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Control of YAG($Y_{3}Al_{5]O_{12}$) Particle Shape prepared by Sol-Gel Process (솔-젤 공정(工程)을 이용(利用)하여 제조(製造)된 YAG($Y_{3}Al_{5}O_{12}$) 분말 입형제어)

  • Park, Jin-Tae;Kim, Chul-Joo;Yoon, Ho-Sung;Sohn, Jung-Soo
    • Resources Recycling
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    • v.17 no.5
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    • pp.52-59
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    • 2008
  • Sol-gel process applied in this study was carried out by chelation of metal ions and citric acid. From the results of thermal gravimetric analysis and XRD analysis of gel powder obtained through sol-gel and heat treatment, gel powders are mostly amorphous, and crystallize completely at $900^{\circ}C$, and the crystalline structure of YAG increases with increasing calcinations temperature. Since YAG prepared by sol-gel & calcinations process was porous, and the sape and size was irregular and nonuniform, the shape and size of YAG powder had to be controlled. Therefore the effects of organic materials such as ethylene glycol and surfactant on the crystalline structure of YAG powder were investigated. Polyesterification of ethylene glycol and citric acid separated reaction area of metal ions in the solution and decreased the size of YAG primary particles. The addition of Igepal 630 as surfactant formed the droplet in the solution, and increased the size of primary particles which forms the aggregate of YAG In order to obtain monodispersed YAG particles of uniform size, gel powder prepared with organic materials had to be milled before calcination. And milling process was very important for obtaining YAG of uniform size.

Genetic Toxicity Test of 8-Hydroxyquinoline by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Lee, Woo-Sun;Kim, Hyun-Joo;Lee, Eun-Mi;Kim, Joo-Hwan;Suh, Soo-Kyung;Kwon, Kyung-Jin;Sheen, Yhun-Yong;Kim, Seung-Hee;Park, Sue-N.
    • Molecular & Cellular Toxicology
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    • v.3 no.2
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    • pp.90-97
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    • 2007
  • 8-Hydroxyquinoline is used as antibacterial agent and antioxidant based on its function inducing the chelation of ferrous ion present in host resulting in production of chelated complex. This complex being transported to cell membrane of bacteria and fungi exerts antibacterial and antifungal action. In this study, we have carried out in vitro genetic toxicity tests and microarray analysis to understand the underlying mechanisms and the mode of action of toxicity of 8-hydroxyquinoline. TA1535 and TA98 cells were treated with 8-hydroxyquinoline to test its toxicity by basic genetic toxicity test, Ames and two new in vitro micronucleus and COMET assays were applied using CHO cells and L5178Y cells, respectively. In addition, microarray analysis of differentially expressed genes in L5178Y cells in response to 8-hydroxyquinoline were analyzed using Affymatrix genechip. The result of Ames test was that 8-hydroxyquinoline treatment increased the mutations in base substitution strain TA1535 and likewise, 8-hydroxyquinoline also increased mutations in frame shift TA98. 8-Hydroxyquinoline increased micronuclei in CHO cells and DNA damage in L5178Y. 8-Hdroxyquinoline resulted in positive response in all three tests showing its ability to induce not only mutation but also DNA damage. 783 Genes were initially selected as differentially expressed genes in response to 8-hydroxyquinoline by microarray analysis and 34 genes among them were over 4 times of log fold changed. These 34 genes could be candidate biomarkers of genetic toxic action of 8-hydroxyquinoline related to induction of mutation and/or induction of micronuclei and DNA damage. Further confirmation of these candidate markers related to their biological function will be useful to understand the detailed mode of action of 8-hydroxyquinoline.

Scutellaria baicalensis Extracts as Natural Inhibitors of Food Browning (천연 갈변저해제로서 황금 추출물의 효소적 갈변 저해 효과)

  • Park, Miji;Chang, Min-Sun;Jeong, Moon-Cheol;Kim, Gun-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.5
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    • pp.792-799
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    • 2013
  • This study was designed to develop natural browning inhibitors. The anti-browning effects of distilled water (SBD) and 80% ethanol extracts (SBE) of Scutellaria baicalensis Georgi in apple slices were investigated by L and ${\Delta}E$ values. Both SBD and SBE were effective in reducing the browning of apple slices and were successively fractionated into chloroform ($CHCl_3$), ethyl acetate (EtOAc), and water ($H_2O$) fractions. These extracts were measured for total phenolic content, flavonoid content, anti-oxidative activity (through free radical scavenging activity and the FRAP assay), ferrous ion chelation, and the inhibition of PPO (polyphenol oxidase) activity. Overall, fractions of SBE were better than fractions of SBD in all measurements. The highest total phenolic and flavonoid content were measured in the EtOAc and $CHCl_3$ fractions of SBE. EtOAc and $CHCl_3$ fractions also exhibited the highest anti-oxidative activities (in DPPH and ABTS free radical scavenging and the FRAP assay). Unusually, the highest ferrous ion chelating capacity was found in the $H_2O$ fraction of SBD, but the other fractions showed more than triple the ascorbic acid already in use. Also, $CHCl_3$ fractions showed a stronger inhibition of PPO activity than ascorbic acid. All of these results suggest that EtOAc and $CHCl_3$ fractions from Scutellaria baicalensis can be used as natural anti-browning agents.

Mitochondrial Ca2+ Uptake Relieves Palmitate-Induced Cytosolic Ca2+ Overload in MIN6 Cells

  • Ly, Luong Dai;Ly, Dat Da;Nguyen, Nhung Thi;Kim, Ji-Hee;Yoo, Heesuk;Chung, Jongkyeong;Lee, Myung-Shik;Cha, Seung-Kuy;Park, Kyu-Sang
    • Molecules and Cells
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    • v.43 no.1
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    • pp.66-75
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    • 2020
  • Saturated fatty acids contribute to β-cell dysfunction in the onset of type 2 diabetes mellitus. Cellular responses to lipotoxicity include oxidative stress, endoplasmic reticulum (ER) stress, and blockage of autophagy. Palmitate induces ER Ca2+ depletion followed by notable store-operated Ca2+ entry. Subsequent elevation of cytosolic Ca2+ can activate undesirable signaling pathways culminating in cell death. Mitochondrial Ca2+ uniporter (MCU) is the major route for Ca2+ uptake into the matrix and couples metabolism with insulin secretion. However, it has been unclear whether mitochondrial Ca2+ uptake plays a protective role or contributes to lipotoxicity. Here, we observed palmitate upregulated MCU protein expression in a mouse clonal β-cell, MIN6, under normal glucose, but not high glucose medium. Palmitate elevated baseline cytosolic Ca2+ concentration ([Ca2+]i) and reduced depolarization-triggered Ca2+ influx likely due to the inactivation of voltage-gated Ca2+ channels (VGCCs). Targeted reduction of MCU expression using RNA interference abolished mitochondrial superoxide production but exacerbated palmitate-induced [Ca2+]i overload. Consequently, MCU knockdown aggravated blockage of autophagic degradation. In contrast, co-treatment with verapamil, a VGCC inhibitor, prevented palmitate-induced basal [Ca2+]i elevation and defective [Ca2+]i transients. Extracellular Ca2+ chelation as well as VGCC inhibitors effectively rescued autophagy defects and cytotoxicity. These observations suggest enhanced mitochondrial Ca2+ uptake via MCU upregulation is a mechanism by which pancreatic β-cells are able to alleviate cytosolic Ca2+ overload and its detrimental consequences.

Study on Synthesis of 68GeO2 and Behavior of 68Ga3+ for Generator Column (Generator 컬럼용 68GeO2 합성 및 68Ga3+의 거동에 관한 연구)

  • Kim, Gun Gyun;Lee, Jun Young;Kim, Sang Wook;Hur, Min Gu;Yang, Seung Dae;Park, Jeong Hoon
    • Journal of Radiation Industry
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    • v.10 no.4
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    • pp.189-192
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    • 2016
  • $^{68}Ga$ has emerged as a promising candidate for non-invasive diagnostic imaging within Positron Emission Tomography (PET) because of its advantageous radiochemical characteristics ($t_{1/2}=68min$, ${\beta}^+$ yield ~89%). $^{68}Ga$ forms a stable chelation with various ligands and it is possible to be quickly and easily study using a $^{68}Ge/^{68}Ga$ generator. Commercial $^{68}Ge/^{68}Ga$ generators are chromatographic system using the inorganic materials such as alumina and tin dioxide which are employed as column matrixes for $^{68}Ge$. In this study, we tried out to make $^{68}Ge/^{68}Ga$ generator system with the $^{68}GeO_2$ microstructures for column matrix. $^{68}Ge$ tends to have stable bond with oxide as $^{68}GeO_2$ microstructures. The $^{68}GeO_2$ has been synthesized by hydrolysis of $GeCl_4$ (sol-gel method) and characterized by X-ray diffraction and scanning electron microscope for geometrical analysis. The stability of $GeO_2$ was tested using eluents with diverse solvents(water, ethanol and 0.1 N HCl). The radioactivity of $^{68}Ga^{3+}$ in eluate through $GeO_2$ was measured to prove a function as column material for a generator.

Kojic Acid Derivatives, Have Tyrosinase Inhibitory Activity to Suppress the Production of Melanin in the Biosynthetic Pathway (생체 내 경로에서 멜라닌 생성을 억제하는 타이로신 억제제로서의 코직산 유도체)

  • Park, Jung Youl;Lee, Ha Neul;Hu, Meng Yang;Park, Jeong Ho
    • Journal of Life Science
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    • v.29 no.7
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    • pp.755-761
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    • 2019
  • Kojic acid (KA) is produced by Aspergillus oryzae-sort of like mushrooms, which is commonly called as koji in Japan. KA is used as a chelation agent and a preservative preventing oxidative browning of fruits. KA also shows antibacterial and antifungal properties. Because KA stops the production of melanin by inhibiting tyrosinase in the biosynthetic pathway from tyrosine to melanin in skin, it has been applied as a skin lightening ingredient in cosmetics. Since some animal studies have shown that high amounts of KA had side effects such as in liver, kidney, reproductive, cardiovascular, gastrointestinal, respiratory, brain, and nervous system, more efficient KA derivatives are needed to be developed in order to safely apply as a skin lightening ingredient. A series of KA derivatives via conjugated with triazole by click reaction were synthesized and their in vitro tyrosinase inhibitory activities were evaluated. Most of all KA derivatives have shown in moderate tyrosinase inhibitory activities. In case of KA-hybrid compound, 1~3 have shown tyrosinase inhibitory activities about 50~10,000 times more effective tyrosinase inhibitor compared to KA itself. Specifically, the $IC_{50}$ value of KA-hybrid compound, 2 was $0.0044{\pm}0.74{\mu}M$ against tyrosinase. It is about 10,000 times more effective tyrosinase inhibitor compared to KA itself ($IC_{50}=45.2{\pm}4.6{\mu}M$).

Oxalate Chelating Activity of Egg White Proteins and Their Hydrolysates

  • Holipitiyage Shyami Rashmiki, Holipitiya;Palihawadanege Iresha Lakmini, Fernando;Ethige Chathura Nishshanka, Rathnapala;Alakolange Gedara Achala Wimukthika, Alakolanga;Edirisinghe Dewage Nalaka Sandun, Abeyrathne;Ki-Chang, Nam
    • Korean Journal of Poultry Science
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    • v.49 no.4
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    • pp.221-228
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    • 2022
  • Major egg white proteins and their hydrolysates serve as functional food ingredients that have certain metal-chelating properties. Employing egg white proteins and their hydrolysates to scavenge dietary oxalates is anticipated to have beneficial effect in the prevention of kidney stones. The objective of this study was to determine the biogenic oxalate-chelating activity of ovalbumin, ovomucin, and ovotransferrin and their hydrolysates. To prepare oxalate extracts, 30 mL of 0.25 N HCl was added to separately to 0.5 g of dried spinach and starfruit powders followed by boiling for 15 min, and after cooling, the addition of a further 20 mL of 0.25 N HCl. Having prepared these extracts, ovalbumin, ovomucin, and ovotransferrin and their hydrolysates were separately mixed with oxalate extracts and incubated at 3℃ for 24 h. Following centrifugation, supernatants were analyzed by HPLC using a reverse-phase C18 column coupled with a diode array detector. We found that all assessed proteins and their hydrolysates showed biogenic oxalate-chelating activity against the oxalates of spinach. In contrast, however, only ovalbumin, ovalbumin-hydrolysate, and ovomucin showed chelating activity (57.10%±8.84%, 85.44%±5.30%, 73.20%±4.13%, respectively) against the oxalates of starfruit (P<0.05). Overall, hydrolyzed ovalbumin was identified as the most effective chelator of the oxalates both spinach and starfruit. In this study, we thus established that the assessed egg white proteins and their hydrolysates have oxalate-chelating activity in vitro, thereby indicating that these compounds have potential utility as nutraceuticals for the chelation of dietary oxalate. However, further research will be necessary to verify their oxalate-chelating activities against different fruits and vegetables and under specific in vivo conditions and against purified oxalate.