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Diagnostic ex vivo assay of glucose in live cell using voltammetry

  • Ly, Suw Young;Leea, Chang Hyun
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.4
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    • pp.1379-1385
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    • 2018
  • The hand held voltammetry systems searched diabetic assay using glucose sensor of fluorine nafion doped carbon nanotube electrode (FCNE). An inexpensive graphite carbon pencil was used as an Ag/AgCl reference and Pt counter electrode. Upon combining and using three electrode systems, optimum square wave (SW) stripping results were attained to 1.0-9.0 ug/L with 8 points. Statistic RSD precision was of 6.02 % with n=15 in 0.1 mg/L glucose. After a total of 200 second accumulation times, analytical detection limit of 0.8 ug/L was obtained. This developed technique was applied to urine samples from diabetic patients urine for fluid analysis, it was determined that the sensor can be used with a diagnostics in the ex vivo of live cells and non treated biological fluid.

Antioxidant effect and iNOS, COX-2 Expression Inhibition on RAW 264.7 Cell of Mangifera indica L. Leafs (애플망고 잎의 추출물의 항산화 및 대식세포(RAW 264.7)에서 iNOS, COX-2 발현 저해 효과)

  • Yoo, Dan-Hee;Lee, In-Chul
    • Journal of Life Science
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    • v.30 no.9
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    • pp.783-790
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    • 2020
  • The present study investigates the antioxidant and anti-inflammatory activities of Mangifera indica L. leaf extract. The total polyphenol content was measured using the Folin-Denis method. Results showed that the M. indica L. leaf extract of water and 70% ethanol showed a content of 440.83±1.02, 475.63±1.3 mg/100 g tannic acid equivalent. To assess antioxidant activity and electron-donating ability, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity were measured, and all extracts were found to be highly efficacious. To assess cell viability of the extract from M. indica L. leaf on macrophage cells (RAW 264.7), a 3-[4,5-dimethyl-thiazol-2- yl]-2,5-diphenyl-tetrazolium-bromide assay was performed. The following experiments were conducted in section where cells was not shown of toxicity. In order to effectively determine anti-inflammatory activity, inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 cells was examined using a Griess assay. The result showed that M. indica L. leaf extract concentration-dependently inhibited NO production. M. indica L. leaf extract was measured using Western blot, reverse transcription- polymerase chain reaction (RT-PCR) that to find the production of pro-inflammatory factor on stimulated RAW 264.7 cells of LPS. According to the results of this study, the M. indica L. leaf extract showed excellent effectiveness in antioxidant and anti-inflammatory activity, thus confirming its usability as a natural material and a functional raw material for cosmetics.

Effect of 5-aza-2'-deoxycytidine on Cell Proliferation of Non-small Cell Lung Cancer Cell Line A549 Cells and Expression of the TFPI-2 Gene

  • Dong, Yong-Qiang;Liang, Jiang-Shui;Zhu, Shui-Bo;Zhang, Xiao-Ming;Ji, Tao;Xu, Jia-Hang;Yin, Gui-Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4421-4426
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    • 2013
  • Objective: The present study employed 5-aza-2'-deoxycytidine (5-Aza-CdR) to treat non-small cell lung cancer (NSCLC) cell line A549 to investigate the effects on proliferation and expression of the TFPI-2 gene. Methods: Proliferation was assessed by MTT assay after A549 cells were treated with 0, 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR, a specific demethylating agent, for 24, 48 and 72h. At the last time point cells were also analyzed by flow cytometry (FCM) to identify any change in their cell cycle profiles. Methylation-specific polymerase chain reaction (MSPCR), real time polymerase chain reaction(real-time PCR) and western blotting were carried out to determine TFPI-2 gene methylation status, mRNA expression and protein expression. Results: MTT assay showed that the growth of A549 cells which were treated with 5-Aza-CdR was significantly suppressed as compared with the control group (0 ${\mu}mol/L$ 5-Aza-CdR). After treatment with 0, 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR for 72h, FCM showed their proportion in G0/G1 was $69.7{\pm}0.99%$, $76.1{\pm}0.83%$, $83.8{\pm}0.35%$, $95.5{\pm}0.55%$ respectively (P<0.05), and the proportion in S was $29.8{\pm}0.43%$, $23.7{\pm}0.96%$, $15.7{\pm}0.75%$, $1.73{\pm}0.45%$, respectively (P<0.05), suggesting 5-Aza-CdR treatment induced G0/G1 phase arrest. MSPCR showed that hypermethylation in the promoter region of TFPI-2 gene was detected in control group (0 ${\mu}mol/L$ 5-Aza-CdR), and demethylation appeared after treatment with 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR for 72h. Real-time PCR showed that the expression levels of TFPI-2 gene mRNA were $1{\pm}0$, $1.49{\pm}0.14$, $1.86{\pm}0.09$ and $5.80{\pm}0.15$ (P<0.05) respectively. Western blotting analysis showed the relative expression levels of TFPI-2 protein were $0.12{\pm}0.01$, $0.23{\pm}0.02$, $0.31{\pm}0.02$, $0.62{\pm}0.03$ (P<0.05). TFPI-2 protein expression in A549 cells was gradually increased significantly with increase in the 5-Aza-CdR concentration. Conclusions: TFPI-2 gene promoter methylation results in the loss of TFPI-2 mRNA and protein expression in the non-small cell lung cancer cell line A549, and 5-Aza-CdR treatment could induce the demethylation of TFPI-2 gene promoter and restore TFPI-2 gene expression. These findings provide theoretic evidence for clinical treatment of advanced non-small cell lung cancer with the demethylation agent 5-Aza-CdR. TFPI-2 may be one molecular marker for effective treatment of advanced non-small cell lung cancer with 5-Aza-CdR.

Determination of Arsenic in Water by ICP-DRC/MS (ICP-DRC/MS를 이용한 수중의 비소 측정)

  • Jeong, Gwan-Jo;Kim, Dok-Chan;Park, Hyeon
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.6
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    • pp.620-625
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    • 2006
  • In this investigation, an ICP-DRC/MS method to measure arsenic with ultra-trace concentration without any interference by the compounds such as $^{40}Ar^{35}Cl^+\;and\;^{40}Ca^{35}Cl^+$, which disturb the precise measurement of arsonic was described. Thus, the oxgen was introduced into the dynamic reaction cell as reaction gas and reacted with arsenic ion created in plasma gas, $AsO^+$ was formed and detected with m/z of 91 by ICP-MS. It resulted in better detection limit than the old method with m/z of 75($As^+$). The optimum condition for oxygen supply as the reaction gas was 0.5 mL/min. The analytical features of the method are as follows: detection limit of $0.02{\mu}g/L$, precision(RSD) of 3.4%, and recovery of 96%. Arsenic in the water samples from the tributary streams to the Han River and the main stream of Paldang were analyzed with this method to identify the characteristics in its distribution. The concentration of As ranged from 0.53 to $1.26{\mu}g/L$. We could measure As with very low concentration, less than $1.0{\mu}g/L$, with excellent reproducibility. The method developed is expected to be applied to analyze As of the samples from sea water, food, and domestic and industrial waste water which have high concentration of Cl and/or Ca.

Neurocognitive Functions in Infants with Malnutrition; Relation with Long-chain Polyunsaturated Fatty Acids, Micronutrients Levels and Magnetic Resonance Spectroscopy

  • Cakir, Murat;Senyuva, Sukran;Kul, Sibel;Sag, Elif;Cansu, Ali;Yucesan, Fulya Balaban;Yaman, Serap Ozer;Orem, Asim
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.22 no.2
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    • pp.171-180
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    • 2019
  • Purpose: Malnutrition may influence neurocognitive development in children by directly affecting the brain structural development, or indirectly by affecting the children's cognition experience. Malnutrition alters the cell numbers, cell migration, synaptogenesis, and neurotransmission due to inadequate availability of necessary micronutrients to support cell growth. We aimed to analyze neurocognitive development in infants with malnutrition and its association with long chain polyunsaturated fatty acids (LC-PUFA), micronutrients levels and magnetic resonance spectroscopy (MRS) findings. Methods: The study included two groups; group 1, infants with malnutrition (n=24), group 2; healthy infants (n=21). Peripheral blood was obtained from the participants for studying micronutrients and LC-PUFA levels. The neurocognitive development was analyzed by the use of an Ankara Developmental Screening Inventory test. MRS were performed on all infants. Results: All parameters of neurocognitive development and serum calcium ($9.6{\pm}0.9mg/dL$ vs. $10.4{\pm}0.3mg/dL$, p<0.05) and magnesium ($2.02{\pm}0.27mg/dL$ vs. $2.2{\pm}0.14mg/dL$, p<0.05) levels were noted as being low in infants with marked malnutrition. No difference was found in LC-PUFA levels between healthy and malnourished infants. Thalamic choline/creatine levels were significantly high in infants with malnutrition ($1.33{\pm}0.22$ vs. $1.18{\pm}0.22$, p<0.05). Total neurocognitive development in infants was positively correlated with serum calcium levels (p<0.05, r=0.381). Conclusion: Calcium supplementation may improve neurocognitive development in malnourished infants.

Mass Production of Poly(3-Hydroxybutyrate) by Fed-Batch Cultures of Ralstonia eutropha with Nitrogen and Phosphate Limitation

  • Ryu, Hee-Wook;Cho, Kyung-Suk;Kim, Beom-Soo;Chang, Yong-Keun;Chang, Ho-Nam;Shim, Hyun-Joo
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.751-756
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    • 1999
  • For mass production of poly(3-hydroxybutyrate) (PHB), high cell density cultures of Ralstonia eutropha were carried out in 2.5-1 and 60-1 fermentors by two fed-batch culture techniques of nitrogen and phosphate limitation. When the nitrogen limitation technique was employed using both an on-line glucose monitoring and control system, a high concentration level of PHB (121g/l) was obtained in the small-scale fermentor of 2.5 1. However, the PHB concentration obtained in a large-scale fermentor of 60 1 only turned out to be 60g/l. In contrast, when another fed-batch culture technique of the phosphate-limitation employing dissolved oxygen (DO) stat glucose feeding was used, a large amount of PHB was successfully produced in both 60-1 and 2.5-1 fermentors. In a 2.5-1 fermentor, concentrations of PHB and cells obtained in 58 h were 175 and 210 g/l, respectively, which corresponded to the PHB productivity level of 3.02 g/l/h. In a 60-1 fermentor, a final cell concentration of 221 g/l and a PHB concentration of 180 g/l with PHB productivity level of 3.75 g/l/h were obtained in 48h. PHB content and yield from glucose were 81% and 0.38g PHB/g glucose, respectively. These data suggest that the phosphate limitation technique is more effective compared to nitrogen limitation in the mass production of PHB by R. eutropha of a large scale.

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Accumulation of inorganic arsenic, and growth rate by changing of phosphate concentration in Hizikia fusiforme (인산염 농도 변화에 따른 톳(Hizikia fusiforme)의 무기비소(As (V)) 축적 및 생장률 변동)

  • Hwang, Un-Ki;Choi, Hoon;Choi, Min-Kyu;Kim, Min-Seob;Choi, Jong-Woo;Heo, Seung;Lee, Ju-Wook
    • Korean Journal of Environmental Biology
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    • v.37 no.2
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    • pp.189-195
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    • 2019
  • In this study, we performed an analysis of the accumulation of inorganic arsenic and growth rate with changes in phosphate concentration in Hizikia fusiforme. When exposed to inorganic arsenic for fourteen days, we found that the collection of inorganic arsenic hardly increased at high phosphate concentrations (2 mg L-1). However, when the phosphate concentration was low (0.02 mg L-1), accumulation of inorganic arsenic increased. Additionally, H. fusiforme decreased in a growth rate of 14.5% in low phosphate concentration (0.02 mg L-1) and fell in a growth rate of 30% when exposed to inorganic arsenic (10 ㎍ L-1). H. fusiforme cannot distinguish between phosphate and inorganic arsenic. Thus, when phosphate concentration was lower, the inorganic arsenic accumulation increased, and accumulated inorganic arsenic inhibited photosynthesis and cell division, reducing the growth rate. H. fusiforme is known to have higher inorganic arsenic accumulation than other seaweeds. Therefore, various studies are needed to secure the food safety of H. fusiforme which is an essential aquaculture species in Korea.

Enzyme Inhibitory and Anti-Proliferation Effects of Peanut Skin Extracts Depending on Cultivar (품종별 땅콩 종피 추출물의 효소저해활성 및 암세포주 증식억제 효과)

  • Kim, Min Young;Kim, Hyun-Joo;Lee, Yu-Young;Kim, Mi Hyang;Lee, Jin Young;Lee, Byoungkyu;Lee, Byong Won
    • The Korean Journal of Food And Nutrition
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    • v.32 no.5
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    • pp.511-521
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    • 2019
  • The purpose of this study was to investigate the inhibitory effect of enzyme activity and anti-proliferation of human cancer cell lines (HCT 116, NCI-H460 and MCF-7) of peanut skin depending on cultivars (Arachis hypogaea L. cv. K-Ol, cv. Sinpalkwang, cv. Daan, cv. Heuksaeng) and extraction solvent. Peanut skin was extracted with 80% ethanol, 80% methanol, 80% acetone, and distilled water, followed by analysis of the enzyme inhibitory activity and anticancer activity. Methanol extract of Daan cultivar most effectively inhibited ${\alpha}$-gluosidase (65.08%, 0.025 mg/mL), tyrosinase (82.49%, 2 mg/mL) and ACE (73.61%, 10 mg/mL). The inhibitory effect of peanut skin extracts on colon cancer cell (HCT-116), lung cancer cell (NCI-H460) and breast cancer cell (MCF-7) growth were investigate using MTT assay. The highest anti-proliferation of cancer cell line of peanut skin extracts was observed in the methanol extract of Daan cultivar. The cell viability on HCT 116, NCI-H460 and MCF-7 cell lines of methanol extracts from peanut skin of Daan cultivar was 48.13%, 41.03%, and 36.02% at $200{\mu}g/mL$, respectively. These results suggest that peanut skin extracts may mediate physiological activity, and provide valuable information for the use of peanut byproduct as a functional food material.

GABA Productivity in Yoghurt Fermented by Freeze Dried Culture Preparations of Lactobacillus acidophilus RMK567 (Lactobacillus acidophilus RMK567의 동결건조 컬쳐로 제조한 요구르트에서 GABA 생성력)

  • Lim, Sang-Dong;Yoo, Sung-Ho;Yang, Hae-Dong;Kim, Sang-Ki;Park, Seung-Yong
    • Food Science of Animal Resources
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    • v.29 no.4
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    • pp.437-444
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    • 2009
  • ${\gamma}-Aminobutyric$ acid (GABA) producing lactic acid bacteria, Lactobacillus acidophilus RMK567 was cultivated in 50 L of sterilized MRS broth using a fermenter at $40^{\circ}C$ for 24 h. The cell number was increased to $10.04{\pm}0.13$ Log CFU/mL with a growth rate constant (k) of 0.454 generation/h and a generation time (g) of 2.303 h after a lapse of a lag phase (L) of 5.16 h. A total of 487 g of cell paste with 40.5% moisture was harvested with viable cell number of 12.48 Log CFU/g cell paste. The cell pastes after preparation with glycerol, glucose, and polydextrose as cryo-protectants were lyophilized under a vacuum of 84 m torr. A total of 408 g of freeze dried (FD) cell powders were mixed with a commercial strain of Streptococcus thermophilus to prepare of three types FD starter cultures with the viable cell numbers of 12.42 (FDA-GY), 12.60 (FDBGG) and 12.91 (FDC-GP) Log CFU/g. During preservation the FD cultures at -$18^{\circ}C$, the cell viability of the FD starter cultures were rapidly dropped to below 3.24% of the day of storage. No significant difference was found in the cell viabilities among three types of FD starters cultures, but significant difference (p<0.01) was found in storage periods. Yoghurts fermented through FD starter culture of L. acidophilus RMK567 were determined to contain $155.16{\pm}8.53$ ppm, $243.82{\pm}4.27$ ppm, and $198.64{\pm}23.46$ ppm of GABA, respectively. This study shows that GABA production activity of L. acidophilus RMK567 is not affected during the freeze drying process and would be available for commercial production of yoghurt containing high GABA content.

Inhibitory Effects of Ethanol Extract of Red Sweet Pepper (Capsicum annuum L.) on Triglyceride Biosynthesis in Rhodosporidium toruloides (Rhodosporidium toruloides에서의 적색 파프리카 에탄올 추출물의 중성지방 억제 효과)

  • Kang, Ju-Won;Lee, Seung-Je;Ahn, Byung-Yong
    • Korean Journal of Plant Resources
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    • v.34 no.2
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    • pp.186-196
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    • 2021
  • In this study, we investigated inhibitory effect of red sweet pepper (Capsicum annuum L.) on triglyceride biosynthesis in Rhodosporidium toruloides. There was no significant difference in the total lipid content of all the experimental groups including 0.02, 0.1 and 0.5% (w/v) of red sweet pepper 70% ethanol extract treatment (RSPE). However, the triglyceride content was significantly decreased in RSPE group campared to the control group. When the formation of lipid droplet in the oleaginous yeast was examined, a small amount of fluorescence was observed compared to the control as the concentration of RSPE increased. The number of cells and free fatty acid increased in a concentration-dependent manner. These results suggest that RSPE has an anti-obesity effect.