Targeting the cystine/glutamate exchange transporter, system xc-, is a promising anticancer strategy that induces ferroptosis, which is a distinct form of cell death mediated by iron-dependent lipid peroxidation. The concentration of ⳑ-cystine in culture medium is higher than the physiological level. This study was aimed to evaluate the effects of ⳑ-cystine concentration on the efficacy of ferroptosis inducers in hepatocellular carcinoma cells. This study showed that treatment with sulfasalazine or erastin, a system xc- inhibitor, decreased the viability of Huh6 and Huh7 cells in a dose-dependent manner, and the degree of growth inhibition was greater in medium containing a physiological ⳑ-cystine concentration of 83 µM than in commercial medium with a concentration of 200 µM ⳑ-cystine. However, RSL3, a glutathione peroxidase 4 inhibitor, decreased cell viability to a similar extent in media containing both ⳑ-cystine concentrations. Sulfasalazine and erastin significantly increased the percentages of propidium iodide-positive cells in media with 83 µM ⳑ-cystine, but not in media with 200 µM ⳑ-cystine. Sulfasalazine- or erastin-induced accumulation of lipid peroxidation as monitored by C11-BODIPY probe was higher in media with 83 µM ⳑ-cystine than in media with 200 µM ⳑ-cystine. In contrast, the changes in the percentages of propidium iodide-positive cells and lipid peroxidation by RSL3 were similar in both media. These results showed that sulfasalazine and erastin, but not RSL3, were efficacious under conditions of physiological ⳑ-cystine concentration, suggesting that medium conditions would be crucial for the design of a bioassay for system xc- inhibitors.
Journal of the Korean Society of Food Science and Nutrition
/
v.42
no.4
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pp.570-576
/
2013
The purpose of this study was to investigate the effect of turmeric on antioxidative systems and oxidative damage in rats fed a high fat and cholesterol diet. A total 40 rats were divided into four experimental groups: a normal diet group (N), a high fat and cholesterol diet group (HF), a high fat and cholesterol diet group supplemented with 2.5% turmeric powder (TPA group) and a high fat and cholesterol diet group supplemented with 5% turmeric powder (TPB group). The serum glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) activity of the turmeric supplemented groups were decreased compared to the HF group. The GPT activity of the TPB group was especially and significantly decreased compared to the HF group. Hepatic superoxide dismutase (SOD) of the TPB group was significantly increased compared to the HF group. However, there were no significant differences in the activities of hepatic glutathione peroxidase (GSHpx) and catalase (CAT) among all experimental groups. Hepatic glutathione S-transferase (GST) activity in the TPA and TPB groups were increased compared to the HF group. Hepatic superoxide radical content in mitochondria of the 5% turmeric supplemented group was significantly decreased compared to the HF group. Hepatic hydrogen peroxide content in the cytosol and mitochondria of the turmeric-supplemented groups were decreased compared to the HF group. Hepatic carbonyl values in the mitochondria of the turmeric supplemented groups were significantly decreased compared to the HF group. Thiobarbituric acid reaction substance (TBARS) values in the liver were significantly reduced in turmeric supplemented groups compared to the HF group. These result suggest that turmeric powder may reduce oxidative damage through the activation of antioxidative defense systems in rats fed high fat and cholesterol diets.
Kim, Kyung-Ran;Choi, Jeong-Hwa;Lee, Sung-Kwon;Woo, Mi-Hee;Choi, Sang-Won
Journal of the Korean Society of Food Science and Nutrition
/
v.35
no.10
/
pp.1356-1362
/
2006
The purpose of the present study was to investigate the effect of enzymatic hydrolysate (extract) of Salicomia herbacea L. (Korean name: Hamcho) on antioxidative defense system in rats fed high cholesterol diet. Rats were divided into six experimental groups which are composed of normal diet group, normal diet group supplemented with 2% Hamcho extract, high cholesterol diet group, high cholesterol diet groups supplemented with 1%, 2% and 4% Hamcho extracts. The activity of serum glutamate oxaloacetae transaminase in rats was not different among all experimental groups, while the activity of glutamate pyruvate transaminase in groups supplemented with Hamcho extract was significantly lower than that of high cholesterol control group. Supplementation of Hamcho extract (SHE) to the high cholesterol fed rats resulted in increased activities of hepatic superoxide dismutase and glutathione peioxidase. However, there was no significant difference in the activity of hepatic catalase among all experimental groups. SHE also resulted in decreased levels of hepatic thiobarbituric acid reactive substances and mitochondrial carbonyl values. Those effects were higher to some extent in 2% and 4% Hamcho extract groups than those of high cholesterol control group. These results suggest that enzymatic hydrolysate of Hamcho may reduce oxidative damage by activation of antioxidative defense system in rats fed high cholesterol diets.
In order to invertigate penicillin acylase produced by a strain of Genus Escherichia, 47 strains of Genus Escherichia were isolated and examined to the extend of the inactivation of penicillins and the ability of 6-amino penicillanic acid (6-APA) production. Among them, 12 strains were found to produce penicillin acylase and to form 6-APA. The strain No. 11 of the isolates was selected to be the most excellent one producing the acylase. Optimum culture conditions for the production of the acylase of the strain were found as follows : pH at 7.6~8.0, time on 18 hrs, temperature at 34 to 38$^{\circ}C$. And effective levels of the medium were found to be contained 0.3% phenylacetic acid, 1.0% yeast extract, 1.0% peptone and 0.3% L-glutamate. And, the production of the acylase by the isolate was strongly inhibited by 1% glycerol and the growth was remarkably retarded by the addition of 1.0% n-butylacetate. The acylase was extracted from the isolate and the crude enzyme of the acylase was prepared and the characteristies of the enzyme were primary examined in optimum pH, temperature, stabilities and reaction time.
A green sulfur bacterium, Chlorobium limicola f. thiosulfatophilum NCIB 8327, was grown in modified Pfennig's medium including glu1amate as a nitrogen source. Glutamine synthetase was isolated through a series of ultracentrifugation. DEAE-Sepharose CL-6B ion exchange chromatography. Sephacryl S-300 gel permeation chromatography, and preparative HPLC. The recovery and purification fold of the enzyme were 2% and 46.3. respectively. The isolated enzyme was homogeneous on UV-Visible spectrum and polyacrylamide gel electrophoretogram. The relative molecular mass of the native enzyme was estimated to be 280,000 by gel permeation chromatography. The enzyme consisted of ten subunits with relative similar molecular mass. 30.000. which was estimated by SDS-polyacrylamide gel electrophoresis. The optimal temperature and pH of the enzyme were $30^{\circ}C$ and 7.0. Km values were 27.9 mM for L-glutamine and 0.92 mM for hydroxylamine-HCr. The enzyme activity was inhibited by alanine. glycine. and tryptophan considerably, but was not affected by asparagine, lysine. leucine. and valine.
Purpose : The purpose of this study was to evaluate the effect of amino acid mixtures on incidence and severity of total parenteral nutrition associated-cholestasis(PNAC) in very low birth weight infants. Methods : Retrospective review of 63 very low birth weight infants(birth weight ${\leq}1,500g$) who received total parenteral nutrition(TPN) in our neonatal intensive care unit from January 2000 to December 2004 was performed. Patients were divided into 2 groups : Group I(n=32, Jan 2000-Jun 2002) and Group II(n=31, Jul 2002-Dec 2004), where infants in Group II received taurine and glutamic acid-rich amino acid mixtures. PNAC was defined as serum direct bilirubin(DB) level greater than 2.0 mg/dL. The incidence and severity of PNAC were compared between these groups. Results : The incidence of PNAC was significantly lower in Group II than in Group I(21.9% vs 6.5%, P<0.148). Maximum and mean DB levels were also significantly lower in Group II(P<0.05). Conclusion : The incidence and severity of PNAC in very low birth weight infants may be reduced with different composition of amino acid mixtures in TPN. Further prospective randomized controlled studies are needed to determine an ideal composition of acid mixtures to prevent the development of PNAC.
LEE Eung-Ho;PARK Hyang-Suk;OH Kwang-Soo;CHA Yong-Jun
Korean Journal of Fisheries and Aquatic Sciences
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v.18
no.4
/
pp.316-324
/
1985
Vacuum-packed and seasoned smoked-dried products of red squid, Ommastrephes bartrami, caught in the Northern Pacific Ocean, were prepared and stored at room temperature for 90 days to test their keeping quality. Defrosted squids were eviscerated, skinned, and cut. The mantle meats were flavored with seasoning powders prepared from sugar, sorbitol, salt, monosodium glutamate, or smoke flavor (Smoke-EZ, Alpha Foods Co., Ltd.). After seasoning, the mantle meats were dried at $45^{\circ}C$ for 7 hours, vacuum packed in plastic film bags, and pasteurized in water at $95^{\circ}C$ for 30 minutes. Three kinds of products were prepared : control products (seasoned-dried), solid smoked seasoned-dried and liquid smoked seasoned-dried. The moisture level, water activity, color value (L, a and b value), texture, and viable cell counts of bacteria in these products were determined during storage at room temperature, $5^{\circ}C\;and\;35^{\circ}C$, respectively. The results showed that the products could be preserved at good condition for 90 days though they developed pale brown color during storage. The contents of free amino acids, nucleotides and their related compounds, and the compositions of fatty acids of raw squid and smoked products were analysed. In the amino acids, arginine, taurine, glycine and proline were abundant in raw and smoked products. The contents of hypoxanthine of raw and smoked products were higher than the other nucleotides and their related compounds. In fatty acid compositions of raw and smoked products, the dominant fatty acids were docosahexaenoic acid (22:6), hexadecanoic acid(16:0) and eicosapentaenoic acid (22:5).
Hwang, Chung Eun;Kim, Su Cheol;Lee, Jin Hwan;Hong, Su Young;Cho, Kye Man
Journal of Applied Biological Chemistry
/
v.61
no.3
/
pp.245-255
/
2018
The research was aimed to analyze the functional constituents (GABA and isoflavone), radical (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and hydroxyl) scavenging activities and enzyme (${\alpha}-glucosidase$ and lipase) inhibitory effects of soypowder milk (SPM) and fermented soy-powder milk (FSPM) with varied Lactobacillus brevis. Ten ${\gamma}-aminobutyric$ acid (GABA) producing lactic acid bacteria that showed 96-99% similarity with L. brevis, according to 16S rRNA gene sequence analysis, were isolated from fermented kimchi. The conversion rates of GABA were obtained 66.96-93.51, 63.76-84.58, and 57.05-69.75% in monosodium glutamate, gluten and soy protein, respectively. The levels of pH and glutamic acid of FSPM were found lower than those of SPM, but the acidity and GABA contents were higher. The GABA conversion rate of FSPM with BMK484 strain was attained the highest 69.97%. The contents of isoflavone glycoside ($1290.93{\mu}g/g$) was higher in SPM, but the content of isoflavone aglycone ($287.27-501.9{\mu}g/g$) was higher in FSPM. The levels of isoflavone aglycone such as daidzein, glycitein and genistein, were found as the highest 240.2, 61.24 and $200.45{\mu}g/g$, respectively, when FSPM was made with BMK484 strain. The DPPH, ABTS and hydroxyl radical scavenging and ${\alpha}-glucosidase$ and pancreatic lipase inhibitory activities of FSPM made with BMK484 strain were the relatively higher 60.31, 88.10, 61.25, 52.71, and 39.37%, respectively. Therefore, the L. brevis can be used as a material capable of simultaneously enhanced GABA and isoflavone aglycone in FSPM.
LEE Eung-Ho;OHSHIMA Toshiaki;WADA Shun;KOIZUMI Chiaki
Korean Journal of Fisheries and Aquatic Sciences
/
v.15
no.1
/
pp.99-106
/
1982
Vacuum-packed and seasoned-dried products of filefish, Navodon modestus, caught in large quantities in the coasts of Japan and Korea, were prepared tentatively and stored at $35^{\circ}C$ for $3\sim4$ months to test their keeping quality. Headed, eviscerated, ana skinned filefish were purchased from Tokyo central wholesale market and filleted. The fillets were seasoned with the seasoning powder prepared from sugar, sorbitol, salt, glutamate, 5'-ribonucleotide, and either beef flavor (Flava-Beef, Kyowa Hakko Kogyo Co. Ltd.), red pepper, or smoke flavor (Smok-EZ, Alpha Foods Co. Ltd.). After seasoning, the fillets were dried at $39\sim41^{\circ}C$ for several hours, vacuum-packed in the bag of plastic film, and cooked in water of $80^{\circ}C$ for 40 min. Two kinds of products different in moisture level were prepared : one containing moisture of $26\sim29\%$ and the other $40\sim46\%$. The moisture level, water activity, color value (L,a, and b values), texture, and viable counts of bacteria of these products were determined during storage at $35^{\circ}C$. From the results obtained, it became clear that the products could be preserved in good condition for $3\sim4$ months at $35^{\circ}C$, though they slightly decreased in the softness with decrease of moisture content and developed a pale brown during storage. Judging from organoleptic evaluation on the flavor, the products containing beef flavor as a seasoning was most desirable.
LEE Eung-Ho;HA Jae-Ho;CHA Yong-Jun;OH Kwang-Soo;KWON Chil-Sung
Korean Journal of Fisheries and Aquatic Sciences
/
v.17
no.4
/
pp.299-305
/
1984
As one trials to utilize sea mussel and anchovy effectively, powdered instant soups were prepared and then their quality stability were examined during storage. Powdered instant soup was made by adding $3\%$ sugar, $20\%$ table salt, $5\%$ monosodium glutamate, $0.2\%$ black pepper and garlic powder to the pulverized dried sea mussel or anchovy. Powdered instant soup products, powderd products, and dried round state sea mussel or anchovy were packed with air in laminated film bag (cellophane/polyester/aluminium foil/polyester: $20{\mu}m/15{\mu}m/7{\mu}m/20{\mu}m,\;13{\times}14cm$). The contents of amino-nitrogen and volatile basic nitrogen of these products were showed little significant variations and also water activity and color value (L, a, b) of these products were little changed during storage. Thiobarbituric acid value increased up to 30 days of storage and then decreased slightly. Comparing the quality of powdered-seasoned products with that of dried round state products, there were no significant differences in stability during storage. Judging from the experimental results, the quality of powdered instant soup of sea mussel and anchovy were stable for 100 days at room temperature($25{\pm}3^{\circ}C$).
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