1.Propionic acid fermentation by Propionibacterium freudenreichii CCTCC M207015 with a fibrous-bed bioreactor.
Xiaohai FENG ; Bo WU ; Xiaobo SHEN ; Hong XU
Chinese Journal of Biotechnology 2008;24(6):1075-1079
The production of propionic acid by Propionibacterium freudenreichii CCTCC M207015 was investigated in a Fibrous-bed bioreactor (FBB). The FBB was constructed by packing spiral cotton fibrous and immobilized into a bioreactor. By applying this bioreactor to propionic acid fermentation, the propionic acid yield had a significant improvement and reached 20.41 g/L, compared with the cell-free culture of 14.58 g/L (40 g/L of glucose). At the same time, the glucose exhausting time decreased from 120 h to 60 h. Batch fermentations at various glucose concentrations were carried out with FBB. Based on the analysis of the time course of production, fed-batch fermentation was also applied to produce propionic acid with FBB, the maximal propionic acid yield reached 45.91 g/L, and the proportion of propionic acid to total acids was about 72.31%.
Bioreactors
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microbiology
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Fermentation
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Glucose
;
metabolism
;
Propionates
;
metabolism
;
Propionibacterium
;
classification
;
metabolism
2.Process development for continuous ethanol fermentation by the flocculating yeast under stillage backset conditions.
Lihan ZI ; Chenguang LIU ; Fengwu BAI
Chinese Journal of Biotechnology 2014;30(2):310-314
Propionic acid, a major inhibitor to yeast cells, was accumulated during continuous ethanol fermentation from corn meal hydrolysate by the flocculating yeast under stillage backset conditions. Based on its inhibition mechanism in yeast cells, strategies were developed for alleviating this effect. Firstly, high temperature processes such as medium sterilization generated more propionic acid, which should be avoided. Propionic acid was reduced significantly during ethanol fermentation without medium sterilization, and concentrations of biomass and ethanol increased by 59.3% and 7.4%, respectively. Secondly, the running time of stillage backset should be controlled so that propionic acid accumulated would be lower than its half inhibition concentration IC50 (40 mmol/L). Finally, because low pH augmented propionic acid inhibition in yeast cells, a higher pH of 5.5 was validated to be suitable for ethanol fermentation under the stillage backset condition.
Biomass
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Ethanol
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metabolism
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Fermentation
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Flocculation
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Propionates
;
chemistry
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Yeasts
;
metabolism
3.Formation of polyhydroxyalkanoates during the dual-nutrient-limited zone by Ralstonia eutropha.
Qun YAN ; Guo-Cheng DU ; Jian CHEN
Chinese Journal of Biotechnology 2003;19(4):497-501
Polyhydroxyalkanoates (PHAs) are a class of polyesters biosynthesized by microorganisms (esp. Ralstonia eutropha) under an unbalanced growth condition, and which are supposed to partly take the place of traditional plastics made from petroleum in the near future since they are harmless to the environment and biodegradable. Organic acids (mainly butyrate, lactate, propionate and acetate) produced from anaerobic digested food wastes, industrial wastes and sewage may be used as cheap carbon sources since the large amounts of the above wastes disposed by industry and family each year. In order to better understand the process of PHAs formation with acids as carbon sources, so as to increase the yields of PHAs. Biosynthesis of PHAs by R. eutropha during the dual nutrient-limitation-zone was investigated with mixed organic acids (the mass ratio of the four component acids was butyrate: propionate: acetate: lactate = 3: 3: 1: 1, which was simulated as once the result of anaerobic digestion of food wastes) as carbon sources and (NH4)2 SO4 as nitrogen source. Two different manners of maintaining the dual-nutrient-limitation zone were adopted by feeding mixed acids and (NH4 )2SO4 at determined rates to the fermentation culture which were free of carbon sources (manner A) or nitrogen sources (manner B) firstly. The results suggest that, first of all, the meaning of the limitation of mixed acids or (NH4)2 SO4 does not mean to limit the supply of them, but mean to feed as more as possible of carbon and nitrogen sources in order to meet the cell growth and PHAs formation of R. eutropha by the largest extent. However, it's indispensable to make the residual concentration of carbon and nitrogen sources as low as possible since organic acids are inhibitive to the cell growth, and most importantly, only under the presence of nitrogen during the PHAS formation period of the fermentation could R. eutropha produce more PHAs than any other unbalanced growth condition. Secondly, with the increase of the width of the dual-nutrient-limitation zone, the yield of PHAs would also increase, it suggest that most of the PHAs were biosynthesized during the dual-nutrient-limitation zone. Finally, in contrast with the dual-nutrient-limitation manner of limiting the nitrogen source at first (manner B), the dual-nutrient-limitation manner of limiting the carbon source at first (manner A) was more favorable for the production of PHAs, and the maximum production of PHAs of these two manners are 3.72 g/L and 2.55 g/L, respectively. It may be because that PHAs formation required enzymes could not be well developed when R. eutropha grow under the state of nitrogen limitation from the beginning of fermentation. Besides, yield of PHAs produced by the dual-nutrient-limitation fermentation is larger than that of the single-nutrient-limitation batch culture. Therefore, it seems that to increase the output of PHAs production, the strategy of maintaining as wide as possible the width of dual-nutrient (C, N)-limitation zone may be effective.
Acetates
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metabolism
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Butyrates
;
metabolism
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Cupriavidus necator
;
metabolism
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Fermentation
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physiology
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Lactates
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metabolism
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Polyhydroxyalkanoates
;
biosynthesis
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Propionates
;
metabolism
4.Cutinase production from short-chain organic acids by Thermobifida fusca.
Gangqiang HE ; Guocheng DU ; Liming LIU ; He LIU ; Guanghua HUO ; Jian CHEN
Chinese Journal of Biotechnology 2008;24(5):821-828
We studied cutinase production from short-chain organic acids by Thermobifida fusca WSH03-11 to evaluate the possibility of converting municipal sludge to high value-added products. The optimum organic acid (8.0 g/L) and nitrogen source (1.5 g/L) concentrations were determined by the single factor experiments with butyric acid, propionic acid and acetic acid as the carbon sources. When lactic acid was used as the carbon source, the optimum organic acid (3.0 g/L) and nitrogen source (1.0 g/L) concentrations were obtained. Cutinase production by T. fusca WSH03-11 was further improved with butyric acid (by 31.0%), propionic acid (by 13.3%), acetic acid (by 43.8%) and lactic acid (by 73.2%) as carbon source, respectively, with the optimized cutin concentrations. Among these four short-chain organic acids, the average specific consumption rate of acetic acid was the highest, higher than that of propionic acid 1.3-folds, butyric acid 2.0-folds and lactic acid 2.2-folds. The highest cutinase activity reached 52.4 u/mL with butyric acid (8 g/L) as the sole carbon source, higher than that of lactic acid (3 g/L) 1.7-folds, acetic acid (8 g/L) 2.5-folds and propionic acid (8 g/L) 3.2-folds. The yield of cutinase activity on lactic acid (12.70 u/mg) higher than that of butyric acid 1.4-folds, propionic acid 3.0-folds and acetic acid 3.8-folds. T. fusca WSH03-11 consumed acetic acid firstly in mixed acids carbon sources, and the consumption of butyric acid was inhibited. Further studies indicated that the consumption rate of butyrate was decreased by 66.7% in the presence of 0.5 g/L acetic acid in the mixed acids. This was the first report concerning the production of cutinase by T. fusca with mixed organic acids as the carbon sources. The results presented here provided a novel and efficient approach to produce high value-add products from municipal sludge, and also established a foundation for the industrial production of cutinase by T. fusca WSH03-11 with cheap carbon sources from the processing of municipal sludge.
Acetates
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metabolism
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Actinomycetales
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growth & development
;
metabolism
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Butyric Acid
;
metabolism
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Carboxylic Ester Hydrolases
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biosynthesis
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Fermentation
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Organic Chemicals
;
metabolism
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Propionates
;
metabolism
5.Enzyme production mechanism of anaerobic fungus Orpinomyces sp. YF3 in yak rumen induced by different carbon source.
Xue'er DU ; Linlin ZHOU ; Fan ZHANG ; Yong LI ; Congcong ZHAO ; Lamei WANG ; Junhu YAO ; Yangchun CAO
Chinese Journal of Biotechnology 2023;39(12):4927-4938
In order to investigate the enzyme production mechanism of yak rumen-derived anaerobic fungus Orpinomyces sp. YF3 under the induction of different carbon sources, anaerobic culture tubes were used for in vitro fermentation. 8 g/L of glucose (Glu), filter paper (Flp) and avicel (Avi) were respectively added to 10 mL of basic culture medium as the sole carbon source. The activity of fiber-degrading enzyme and the concentration of volatile fatty acid in the fermentation liquid were detected, and the enzyme producing mechanism of Orpinomyces sp. YF3 was explored by transcriptomics. It was found that, in glucose-induced fermentation solution, the activities of carboxymethyl cellulase, microcrystalline cellulase, filter paper enzyme, xylanase and the proportion of acetate were significantly increased (P < 0.05), the proportion of propionate, butyrate, isobutyrate were significantly decreased (P < 0.05). The results of transcriptome analysis showed that there were 5 949 differentially expressed genes (DEGs) between the Glu group and the Flp group, 10 970 DEGs between the Glu group and the Avi group, and 6 057 DEGs between the Flp group and the Avi group. It was found that the DEGs associated with fiber degrading enzymes were significantly up-regulated in the Glu group. Gene ontology (GO) function enrichment analysis identified that DEGs were mainly associated with the xylan catabolic process, hemicellulose metabolic process, β-glucan metabolic process, cellulase activity, endo-1,4-β-xylanase activity, cell wall polysaccharide metabolic process, carbohydrate catabolic process, glucan catabolic process and carbohydrate metabolic process. Moreover, the differentially expressed pathways associated with fiber degrading enzymes enriched by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were mainly starch and sucrose metabolic pathways and other glycan degradation pathways. In conclusion, Orpinomyces sp. YF3 with glucose as carbon source substrate significantly increased the activity of cellulose degrading enzyme and the proportion of acetate, decreased the proportion of propionate, butyrate and isobutyrate. Furthermore, the degradation ability and energy utilization efficiency of fungus in the presence of glucose were improved by means of regulating the expression of cellulose degrading enzyme gene and participating in starch and sucrose metabolism pathway, and other glycan degradation pathways, which provides a theoretical basis for the application of Orpinomyces sp. YF3 in practical production and facilitates the application of Orpinomyces sp. YF3 in the future.
Animals
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Cattle
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Neocallimastigales/metabolism*
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Anaerobiosis
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Rumen/microbiology*
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Propionates/metabolism*
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Isobutyrates/metabolism*
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Cellulose/metabolism*
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Fungi
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Starch/metabolism*
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Glucose/metabolism*
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Acetates
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Sucrose/metabolism*
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Cellulases
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Cellulase
6.Potential use of the herbicide quizalofop-p-ethyl for eicosapentaenoic acid overproduction by the diatom Nitzschia laevis.
Xiao-Hong CAO ; Song-Yao LI ; Chun-Ling WANG ; Mei-Fang LU
Chinese Journal of Biotechnology 2007;23(5):885-890
The diatom Nitzschia laevis is a good alternative source of eicosapentaenoic acid (EPA). Besides strategies for high cell density culture, EPA productivity may be further improved by herbicides. The effect of the herbicide quizalofop-p-ethyl on the growth and EPA production was studied in this paper. As the solvent of the herbicide, DMSO was proved to inhibit the growth and EPA production of N. laevis. The concentration of DMSO in the medium should not exceed 0.2%. Quizalofop-p-ethyl could cause morphology damage to the N. laevis cells. With the increasing concentration of quizalofop-p-ethyl from 0 mmol/L to 0.4 mmol/L, the dry cell weight production decreased, while at the same time, the lipid content of the dry cell mass increased. When treated with 0.1 mmol/L quizalofop-p-ethyl, the EPA content increased from 3.00% to 3.58% (of dry cell weight, DW), and the proportion of EPA (20:5) in total fatty acids (TFA) increased from 25.15% to 32.88% . These results indicated that the herbicide quizalofop-p-ethyl could stimulate the accumulation of EPA; therefore it might be useful for selecting algae colonies that overproduce EPA.
Culture Media
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Culture Techniques
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Diatoms
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growth & development
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metabolism
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Eicosapentaenoic Acid
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biosynthesis
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Herbicides
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pharmacology
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Propionates
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pharmacology
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Quinoxalines
;
pharmacology
7.Effect of nitrogen, phosphorus and potassium deficiency on content of phenolic compounds in exudation of American ginseng.
Du JING ; Jiaxue YANG ; Xiaolin JIAO ; Weiwei GAO
China Journal of Chinese Materia Medica 2011;36(3):326-329
OBJECTIVESome of the phenolic compounds detected in the soil of commercially cultivated American ginseng could inhibit the seed germination and seedling growth of American ginseng. In this paper we studied the root exudation of American ginseng induced by deficiency of nitrogen, phosphorus and potassium on the content of phenolic compounds.
METHODTwo years old American ginsengs were cultured in hydroponic culture with different nutrient solution. The culture solution was collected after 14 days. The exudations of different polarities in the culture solution were enriched by the amberlite XAD4 and XAD7. The content of the total phenolic acids in the exudation was analyzed by Folin-Ciocalteu colorimetry; the contents of vanillic acid, p-coumaric acid and trans-cinnamic acid were detected and quantified by HPLC.
RESULTBoth in the situation of nitrogen and potassium deficiency, the concentration of total phenolic compounds increased significantly in the exudation of American ginseng comparing with the complete nutrient solution (P < 0.05) , while decreased significantly under phosphorus deficient conditions (P < 0.05). The contents of the 3 autotoxic phenolic acids decreased significantly under nitrogen, phosphorus and potassium deficient conditions (P < 0.05).
CONCLUSIONThe contents of total phenolic compounds and the 3 autotoxic phenolics in the root exudation of American ginseng altered variously in the deficiency of nitrogen, phosphorus and potassium.
Coumaric Acids ; analysis ; Hydroxybenzoates ; analysis ; Nitrogen ; deficiency ; Panax ; chemistry ; metabolism ; Phosphorus ; deficiency ; Plant Exudates ; chemistry ; Plant Roots ; chemistry ; metabolism ; Potassium ; chemistry ; Propionates ; Soil ; chemistry ; Vanillic Acid ; analysis
8.pH-dependent modulation of intracellular free magnesium ions with ionselective electrodes in papillary muscle of guinea pig.
Shang Jin KIM ; In Gook CHO ; Hyung Sub KANG ; Jin Shang KIM
Journal of Veterinary Science 2006;7(1):31-36
A change in pH can alter the intracellular concentration of electrolytes such as intracellular Ca2+ and Na+ ([Na+]i) that are important for the cardiac function. For the determination of the role of pH in the cardiac magnesium homeostasis, the intracellular Mg2+ concentration ([Mg2+]i), membrane potential and contraction in the papillary muscle of guinea pigs using ion-selective electrodes changing extracellular pH ([pH]o) or intracellular pH ([pH]i) were measured in this study. A high CO2-induced low [pH]o causes a significant increase in the [Mg2+]i and [Na+]i, which was accompanied by a decrease in the membrane potential and twitch force. The high [pH]o had the opposite effect. These effects were reversible in both the beating and quiescent muscles. The low [pH]o-induced increase in [Mg2+]i occurred in the absence of [Mg2+]o. The [Mg2+]i was increased by the low [pH]i induced by propionate. The [Mg2+]i was increased by the low [pH]i induced by NH4Cl-prepulse and decreased by the recovery of [pH]i induced by the removal of NH4Cl. These results suggest that the pH can modulate [Mg2+]i with a reverse relationship in heart, probably by affecting the intracellular Mg2+ homeostasis, but not by Mg2+ transport across the sarcolemma.
Animals
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Cations, Divalent
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Female
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Guinea Pigs
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Heart Ventricles/metabolism
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Hydrogen-Ion Concentration
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Ion Transport/physiology
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Ion-Selective Electrodes/veterinary
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Magnesium/*metabolism
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Male
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Membrane Potentials/physiology
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Papillary Muscles/*metabolism
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Propionates/pharmacology
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Sodium/*metabolism
9.Analysis of volatile fatty acids in gingival crevicular fluid of patients with chronic periodontitis.
Qi-qiang LI ; Huan-xin MENG ; Xue-jun GAO ; Zu-hua WANG
Chinese Journal of Stomatology 2005;40(3):208-210
OBJECTIVETo investigate the volatile fatty acids in gingival crevicular fluid (GCF) and to analyze the relationship between the levels of the volatile fatty acids and chronic periodontitis.
METHODSGCF samples taken from 37 patients with chronic periodontitis and 16 volunteers with healthy periodontal status were analyzed by capillary electrophoresis.
RESULTSThe detection frequencies and concentrations of succinic acid, butyric acid and valeric acid were significantly higher in GCF of chronic periodontitis than in that of healthy group. The detection frequencies of propionic acid had no statistic difference between the two groups, but the concentrations of it was significantly higher in inflammation group. We also found that the concentrations of succinic acid, propionic acid and butyric acid were significantly lower in shallow pockets than that in deep pockets.
CONCLUSIONSThe volatile fatty acids, especially succinic acid, propionic acid, butyric acid and valeric acid were associated significantly with the severity and inflammation of periodontal disease. The levels of succinic acid, propionic acid and butyric acid in GCF were related to pocket depth.
Adult ; Aged ; Butyrates ; analysis ; Case-Control Studies ; Chronic Periodontitis ; metabolism ; Electrophoresis, Capillary ; Fatty Acids, Volatile ; analysis ; Female ; Gingival Crevicular Fluid ; metabolism ; Humans ; Male ; Middle Aged ; Periodontium ; metabolism ; Propionates ; analysis ; Succinic Acid ; analysis
10.The Toxicity of Nonsteroidal Anti-inflammatory Eye Drops against Human Corneal Epithelial Cells in Vitro.
Jong Soo LEE ; Young Hi KIM ; Young Min PARK
Journal of Korean Medical Science 2015;30(12):1856-1864
This study investigated the toxicity of commercial non-steroid anti-inflammatory drug (NSAID) eye solutions against corneal epithelial cells in vitro. The biologic effects of 1/100-, 1/50-, and 1/10-diluted bromfenac sodium, pranoprofen, diclofenac sodium, and the fluorometholone on corneal epithelial cells were evaluated after 1-, 4-, 12-, and 24-hr of exposure compared to corneal epithelial cell treated with balanced salt solution as control. Cellular metabolic activity, cellular damage, and morphology were assessed. Corneal epithelial cell migration was quantified by the scratch-wound assay. Compared to bromfenac and pranoprofen, the cellular metabolic activity of diclofenac and fluorometholone significantly decreased after 12-hr exposure, which was maintained for 24-hr compared to control. Especially, at 1/10-diluted eye solution for 24-hr exposure, the LDH titers of fluorometholone and diclofenac sodium markedly increased more than those of bromfenac and pranoprofen. In diclofenac sodium, the Na+ concentration was lower and amount of preservatives was higher than other NSAIDs eye solutions tested. However, the K+ and Cl- concentration, pH, and osmolarity were similar for all NSAIDs eye solutions. Bromfenac and pranoprofen significantly promoted cell migration, and restored wound gap after 48-hr exposure, compared with that of diclofenac or fluorometholone. At 1/50-diluted eye solution for 48-hr exposure, the corneal epithelial cellular morphology of diclofenac and fluorometholone induced more damage than that of bromfenac or pranoprofen. Overall, the corneal epithelial cells in bromfenac and pranoprofen NSAID eye solutions are less damaged compared to those in diclofenac, included fluorometholone as steroid eye solution.
Anti-Inflammatory Agents, Non-Steroidal/administration & dosage/*toxicity
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Benzophenones/administration & dosage/toxicity
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Benzopyrans/administration & dosage/toxicity
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Bromobenzenes/administration & dosage/toxicity
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Cell Movement/drug effects
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Cells, Cultured
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Diclofenac/administration & dosage/toxicity
;
Epithelial Cells/drug effects/metabolism/ultrastructure
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Epithelium, Corneal/cytology/*drug effects/metabolism
;
Fluorometholone/administration & dosage/toxicity
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Humans
;
L-Lactate Dehydrogenase/metabolism
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Microscopy, Electron, Transmission
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Ophthalmic Solutions
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Propionates/administration & dosage/toxicity