• Animal Bioscience
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• v.36 no.11
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• pp.1693-1699
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• 2023

Objective: Cows that are nursing get around 80% of their glucose from liver gluconeogenesis. Propionate, a significant precursor of liver gluconeogenesis, can regulate the key genes involved in hepatic gluconeogenesis expression, but its precise effects on the activity of enzymes have not yet been fully elucidated. Therefore, the aim of this study was to investigate the effects of propionate on the activity, gene expression, and protein abundance of the key enzymes involved in the gluconeogenesis of dairy cow hepatocytes. Methods: The hepatocytes were cultured and treated with various concentrations of sodium propionate (0, 1.25, 2.50, 3.75, and 5.00 mM) for 12 h. Glucose content in the culture media was determined by an enzymatic coloring method. The activities of gluconeogenesis related enzymes were determined by enzyme linked immunosorbent assay kits, and the levels of gene expression and protein abundance of the enzymes were detected by real-time quantitative polymerase chain reaction and Western blot, respectively. Results: Propionate supplementation considerably increased the amount of glucose in the culture medium compared to the control (p<0.05); while there was no discernible difference among the various treatment concentrations (p>0.05). The activities of cytoplasmic phosphoenolpyruvate carboxylase (PEPCK1), mitochondrial phosphoenolpyruvate carboxylase (PEPCK2), pyruvate carboxylase (PC), and glucose-6-phosphatase (G6PC) were increased with the addition of 2.50 and 3.75 mM propionate; the gene expressions and protein abundances of PEPCK1, PEPCK2, PC, and G6PC were increased by 3.75 mM propionate addition. Conclusion: Propionate encouraged glucose synthesis in bovine hepatocytes, and 3.75 mM propionate directly increased the activities, gene expressions and protein abundances of PC, PEPCK1, PEPCK2, and G6PC in bovine hepatocytes, providing a theoretical basis of propionate-regulating gluconeogenesis in bovine hepatocytes.

• Food Science and Preservation
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• v.30 no.6
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• pp.929-943
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• 2023

This study analyzed the variation in vitamin K content in conger eel (Conger myriaster) caught off Tongyeong, Gyeongsangnam-do, Korea, focusing on the influence of size (large and small) and harvest period (monthly throughout 2021). We applied enzymatic extraction and HPLC-fluorescence methods for the analysis of vitamin K₁ (phylloquinone) and K₂ (menaquinone). The vitamin K content in conger eel varied significantly with size and harvest season (p<0.05). In large-sized samples, the phylloquinone content peaked in July (0.80±0.09 ㎍/100 g), while the highest menaquinone content was in May (0.79±0.11 ㎍/100 g). Conversely, in small-sized conger eels, the highest phylloquinone was found in December (1.94±0.15 ㎍/100 g), and the peak menaquinone level was in January (0.66±0.02 ㎍/100 g). The fat content was highest in July for large samples and in January for smaller ones. There was a positive correlation between fat and total vitamin K contents in conger eel (r=0.631, 0.667). Method validation and quality control measures ensured data reliability for vitamin K₁ and K₂ analyses. This study provides reliable information on the size and seasonal variations of vitamin K in conger eels, a staple in the Korean diet. This information is valuable for inclusion in Korea's national food nutrition database and for formulating future national health and nutrition policies.

• Journal of Periodontal and Implant Science
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• v.53 no.5
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• pp.321-335
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• 2023

Purpose: The aim of this study was to investigate the efficacy of photo-crosslinked gelatin methacryloyl (GelMa) hydrogel containing calcium phosphate nanoparticles (CNp) when applying different fabrication methods for bone regeneration. Methods: Four circular defects were created in the calvaria of 10 rabbits. Each defect was randomly allocated to the following study groups: 1) the sham control group, 2) the GelMa group (defect filled with crosslinked GelMa hydrogel), 3) the CNp-GelMa group (GelMa hydrogel crosslinked with nanoparticles), and 4) the CNp+GelMa group (crosslinked GelMa loaded with nanoparticles). At 2, 4, and 8 weeks, samples were harvested, and histological and micro-computed tomography analyses were performed. Results: Histomorphometric analysis showed that the CNp-GelMa and CNp+GelMa groups at 2 weeks had significantly greater total augmented areas than the control group (P<0.05). The greatest new bone area was observed in the CNp-GelMa group, but without statistical significance (P>0.05). Crosslinked GelMa hydrogel with nanoparticles exhibited good biocompatibility with a minimal inflammatory reaction. Conclusions: There was no difference in the efficacy of bone regeneration according to the synthesized method of photo-crosslinked GelMa hydrogel with nanoparticles. However, these materials could remain within a bone defect up to 2 weeks and showed good biocompatibility with little inflammatory response. Further improvement in mechanical properties and resistance to enzymatic degradation would be needed for the clinical application.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1265-1265
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• 2001

The organic materials included in excreta of livestock are important resources for organic manure and for improving soil quality, although there is still far from effective using. One reason for this is still unclearly standard of quality for evaluation of manure made from excreta of livestock. Therefore, the objective of this study is to develop rapid and accurate analytical method for analyzing organic compositions of manure made from excreta of livestock, and to establish quality evaluation method based on the compositions predicted by near infrared reflectance spectroscopy (NIRS). Sixteen samples of manure, each eight samples prepared from two treatments, were used in this study. The manure samples were prepared by mixing 560 kg feces of swine,60 kg sawdust with moisture content was adjusted to be 65%. The mixture was then keep under two kinds of shelter, black and clear sheets, as a treatment on the effect of sunlight. Samples were taken in every week (form week-0 to 7) during the process of manure making. Samples were analyzed to determine neutral detergent fiber (NDF), acid detergent fiber (ADF) and acid detergent lignin (ADL) by detergent methods, and organic cell wall (OCW) and fibrous content of low digestibility in OCW (Ob) by enzymatic methods. Biological oxygen demand (BOD) was analyzed by coulometric respirometer method. These compositions were carbohydrateds and lignin that were hardly digested. Spectra of samples were scanned by NIR instrument model 6500 (Pacific Scientific) and read over the range of wavelength between 400 and 2500nm. Calibration equations were developed using eight manure samples collected from black sheet shelter, while prediction was conducted to the other eight samples from clear sheet shelter. Accuracy of NTRS prediction was evaluated by correlation coefficients (r), standard error of prediction (SEP) and ration of standard deviation of reference data in prediction sample set to SEP (RPD). The r, SEP and RPD value of forage were 0.99, 0.69 and 7.6 for ADL, 0.96, 1.03 and 4.1 for NDF, 0.98, 0.60 and 4.9 for ADF, 0.92, 1.24 and 2.6 for Ob, and 0.91, 1.02 and 7.3 for BOD, respectively. The results indicated that NIRS could be used to measure the organic composition of forage used in manure samples.

• Journal of Ginseng Research
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• v.42 no.2
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• pp.149-157
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• 2018

Background: Panax notoginseng leaves (PNL) exhibit extensive activities, but few analytical methods have been established to exclusively determine the dammarane triterpene saponins in PNL. Methods: Ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC/Q-TOF MS) and HPLC-UV methods were developed for the qualitative and quantitative analysis of ginsenosides in PNL, respectively. Results: Extraction conditions, including solvents and extraction methods, were optimized, which showed that ginsenosides Rc and Rb3, the main components of PNL, are transformed to notoginsenosides Fe and Fd, respectively, in the presence of water, by removing a glucose residue from position C-3 via possible enzymatic hydrolysis. A total of 57 saponins were identified in the methanolic extract of PNL by UPLC/Q-TOF MS. Among them, 19 components were unambiguously characterized by their reference substances. Additionally, seven saponins of PNL-ginsenosides Rb1, Rc, Rb2, and Rb3, and notoginsenosides Fc, Fe, and Fd-were quantified using the HPLC-UV method after extraction with methanol. The separation of analytes, particularly the separation of notoginsenoside Fc and ginsenoside Rc, was achieved on a Zorbax ODS C8 column at a temperature of $35^{\circ}C$. This developed HPLC-UV method provides an adequate linearity ($r^2$ > 0.999), repeatability (relative standard deviation, RSD < 2.98%), and inter- and intraday variations (RSD < 4.40%) with recovery (98.7-106.1%) of seven saponins concerned. This validated method was also conducted to determine seven components in 10 batches of PNL. Conclusion: These findings are beneficial to the quality control of PNL and its relevant products.

• Journal of Korean Society of Forest Science
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• v.39 no.1
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• pp.57-63
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• 1978

Enzymatic hydrolysis of the substrate from Alnus hirsuta (Spach) Rupr (8-14years) was investigated using cellulase preparations of Trichoderma viride Pers. ex. Fr. SANK 16374 and conduced on the optimum reaction conditions of the cellulase on saccharification. The crude cellulase was produced by the submerged culture process and produced in the culture fluid was salted out quantitatively by the use of ammonium sulfate. The method of delignification from wood(Saw dust) was treated by the peracetic acid (PA) method. Reducing sugar was determined by the dinitrosalicylic acid (DNS) method. The results were summerized as follows; 1. The optimum pH of cellulase was 5.0 and the range of stability with respect to pH was generally from 4.0 to 6.0 2. The optimum temperature of cellulase was generally $40^{\circ}C$, but reducing sugar formation did not show significent differences at 5% levels in the reaction temperature from $40^{\circ}C$ to $50^{\circ}C$. 3. The redusing sugar were increased with increase of cellulase concentration. 4. The reducing sugar were decreased with increase of substrate concentration. 5. Fructose was a very good inhibitor of the enzyme from Trichoderma viride, but glucose inhibition was generally weak.

• Tuberculosis and Respiratory Diseases
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• v.42 no.6
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• pp.838-845
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• 1995

The determination of ADA(adenosine deaminase) activity in pleural fluid is useful in differental diagnosis of pleural effusion. The conventional method of determining ADA activity used by Giusti was influenced by contamination of ammonia. Additionally, because Giusti's method was mannual method a determining the ADA activities in sample, was not easily automated. In 1993, Oosthuizen HM with collegues developed simple kinetic method for determining ADA activity. It was reliable and suiable method for automation. In this study, we have measured ADA activity in 162 patients with various pleural effusion by Hitachi 747 autoanalyser using the Oosthuizen kinetic method for the purpuse of determination of new diagnostic cut-off value for the tuberculous effusion and evaluation of the correlation between the conventional method and new automated method. This new method of an enzymatic reaction involves 2, 6-dichlorophenolindophenol dye(DICP), adenosine, xanthine oxidase(XO), and nucleoside phosphorylase(NP). The results were as follows: 1) The mean pleural ADA activity of the tuberculous effusion was $52.53{\pm}16.43\;U/L$ and significantly higher than that of other groups(p<0.001). If the diagnostic cut-off value of pleural ADA activity for tuberculous effusion is above 30 U/L, the sensitivity is 96% and the specificity is 90%. 2) The mean pleural to serum ADA activity ratio of the tuberculous effusion was $2.29{\pm}0.96$ and it was also significantly higher than that of other pleural groups(p<0.001). If the diagnostic cut-off value of pleural to serum ADA activity ratio is 1.5, the sensitivity is 80% and the specificity is 88% in the diagnosis of tuberculous pleural effusion. 3) The new kinetic method is correlates well to Giuisti's conventional method(r=0.971). In conclusion, the new kinetic method described is easily automated and seems to be suitable for the routine determination of ADA activity.

• Korean journal of applied entomology
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• v.16 no.4 s.33
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• pp.199-208
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• 1977

Activities of various hydrolytic enzymes produced by three plant pathogenic fungi, Sclerotium rolfsii Sacc., Sclerotinia sclerotiorum (Lieb.) deBary and Helminthosporium sigmoideum var. irregulare Crallery et Tullius, were measured. Activties and amounts of the enzymes in mycelia, cultural filtrates, and sclerotia(except of sclerotia of H. sigmoideum var. irregulare) were estimated at various pH levels in order to find out optimal pH for their enzymatic activities. Enzymes such as cellulase (ex), invertase, xylanase, $\beta-amylase$, polymethylgalacturonase, polygalacturonase, phosphatase and protease were estimated. Culture solution for production of enzymes was prepared by adding of 10g, D-glucose, 1.3g $NH_4NO_3,\; 0.5g\; MgSO_4,\;7H_2O,\; and\; 1.0g\; KH_2PO_4$ into 1 liter of potato decoction plus 2ml of micro element solution consisting of 0.2mg. Fe, 0.2mg Zn, and 0.1mg Mn as the sulphates into 1 liter of distilled water. All tested mycelia and cultural filtrates were obtained from the cultures incubarted in previous solution for ten days at $25^{\circ}C$, and sclerotia were harvested from PDA plates of 3. days old, The crude enzyme solutions were prepared according to the method of Miyazaki etal. Ten days after incubation, activities of Cx produced by Scl. sclerotiorum were higher than those of the other fung and each of Cx from three fungi showed different pH optima, such as S. rolfsii and Scl. schlerotiorum in acid side (around pH 3.0), H. sigmoideum var. irregulare in neutral side (around pH 6.3). Invertase activities of S. rolfsii were 20 times higher than those of the other fungi in all samples. All tested fungi, however, showed no significant difference between the enzymatic activities of their cultural filtrate and mycelia and the activities in sclerotia of S. rolfsii and Scl. sclerotiorum were hardly recognized. There were multiple peaks on the xylanase activity curves of three fungi in terms of pH values. High activities of the xylanase were revealed in sclerotia of S. rolfsii and Scl. sclerotiorum, and in mycelia of H. sigmoideum var. irregulare. The highest activities of $\beta-amylase$ were shown both in mycelia and cultural filtrate of H. sigmoideum var. irregulae among the tested fungi, and their optimal pH was 6.2 in both mycelia and cultural filtrate. In the S. rofsii and Sel. sclerotiorum, however, the activities of cultural filtrates were higher than those of the other fungi, and optimal pH was 3.0 and 6.2 for cultural filtrate and both mycelia and sclerotia, respectively. Activities of PMG were high in cultural filtrates of all tested fungi, especially in Scl. sclerotiorum and H. sigmoideum var. irregulare. Mycelia of themalso showed the considerable activities. Optimal pH for enzymatic activities were variable with thekind of fungi or with the samples measured. The highest activities of PG were presented by mycelia of S. rolfsii and Scl. sclerotiorum. $9.l\mu /min.\; and\; 9.5\mu g/min.$, respectively. Optimal pH for activity of PG in mycelia was around 4.5 in S. rolfsii and around 3.0 in Scl. sclerotiorum. Phosphatase of S. rolfsii and Scl. sclerotiorum was more active in acid side (optimal PH3. 5) and that of H. sigmoideum var. irregulare showed one peak each in acid, neutral and alkaline side. But the highest peak was at pH 9.5. Protease of all tested fungi was more active at pH 10.0, especially that of the cultural filtrate of H. sigmoideum var. irregualre.

• KSBB Journal
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• v.23 no.4
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• pp.285-290
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• 2008

Various methods (OECD 301B, ISO 9439 and ASTM 5864) for biodegradability test of lubricants were reviewed, and a standard procedure was developed. Most lubrication products are released in rivers or sea then is degraded by microbial action in aerobic condition. Most international method are based on $CO_2$ evolution test. Inoculum obtained from a sewage disposal plant and test compound are cultivated in a mineral medium. Organic carbon of the test compound is degraded and oxidized through the enzymatic actions of inoculum, and ultimately mineralized to carbon dioxide. Biodegradability test conditions of lubricant oils were optimized. The highest biodegradability was achieved when the same medium as in ASTM 5864 and inoculum concentration of $10^4{\sim}10^5$ cell/L were used. The optimum standard materials were selected as aniline and sodium acetate. Additionally the effects of inoculum type on microbial growth and biodegradability were examined. Finally the standard operating procedure (SOP) for biodegradability test method was proposed.

• Korean Journal of Microbiology
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• v.28 no.2
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• pp.91-97
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• 1990

Complementary DNA (cDNA) coding for human cytoplasmic superoxide dismutase (SOD1) (superoxide: superoxide oxidoreductase E.C.1.15.1.1) was isolated from human liver cDNA library of $\lambda$gt11 by in situ plaque hybridization. The insery cDNA gas the 5' untranslational region (UTR) and 3'UTR of SOD1 gene. Polymerase Chain Reaction (PCR) method was used fro subcloning of SOD1 structural gene. Using synthetic sense strand primer (24mer) containing a start codon and antisense strand primer (24mer), SOD1 structural gene was selectively amplified. Amplified DNA was directly cloned into the HincII site of pUC19 plasmid. Insery cDNA was subcloned into M13 mp19 and sequenced by dideowy chain termination method with Sequenase. The nucleotide sequence of insert cDNA had an open reading frame (ORF) coding for 153 amino acid residues. The structural gene of cytoplasmic SOD was placed under the control of bacteriophage $\lambda P_{L}$ regulatory sequences, generating a highly efficient expression plasmid. The production of human SOD1 in E. coli cells was about 7% of total cellular proteins and recombinant human SOD1 possessed its own enzymatic acitivity.