• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1426-1431
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• 1998

The research was undertaken to characterize enzymatic properties of Streptomyces albus amylase expressed in recombinant Bacillus subtilis. Molecular weight and pI of the purified enzyme were estimated to be 50 kD by SDS-PAGE and 4.3 by isoelectric focusing. The optimum temperature and optimum pH were $45^{\circ}C$ and 6.0, respectively. D-and Z-value were estimated to measure thermostability of the purified enzyme. The Z-value was estimated $17.7^{\circ}C$, which is lower than typical amylase. Maltotetraose was produced as a major component from soluble starch in the early state of reaction but gradually degraded to maltose. Thin layer chromatography was also performed to analyze the reaction products. The parameters involved in Michaelis-Menten enzyme kinetics were found to be the maximum velocity of 0.37 mM/min and the Michaelis constant of 0.13%, respectively.

• The Journal of the Korean bone and joint tumor society
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• v.3 no.1
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• pp.18-25
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• 1997

Prosthetic replacement is one of the most common methods of reconstruction after resection of malignant tumor around the knee. Gait analysis provides a relative objective data about the gait function of patients with prosthesis. The purpose of this study was to compare the gait pattern of the patients who underwent limb salvage surgery with prosthesis for distal femur and that of patients with prosthesis for proximal tibia. This study included ten patients (4 males, 6 females, mean age 22.7 years, range 14-36) who underwent a wide resection and Kotz hinged modular reconstruction prosthesis replacement and six normal adult(Control). The site of bone tumor was the distal femur (Group 1) in six patients and proximal tibia (Group 2) in 4 patients. The follow-up period ranged from 15 to 82 months (mean : 33 months). The evaluation consisted of clinical assessment, radiographic assessment, gait analysis using VICON 370 Motion Analysis System. The gait analysis included the linear parameters such as, walking velocity, cadence, step length, stride length, stance time, swing time, single support and double support time and the three-dimensional kinematics (joint rotation angle, velocity of joint rotation) of ankle, knee, hip and pelvis in sagittal, coronal and transverse plane. For the kinetic evaluation, the moment of force (unit: Nm/kg) and power (unit: Watt/kg) of ankle, knee and hip joint in sagittal, coronal and transverse plane. In the linear parameters, cadence, velocity, step time and single support were decreased in both group 1 and group 2 compared with control. Double support decreased in group 2 compared with control significantly(p<.05). In contrast to our hypothesis, there was no significant difference between group 1 and group 2. In Kinematics, we observed significant difference (p<.05) of decreased knee flexion in loading response (G2

• The Journal of Advanced Prosthodontics
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• v.3 no.3
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• pp.152-160
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• 2011

PURPOSE. This study was to investigate the effects of recombinant human platelet-derived growth factor (rhPDGF-BB) and heparin to titanium surfaces for enhancement of osteoblastic functions and inhibition of inflammation activity. MATERIALS AND METHODS. The anodized titanium discs, not coated with any material, were used as a control group. In heparinized-Ti group, dopamine was anchored to the surface of Ti substrates, and coated with heparin. In PDGF-Ti group, rhPDGF-BB was immobilized onto heparinized Ti surface. The surface morphologies were investigated by the scanning electron microscope in each group. The release kinetics of rhPDGF-BB were analyzed, and cytotoxicity tests for each group were conducted. The biocompatibilities were characterized by measuring cell proliferation, alkaline phosphatase activity, and calcium deposition using MG-63 cells. Statistical comparisons were carried out by one-way ANOVA tests. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The combination of rhPDGF-BB and heparin stimulated alkaline phosphatase activity and OCN mRNA expression in osteoblastic cells ($^*$P<.05 and $^{**}$P<.001). MG-63 cells grown on PDGF-Ti had significantly higher amounts of calcium deposition than those grown on anodized Ti ($^{**}$ P<.001). Heparinized Ti was more anti-inflammatory compared to anodized Ti, when exposed to lipopolysaccharide using the transcript levels of TNF-${\alpha}$ and IL-6 of proinflammatory cytokine ($^*$P<.05 and $^{**}$P<.001). CONCLUSION. The result of this study demonstrated that the incorporation of rhPDGF-BB and heparin onto Ti surface enhanced osteoblastic functions and inhibited inflammation.

• The Journal of Advanced Prosthodontics
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• v.3 no.3
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• pp.145-151
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• 2011

PURPOSE. This study was performed to investigate the ability of recombinant human-bone morphogenic protein-2 immobilized on a heparin-grafted bone substrate to enhance the osteoblastic functions. MATERIALS AND METHODS. The Bio-$Oss^{(R)}$, not coated with any material, was used as a control group. In rhBMP-2-Bio-$Oss^{(R)}$ group, rhBMP-2 was coated with Bio-$Oss^{(R)}$ using only deep and dry methods (50 ng/mL, 24 h). In heparinized rhBMP-2-Bio-$Oss^{(R)}$ group, dopamine was anchored to the surface of Bio-$Oss^{(R)}$, and coated with heparin. rhBMP-2 was immobilized onto the heparinized- Bio-$Oss^{(R)}$ surface. The release kinetics of the rhBMP-2-Bio-$Oss^{(R)}$ and heparinized rhBMP-2-Bio-$Oss^{(R)}$ were analyzed using an enzyme-linked immunosorbent assay. The biological activities of the MG63 cells on the three groups were investigated via cytotoxicity assay, cell proliferation assay, alkaline phosphatase (ALP) measurement, and calcium deposition determination. Statistical comparisons were carried out by one-way ANOVA test. Differences were considered statistically significant at $^*$P<.05 and $^{**}$P<.001. RESULTS. The heparinized rhBMP-2-Bio-$Oss^{(R)}$ showed more sustained release compared to the rhBMP-2-Bio-$Oss^{(R)}$ over an extended time. In the measurement of the ALP activity, the heparinized group showed a significantly higher ALP activity when compared with the non-heparinized groups (P<.05). The MG63 cells cultivated in the group with rhBMP-2 showed increased calcium deposition, and the MG63 cells from the heparinized group increased more than those that were cultivated in the non-heparinized groups. CONCLUSION. Heparin increased the rhBMP-2 release amount and made sustained release possible, and heparinized Bio-$Oss^{(R)}$ with rhBMP-2 successfully improved the osteoblastic functions.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.4
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• pp.536-542
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• 2007

The responses of whole body protein and glucose kinetics and of nitrogen (N) metabolism to non-protein energy intake (NPEI) were determined using an isotope dilution approach and measurement of N balance in three adult male goats. The diets containing 1.0, 1.5 and 2.0 times ME maintenance requirement, with fixed intake of CP (1.5 times maintenance) and percentage of hay (33%), were fed twice daily for each 21 d experimental period. After an adaptation period of 11 d, N balance was determined over 3 d. On day 17, whole body protein synthesis (WBPS) and glucose irreversible loss rate (ILR) were determined during the absorptive state by a primed-continuous infusion of [$^2H_5$]phenylalanine, [$^2H_2$]tyrosine, [$^2H_4$]tyrosine and [$^{13}C_6$]glucose, with simultaneous measurements of plasma concentrations of metabolites and insulin. Ruminal characteristics were also measured at 6 h after feeding over 3 d. Nitrogen retention tended to increase (p<0.10) with increasing NPEI, although digestible N decreased linearly (p<0.05). Increasing NPEI decreased (p<0.01) ammonia N concentration, but increased acetate (p<0.05) and propionate (p<0.05) concentrations in the rumen. Despite decreased plasma urea N concentration (p<0.01), increased plasma tyrosine concentration (p<0.05), and trends toward increased plasma total amino N (p<0.10) and phenylalanine concentrations (p<0.10) were found in response to increasing NPEI. Increasing NPEI increased ILR of both glucose (p<0.01) and phenylalanine (p<0.05), but did not affect ($p{\geq}0.10$) that of tyrosine. Whole body protein synthesis increased (p<0.05) in response to increasing NPEI, resulting from increased utilization rate for protein synthesis (p<0.05) and unchanged hydroxylation rate of phenylalanine ($p{\geq}0.10$). These results suggest that increasing NPEI may enhance WBPS and glucose turnover at the absorptive state and improve the efficiency of digestible N retention in goats, with possibly decreased ammonia and increased amino acid absorption. In addition, simultaneous increases in WBPS and glucose ILR suggest stimulatory effect of glucose availability on WBPS, especially when sufficient amino acid is supplied.

• KSCE Journal of Civil and Environmental Engineering Research
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• v.10 no.3
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• pp.127-136
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• 1990

In this paper, the experimental study was carried out for the removal of olive oil in wastewater by the use of Biological Fluidized Bed(BFB) with the reticulated polypropylene sheets as media. The nonbiodegradable olive oil, one of the animal and vegitable oil, was used bacause of the relative simplicity of constitution. Biological degraciability and removal characteristics of emulsified olive oil were studied by batch and continuous experiments respectively. From the results of batch experiments, it was observed that the emulsified olive oli used in BFB reactor was absorbed by media and sludge in about 12 hours, and degradation of the absorbed olive oli was mostly completed for 24 hours. The functional relationship of Michaelis-Menten's Enzyme reaction equation exists between oil concentration and maximum specific rate of olive oil. From the continuous experiments for the removal of olive oil using BFB reactor, it was proved that the substrate removal rate coefficient $k=0.004d^{-1}$, which is the first order kinetics. It was apperared that oxygen utlization coefficients for synthesis(a') and endogeneous respiration(b') of microorganisms in the reactor are respectively 0.85mg $O_2/mg$ $COD_{cr}$ and 0.011mg $O_2/mg$ BVS. day.

• KSCE Journal of Civil and Environmental Engineering Research
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• v.6 no.2
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• pp.35-44
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• 1986

This paper is a basic study of the experimental results for the treatment of ammonia-nitrogen in the septic tank effluent. The substrates in this experiment are actual septic tank effluent and synthetic waste-water which is similar to septic tank effluent containing a considerable amount of nitrogenous component. Experiments were conducted for organic removal and nitrification using various recycle ratio and hydraulic retention time at each stage. The results obtained show that organic removal rate was above 80% in the 1st and 2nd stage, but as nitrification process was proceeded, above 90% of ammonia-nitrogen was removed in the 3rd and 4th stage. In these cases, the recycle ratio and HRT were found 30 and 7 hrs respectively. In the relation of $NH_4{^+}-N$ removal to $NO_3{^-}-N$ formation in the synthetic waste-water and septic tank effluent, when $1mg/{\ell}$ of $NH_4{^+}-N$ was removed, $NO_3{^-}-N$ formations were $0.95mg/{\ell}$ and $0.82mg/{\ell}$ respectively. And kinetics of nitrification using Biological Fluidized Bed was discussed also.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.9
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• pp.1278-1285
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• 2008

Measurement of gas produced during in vitro fermentation was used to investigate the fermentation characteristics and interactions of rice straw, wheat straw or maize stover mixed with alfalfa or clover at proportions of 100:0, 75:25, 50:50, 25:75 and 0:100, respectively. Cumulative gas production was recorded at 2, 4, 8, 12, 16, 24 and 48 h of incubation, and the Gompertz function was used to describe the kinetics of gas production. In vitro dry matter and organic matter disappearances (IVDMD and IVOMD) were determined after 48 h incubation. The rate of gas production of clover was higher (p<0.05) than that of rice straw, wheat straw, maize stover and alfalfa when straws and hays were incubated separately. Increasing the proportion of alfalfa in the straw-alfalfa mixtures increased (p<0.05) the rates, but not the maximum volume of gas production. However, both rate and the maximum volume of gas production were increased (p<0.01) as the proportions of clover increased in the straw-clover mixtures. The cumulative gas production at 48 h, IVDMD and IVOMD showed no consistent interaction effects between different mixtures of cereal straws and hays. The extent of interactive effects was affected by the types of cereal straw, legume hay and their proportions in the mixture. The appropriate combination for the mixture of rice straw or maize stover with leguminous hays was 75:25 and 25:75, respectively. The better combination occurred at a proportion of 50:50 for the mixture of wheat straw and alfalfa. We conclude that the suitable proportion of low-quality straw and high quality legume hay combination should be considered in the ration formulation system of ruminants according to the extent of positive interactive effects.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.1
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• pp.127-131
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• 2020

Objective: This study was conducted to determine catalytic properties of wheat phytase with exopolyphosphatase activity toward medium-chain and long-chain inorganic polyphosphate (polyP) substrates for comparative purpose. Methods: Exopolyphosphatase assay of wheat phytase toward polyP75 (medium-chain polyP with average 75 phosphate residues) and polyP1150 (long-chain polyP with average 1150 phosphate residues) was performed at pH 5.2 and pH 7.5. Its activity toward these substrates was investigated in the presence of Mg²⁺, Ni²⁺, Co²⁺, Mn²⁺, or ethylenediaminetetraacetic acid (EDTA). Michaelis constant (K_m) and maximum reaction velocity (V_max) were determined from Lineweaver-Burk plot with polyP75 or polyP1150. Monophosphate esterase activity toward p-nitrophenyl phosphate (pNPP) was assayed in the presence of polyP75 or polyP1150. Results: Wheat phytase dephosphorylated polyP75 and polyP1150 at pH 7.5 more effectively than that at pH 5.2. Its exopolyphosphatase activity toward polyP75 at pH 5.2 was 1.4-fold higher than that toward polyP1150 whereas its activity toward polyP75 at pH 7.5 was 1.4-fold lower than that toward polyP1150. Regarding enzyme kinetics, K_m for polyP75 was 1.4-fold lower than that for polyP1150 while V_max for polyP1150 was 2-fold higher than that for polyP75. The presence of Mg²⁺, Ni²⁺, Co²⁺, Mn²⁺, or EDTA (1 or 5 mM) exhibited no inhibitory effect on its activity toward polyP75. Its activity toward polyP1150 was inhibited by 1 mM of Ni²⁺ or Co²⁺ and 5 mM of Ni²⁺, Co²⁺, or Mg²⁺. Ni²⁺ inhibited its activity toward polyP1150 the most strongly among tested additives. Both polyP75 and polyP1150 inhibited the monophosphate esterase activity of wheat phytase toward pNPP in a dose-dependent manner. Conclusion: Wheat phytase with an unexpected exopolyphosphatase activity has potential as a therapeutic tool and a next-generational feed additive for controlling long-chain polyP-induced inappropriate inflammation from Campylobacter jejuni and Salmonella typhimurium infection in public health and animal husbandry.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.1
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• pp.24-34
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• 2020

Objective: The effects of Pleurotus ostreatus on the feed utilization of broad bean stalks (BBS), rape straw (RS), paddy straw (PS), and corn stalk (CS) was examined. Methods: The four roughages were co-cultured with Pleurotus ostreatus. The chemical composition; enzyme activities of laccase, carboxymethylcellulase (CMCase) and xylanase; carbohydrate and protein fractions (based on The Cornell Net Carbohydrate and Protein System [CNCPS]) were assessed at different days after inoculation (7, 14, 21, 28 d) and un-inoculated roughages (control, 0 d). The digestibility of nutrient components and the gas production of roughage with various incubation times were monitored at 0, 2, 4, 6, 9, 12, 24, 36, 48, 60, and 72 h using an in vitro ruminal fermentation method. Results: A higher CMCase activity (0.1039 U/mL) and earlier time to peak (14 d) were detected in Pleurotus ostreatus cultured with CS (p<0.05). Significantly, the incubation length-dependent responses of cumulative gas production were observed from 24 to 72 hours post fermentation (p<0.05), and these incubation length-dependent effects on cumulative gas production of PS and CS appeared earlier (24 h) for PS and CS than those (48 h) for BBS and RS (p<0.05). The fast-degradable carbohydrate (CA) content for all four roughages significantly increased over time (p<0.05). Nonetheless, increased degradation efficiency for CA treated with Pleurotus ostreatus was detected at both 21 and 28 days of incubation (p<0.05). With the exception of PS (p<0.05), there were no significant difference among the roughages (p>0.05) in slowly-degradable carbohydrate (CB2) at different incubation times (p<0.05). Conclusion: Assessment of the alterations in chemical composition, CNCPS system fractions, and the fermentation kinetics after biological pretreatment may yield a valuable database for evaluating the biological pretreatment of Pleurotus ostreatus in ruminant feed.