Previously, we discovered a new MMP-2 isoform GA110, of which appearance in human follicular fluid(FF) and serum was increased by EDTA. The present study was conducted to investigate how GAI 10 can appear by EDTA. To examine possible involvement of protein disulfide isomerase(PDI), an enzyme responsible for the dimerization of protein via disulfide formation, effect of PDI inhibitor on the appearance of GA110 by EDTA was investigated. When PDI inhibitor added to FF before EDTA treatment, the gelatinolytic activity of GA110 was abolished in a concentration dependent manner. By contrast, the activity of 72 kDa gelatinase increased. However, the PDI inhibitor added to FF after EDTA treatment, the gelatinolytic activity of GA110 was unaffected. To find out the nature of the enzyme which converts 72 kDa gelatinase into GAI 10, chromatographic separation method of FF proteins was done. Using hydroxyapatite column, fractions rich in 72 kDa gelatinase were isolated and pooled. By using this pool as substrate for the 72 kDa converting enzyme, protein fractions containing the converting activity were obtained from chromatographic separation of FF onto glutathione sepharose fast flow column. When immunoblotting was performed on this enzymatically active protein fractions against polyclonal anti-PDI antibody, distinct immunoreactivity was observed, although appeared in smaller molecular weight region. Based on these observations, it is suggested that the appearance of GAI 10 in FF by EDTA treatment could be due to an activation of PDI-like enzyme, which dimerizes 72 kDa gelatinase into GAI 10 via the formation of disulfide bond between molecules.
Kim, Hee-Kyong;Noh, Hye-Ji;Cho, Hyang-Hyun;Koh, Hong Bum
Journal of Dairy Science and Biotechnology
/
v.34
no.2
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pp.137-144
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2016
The ultimate research goal of the current study was a development of hydrolyzed whey protein powder (7%-GNANA) manufactured with normal content of sialic acid, a marker compound, that is naturally occurring at 7% concentration in GMP obtained from the milk protein. GMP is a safe food, used worldwide in infant and baby foods, etc. The test substance was prepared using (7% sialic acid containing) GMP as a raw material, and then using alcalase, an enzyme approved as a food additive, after separation of sialic acid with 100% efficiency and 7%-GNANA (containing 7% sialic acid and protein; product name: HELICOBACTROL-7) provided by MEDINUTROL Inc. (Korea). Bacterial reverse mutation (Ames) test was conducted in accordance with GLP Guideline using the test substance specified above. To identify its mutagenic potential against microorganisms, histidine auxotrophic strains of Salmonella Typhimurium, TA98, TA100, TA1535, and TA1537, and tryptophan auxotrophic strain of Escherichia coli, WP2uvrA, were used. The bacterial reverse mutation (Ames) test was performed by dividing the test substances into five different concentration groups (0, 61.7, 185, 556, 1,670, $5,000{\mu}g/plate$). Results of this experiment did not reveal repetitive increase of colony generating values or positive criteria for reverse mutagenicity for any concentration of test substances in any of the five strains, regardless of the presence of a metabolic activation system, and no dose-dependency was identified. In conclusion, the safety of 7%-GNANA test substance was verified by bacterial reverse mutation test conducted before registration of 7%-GNANA as a food additive.
The goal of this study was to develop hydrolyzed whey protein powder (23%-GNANA) manufactured with high content of sialic acid, a marker compound that is usually present at 7% concentration in GMP obtained from the milk protein. It is a safe food, used worldwide in infant and baby foods, etc. The test substance was prepared using (7% sialic acid containing) GMP as a raw material. Alcalase, an enzyme approved as a food additive, was used after separating sialic acid, with 100% efficiency, and 23%-GNANA (composed of 23% sialic acid and protein; product name: HELICOBACTROL-23), provided by MEDINUTROL Inc. (Korea), manufactured to have high (23%) content through ethanol soaking and enrichment. Bacterial reverse mutation (Ames) test was conducted in accordance with the GLP Guideline using the test substance specified above. To detect its mutagenicity potential in microorganisms, histidine auxotrophic strains of Salmonella typhimurium, TA98, TA100, TA1535, and TA1537, and tryptophan auxotrophic Escherichia coli strain, WP2uvrA, were used. The bacterial reverse mutation (Ames) test was performed using five concentrations of the test substances (0, 61.7, 185, 556, 1,670, $5,000{\mu}g/plate$). The evaluation did not reveal repetitive increase of colony generating values and positive criteria for reverse mutagenicity for any tested concentration in the five strains regardless of the presence of metabolic activation system, and no dose-dependency. In conclusion, the safety of 23%-GNANA test substance was verified by the bacterial reverse mutation test conducted before registration of 23%-GNANA as a food additive.
Journal of The Korean Society of Integrative Medicine
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v.8
no.1
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pp.137-146
/
2020
Purpose : The enhancement of abdominal muscles increases the activation and contraction of respiratory muscles, including the diaphragm. Generally, diaphragm exercises are applied to increase the breathing ability of patients with respiratory disease. Previous studies have shown that breathing capacity can be increased through abdominal muscle strengthening exercises. However, studies on breathing ability are rare and it is doubtful whether these affect respiratory ability more than diaphragm exercises. Therefore, this study seeks to compare whether abdominal exercises can improve breathing ability and whether any increase is comparable to diaphragm exercises. Methods : After selecting subjects, the place of intervention was separated for blindness. The plank group was allowed to relax for 30 seconds after 30 seconds of planking; this was set at three and increased by one set each week. Subjects in the draw-in group were allowed to relax for 30 seconds after maintaining the draw-in contraction state for 30 seconds and this was done for 15 minutes. Subjects in the control group underwent abdominal dilation for five seconds of inspiration time and expired air for five seconds by exposing the lips; breathing was performed repeatedly for 15 minutes. Subjects in each group measured their respiration function three times before intervention, three weeks after the commencement of intervention and after intervention. Spirovit SP-1 was used to measure respiratory function. In each group, repeated ANOVA was used to compare the respiratory function over time and one-way ANOVA was used to compare the respiratory function between groups. The post hoc was conducted using the LSD method. Results : There was a significant increase in respiratory ability between the forced vital capacity (FVC), forced expiratory volume in one second (FEV1) and peak expiratory flow (PEF), forced expiratory volume in one second (FEV1)/forced vital capacity (FVC) according to the six-week period. However, there was no difference between each group. Conclusion : For patients with low respiratory muscle strength, plank exercises and abdominal draw-in are beneficial exercises for improving respiratory function. These are expected to be widely used in clinical practice for patients with weak respiratory muscles.
The spherical Al/RDX/AP composites with an average size of $550{\mu}m$ were successfully prepared by drowning-out/agglomeration (D/A) process. The surface morphology and dispersion of Al particles of those composites were investigated using SEM and EDS (energy dispersive spectrometry). As a result of thermal analysis, the onset temperature of thermal decomposition of the Al/RDX/AP composites by the D/A process was found to decrease about $50^{\circ}C$ and their thermal stability was shown to be relatively enhanced due to the increase of activation energy compared to those of using the physical mixing method. In the first decomposition region of AP, Prout-Tompkins model was shown to describe well the thermal decomposition of both composites by the physical mixing and D/A process. On the other hand, in the second decomposition region of AP, the decomposition mechanisms of composites by the physical mixing and D/A process were explained by the zero-order and contracting volume model, respectively.
Effects of elevated carbon dioxide ($CO_2$) on soil microbial processes were studied in a northern peatland. Intact peat cores with surface vegetation were collected from a northern Welsh fen, and incubated either under elevated carbon dioxide (700 ppm) or ambient carbon dioxide (350 ppm) conditions for 4 months. Higher algal biomass was found under the elevated $CO_2$ condition, suggesting $CO_2$ fertilization effect on primary production, At the end of the incubation, trace gas production and consumption were analyzed using chemical inhibitors. For methane ($CH_4$ ), methyl fluoride ($CH_3F$) was applied to determine methane oxidation rates, while acetylene ($C_2H_2$) blocking method were applied to determine nitrification and denitrification rates. First, we have adopted those methods to optimize the reaction conditions for the wetland samples. Secondly, the methods were applied to the samples incubated under two levels of $CO_2$. The results exhibited that elevated carbon dioxide increased both methane production (210 vs. $100\;ng\;CH_4 g^{-1}\;hr^{-1}$) and oxidation (128 vs. $15\;ng\;CH_4 g^{-1}\;hr^{-1}$), resulting in no net increase in methane flux. For nitrous oxide ($N_2O$) , elevated carbon dioxide enhanced nitrous oxide emission probably from activation of nitrification process rather than denitrification rates. All of these changes seemed to be substantially influenced by higher oxygen diffusion from enhanced algal productivity under elevated $CO_2$.
Objectives : In this study, we made an effort to investigate the protective mechanism of Ukgan-san (UGS) extracts on hypoxia-induced C6 glial cell death. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were analysed with microscope after staining with crystal violet (CV). Reactive oxygen species (ROS) formation was assessed by flow cytometer after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). We also analyzed expression of hypoxia-inducible factor-1 alpha (HIF-$1{\alpha}$) and p53, processing of procaspase-3 and procyclic acidic repetitive protein (PARP) by western blot method. Results : We estimated the elevated cell viability by UGS extract on $CoCl_2$-induced C6 glial cells. UGS attenuated $CoCl_2$-induced ROS formation in C6 glial cells and also showed a protective activity compared to antioxidants and exhibited abrogation of LDH-released by $CoCl_2$. UGS suppressed the typical apoptotic cell death markers, caspase-3 and PARP activation. UGS inhibited $CoCl_2$-induced HIF-1${\alpha}$ expression which is known as a major regulator for hypoxia-induced cell death, and suppressed p53 expression. Conclusions : These results suggest that UGS extract contains protective constituents for hypoxia-induced C6 glial cell death.
Recently, various public transportation activation policies are being implemented in order to mitigate traffic congestion in metropolitan areas. Especially in the metropolitan area, the bus information system has been introduced to provide information on the current location of the bus and the estimated arrival time. However, it is difficult to predict the travel time due to repetitive traffic congestion in buses passing through complex urban areas due to repetitive traffic congestion and bus bunching. The previous bus travel time study has difficulties in providing information on route travel time of bus users and information on long-term travel time due to short-term travel time prediction based on the data-driven method. In this study, the path based long-term bus travel time prediction methodology is studied. For this purpose, the training data is composed of 2015 bus travel information and the 2016 data are composed of verification data. We analyze bus travel information and factors affecting bus travel time were classified into departure time, day of week, and weather factors. These factors were used into clusters with similar patterns using self organizing map. Based on the derived clusters, the reference table for bus travel time by day and departure time for sunny and rainy days were constructed. The accuracy of bus travel time derived from this study was verified using the verification data. It is expected that the prediction algorithm of this paper could overcome the limitation of the existing intuitive and empirical approach, and it is possible to improve bus user satisfaction and to establish flexible public transportation policy by improving prediction accuracy.
Kim, Sung-Min;Kim, Hye-Ree;Ozkaya, Gizem;Shin, Sung-Hoon;Kong, Se-Jin;Kim, Eon-Ho;Lee, Ki-Kwang
Korean Journal of Applied Biomechanics
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v.25
no.3
/
pp.323-333
/
2015
Objective : The purpose of this study was to investigate the changes of gait patterns and muscle activations with increased loads during stair walking. Also, it can be used as descriptive data about continuous stair walking in a real life setting. Method : Twelve sedentary young male adults(Age: $27.0{\pm}1.8yrs$, Weight: $65.8{\pm}9.9kg$) without any lower extremity injuries participated in this study. Participants performed stair walking up 7 floors and their ascending and descending motion on each floor was analyzed. A wireless electromyography(EMG) were attached on the Rectus Femoris(RF), Biceps Femoris(BF), Gastrocnemius(GN), Tibialis Anterior(TA) muscle to calculate integrated EMG(iEMG), median frequency(MDF) and co-contraction index(CI). Chest and left heel accelerometer signal were recorded by wireless accelerometer and those were used to calculate approximate entropy(ApEn) for analyzing gait pattern. All analyses were performed with SPSS 21.0 and for repeated measured ANOVA and Post-hoc was LSD. Results : During ascending stairs, there were a statistically significant difference in Walking time between 1-2nd and other floors(p=.000), GN iEMG between 2-3th and 6-7th(p=.043) floor, TA MDF between 1-2nd and 5-6th(p=.030), 6-7th(p=.015) floor and TA/GN CI between 2-3th and 6-7th(p=.038) floor and ApEn between 1-2nd and 6-7th(x: p=.003, y: p=.005, z: p=.006) floor. During descending stairs, there were a statistically significant difference in TA iEMG between the 6-5th and 3-2nd(p=.026) floor, and for the ApEn between the 1-2nd and 6-7th(x: p=.037, y: p=.000, z: p=.000) floor. Conclusion : Subjects showed more regular pattern and muscle activation response caused by regularity during ascending stairs. Regularity during the first part of stair-descending could be a sign of adaptation; however, complexity during the second part could be a strategy to decrease the impact.
Objective : The purpose of this study was to investigate the effects in the muscle function following 8-week dead-lift training with Whole-body Vibration(WBV) in rehabilitation for sports players. Method : Twenty young sports players. Each subjects were randomly assigned to a resistance training with Whole-body vibration group(TG, n=10), a resistance training without Whole-body vibration group(CG, N=10). The measurements which physical fitness test and joint torque test were performed before the randomization and after the 4-week and 8-week. The WBV group performed the dynamic Dead-lift exercise on a vibration platform during one minute. The CG group performed the equal training without vibration. The WBV and CON group repeated 5 set and trained two times weekly for 8-weeks. Paired t-test was used to test for differences between the groups at baseline and after 4-weeks and 8-Weeks. And independent t-test was used to test for differences between the groups at TG and CG. All analyses were executed using SPSS software 18.0. The level of significance was set at p<.050. Results : Following the 8-Weeks training sessions, an increase in the back-muscle strength was found to be greater for the TG compared with the CG group(p<.05). Muscle endurance was significantly decreased after training than before training only for the CG(p<.05). Isometric Hip/Lumber Extension/Flextion measurement was found to be significantly greater for the TG compared with the CG group(p<.05). The finding indicates that WBV effects as an efficient training stimulus to enhance muscle function by facilitating neural control trail. Following muscle activation in motor unit synchronization of the co-contraction of the muscles. Conclusion : The results imply that the WBV training may have enhanced muscle function in rehabilitation for sports players.
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