Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.
Carvedilol is an antihypertensive and antianginal compound that combines nonselective beta-adrenoceptor blocking and vasodilation properties and is devoid of intrinsic sympathomimetic activity. The purpose of the present study was to evaluate the bioequivalence of two carvedilol tablets, $Dilatrend^{TM}$ (Chong Kun Dang Pharmaceutical Co., Ltd.) and $Carvelol^{TM}$ (Dae Won Pharmaceutical Co., Ltd.), according to the prior and revised guidelines of Korea Food and Drug Administration (KFDA). The carvedilol release from the two carvedilol tablets in vitro was tested using KP VII Apparatus II method with various different kinds of dissolution media (pH 1.2, 4.0, 6.8 buffer solution, water and blend of PSB80 into water). Eighteen normal male volunteers, $24.22{\pm}1.86$ years in age and $64.81{\pm}4.56\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After one tablet containing 25 mg of carvedilol was orally administered, blood was taken at predetermined time intervals and the concentrations of carvedilol in serum were determined using HPLC method with fluorescence detector. The dissolution profiles of two carvedilol tablets were very similar at all dissolution media. Besides, the pharmacokinetic parameters such as $AUC_t$, $C_{max}$ and $T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using non-transformed and logarithmically transformed $AUC_t$ and $C_{max}$. The results showed that the differences in $AUC_t$, $C_{max}$ and $T_{max}$ between two tablets based on the $Dilatrend^{TM}$ were 2.23%, -2.00% and 0.00%, respectively. Minimum detectable differences $({\Delta})$ at ${\alpha}=0.05$ and $1-{\beta}=0.8$ were less than 20% (e.g., 13.55% and 17.61% for $AUC_t$ and $C_{max}$, respectively). The powers $(l-{\beta})$ at ${\alpha}=0.05$, ${\Delta}=0.2$ for $AUC_t$ and $C_{max}$ were 98.08% and 88.81%, respectively. The 90% confidence intervals were within ${\pm}20%$ (e.g., $-5.69{\sim}10.16$ and $-12.30{\sim}8.30$ for $AUC_t$ and $C_{max}$, respectively). There were no sequence effect between two tablets in logarithmically transformed $AUC_t$ and $C_{max}$. The 90% confidence intervals using logarithmically transformed were within the acceptance range of log(0.8) to log(1.25) (e.g., $0.95{\sim}1.11$ and $0.89{\sim}1.09$ for $AUC_t$ and $C_{max}$, respectively). Two parameters met the criteria of prior and revised KFDA guideline for bioequivalence, indicating that $Carvelol^{TM}$ tablet is bioequivalent to $Dilatrend^{TM}$ tablet.
Lee, H.J.;Lee, S.H.;Cho, Y.M.;Yoon, H.B.;Jeon, B. K.;Oh, S.J.;Kwon, M.S.;Yoon, D.H.
Journal of Animal Science and Technology
/
v.46
no.6
/
pp.947-956
/
2004
The primary role of lipoprotein lipase(LPL) is the hydrolysis of triglycerides(TG) from the core of triglyceride-rich lipoproteins such as chylomicrons and very low density lipoproteins in plasma. Fatty acids liberated by LPL on capillary endothelial surfaces are available for tissues as energy sources especially in muscles or for storage in the form of TG in adipose tissues. Therefore, as the candidate gene related to the carcass traits of the beef cattle, we have directly sequenced the exon 5${\sim}$exon 6 region in the bovine LPL gene for discovery of single nucleotide polymorphism(SNP) with 24 unrelated Hanwoo(Korean cattle). Novel eight sequence variants were detected: three loci on exon 5, three on intron 5 and two on exon 6. All SNPs identified were strongly linked each other, and one hundred twenty eight Hanwoo samples were genotyped one SNP on intron 5 using PCR-restriction fragment length polymorphism method by digestion with Hae III restriction enzyme. The allele frequency of the polymorphism was 0.76 and 0.24. The effects of this polymorphism on the breeding values of the carcass weight, loin muscle area, back fat thickness and marbling score were analyzed using least square methods of SAS GLM. The marbling score of BB genotype was significantly higher than those of AA and AB genotypes(P<0.05). This result indicates that this polymorphism may be associated with the variation of marbling score. Further study is warranted to investigate the phenotypic association in Hanwoo.
Kim, Mi-Hyung;Choi, Nam-Ki;Kim, Seon-Mi;Oh, Jung-Suk;Yang, Kyu-Ho
Journal of the korean academy of Pediatric Dentistry
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v.32
no.2
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pp.344-356
/
2005
There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic acid bacteria were isolated from inhabitants of caries-free children's oral cavity, which inhibited the formation of artificial plaque by Streptococcus mutans and the production of volatile sulfur compounds by anaerobic bacteria. The isolates were identified by the test using API 50 CHL medium kit and 16S rDNA partial sequencing. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. When Streptococcus mutans was cultured in the media, the mean weight of formed artificial plaque on the orthodontic wires was $124.4{\pm}30.4\;mg$, whereas being reduced to $5.2{\pm}2.0mg$ and $10.6{\pm}6.6mg$ in the media cultured with Streptococcus mutans and each isolate, respectively (p<0.05) 3. The number of viable cells of Streptococcus mutans was $3.4{\times}10^9$ per ml in the cultured solution, whereas those of Streptococcus mutans in the combined culture with each of isolates were $4.6{\times}10^8\;and\;2.4{\times}10^8$ per ml. 4. The optical density was 1.286 in the supernatant of Fusobacterium nucleatum after vortexing for 30minutes, whereas in the supernatant of combined Fusobacterium nucleatum and each isolate, they were reduced to 0.628 and 0.497, which the percentages of coaggregation between them were 29.4% and 57.8%, respectively 5. The optical density of Fusobacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$, being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusobacterium nucleatum and each isolate. The optical density of Porphyromonas gingivalis precipitate was 1.932 in the culture media, being reduced to 1.170 and 1.266 in the coaggregated precipitates of Porphyromonas gingivalis and each isolate. 6. When two isolates were tested with API 50 CHL medium kit, those were identified Lactobaciallius salivarius and Lactobaciallius delbrueckii subsp. lactis. 7. The similarity values of 16S rDNA sequence between each of isolates and Lactobaciallius salivarius subsp. salicinius were 99.60% and 99.73%, respectively, meaning that isolates were Lactobaciallius salivarius subsp. salicinius. These results indicated that two strains isolated from caries-free children's saliva, which inhibited the formation of artificial plaque and the production of volatile sulfur compounds, were identified as Lactobaciallius salivarius subsp. salicinius.
Kim Soo-Youn;Choi Gang Guk;Park Youn Il;Park Young Mok;Yang Young Ki;Rhee Young Ha
Korean Journal of Microbiology
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v.41
no.1
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pp.67-73
/
2005
Photoautotrophic bacteria are promising candidates for the production of poly(3-hydroxybutyrate) (PHB) since they can address the critical problem of substrate costs. In this study, we isolated 25 Tn5-inserted mutants of the Synechocystis sp. PCC 6803 which showed enhanced PHB accumulation compared to the wild-type strain. After 5-days cultivation under nitrogen-limited mixotrophic conditions, the intracellular levels of PHB content in these mutants reached up to $10-30\%$ of dry cell weight (DCW) comparable to $4\%$ of DCW in the wild-type strain. Using the method of inverse PCR, the affected genes of the mutants were mapped on the completely known genome sequence of Synechocystis sp. PCC 6803. As a result, the increased PHB accumulation in 5 mutants were found to be resulted from defects of genes coding for NADH-ubiquinone oxidoreductase, O-succinylbenzoic-CoA ligase, photosystem II PsbT protein or histidine kinase, which are involved in photosystem in thylakoid inner membrane of the cell. The values of $NAD(P)H/NAD(P)^+$ ratio in the cells of these mutants were much higher than that of the wild-type strain as measured by using pulse-amplitude modulated fluorometer, suggesting that PHB synthesis could be enhanced by increasing the level of cellular NAD(P)H which is a limiting substrate for NADPH-dependent acetoacetyl-CoA reductase. From these results, it is likely that NAD(P)H would be a limiting factor for PHB synthesis in Synechocystis sp. PCC 6803.
Journal of Korean Society of Environmental Engineers
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v.34
no.7
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pp.473-479
/
2012
Water reuse has been highlighted as a representative alternative to solve the lacking water resource. This study carried out a study on the pipe corrosion and water quality change which can occur through the supply of reclaimed water, using a simulated reclaimed water distribution pipeline. Galvanized steel pipe (GSP), cast iron pipe (CIP), stainless steel pipe (STSP) and PVC pipe (PVCP) were used for the pipe materials. Reclaimed water(RW) and tap water(TW) were respectively supplied into simulated reclaimed water distribution pipelines. As a result of performing a loop test to supply reclaimed water to simulated reclaimed water distribution pipelines, the weight reduction of pipe coupons showed the sequence of CIP > GSP > STSP ${\approx}$ PVCP. In addition, reclaimed water showed a high corrosion rate comparing to that of tap water. In case of CIP, the initial corrosion rate showed 3.511 mdd(milligrams per square decimeter per day) for reclaimed water and 2.064 mdd for tap water and the corrosion rate for 90 days showed 0.833 mdd for reclaimed water and 0.294 mdd for tap water. Also in case of GSP, the initial corrosion rate showed 2.703 mdd for reclaimed water and 2.499 mdd for tap water and the corrosion rate for 90 days showed 0.349 mdd for reclaimed water and 0.248 mdd for tap water, which was a tendency similar to that appeared in CIP with a tendency to reduce the corrosion rate. As a result of water quality changes of reclaimed water at pipe materials to carry out the loop test, there was higher conversion ratio of ammonia into nitrate in CIP and GSP with higher corrosion rate than that in STSP and PVCP where no corrosion has occurred. The highest denitrification rate of nitrate could be observed from CIP with the most particles generated from corrosion. In CIP, it could be confirmed that there was MIC (Microbiologically Induced Corrosion) as a result of EDS (Energy Dispersive X-ray spectrometer System) analysis results.
This study was done to determine if there is any difference in microleakage between experimental composite resins, in which various proportions of three component photoinitiators (Camphoroquinone, OPPI, Amine) were included. Four kinds of experimental composite resin were made by mixing 3.2% silanated barium glass (78 wt.%, average size; 1 ${\mu}m$) with each monomer system including variously proportioned photoinitiator systems used for photoinitiating BisGMA/BisEMA/TEGDMA monomer blend (37.5:37.5:25 wt.%). The weight percentage of each component were as follows (in sequence Camphoroquinone, OPPI, Amine): Group A - 0.5%, 0%, 1% / Group B - 2%, 0.2%, 2% / Group C - 0.2%, 1%, 0.2% / Group D - 1%, 1%, 2%. Each composite resin was used as a filling material for round class V cavities (diameter: 2/3 of mesiodistal width; depth: 1.5 mm) made on extracted human premolars and they were polymerized using curing light unit (XL 2500, 3M ESPE) for 40 s with an intensity of 600 mW/$cm^2$. Teeth were thermocycled fivehundred times between $50^{\circ}C$and $550^{\circ}C$for 30s at each temperature. Electrical conductivity (${\mu}A$) was recorded two times (just after thermocycling and after three-month storage in saline solution) by electrochemical method. Microleakage scores of each group according to evaluation time were as follows [Group: at first record / at second record; unit (${\mu}A$)]: A: 3.80 (0.69) / 13.22 (4.48), B: 3.42 (1.33) / 18.84 (5.53), C: 4.18 (2.55) / 28.08 (7.75), D: 4.12 (1.86) / 7.41 (3.41). Just after thermocycling, there was no difference in microleakage between groups, however, group C showed the largest score after three-month storage. Although there seems to be no difference in microleakage between groups just after thermocycling, composite resin with highly concentrated initiation system or classical design (Camphoroquinone and Amine system) would be more desirable for minimizing microleakage after three-month storage.
Park Hyun-Jeong;Ko Sung-Min;Kim Yong-Sun;Chang Yongmin
Investigative Magnetic Resonance Imaging
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v.8
no.1
/
pp.32-41
/
2004
Purpose : To measure the NMR relaxation properties of MnPC, to observe the characteristics of liver enhancement patterns on MR images in experimentally implanted rabbit VX2 tumor model, and to estimate the possibility of tissue specific contrast agent for MnPC in comparison with the hepatobiliary agent. Materials and Methods : Phthalocyanine (PC) was chelated with paramagnetic ions, manganese (Mn). 2.01 g (5.2 mmol) of phthalocyanine was mixed with 0.37 g (1.4 nlmol) of Mn chloride at $310^{\circ}C$ for 36 hours and then purified by chromatography ($CHCl_3:\;CH_3OH=98:2$, volume ratio) to obtain 1.04 g $(46\%)$ of MnPC (molecular weight = 2000 daltons). The T1/T2 relaxivity (R1/R2) for MnPC were determined at a 1.5 T (64 MHz) MR spectrometer. VX2 tumor model was experimentally implanted in the liver parenchyma of rabbits. All MR studies were performed on 1.5 T. The human extremity radio frequency coil of a bird cage type was employed. MR images were acquired at 17 to 24 days after VX2 carcinoma implantation.4 mmol/kg MnPC and 0.01 mmol/kg Mn-DPDP were injected via the ear vein of rabbits. T1-weighted images were obtained with spin-echo (TR/TE=516/14 msec) and fast multiplanar spoiled gradient recalled (TR/TE : 80/4 msec, $60^{\circ}$ flip angle) pulse sequence. Fast spin-echo (TR/TE=1200/85 msec) was used to obtain the T2-weighted images. Results : The value of T1/T2 relaxivity (R1/R2) of MnPC was $7.28\;mM^{-1}S^{-1}$ and $55.56\;mM^{-1}S^{-1}$ respectively at 1.5 T (64 MHz). Because the T2 relaxivity of MnPC that bonded strongly, covalently manganese with phthalocyanine was very high, the signal intensity of liver parenchyma was decreased on postcontrast T2-weighted images and we could easily distinguish the VX2 carcinoma within the liver parenchyma. When MnPC was administrated intravenously, the tumor margin delineation was more remarkable than Mn-DPDP-enhanced images. The enhancement of liver parenchyma with MnPC persisted at relatively high levels over at least one hour after injection of the contrast agents. Conclusion : The hepatic uptake and biliary excretion of MnPC, which are similar to Mn-DPDP, suggest that this agent is a new liver-specific agent. Also, MnPC seems to be used as a dual contrast agent (T1 and T2) with high T2 relaxivity. However, it is warranted that MnPC needs further investigation as a potential contrast agent for MR imaging of the liver. That is, further characterizations of MnPC are needed in vivo and in vitro before clinical trials. The diagnostic potential of MnPC will also have to be examined more in the animal models of additional types.
This study uses Node2vec graph embedding method and Light GBM link prediction to explore undeveloped export candidate countries in Korea's food and beverage industry. Node2vec is the method that improves the limit of the structural equivalence representation of the network, which is known to be relatively weak compared to the existing link prediction method based on the number of common neighbors of the network. Therefore, the method is known to show excellent performance in both community detection and structural equivalence of the network. The vector value obtained by embedding the network in this way operates under the condition of a constant length from an arbitrarily designated starting point node. Therefore, it has the advantage that it is easy to apply the sequence of nodes as an input value to the model for downstream tasks such as Logistic Regression, Support Vector Machine, and Random Forest. Based on these features of the Node2vec graph embedding method, this study applied the above method to the international trade information of the Korean food and beverage industry. Through this, we intend to contribute to creating the effect of extensive margin diversification in Korea in the global value chain relationship of the industry. The optimal predictive model derived from the results of this study recorded a precision of 0.95 and a recall of 0.79, and an F1 score of 0.86, showing excellent performance. This performance was shown to be superior to that of the binary classifier based on Logistic Regression set as the baseline model. In the baseline model, a precision of 0.95 and a recall of 0.73 were recorded, and an F1 score of 0.83 was recorded. In addition, the light GBM-based optimal prediction model derived from this study showed superior performance than the link prediction model of previous studies, which is set as a benchmarking model in this study. The predictive model of the previous study recorded only a recall rate of 0.75, but the proposed model of this study showed better performance which recall rate is 0.79. The difference in the performance of the prediction results between benchmarking model and this study model is due to the model learning strategy. In this study, groups were classified by the trade value scale, and prediction models were trained differently for these groups. Specific methods are (1) a method of randomly masking and learning a model for all trades without setting specific conditions for trade value, (2) arbitrarily masking a part of the trades with an average trade value or higher and using the model method, and (3) a method of arbitrarily masking some of the trades with the top 25% or higher trade value and learning the model. As a result of the experiment, it was confirmed that the performance of the model trained by randomly masking some of the trades with the above-average trade value in this method was the best and appeared stably. It was found that most of the results of potential export candidates for Korea derived through the above model appeared appropriate through additional investigation. Combining the above, this study could suggest the practical utility of the link prediction method applying Node2vec and Light GBM. In addition, useful implications could be derived for weight update strategies that can perform better link prediction while training the model. On the other hand, this study also has policy utility because it is applied to trade transactions that have not been performed much in the research related to link prediction based on graph embedding. The results of this study support a rapid response to changes in the global value chain such as the recent US-China trade conflict or Japan's export regulations, and I think that it has sufficient usefulness as a tool for policy decision-making.
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