Trichoplusia ni cells are used as a host permissive cell line in the baculovirus expression system, which is useful for large-scale production of human sugar transport proteins. However, the activity of endogenous sugar transport systems in insect cells is extremely high. Therefore, the transport activity resulting from the expression of exogenous transporters is difficult to detect. Furthermore, very little is known about the nature of endogenous insect transporters. To exploit the expression system further, the effect of D-fructose on 2-deoxy-D-glucose (2dGlc) transport by T. ni cells was investigated, and T. ni cell-expressed human transporters were photolabeled with [$^3H$] cytochalasin B to develop a convenient method for measuring the biological activity of insect cell-expressed transporters. The uptake of 1 mM 2dGlc by uninfected- and recombinant AcMPV-GTL infected cells was examined in the presence and absence of 300 mM of D-fructose, with and without $20{\mu}M$ of cytochalasin B. The sugar uptake in the uninfected cells was strongly inhibited by fructose but only poorly inhibited by cytochalasin B. Interestingly, the AcMPV-GTL-infected cells showed an essentially identical pattern of transport inhibition, and the rate of 2dGlc uptake was somewhat less than that seen in the non-infected cells. In addition, a sharply labeled peak was produced only in the AcMPV-GTL-infected membranes labeled with [$^3H$] cytochalasin B in the presence of L-glucose. No peak of labeling was seen in the membranes prepared from the uninfected cells. Furthermore, photolabeling of the expressed protein was completely inhibited by the presence of D-glucose, demonstrating the stereoselectivity of labeling.
Purpose : In order to understand in vivo radiation damage modifying of bFGF on jejunal mucosa, bone marrow and the effect of bFGF on the growth of transplanted mouse sarcoma 180 tumor in mice. Materials and Methods : Mice were treated with $6\;{\mu}g$ of bFGF at 24 hours and 4 hours before exposing to 600 cGy, 800 cGy and 1,000 cGy total body irradiation (TBI), and then exposed to 3,000 cGy local radiation therapy on the tumor bearing thigh. Survival and tumor growth curve were plotted in radiation alone group and combined group of bFGF and irradiation (RT). Histologic examination was performed in another experimental group. Experimental groups consisted of normal control, tumor control, RT (radiation therapy) alone, $6\;{\mu}g$ bFGF alone, combined group of $3\;{\mu}g$ bFGF and irradiation (RT), combined group of $6\;{\mu}g$ bFGF and irradiation (RT). Histologic examination was peformed with H-E staining in marrow, jejunal mucosa, lung and sarcoma 180 bearing tumor. Radiation induced apoptosis was determined in each group with the DNA terminal transferase nick-end labeling method ($ApopTag^{\circledR}$ S7100-kit, Intergen Co.) Results : The results were as follows 1) $6\;{\mu}g$ bFGF given before TBI significantly improved the survival of lethally irradiated mice. bFGF would protect against lethal bone marrow syndrome. 2) $6\;{\mu}g$ bFGF treated group showed a significant higher crypt depth and microvilli length than RT alone group (p<0.05). 3) The bone marrow of bFGF treated group showed less hypocellularity than radiation alone group on day 7 and 14 after TBI (p<0.05), and this protective effect was more evident in $6\;{\mu}g$ bFGF treated group than that of $3\;{\mu}g$ bFGF treated group. 4) bFGF protected against early radiation induced apoptosis in intestinal crypt cell but might have had no antiapoptotic effect in bone marrow stem cell and pulmonary endothelial cells. 5) There was no significant differences in tumor growth rate between tumor control and bFGF alone groups (p>0.05). 6) There were no significant differences in histopathologic findings of lung and mouse sarcoma 180 tumor between radiation alone group and bFGF treated group. Conclusions : Our results suggest that bFGF protects small bowel and bone marrow from acute radiation damage without promoting the inoculated tumor growth in C3H mice. Improved recovery of early responding normal tissue and reduced number of radiation induced apoptosis may be possible mechanism of radioprotective effect of bFGF.
This study was conducted to establish the standard method for the contents of biotin in milk formulas. To optimize the method, we compared several conditions for liquid extraction, purification and instrumental measurement using spiked samples and certified reference material (NIST SRM 1849a) as test materials. LC-MS/MS method for biotin was established using $C_{18}$ column and binary gradient 0.1% formic acid/acetonitrile, 0.1% formic acid/water mobile phase is applied for biotin. Product-ion traces at m/z 245.1 ${\rightarrow}$ 227.1, 166.1 are used for quantitative analysis of biotin. The linearity was over $R^2=0.999$ in range of $5{\sim}60{\mu}g/L$. For purification, chloroform was used as a solvent for eliminating lipids in milk formula. The linearity was over 0.999 in range of 5~60 ng/mL. The detection limit and quantification limit were 0.10, 0.31 ng/mL. The accuracy and precision of LC-MS/MS method using CRM were 103%, 2.5% respectively. Optimized methods were applied in sample analysis to verify the reliability. All the tested milk formulas were acceptable contents of biotin compared with component specification and standards for nutrition labeling. The standard operating procedures were prepared for biotin to provide experimental information and to strengthen the management of nutrient in milk formula.
Clustering is a process of grouping similar or relevant documents into a cluster and assigning a meaningful concept to the cluster. By this process, clustering facilitates fast and correct search for the relevant documents by narrowing down the range of searching only to the collection of documents belonging to related clusters. For effective clustering, techniques are required for identifying similar documents and grouping them into a cluster, and discovering a concept that is most relevant to the cluster. One of the problems often appearing in this context is the detection of a complex concept that overlaps with several simple concepts at the same hierarchical level. Previous clustering methods were unable to identify and represent a complex concept that belongs to several different clusters at the same level in the concept hierarchy, and also could not validate the semantic hierarchical relationship between a complex concept and each of simple concepts. In order to solve these problems, this paper proposes a new clustering method that identifies and represents complex concepts efficiently. We developed the Hierarchical Overlapping Clustering (HOC) algorithm that modified the traditional Agglomerative Hierarchical Clustering algorithm to allow overlapped clusters at the same level in the concept hierarchy. The HOC algorithm represents the clustering result not by a tree but by a lattice to detect complex concepts. We developed a system that employs the HOC algorithm to carry out the goal of complex concept detection. This system operates in three phases; 1) the preprocessing of documents, 2) the clustering using the HOC algorithm, and 3) the validation of semantic hierarchical relationships among the concepts in the lattice obtained as a result of clustering. The preprocessing phase represents the documents as x-y coordinate values in a 2-dimensional space by considering the weights of terms appearing in the documents. First, it goes through some refinement process by applying stopwords removal and stemming to extract index terms. Then, each index term is assigned a TF-IDF weight value and the x-y coordinate value for each document is determined by combining the TF-IDF values of the terms in it. The clustering phase uses the HOC algorithm in which the similarity between the documents is calculated by applying the Euclidean distance method. Initially, a cluster is generated for each document by grouping those documents that are closest to it. Then, the distance between any two clusters is measured, grouping the closest clusters as a new cluster. This process is repeated until the root cluster is generated. In the validation phase, the feature selection method is applied to validate the appropriateness of the cluster concepts built by the HOC algorithm to see if they have meaningful hierarchical relationships. Feature selection is a method of extracting key features from a document by identifying and assigning weight values to important and representative terms in the document. In order to correctly select key features, a method is needed to determine how each term contributes to the class of the document. Among several methods achieving this goal, this paper adopted the $x^2$�� statistics, which measures the dependency degree of a term t to a class c, and represents the relationship between t and c by a numerical value. To demonstrate the effectiveness of the HOC algorithm, a series of performance evaluation is carried out by using a well-known Reuter-21578 news collection. The result of performance evaluation showed that the HOC algorithm greatly contributes to detecting and producing complex concepts by generating the concept hierarchy in a lattice structure.
Hwang, Kyung Mi;Ham, Hyeon Suk;Lee, Hwa Jung;Kang, Yoon Jung;Yoon, Hae Seong;Hong, Jin Hwan;Lee, Hyoun Young;Kim, Cheon Hoe;Oh, Keum Soon
Journal of Food Hygiene and Safety
/
v.32
no.5
/
pp.411-417
/
2017
This study was conducted to establish the analysis method for the contents of choline in infant formulas and follow-up formulas by ion chromatograph (IC). To optimize the method, we compared several conditions for extraction, purification and instrumental measurement using spiked samples and certified reference material (CRM; NIST SRM 1849a) as test materials. IC method for choline was established using Ion Pac CG column and 18 mM $H_2SO_4$ mobile phase. The parameters of validation were specificity, linearity, LOD, LOQ, recovery, accuracy, precision and repeatability. The specificity was confirmed by the retention time and the linearity, $R_2$ was over 0.999 in range of 0.5~10 mg/L. The detection limit and quantification limit were 0.14, 0.43 mg/L. The accuracy and precision of this method using CRM were 95%, 2.1% respectively. Optimized methods were applied in sample analysis to verify the reliability. All the tested products were acceptable contents of choline compared with component specification for nutrition labeling. The standard operating procedures were prepared for choline to provide experimental information and to strengthen the management of nutrient in infant formula and follow-up formula.
The compulsory beef labelling system has launched from January 1st 2007 by the amended Food Hygiene Law, we were checked the actual conditions of beef origin with a nationwide scale by the Hanwoo differentiation specific test method which was developed by Korea FDA using 90 SNP biomarkers. The test method is useful tool to differentiate the beef origin carrying out the mission of KFDA's annual food safety management guidance. Also we have technically transferred the Hanwoo differentiation specific test method to other institutes as well regional KFDA and established the training program as a regular course in Korea Human Resource Development Institute for Health and Welfare. The beef used in this study were collected according to the 2009 Food safety guidance in roast beef restaurants where business site area greater than 100 $m^2$. Total 216 samples were consisted of 48 samples of the Seoul area and 168 of the region. The monitoring result from restaurants in all the region of Korea showed that 3 of 216 Hanwoo-labelled beefs were found out as a non-Hanwoo (1.3%). This results are gradually deceasing trend compared with 34.0% in 2005, 30.1% in 2006, 3.2% in 2007 and 5.14% in 2008. From these data, the Hanwoo differentiation specific test method on the settlement of the compulsory beef labelling system has an important role. As a outcome of this project, we might be considered the early settlement of the compulsory beef labelling system, technically transferred to other institutes and the establishment of regular training program of the test method.
Kim, Do-Woo;Lee, Moo-Sik;Na, Baeg-Ju;Hong, Jee-Young
Journal of the Korea Academia-Industrial cooperation Society
/
v.14
no.7
/
pp.3326-3337
/
2013
This study was performed to determine health-related dietary and lifestyle of characteristic factors associated with sodium hyperingestion of korean adults by sexual difference. Study subjects were 30~69years (n=1,622) adults who participated in the 2010 Korean National Health and Nutrition Examination Survey (KNHANES V-1). Subjects were divided into the hyperingestion group (HG, n=1,494) and normal ingestion group (NG, n=128) by 'WHO growth standard sodium intakes(2,000mg/day)' and the relationships between Comparison of demographics and physical measurement, eating behaviors and health-related factors were analyzed. By logistic regression method, risk factors for sodium hyperingestion group revealed in this study were gender(female), number of family', 'breakfast', 'lunch', 'dinner', 'nutrition labeling use'. and In case of male, risk factors for sodium hyperingestion group were 'lunch', 'dinner', 'self-rated body image' of eating behaviors. In case of female, Which were number of family' of demographics, 'breakfast', 'lunch', 'dinner' of eating behaviors. In these results, according to different sexual aspect of sodium hyperingestion, we suggest that nutrition label use and proper nutrition intake criteria among sexual, recognition promotion campaign along with education program for repressed sodium hyperingestion related to hypertension prevention of korean adults should include eating habits improvement and demographics lifestyle modification.
Park, Yeon Jung;Lee, Mi Nan;Kim, Eun Mi;Noh, Eun Soo;Noh, Jae Koo;Park, Jung Youn;Kang, Jung-Ha
Journal of Life Science
/
v.28
no.9
/
pp.1062-1067
/
2018
Reliable labeling of fish products can reassure consumers regarding the identity and quality of seafoods. Therefore, techniques that can identify adulteration or mislabeling are valuable. To rapidly identify two Trachurus species, Trachurus japonicus and Trachurus novaezelandiae, a highly efficient, rapid, duplex polymerase chain reaction (PCR) having two species-specific primers simultaneously was identified. This species-specific primer focused on a single nucleotide mismatch in the 3'-terminal base of a primer designed in the mitochondrial cytochrome c oxidase (COI) subunit I DNA. To optimize the duplex PCR condition, gradient PCR reactions were conducted to determine the primer annealing temperature and the primer concentration. The PCR's product was observed on the gel, suggesting that DNA molecules may be useful in differentiating the two species. The length of the amplification fragments were 103 bp for Trachurus japonicus and 214 bp for Trachurus novaezelandiae, which, along with the species-specific primer visualized by agarose gel electrophoresis, enabled accurate distinction of the species of the Trachurus genus. These results indicate that the duplex PCR, which has a species-specific primer based on single nucleotide polymorphism (SNP), can be useful for rapidly differentiating the two species of Trachurus. This duplex PCR analysis is simple, rapid, and reliable, and could be beneficial to protecting consumers' rights.
Kim, Dae Yong;Son, Hyun Jin;Chung, Myoung Ja;Kang, Myoung Jae
Journal of Korean Neurosurgical Society
/
v.30
no.4
/
pp.443-450
/
2001
Objective : The cyclin-dependent kinase inhibitor $p27^{kip1}$ protein is a negative regulator of the cell cycle, and its degradation is required for entry into the S phase. Loss of $p27^{kip1}$ expression has been reported to be associated with aggressive behavior in a variety of tumors of epithelial and lymphoid origin. However, its association with various astrocytic tumors has not been clearly demonstrated. We studied to investigate the relationship of $p27^{kip1}$ expression with the biological behavior of astrocytic tumors in addition to study on the role of $p27^{kip1}$ in the tumorigenesis of these tumors. Patients and Methods : From 1990 to 1998, a total of 29 astrocytic tumor of all grades obtained by operative resection were included for evaluation. We studied the expression of $p27^{kip1}$ protein immunohistochemical assay in astrocytic tumors and compared the findings with the clinicopathologic parameters. Immunohistochemical staining was performed on formalin-fixed paraffin-embedded sections by the avidin-biotin-peroxidase complex method. According to WHO classification, all cases were divided into astrocytomas(4 cases), anaplastic astrocytomas(9 cases), and glioblastomas(16 cases) by 3 pathologists. Clinical information was obtained from medical records, and others such as location and size of tumors from imaging studies. Results : Mean $p27^{kip1}$ protein labeling indexes(LI, mean${\pm}$standard deviation) of astrocytomas, anaplastic astrocytomas, and glioblastomas were $80.6{\pm}9.1$, $63.6{\pm}21.0$, and $28.9{\pm}18.7$, respectively, and were inversely correlated with grade of glial tumors(p<0.0001). Mean $p27^{kip1}$ protein LI in the recurrent group was lower than that in the nonrecurrent group, but there was no significant difference statistically(p=0.464). Additionally, $p27^{kip1}$ protein expression did not show any significant relationship to other prognostic factors such as age(p=0.1643), tumor size(p=0.8), or location(p=0.8). Conclusion : These results suggested that reduced expression of $p27^{kip1}$ protein may play a important role in the malignant transformation process of astrocytic tumor cells.
In this paper, we propose a highly precise vehicle detection method with low false alarm using billboard sweep stereo matching and multi-stage hypothesis generation. First, we capture stereo images from cameras established in front of the vehicle and obtain the disparity map in which the regions of ground plane or background are removed using billboard sweep stereo matching algorithm. And then, we perform the vehicle detection and tracking on the labeled disparity map. The vehicle detection and tracking consists of three steps. In the learning step, the SVM(support vector machine) classifier is obtained using the features extracted from the gabor filter. The second step is the vehicle detection which performs the sobel edge detection in the image of the left camera and extracts candidates of the vehicle using edge image and billboard sweep stereo disparity map. The final step is the vehicle tracking using template matching in the next frame. Removal process of the tracking regions improves the system performance in the candidate region of the vehicle on the succeeding frames.
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