The baculovirus/insect cell expression system is of great value in the study of structure-function relationships in mammalian glucose-transport proteins by site-directed mutagenesis and for the large-scale production of these proteins for mechanistic and biochemical studies. Spodoptera frugiperda Clone 21 (Sf2l) cells grow well on TC-100 medium that contains $0.1\%$ D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, very little is known about the properties of the endogenous sugar transporter(s) in Sf2l cells, although a saturable transport system for hexose uptake has been previously revealed in the Sf cells. In order to further examine the substrate and inhibitor recognition properties of the Sf2l cell transporter, the ability of hexoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. The $K_i's$ for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. Transport was effectively inhibited by D-mannose and D-glucose. Of the hexoses tested, L-glucose had the least effect on 2dGlc transport in the Sf2l cells, indicating that the transport is stereoselective. Unlike the human HepG2 type glucose transport system, D-mannose had a somewhat greater affinity for the Sf2l cell transporter than D-glucose, implying that the hydroxyl group at the C-2 position is not necessary for strong binding. However, epimerization at the C-4 position of D-glucose (D-galactose) resulted in a dramatic decrease in affinity of the hexose for the Sf2l cell transporter. Such a lowering of affinity might be the result of the involvement of the C-4 hydroxyl in hydrogen bonding. It is therefore suggested that Sf2l cells were found to contain an endogenous sugar transport activity that in several aspects resembles the human HepG2 type glucose transporter, although the insect and human transporters do differ in their affinity for cytochalasin B.
Insect cells such as Spodoptera frugiperda Clone 9 (Sf9) cells are widely chosen as the host for heterologous expression of a mammalian sugar transport protein using the baculovirus expression system. Characterization of the expressed protein is expected to include assay of its function, including its ability to transport sugars and to bind inhibitory ligands such as cytochalasin B. It is therefore very important first to establish the transport characteristics and other properties of the endogenous sugar transport proteins of the host insect cells. However, very little is known of the transport characteristics of Sf9 cells, although their ability to grow on TC-100 medium strongly suggested the presence of endogenous glucose transport system. In order to investigate the substrate and inhibitor recognition properties of the Sf9 cell transporter, the ability of pentoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. To determine the time period over which of sugar into the Sf cells was linear, the uptake of 2dGlc 0.1mM extracellular concentration was measured over periods ranging from 30 seconds to 30 minutes. The uptake was linear for at least 2 minutes at the concentration, implying that uptake made over a 1 minute time course would reflect initial rates of the sugar uptake. The data have also revealed the existence of a saturable transport system for pentose uptake by the insect cells. The transport was inhibited by D-xylose and D-ribose, although not as effective as hexoses. However, L-xylose had a little effect on 2dGlc transport in the Sf9 cells, indicating that the transport is stereoselective. Unlike the human erythrocyte-type glucose transport system, D-ribose had a somewhat greater apparent affinity for the Sf9 cell transporter than D-xylose. It is therefore concluded that Sf9 cells contain an endogenous sugar transport activity that in some aspects resembled the human erythrocyte-type counterpart, although the Sf9 and human transport systems do differ in their affinity for cytochalasin B.
Kim, Moo-Seong;Kim, Kyoung-Ryong;Ahn, Do-Whan;Park, Yang-Saeng
The Korean Journal of Physiology and Pharmacology
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v.4
no.1
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pp.63-72
/
2000
Chronic exposure to cadmium (Cd) results in an inhibition of protein endocytosis in the renal proximal tubule, leading to proteinuria. In order to gain insight into the mechanism by which Cd impairs the protein endocytosis, we investigated the effect of Cd on the acidification of renal cortical endocytotic vesicles (endosomes). The endosomal acidification was assessed by measuring the pH gradient-dependent fluorescence change, using acridine orange or FITC-dextran as a probe. In renal endosomes isolated from Cd-intoxicated rats, the $V_{max}$ of ATP-driven fluorescence quenching ($H^+-ATPase$ dependent intravesicular acidification) was significantly attenuated with no substantial changes in the apparent $K_m,$ indicating that the capacity of acidification was reduced. When endosomes from normal animals were directly exposed to free Cd in vitro, the $V_{max}$ was slightly reduced, whereas the $K_m$ was markedly increased, implying that the biochemical property of the $H^+-ATPase$ was altered by Cd. In endosomes exposed to free Cd in vitro, the rate of dissipation of the transmembrane pH gradient after $H^+-ATPase$ inhibition appeared to be significantly faster compared to that in normal endosomes, indicating that the $H^+-conductance$ of the membrane was increased by Cd. These results suggest that in long-term Cd-exposed animals, free Cd ions liberated in the proximal tubular cytoplasm by lysosomal degradation of cadmium-metallothionein complex (CdMT) may impair endosomal acidification 1) by reducing the $H^+-ATPase$ density in the endosomal membrane, 2) by suppressing the intrinsic $H^+-ATPase$ activity, and 3) possibly by increasing the membrane conductance to $H^+$ ion. Such effects of Cd could be responsible for the alterations of proximal tubular endocytotic activities, protein reabsorption and various transporter distributions observed in Cd-exposed cells and animals.
The purpose of this study was to develop an effective horticultural activity program for the vocational rehabilitation of people with mental disabilities. The subjects of this study were seven persons with mental disabilities in the vocational rehabilitation team of a public health center located in G city, Gyeongsangbuk-do. A total of 10 horticultural programs were held from October 10 to December 11, 2018. Five programs were selected among those that were easy for the persons with mental illness to handle and their preference was high. In each session, the same task was repeated 10 times and finally changes in hand functions and working speed were measured. As a result, the assembly task of the Purdue Pegboard test was 23.6 (SD = 10.5) before the program, 26.6 (SD = 11.5) after planting, 27.7 (SD = 12.2) after wrapping pots, and 28.0 (SD = 13.9) after making mini flower baskets, 26.9 (SD = 12.0) after wrapping a single flower, and 29.1 (SD = 11.9) after making corsages. There was a significant difference between the five programs (p = .016). As a result of measuring the time taken to produce the same 10 products, the average planting time was 9.9 (SD = 4.0) minutes in the first class and 6.0 (SD = 2.3) minutes in the second class (p = .018), and making mini flower baskets was reduced from 35.2 (SD = 10.1) minutes to 21.0 (SD = 7.7) minutes (p = .018), wrapping a single flower from 23.3 (SD = 7.9) minutes to 16.2 (SD = 7.3) minutes (p = .043), and making corsages from 53.6 (SD = 15.9) to 40.8 (SD = 16.8) minutes (p = .043). The working time was significantly shortened in the second class compared to the first class. The class for wrapping pots was shortened from 52.7 (SD = 11.7) to 49.6 (SD = 17.8) minutes, but there was no significant difference (p = .398).
Journal of The Korean Association For Science Education
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v.31
no.5
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pp.709-719
/
2011
The purpose of this study was to identify the differences in classification skills shown in classification activities between the gifted and regular students in elementary schools. The subjects for the research consisted of six gifted students in an institute for the gifted for science annexed to P school district in Gangwon-do and 6 students at B and M general elementary schools. Results were as follows: The time taken for classification activities of the gifted was shorter than regular regardless of subjects for classifying. The number of standards for classifying for the gifted was more than regular students. Coefficient for measuring classification skills of the gifted was higher than regulars regardless of age. Consequently, there was a difference in the time taken for classifying and generating the number of standards and in a numerical index of classification activities performed at science classes between the science gifted and the regular students.
The purpose of the study was to identify disease characteristics and behavior pattern of treatment for patients with Fibromyalgia. This study was carried out between May to Aug. in 1998 through direct interview in Rheumatism clinic at H. University Hospital and subject in this study were 125 outpatients diagnosed with Fibromyalgia. Collected data were analyzed by descriptive statistics and t-test, ANOVA using SPSS Window program. The results of this study are as follows. 1. General Characteristics : All of the persons with Fibromyalgia were female who were mostly in their forties(37.5%). A third of them(38.4%) were graduated from high school. The greatest part of them(54.4%) were christians but little part of them(16.8%) were employed 2. Disease Characteristics : They have struggled with Fibromyalgia for 10 years on an average. About half of them(56.9%) suffered from Fibromyalgia only but the others had another diseases which were in greatest part occupied by Osteoarthritis. The number of tender point which is a feature of Fibromyalgia differed according to measuring criteria. Yunus criteria. however, was proved to be the most proper measuring criteria than any other method as it showed high correlations between symptoms and physical activities. The most serious symptoms that complained the patients among subjective symptoms are pain, sleep disorder, and fatigue in sequence, and activities most hard to do among physical activities are washing by hand, scrubbing by hand, and shopping in sequence. 3. Behavior Pattern of Treatment : The largest part of them(42.4%) had received medical treatment after they were determined to have the disease and most of them were taking medicine as prescribed by physician(88.8%) or other medicines(16.8%), Of them, two thirds stated that the medicine they took were effective. Around a third of them took exercises mostly composed of swimming. The medical institution they visited in the past were orthopedic surgical department, Oriental hospital, physical therapy department in sequence and, in a slight percent(11.4%), psychiatric department. 4. Relations between Disease Characteristics and Behavior pattern of Treatment : It shows that number of tender point and level of symptom are significantly different according to duration of disease and medical intervention. As a results, it can be suggest that persons with Fibromyalgia need to take exercises and medicine continually regardless of medical treatment as it is a chronic disease whose symptoms are hardly mitigated.
Park, Kyung-Moo;Yeom, Do-Sung;Kim, Hak-Ju;Ko, Seong-Gyu;Lim, Hyung-Ho;Song, Yun-Kyung
Journal of Korean Medicine for Obesity Research
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v.12
no.2
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pp.44-55
/
2012
Objectives: The purpose of this study is to investigate biological effects of Euiiin-tang(Y$\grave{i}$y$\check{i}$r$\acute{e}$n-t$\bar{a}$ng) extracts by extraction methods. Methods: To investigate the effects of anti-oxidative and anti-inflammatory activity, we measured DPPH radical scavenging assay, collagenase inhibition assay and COX-2 inhibition assay. Results: In case of measuring anti-oxidative and anti-inflammatory activities of Euiiin-tang(Y$\grave{i}$y$\check{i}$r$\acute{e}$n-t$\bar{a}$ng) by collagenase and COX-2 inhibition activity, it came out that Euiiin-tang(Y$\grave{i}$y$\check{i}$r$\acute{e}$n-t$\bar{a}$ng) was unable to inhibit collagenase, but was effective to inhibit COX-2. Cytotoxicity analysis of Euiiin-tang(Y$\grave{i}$y$\check{i}$r$\acute{e}$n-t$\bar{a}$ng) extracts by MTT assay principles, there was no cytotoxicity in aqueous and 50% ethanol extracts in any level of concentration. Conclusions: This results suggest that Euiiin-tang(Y$\grave{i}$y$\check{i}$r$\acute{e}$n-t$\bar{a}$ng) extracts have anti-oxidative and anti-inflammatory activities.
Kim, Seok-Gi;An, Guk-Hwan;Yoon, Seung-Won;Lee, Young-Chun;Ha, Sang-Do
Korean Journal of Food Science and Technology
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v.35
no.3
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pp.519-526
/
2003
Pancreatic lipase and ${\alpha}$-amylase inhibitory activities in purified extracts of pumpkin and job's tear were studied. Pancreatic lipase inhibitory activity was determined by measuring the rate of releasing oleic acid from triolein, and ${\alpha}$-amylase inhibitory activity was determined by iodometric method. The extracts of pumpkin and job's tear were purified using silica gel and C-18 gel column chromatographies. Treatment of pumpkin extract $(120\;{\mu}g/mL)$ in 3T3-L1 preadipocyte decreased differentiation about 95% and blocked accumulation of lipid. Body weights of rats fed high-fat diet containing dietary supplement decreased about 13% as compared with those fed only highfat diet. These results revealed dietary supplement is a good obesity-reducing material for decreasing lipid and carbohydrate absorptions.
Nam, Sohyun;Rhee, Young Kyoung;Hong, Hee-Do;Lee, Young-Chul;Kim, Young-Chan;Shin, Kwang-Soon;Cho, Chang-Won
The Korean Journal of Food And Nutrition
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v.25
no.4
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pp.755-761
/
2012
In this study, we examined immuno-modulatory activities of crude polysaccharides from wild ginseng adventitious roots (WGAR). The crude polysaccharide (WGAR-CP) was isolated from WGAR by hot water extraction, ethanol precipitation, and dialysis. The major constituents in WGAR-CP were neutral sugar (64.77%), and uronic acid (34.32%). WGAR-CP demonstrated anti-complementary activity dose-dependently. The immuno-modulatory effects of WGAR-CP were also analyzed by measuring nitric oxide and cytokines in the supernatants of mouse peritoneal macrophages. Mouse peritoneal macrophages stimulated with WGAR-CP produced nitric oxide and various cytokines such as interleukin (IL)-6 and IL-12 in a dose-dependent manner. In conclusion, WGAR-CP may have immuno-modulatory activities by activating a complementary system and macrophages, which produces cytokines.
Huh Hyun Do;Park Sung Yong;Lee Rena J;Shin Dong Oh;Kwon Soo Il;Loh John J K;Choi Jinho
Progress in Medical Physics
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v.15
no.4
/
pp.192-196
/
2004
The activity of Ir-192 sources for high dose rate (HDR) Brachytherapy in Korea were measured by using the well-type chamber and using the calibration Jig with the Farmer-type ionization chamber to compare the manufacturer certificated source strength which is supplied with each new Ir-192 source. The activity of two different source models used in six hospitals were measured. The range of measured activities to the manufacturer's suggested ones was -2.40% to +3.31% for the calibration Jig and -3.12% to 0.00% for the well-type chamber system. The source strength values given by the manufacturer for the 6 sources were within ${\pm}5%$ for the two different measuring equipment. Our results demonstrate that well-type chamber as wall as Farmer-type chamber system are appropriate system for the routine source calibration procedures in HDR brachytherapy. Whenever a new source is installed to use in clinics, a source calibration should be carried out.
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