Tryptic Digestion and Cytochalasin B Binding Assay of the Human HepG2-Type Glucose Transporter Expressed in Spodoptera frugiperda Clone 21-AE Cells

이종기 대한의생명과학회 2005 Biomedical Science Letters Vol.11 No.1

The number of sites at which a protein can be readily cleaved by a proteolytic enzyme is greatly influenced by its three-dimensional structure. For native, properly-folded proteins both the rate of cleavage and number of sites at which cleavage takes place are usually much less than for the denatured protein. In order to compare the tertiary structure of recombinant HepG2 type glucose transporter with that of its native counterpart in the erythrocyte, the pattern of tryptic cleavage of the protein expressed in insect cell membranes was therefore examined. After 30 minutes digestion, a fragment of approximate Mr 19,000~21,000 was generated. In addition to this, there were two less intensely stained fragments of apparent Mr 28,000 and 17,000. The pattern of labelling was similar up to 2 hours of digestion. However, the fragments of Mr 19,000~21,000 and Mr 17,000 were no longer detectable after 4 hours digestion. The observation of a very similar pattern of fragments yielded by tryptic digestion of the HepG2 type transporter expressed in insect cells suggests that the recombinant protein exhibits a tertiary structure similar if not identical to that of its human counterpart. Also, the endogenous sugar transporter(s) present in Sf21 cells did not bind cytochalasin B, the potent transporter inhibitor. Therefore, the baculovirus/Spodoptera frugiperda (Sf) cell expression system could be very useful for production of large amounts of human glucose transporters, heterologously.

일본 근대작가의 자살에 관한 연구 : 北村透谷와 川上眉山을 중심으로 Of Kita Mura, To-koku and Kawa Kami, Bi-Zan

정병남,이기섭 진주여자전문대학 1991 論文集 Vol.14 No.-

The purpose of this study was to introduce the writers, such as Kita Mura, To-Koku(北村透谷)and Kawa Kami, Bi-Zan(川上眉山), who committed suicide in the modern age of Japan, and to investigate their suicide. The result of this study suggested that the suicide motives of Kita Mura were the frustration and the loss of life activity come from the gap between his ideal and reality. The factors which Kawa Kami committed suicide were considered as the maladjustment to rapidly changing socicety, the pain of creative writing, and that of living affairs. By the way, as the motive of suicide is very complex and its behavior is individual, a more scientific and objective investigation should be conducted in further reseatrch.

연산동에서 집단발생한 장티브스환자에 대한 세균학적 관찰

李潤日,金慶淑,金明淑,黃基銑,李鍾訓 中央醫學社 1973 中央醫學 Vol.25 No.3

An attempt was made to isolate the causative agent from a group of febrile patients in a construction camp, where the endemic was occurred in a fashion of explosive, and it seemed to be a salmonellosis on the clinical symptoms. Direct rectal swabs and' food samples were taken repeatedly from febrile patients, and only the rectal swabs from the healthy persons to obtain the bacteriological specimens. Sera were also obtained from both healthy persons and febrile patients. Water of glen which penetrates through the camp could be worthy of notice as an infectious source, because one chronic typhoid carrier was found in a slum which was located on a steep ascent just outside of the camp. Typhoid bacilli was isolated from the specimens of 17 patients (58.8%) out of 29 febrile patients, and from 5 out of 92 healthy persons. Widal agglutinin rise was so delayed that it was found not so worthy of examination in the situation of the early management of the patients for prevention of the endemic.

β-CuZn 합금의 항복강도 이상거동에 미치는 변형률 속도와 온도의 영향

이기안,장영원,이종수 대한금속재료학회 2003 대한금속·재료학회지 Vol.41 No.3

A series of tensile and load relaxation behaviors in B2 type β-CuZn alloy have been examined to clarify the strain rate & the temperature dependence of yield stress anomaly. It has been found in tensile tests that the region showing anomalous peak of yield stress moves to a higher stress and higher temperature region with increasing strain rate. The anomalous yielding region below peak temperature(T_p) revealed slightly positive strain rate dependence, while the region above T_p showed considerably positive strain rate dependence. The slip lines formed below peak temperatures (T_P) were clearly visible with naked eye(coarse slip lines), and became finer near T_p(fine slip lines), while they disappeared completely above T_p. Based on the different tendency of temperature and strain rate dependence of anomalous yielding as the above-mentioned results, two types of deformation modes were suggested and incorporated into the flow curve analysis, which was in accord well with the experimental results. The slightly positive strain rate dependence of yield stress anomaly indicated that the yield stress anomaly in β-CuZn alloy mainly seemed to be attributed to the vacancy hardening.

β-CuZn 합금의 인장 및 파괴 거동에 미치는 변형률 속도와 온도의 영향

이기안,이종수 대한금속재료학회 2003 대한금속·재료학회지 Vol.41 No.12

A series of tensile tests were carried out on a B2-type b-CuZn alloy in the temperature range, room temperature to 400oC at three different strain rates (10^(-5) sec^(-1), 10^(-4) sec^(-1), 5.2x10^(-2) sec^(-1)). The stress-strain curves could be divided into three types. Type I indicates a remarkably linear sustained work hardening to fracture, type II work hardening and subsequent necking, type III yield drop behavior and stress saturation, respectively; corresponding to the insensitive yield stress region, anomalous yield stress region, and yield stress decreasing region, in the yield strength vs. temperature graph. The ultimate tensile strength basically decreased as temperature increased, only showing characterized strength hump-up region just below T_(p). The fracture mode was changed mainly from transgranular at room temperature to intergranular at high temperatures. The elongation increases with increasing temperature in region I, and then slightly decreases in region II (anomalous yield stress region). When the specimens were tensile-fractured at high temperatures (region III, above Tp), the elongation drastically increased with decreasing strain rate, for instance from 12% at 5.2x10^(-2) sec^(-1) to 87% at 10^(-5) sec^(-1). The dependence of ductility on strain rate was mainly attributed to the thermally activated <100> dislocation climb, inducing stress relief at the grain boundaries, as evidenced by microvoids at grain facets.

耳性側頭葉膿瘍의 1剖檢例

姜基澤,李鍾元 全南大醫大 1964 전남의대학술지 Vol.1 No.1

貧血症과 赤血球過多症에 있어서 血淸 Erythropoietin 活性度에 關한 硏究

尹起英,李夏白,朴鍾茂 한양대학교 의과대학 1985 한양의대 학술지 Vol.5 No.2

Erythropoietin (Ep) is a glycoprotein hormone of mostly renal origin that stimulates red cell production in response to tissue hypoxia. Serum Ep activities provide important clinical clues in making the differential diagnosis of some types of anemia and polycythemia. The study was performed in order to evaluate the diagnostic value of serum Ep activities in 29 normal subjects, 43 anemic patients and 15 secondary polycythemia patients and to find out the relationship between hemoglobin concentration and serum Ep activities in these patients using the Enzyme Immunoassay with Second International Preparation for Ep. The results obtained were as follows: 1. Ep activities in sera from 29 normal subjects among children ranged from 11 to 72 mU/ml, with a mean of 37.10±16.54 mU/ml. There was no significant difference between the serum Ep activities in male and female subjects (p>0.05). 2. Ep activities in sera from 22 patients with iron deficiency anemia ranged from 48 to 400 mU/ml, with a mean of 153.5 mU/ml which was about 4 times higher than normal value. Ep activities in sera from 18 anemic patients with acute infection ranged from 35 to 290 mU/ml, with a mean of 134.05 mU/ml which was about 3.5 times higher than normal value. Ep activities in sera from 3 patients with aplastic anemic ranged from 620 to 840 mU/ml, with a mean of 720 mU/ml which was about 20 times higher than normal value. 3. Ep activities in sera from 15 patients with secondary polycythemia ranged from 12 to 140 mU/ml. In 10 of 15 patients, the mean Ep activities was 87.50 ± 25.60 mU/ml, which was about 2 times higher than normal value. In 5 of 15 patients, Ep activities were within normal range as a mean of 35.80 ± 17.23 mU/ml. In summary, a significant negative correlation was found between the hemoglobin concentration and the logarithm of the serum Ep activities in 43 patients with anemia (r=0.73, Y=3.6766-0.1802x) and Ep activities in patients with aplastic anemia was higher than those in patients with iron deficiency anemia at comparable hemoglobin concentration. From the above results, it was concluded that serum Ep activities in anemic patients are not only inversely related to the hemoglobin concentration but also to the activity of the erythroid bone marrow. While no significant correlation was found between hemoglobin concentration, hematocrit percentages and Ep activities in 15 patients with secondary polycythemia (r=0.18, 0.39 respectively).

3차원 전산화단층촬영 영상을 이용한 안면 연조직 두께 계측의 임상적 유용성

정호걸,김기덕,한승호,허경석,이제범,박혁,최성호,김종관,박창서 대한구강악안면방사선학회 2006 Imaging Science in Dentistry Vol.36 No.2

Purpose : To evaluate clinical usefulness of facial soft tissue thickness measurement using 3D computed tomographic images. Materials and Methods : One cadaver that had sound facial soft tissues was chosen for the study. The cadaver was scanned with a Helical CT under following scanning protocols about slice thickness and table speed; 3 mm and 3 mm/sec, 5 mm and 5 mm/sec, 7 mm and 7 mm/sec. The acquired data were reconstructed 1.5, 2.5, 3.5 mm reconstruction interval respectively and the images were transferred to a personal computer. Using a program developed to measure facial soft tissue thickness in 3D image, the facial soft tissue thickness was measured. After the ten-time repeation of the measurement for ten times, repeated measure analysis of variance (ANOVA) was adopted to compare and analyze the measurements using the three scanning protocols. Comparison according to the areas was analyzed by Mann-Whitney test. Results : There were no statistically significant intraobserver differences in the measurements of the facial soft tissue thickness using the three scanning protocols (p>0.05). There were no statistically significant differences between measurements in the 3 mm slice thickness and those in the 5 mm, 7 mm slice thickness (p>0.05). There were statistical differences in the 14 of the total 30 measured points in the 5 mm slice thickness and 22 in the 7mm slice thickness. Conclusion : The facial soft tissue thickness measurement using 3D images of 7 mm slice thickness is acceptable clinically, but those of 5 mm slice thickness is recommended for the more accurate measurement.

Effects of Phloretin, Cytochalasin B, and D-Fructose on 2-deoxy-D-Glucose Transport of the Glucose Transport System Present in Spodoptera frugiperda Clone 21-AE Cells

Lee Chong-Kee 대한의생명과학회 2006 Biomedical Science Letters Vol.12 No.1

The baculovirus expression system is a powerful method for producing large amounts of the human erythrocyte-type glucose transport protein, heterologously. Characterization of the expressed protein is expected to show its ability to transport sugars directly. To achieve this, it is a prerequisite to know the properties of the endogenous sugar transport system in Spodoptera frugiperda Clone 21 (Sf21) cells, which are commonly employed as a host permissive cell line to support the baculovirus replication. The Sf21 cells can grow well on TC-100 medium that contains 0.1% D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transport system. However, unlike the human glucose transport protein that has a broad substrate and inhibitor specificity, very little is known about the nature of the endogenous sugar transport system in Sf21 cells. In order to characterize further the inhibitor recognition properties of the Sf21 cell transporter, the ability of phloretin, cytochalasin B and D-fructose to inhibit 2-deoxy-D-glucose (2dGlc) transport was examined by measuring inhibition constants $(K_i)$. The $K_i's$ for reversible inhibitors were determined from plots of uptake versus inhibitor concentration. The 2dGlc transport in the Sf21 cells was very potently inhibited by phloretin, the aglucone of phlorizin with a $K_i$ similar to the value of about $2{\mu}M$ reported for inhibition of glucose transport in human erythrocytes. However, the Sf21 cell transport system was found to differ from the human transport protein in being much less sensitive to inhibition by cytochalasin B (apparent $K_i$ approximately $10\;{\mu}M$). In contrast, It is reported that the inhibitor binds the human erythrocyte counterpart with a $K_d$ of approximately $0.12\;{\mu}M$. Interestingly, the Sf21 glucose transport system also appeared to have high affinity for D-fructose with a $K_i$ of approximately 5mM, contrasting the reported $K_m$ of the human erythrocyte transport protein for the ketose of 1.5M.

1972년 유행한 유행성 출혈성 결막염 병원체의 항원분석 : Ⅰ.분리 병원체의 성상과 교차중화반응상의 항원성

이종훈,이연태,황기선,최상규,고광균,이윤일,면지선,기용숙 中央醫學社 1974 中央醫學 Vol.26 No.5

An unusual type of conjunctivitis was prevailing in an epidemic fashion in Korea during 1971 and 1972. The conjunctivitis was characterized by severe subconjunctival hemorrhage, lid-swelling and tearing. Its clinical course, however,. was usually shorter than the ordinary epidemic keratoconjunctivitis and completely recovered within 2 weeks after the onset. This disease seemed to' be highly contagious in unhygienic and crowded public places. An attempt was made to isolate the causative agent of the epidemic hemorrhagic conjunctivitis by HeLa and LLCMK2 cell cultures. The cytopathic agent was filtered through millipore membrane filters to get rough estimates of the size. Nucleic acid type, and ethyl ether sensitivity and acid sensitivity were also examined. Sections of LLCMK2 cells infected with the isolated agent were stained with uranylacetate and lead citrate and subjected to electron microscopy to determine the intracellular position and structure of the agents. Cross-neutralization test was done for comparison of antigenic similarity between the isolates and YC-71648 versus with their immune guinea pig sera. In addition the neutralization test was. carried out on 41 sera 6 months after complete recovery from the epidemic hemorrhagic conjunctivitis, 77 healthy sera. The amount of 100 TCID50 of virus was Tested' against 2 fold-serial-dilution of these sera. Virus serum mixtures Were incubated at 37°C for 60 minutes and 0.2 ml of the each mixtures was inoculated into 5 LLCMK2 tube cultures. The inoculated cell cultures were adsorbed at 37°C for 2 hours and thereafter kept at 32°C, and a cytopathic effect was observed daily for a week. The results obtained were as follows: Two strains of cytopathic agents were isolated by LLCMK2 cell cultures out of 7 conjunctival swabs taken within 2 days after the onset from patients with the -epidemic hemorr hagic conjunctivitis in an outbreak' 1972. At the first to third passages by LLCMKZ cell cultures, the cytopathic effect-was often indistinct on the cell monolayer, but it became more clear cut on the 3rd or 4th day of cultivation at the 4th passage, whereas the cytopathic effect was not distinct even after repeated passages on HeLa cell cultures. Two strains of the isolates could have been gradually adapted to LLCMK2 by passages, as the infectivity titer increased up to 10'-7. ' TCIDSo per ml at 9th to 10th passages from the titer below 104'° TCID$° per ml until the 8th passages. The cytopathic agent was very small particle which passed through a membrane filter of pore size 50nm, and spherical in form by elects onphotomicrography. It was resistant to ethyl ether and acid (pH 3.0) treatment, and the infectivity titer did not drop in the presence of 10-4-°' Mole per ml of 5-iododeoxyur dine in the culture. There was a definite cross-neutralization between the YC-71-648 (virus strain isolated in. Japan) and the two isolates with their immune guinea pig sera. A -definite rise in neutralizing antibody titer against the two isolates as well as .Japan strain, was observed on 41 sera obtained from persons 6 months after recovery from the epidemic hemorrhagic conjunctivitis. 'On the other hand, the neutralizing antibody was not detected on 65-66 (84.5-'85.7%) out of 77 healthy sera, except 11-12 sera (14.315.5%) responded to 1 : 4 serum dilution.