Journal of Yeungnam Medical Science

Jung Hye Kim

19 Articles

Coordination and Activation of Biotechnology-Related R&D Budget of Korea: Jung Hye Kim; Yeungnam Univ J Med. 2007;24(2 Suppl):S75-86. Published online December 31, 2007; DOI: https://doi.org/10.12701/yujm.2007.24.2S.S75

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Abstract PDF: Importance of science and technology is based on modern society which means knowledge based society. Now, today’s quality of science and technology in Korea says that is ordered as 8th powerful nationality in the world. Research and development budget of our government is over KRW 9.8 trillion dollars in 2007, it is ordered amount of 8th national in the world. First nation is America, followed Japan, Germany, France, England, Italy, China. By the Swiss IMD report, it is evaluated as competitive power of science of our country is 7th order and technology is 6th order in the world. These show brilliant achievement by recent many research products. 1) In order to increase investment efficiency of research & development of government, reorganization of government setups were carried out national innovation system (NIS) in Oct. 2004. Policy of coordination and division of government R&D budget is in force strategy by selection and convergence, total roadmap in mid- and long-term by technological field. The priority order by total roadmap take biological field as first. In the present 2007, government budget of biological field occupied 16.5%, it is developed 17% in next year. Next order is followed the Environment, the Space and the Universe. Strategy of R&D investment of fields of information and electron and field of machinery and the manufacturing industry will be decrease. National R&D investment will be used as an advanced country form. It will be induced to increase a private enterprise. These field will be decrease a weight. Especially, a scientist for life have to effort to development of global new drug by Korea-America FTA (Free Trade Agreement) and we will see what we can do a sense of responsibility and good results.

Anticancer Effects of Gleditsin in Human Mammary Cancer Cells: Hyun Sook Ko, Kyung Won Kang, Jung Hye Kim; Yeungnam Univ J Med. 2007;24(2 Suppl):S580-590. Published online December 31, 2007; DOI: https://doi.org/10.12701/yujm.2007.24.2S.S580

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Abstract PDF: Background
：Gleditsin is a herb medicine from extracted by Gleditschia spina. In oder to investigate anticancer effects of gleditsin in human various breast cancer cells, we tested with gleditsin on cytotoxicity of materials, observed to cell survival and cell cycle progression, and analyzed in starvation condition. Materials and Methods：The cytoxicity and cell cycle progression were analyzed in human breast cells, MCF-10A and human breast cancer cell lines, MDA-MB-231, MDA-MB-361, and MDA-MB-435. IC50s of breast cancer cell lines were measured by MTT assay. The cell cycle were showed by flow cytometric analysis in cells treated with gleditsin. We analyzed DNA content of sub-Go/G1 phase, it was detected apoptosis.
Results
：Cell survivals were decreased in a dose-dependent manner by the treatment of cells with gleditsin. IC50s were 4.11-fold higher in MDA-MB-435, 2.53-fold higher in MDAMB- 231, and 2.55-fold higher in MDA-MB-361 than in normal breast cells. Flow cytometric analysis showed that sub-G0/G1 fractions in cancer cells treated with gleditsin were higher than that normal cells, suggesting that increases in cytotoxicity of cancer cells by gleditsin were resulted from apoptosis. Cell cycle progression was also changed by the treatment of gleditsin. The treatments of gleditsin resulted in a decrease in G1 phase and an increase in G2/M phase in normal breast cells as well as cancer cell lines. Apoptotic cell death was synergistically increased by cell starvation and gleditsin treatments in cancer cells. MDAMB- 435 cells were more sensitive to apoptotic cell death by gleditsin than other cells.
Conclusion
：An anti-tumor effect of gleditsin was selectively higher in hman breast cancer cells than in normal human breast cells, and was mediated by apoptotic cell death.

Association Analyses of beta3AR Trp64Arg and UCP-2 -866G/A Polymorphisms with Body Mass Index in Korean.: Hong Soo Jung, Joo Hyun Lee, Jun Sakong, Sung Wook Bae, Jung Hye Kim, Jae Ryong Kim; Yeungnam Univ J Med. 2007;24(2):252-261. Published online December 31, 2007; DOI: https://doi.org/10.12701/yujm.2007.24.2.252

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Abstract PDF

BACKGROUND
Obesity is the most common nutritional disorder in Western society as well as in Korea. Obesity results from a combination of genetic, environmental, and behavioral factors. MATERIALS AND METHODS: In an attempt to investigate the association of obesity with its candidate genes, beta3 adrenergic receptor (beta3AR) and uncoupling protein 2 (UCP2), we analyzed polymorphisms of beta3AR Trp64Arg and UCP2 -866G/A by PCR-RFLP analysis and the obesity-related phenotypes, including body mass index (BMI), fasting glucose concentration, and plasma lipid profiles in 750 subjects. RESULTS: The Trp64Arg polymorphism in the beta3AR gene was not statistically associated with the BMI. The UCP2 -866G/A polymorphism was significantly higher in obese than in non-obese subjects (P<0.05). However, the UCP2 -866A/A polymorphism was higher in the non-obese subjects. CONCLUSION: These results suggest that the UCP2 -866G/A polymorphism might be more useful for the prediction of obesity and obesity-associated diseases in Korean patients than the beta3AR Trp64Arg polymorphism.

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Clinical Course of Bipolar Disorder in Children and Adolescents
Na-Ri Kang, Young-Sook Kwack
Journal of korean Academy of Child and Adolescent Psychiatry.2012; 23(1): 3. CrossRef

Profile of Gene Expression Changes During Doxorubicin Induced Apoptosis of Saos-2.: Jeong Sook Lim, Min Jae Bae, Suk Hwan Baek, Jae Ryong Kim, Jung Hye Kim, Seong Yong Kim; Yeungnam Univ J Med. 2005;22(2):221-240. Published online December 31, 2005; DOI: https://doi.org/10.12701/yujm.2005.22.2.221

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Abstract PDF: BACKGROUND
Doxorubicin has proved to be a useful chemotherapeutic agent especially for osteogenic sarcoma. It induces cancer cell death via apoptosis. MATERIALS AND METHODS: To explore and analyze the changes of gene expression during doxorubicin induced apoptosis on human osteogenic sarcoma, Saos-2 cell, cDNA microarray was performed. After treatment with doxorubicin, total RNA was purified and expressed genes were investigated with a 17k human cDNA microarray. RESULTS: For analysis of the cDNA microarray, the genes were filtered using the sum of the median value of Cy3 and Cy5 signal intensity of greater than 800. Expression of 264 genes was changed by more than 2 fold, and the expression of 35 genes was changed more than 3 fold after treatment with doxorubicin. The genes were primarily related to cell death, cell growth and maintenance, signal transduction, cellular component, transport, and metabolism. CONCLUSION: Treatment with doxorubicin induced expressional change of many genes. Some of the genes might be related with apoptosis directly or indirectly. Further study is now needed to characterize these genes.

Multidrug Resistance in Cancer Chemotherapy.: Jung Hye Kim; Yeungnam Univ J Med. 1996;13(1):11-21. Published online June 30, 1996; DOI: https://doi.org/10.12701/yujm.1996.13.1.11

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Abstract PDF: No abstract available.

Gene Expression of Enzymes Related to Glutathione Metabolism in Anticancer Drug-resistant L1210 Sublines: Seong Yong Kim, Jae Ryong Kim, Jung Hye Kim; Yeungnam Univ J Med. 1995;12(1):32-47. Published online June 30, 1995; DOI: https://doi.org/10.12701/yujm.1995.12.1.32

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Abstract PDF: Glutathione(GSH) has a very important role in detoxification of cells and is closely related to antitumor drug-resistance of cancer cells. In order to evaluate the importance of glutathione metabolism in the drug-resistant cancer cells, the concentration of celluar GSH and activities of y-glutamylcysteine synthetase(GCS), y-glutamyl transpeptidase (GGT) and glutathione S-transferases(GST) in the adriamycin, vincristine, or cisplatin resistant L1210 (L1210AdR; L1210VcR, or L12100s) sublines were measured. Expression and amplification of GCS, GGT, and GST-i7 genes were also observed in the parent L1210 and the drug-resistant L1210 sublines. The concentration of GSH was increased 5.34 fold in L12100s, 2.83 fold in L1210VcR, and 1.78 fo-d in L1210AdR, compared to L1210. The activities of GCS and GGT were -increased in drug-resistant L1210 sublines. The GST activity was increased in L1210VcR and L1210Cis but decreased in L1210AdR compared to L1210. Expression of GCS, GGT, and GST-rr genes were increased in the resistant L1210 sublines compare to the parent L1210 in northern blot analyses. Overexpression of GCS, GGT, and GST-77 were observed in the resistant sublines, and the increases of the concentration of glutathione and the activities of GCS and GGT in the resistant sublines may be involved in a part of the drug-resistance in the resistant sublines.

The Change of Glutathione Metabolism in Liver and Kidney of Cisplatin treated Rats.: Seong Yong Kim, Jae Yong Chung, Jae Ryong Kim, Jung Hye Kim; Yeungnam Univ J Med. 1994;11(2):262-269. Published online December 31, 1994; DOI: https://doi.org/10.12701/yujm.1994.11.2.262

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Abstract PDF: Glutathione (GSH) is a well-known antioxidative cellular component which is ubiquitous in nature. Several enzymes involved in GSH metabolism and recycling have been found to play important roles in detoxification of xenobiotics and free radicals. In this study, total GSH content, activity of GSH peroxidase and GSH reductase were measured in liver and kidney of cisplatin treated rats. Total GSH content (mM/g protein) of liver was higher in cisplatin treated rats (1.51±0.28) than of nontreated control (0.95±0.28), and in kidney, it was also higher in cisplatin treated rats (0.87±0.20) than that of control (0.68±0.14). The activity of GSH peroxidase (µM/mg protein/min) was lower in liver of cisplatin treated rats (348.0±18.54) than that of control (415.5±53.15), in kidney it was increase din cisplatin treated rats (380.5±51.86) compared to control (327.3±20.36). The activity of GSH reductase (µM/mg protein/min) was higher in liver of cisplatin treated rats (3.09±0.88) than that of control (2.28±0.61), in kidney it was also higher in cisplatin treated rats (8.50±2.62) than that of control (3.30±1.10). In summary, detoxification of ciplatin was revealed lesser effect in kidney as show increasion of GSH peroxidase and reductase and detoxification of cisplatin was expressed effectively in liver by increasing of GSH content and decreasing GSH peroxidase.

Membrane protein alterations associated with anticancer drug resistance in mouse lymphoblastic leukemia L1210 cells: Seong Yong Kim, Sung Kweon Son, Jae Ryong Kim, Jung Hye Kim; Yeungnam Univ J Med. 1993;10(2):432-444. Published online December 31, 1993; DOI: https://doi.org/10.12701/yujm.1993.10.2.432

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Abstract PDF

Multidrug resistance (MDR) phenotype is frequently observed in animal and human cancer cell lines selected for in vitro resistance to a single chemotherapeutic agent. It is characterized by the diminished j drug accumulation and is related to the drug efflux mechanism in resistant cells. In the present study, adriamycin resistant cells (L1210-AdR6: 10-6M adriamycin, -AdR5: 10-5M) and vincristine resistant cells (L1210-VcR7: 10-7M vincristine, -VcR6: 10-6M) were produced from mouse lymphoblastic leukemia cell line L1210. Growth profiles of survived cells were observed for 5 days with MTT (thiazolyl blue) assay and resistance was compared with IC50 (drug concentration of 50% survival reduction in absorbance). Resistant cells proliferated more slowly than sensitive cell. Doubling times were 29.7hr in L1210, 68.7 hr in L1210-AdR5 and 58.2 hr in -VcR6. MDRs expressed as resistance factor were as follows, L1210-AdR5 was 76.4 times for vincristine, L1210-VcR6 was 96.4 times for adriamycin. The cell membrane proteins with three different M.W. were recognized to be related resistance, 220, 158, and 88 Kd in L1210-AdR5, 158, 140 and 88 Kd in L1210-VcR6 by SDS-PAG electrophoresis. Cell surface membrane proteins were identified by radio-iodination and autoradiogram. Their molecular weights were 158, 72.8, and 42.4 Kd in L1210-VcR6.

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Suppression of P-glycoprotein expression by antipsychotics trifluoperazine in adriamycin-resistant L1210 mouse leukemia cells
Soon Young Shin, Byeong Hyeok Choi, Jae-Ryong Kim, Jung-Hye Kim, Young Han Lee
European Journal of Pharmaceutical Sciences.2006; 28(4): 300. CrossRef

Isolation of GTP binding from bovine brain.: Jung Hye Kim; Yeungnam Univ J Med. 1993;10(2):360-368. Published online December 31, 1993; DOI: https://doi.org/10.12701/yujm.1993.10.2.360

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Abstract PDF: GTP binding protein (G-protein) associated with membrane and involved in signal transduction was isolated from bovine brain, and molecular weight of G protein was observed. As the results, cell membranes were homogenized from bovine brain tissues and proteins of membrane were gained using 1% cholate, and progressed the chromatography. The purification process was performed by step, DEAE-Sephacel, Ulttrogel AcA 34 and heptylamine-Sepharose column chromatography. The chromatographic fractions were confirmed by GTP binding assay and SDS-polyacrylamide gel electrophoresis. Molecular weight of Goa was revealed 39,000 dalton and GR 36,000 dalton. One more step of heptylamine-Sepharose was enforced to purify the GTP binding protein. Finally I gained the GTP binding protein isolated subtype of Goalpha and Gbeta.

The role of G protein in the activation of phospholipase C from bovine brain.: Jung Hye Kim, Dong Jin Lee, Yeung Ju Byun; Yeungnam Univ J Med. 1992;9(2):288-301. Published online December 31, 1992; DOI: https://doi.org/10.12701/yujm.1992.9.2.288

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Abstract PDF: The objective of the present study was to identify the characteristics of phospholipase C (PLC) isozymes purified from bovine brain and to investigate their interrelationship with G protein. The purified PLC isozymes β, γ and δ were obtained and the characteristics of PLC activity on various concentrations of free Ca²⁺ were observed. The activity of PLC was increased with increasing Ca²⁺ concentration and the activity PLC δ was increased higher in the presence of phosphatidyl choline (PC) than in the absence of PC. For vesicle formation as the structure of cell membrane, cholic acid and deoxycholic acid as detergent on phosphatidylinositol bisphosphate (PIP₂) substrate containing PC were used, and then the activity of PLC isozymes were increased with increasing concentration of cholate, from 0.2% to 1% and were increased slightly in deoxycholate. In the PIP₂ containing phospholipid and glycolipid as brain extract, the activity of PLC isozymes were checked in 0.2-1% cholic acid. The activities of PLC isozymes were continuously increased up to 1% cholic acid. The quantitation of PLC isozymes from several bovine organs by radioimmunoassay was made. Brain was the most sufficient organ in terms of amount of PLC β and δ. A large amount of PLC δ was existed in adrenal gland. The binding capacity of GTPrS and G protein was observed and other observations of the binding effect of GTPrS-G protein and PLC monoclonal Ab-Protein A from tissue homogenate with PLC were made. From the observation the binding capacity was revealed the range of 0.11-1.49%. The effects of each type of G protein on the percent activity of purified PLC isozymes were observed. From the observation, activities of isozymes were increased in Goa & Gmix, and the activities of PLC β and δ were increased in Gβγ and Gia. Activities of PLC β and γ were decreased in Gta but PLC δ increased.

Analysis of nucleotides and their derivatives in renal tissue of rat during ischemia by HPLC.: Seong Yong Kim, Jung Hye Kim; Yeungnam Univ J Med. 1992;9(1):90-101. Published online June 30, 1992; DOI: https://doi.org/10.12701/yujm.1992.9.1.90

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Abstract PDF: In rat kidney, the changes in concentrations of nucleotides and their derivatives during ischemia induced by renal artery ligation was measured quantitatively with high performance liquid chromatography (HPLC). After the ligation of renal artery for 60minutes, the concentrations of the nucleotides and derivatives to 80.7±18.39 µg (p<0.01); ATP, 307.2±56.68 µg to 47.6±5.95µg (p<0.01); ADP+AMP, 227.1±7.98 µg to 61.4±3.92 µg (P<0.01); NAD+, 217.9±4.49 µg to 126.6±10.44 µg (P<0.01); GTP, 202.5±23.76 µg to 117.7±14.24 µg (P<0.05); GMP, 54.5±9.03µg to 23.7±0.46 µg (p<0.05), and inosine, 16.6±3.45 µg to 7.8±0.87 µg (P<0.05). But hypoxanthine and xanthine were significantly increased from 113.0±15.58µg to 159.7±12.97µg (P<0.05) and from 87.7±6.77µg to 173.1±12.52µg (P<0.01). In ischemic kidney, concentration of ATP was decreased to 39.9% of control at 10 minutes, 19.8% at 30 minutes, and 15.5% at 60 minutes, and ADP+AMP were decreased to 70.3% of control at 10 minutes, 67.3% at 30 minutes, and to 27.0% at 60 minutes, but hypoxanthine and xanthine were increased to 121.5% and 127.1% at 10 minutes, 126.0% and 174.4% at 30 minutes, and 141.4% and 197.3% at 60 minutes. Total adenosine nucleotides were decreased to 20.3% of control during 60 minutes of ischemia, but hypoxanthine and xanthine were increased to 157.5% of control. These results suggest that the changes in the concentration of nucleotides and their metabolic derivatives are useful indices of the extents of tissue ischemia in rat kidney.

Multidrug resistance and cytotoxicity of anticancer drug by verapamil in cisplatin resistant human stomach cancer cell.: Seong Kweon Son, Jung Hye Kim; Yeungnam Univ J Med. 1992;9(1):75-89. Published online June 30, 1992; DOI: https://doi.org/10.12701/yujm.1992.9.1.75

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Abstract PDF: The development of multidrug-resistant tumor cell population is a major problem in the chemotherapy of human cancer. These cells are often cross resistant to unrelated drugs and the precise mechanisms of multidrug resistant phenotype of tumor cells has not been fully elucidated. Cisplatin resistant tumor cell (SNU-1/Cis₅) was induced from human stomach cancer cell line (SNU-1) in vitro. Growth profiles of survival cells were observed during 5 days by thiazolyl blue (MTT) assay. To investigate the cross resistance of various anticancer drugs in SNU-1 and SNU-1/Cis5, We compared the value of IC₅₀-drug concentration at 50% survival of control and gained relative resistances (RR). The RR for SNC-1/Cis₅ were as follows; vinblastine, > 43.0; epirubicin, 22.9; dactinomycin, 16.0; etoposide, 15.0; vincristine, 9.2; adriamycin, 5.7; aclarubicin, 5.3. But 5-fluorouracil, methotrexate, daunorubicin have not cross resistance with cisplatin. Resistant inhibition values of 10µM verapamil for SNU-1/Cis₅ were as follows; vincristine, 13.1; epirubicin, 10.0; etoposide, 6.3; vinblastine, 4.4; dactinomycin, 3.6; daunorubicin, 2.4. Membrane proteins of 51,400 and 81,300 daltons were identified by radioiodination with SDS-PAGE, which might represented the drug resistance.

The Affinity of Calmodulin-Affigel for Inositol Triphosphate Kinase From Bovine Brain.: Sung Woo Lim, Jung Hye Kim; Yeungnam Univ J Med. 1990;7(1):39-50. Published online June 30, 1990; DOI: https://doi.org/10.12701/yujm.1990.7.1.39

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Abstract PDF: The one event on signaling mechanism is the cleavage by adenyl cyclase of ATP into second messenger, cyclic AMP. The other transfer system of inositol metabolism, it is widely recognized that hydrolysis of the minor membrane lipid phosphoinositide bisphosphate (PIP₂) initiated by occupation of certain receptors and catalyzed by phospholipase C, lead to toe generation of the two intracellular messengers, inositol triphosphate (IP₃) and diacylglycerol (DG). IP₃ is converted to inositol tetrakisphosphate (IP₄) by IP₃ kinase. In the present study, it is that purification of calmodulin is used by phenyl-Sepharose CL-4B chromatography, it's molecular weigh, 17,000 in SDS-polyacrylamide gel electrophoresis. In order to observe the affinity between calmodulin (CaM)-Affigel 15 and IP₃ kinase, and isolated IP₃ kinase, was applied in CaM-Affigel with Ca²⁺ equilibrium buffer and EGTA equilibrium buffer. We compared with binding and elution effect of IP₃ kinase in several condition of buffer. In affinity of binding, Ca²⁺ equilibrium buffer was in the most proper condition, and elution, CaM/Ca²⁺buffer (CE 1 10.36, CE2 12.76pM/min/mg of protein) was effected much more than EGTA buffer (E2 1.48, E 2.43pM/min/mg of protein), but CaM/Ca²⁺stimulate the activity of IP₃ kinase. And then, several detergents such as sodium deoxycholate, tween 20, cholic acid, polyethylene glycol, chaps were applied. The 0.2% chaps buffer (E2 23.19, E3 8.05pnM/min/mg of protein) was the most effective in elution of IP3 kinase.

Study for Metabolism of Resistant Production in Anticancer drug Resistant Stomach Cancer Cell SNU-1.: Jung Hye Kim, Mi Wha Kang, Jae Ryong Kim; Yeungnam Univ J Med. 1989;6(2):195-205. Published online December 31, 1989; DOI: https://doi.org/10.12701/yujm.1989.6.2.195

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Abstract PDF: Development of drug resistance in tumors during treatment is a major factor limiting the clinical use of anticancer agents. When tumor cells acquire resistance to anticancer drug, they show cross-resistance to other antitumor agents. In the present study, SNU-1 cell was induced adriamycin 10-7 drug resistance, SNU-1/ADR, in vitro culture system. We got the doubling time and number for viability test during 96 hours by MTT assay. To investigate the cross resistance of various anticancer drugs in human stomach cancer cell SNU-1 and SNU-1/ADR, We compared IC50 (drug concentration of 50% reduction) and the relative resistance (RR). SNU-1/ADR was expressed multidrug resistant with vinblastine (RR;>31.62), vincristine (RR;29.50), dactinomycin (RR;21.37), epirubicin (RR;17.78), daunorubicin (RR;14.12), adriamycin (RR;7.76), and etoposide (RR;4.46), and other drugs, 5-fluorouracil, cisplatin, cyclophosphamide, methotrexate, and calarubicin, have not cross resistant with adriamycin. There was double minute chromosome in SNU-1/ADR by karyotyping although this change was not seen in SUN-1.

Homogeneity of Phospholipase C of Bovine Uterus and Seminal Vesicle Compared with Brain Isozymes.: Jung Hye Kim, Ki Yung Lee, Sue Goo Rhee; Yeungnam Univ J Med. 1988;5(2):37-45. Published online December 31, 1988; DOI: https://doi.org/10.12701/yujm.1988.5.2.37

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Abstract PDF: Phosphoinositide-specific Phospholipase C (PI-PLC) is a second messenger of signal transducer on cell membrane. In the previous study, PLC of bovine brain has been purified three isozymes. In this paper, uterus and seminal vesicle have been purified. Two peaks of PI-PLC activity were resolved when bovine uterus and seminal vesicle proteins were chromatographed on a DEAE and phenyl TSK 5PW HPLC column. Each two peak was compared with PI-PLC I, II and III from bovine brain and we got the retention time on HPLC. The peak fractions with PLC activity were tested homogeneity with brain PLC monoclonal antibodies (Mab). Mab-labeled affigels were bounded in the range of 73.8%~97.5% with PLC I, II and III. Homogeneity of fractions were revealed that DEAE F-1 and phenyl F-1-I were highest level of PLC III in uterus and seminal vesicle and DEAE F-2 and phenyl F-2-I were mixed PLC I and II.

Production of Monoclonal Antibody to Polychlorinated Biphenyl Induced Cytochrome P-450 LMII in Rat Liver.: Jung Hye Kim, Jae Ryong Kim, Ki Yung Lee; Yeungnam Univ J Med. 1986;3(1):103-110. Published online December 31, 1986; DOI: https://doi.org/10.12701/yujm.1986.3.1.103

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Abstract PDF: Cytochrome P-450 (CP-450) is one of the three components of the liver microsomal enzyme system which hydroxylates fatty acids, hydrocarbons and a variety of drugs and other foreign compounds. Female Balb/c mice were immunized with purified polychlorinated biphenyl (PCB) – induced CP-450 LMII. The spleen cells derived from immunized mice were fused with SP2 myeloma cells using polyethylene glycol (PEG 3500). The hybrid cells were selected by hypoxanthine-aminopterin-thymidine (HAT) medium and the culture fluid were screened by enzyme-linked immunosorbent assay to CP450 LMII. The hybrid cells(×107) were inoculated into intraperitoneal cavity of Balb/c mice for the purpose of production of ascetic fluids. Monoclonal antibody (Mab) was purified from ascitic fluid by DEAE cellulose ion exchange chromatography and I¹²⁵ labeled Mab was also confirmed by autoradiography and SDS-polyacrylamide gel electrophoresis (MW:55,000 and 110,000)

Purification of Band 3 from the Human Erythrocyte Membrane and its Incorporation into Liposome.: Jae Ryong Kim, Jung Hye Kim, Ki Yung Lee; Yeungnam Univ J Med. 1986;3(1):41-48. Published online December 31, 1986; DOI: https://doi.org/10.12701/yujm.1986.3.1.41

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Abstract PDF: Band 3, the predominant 95,000 dalton anion transport protein, is the major intrinsic glycoprotein of the human erythrocyte membrane. This anion carrier exists as a dimer and binds the cytoskeletons such as spectrin, ankyrin and actin. And the liposomes are vesicular structures which form spontaneously upon hydration of phospholipids. These artificial lipid vesicles have been investigated as model of the biological membranes and as a mean of improving the delivery of nucleic acids, drugs, proteins and biological substances to specific target tissues and cells. In this study, we were purified Band 3 from the human erythrocyte membrane (ghost) was prepared by hemolysis of intact human erythrocyte with weak alkali-hypotonic solution. Band 6 was removed from ghost by extracting with solution of an ionic strength of 0.15. Band 3 and Band 4 were solubilized selectively by extracting Band 6-depleted ghosts with Triton X-100 under nondenaturing conditions. Band 3 was then purified from Triton X-100 extract treated with p-chloromercuribenzoate by sucrose density gradient ultracentrifugation. This purified Band 3 was incorporated into liposomes prepared by reverse-phase evaporation. Phosphatidyl L-serine and cholesterol (1:1 molar ratio) were dissolved in chloroform and the chloroform was removed by rotator evaporation under reduced pressure. Band 3 solution without Triton X-100 was introduced into a mixture of lipids and diethylether. Diethylether was subsequently removed by evaporation. This purified Band 3 and its incorporation into liposomes were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Cytological Study of the Introduction of Agrobacterium tumefaciens Spheroplasts into Nicotiana tabacum Protoplasts.: Jung Hye Kim, Yong Bum Koo, Ki Yung Lee; Yeungnam Univ J Med. 1985;2(1):175-181. Published online December 31, 1985; DOI: https://doi.org/10.12701/yujm.1985.2.1.175

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Abstract PDF: Agrobacterium tumefaciens induces cancerous growths called crown galls at wound sites on dicotyledonous plants. A large plasmid called T1 plasmid is responsible for virulence. Upon tumor induction, part of the plasmid, termed T-DNA, becomes integrated into plant genome and its genetic sequences are expressed. These properties allow T1 plasmids to be used as gene vectors in plants. Several in vitro methods for the transfer of T1 plasmid into plant cell have been developed. One of them is the treatment of bacterial spheroplasts and plant protoplasts mixture with polyethylene glycol that is generally used as fusogen in cell-to-cell fusion. Several workers investigated the interaction of bacterial spheroplasts with plant protoplasts in the presence of polyethylene glycol and suggested that the interaction is not fusion but endocytosis. In this report we observed the interaction of Agrobacterium tumefaciens spheroplasts with Nicotiana tabacum protoplasts by electron microscope. Agrobacterium tumefaciens spheroplasts with Nicotiana tabacum protoplasts were prepared and mixed in the presence of polyethylene glycol and high pH-high Ca²⁺ buffer. Then the interaction of the spheroplasts with the protoplasts was examined by transmission electron microscope. After the treatment of polyethylene glycol the spheroplasts adhered to the surface of the protoplasts and then they were engulfed by the protoplasts. After the high pH-high Ca²⁺ buffer treatment the engulfed spheroplasts lost their cell integrity. No fusion process was observed. Thus all these observation suggest that the introduction process of Agrobacterium tumefaciens spheroplasts into Nicotiana tabacum protoplasts with the aid of polyethylene glycol is endocytosis.

Comparative Genetic Characterization of Plasmids of Agrobacterium Species Isolated in Korea.: Jung Hye Kim, Yong Bum Koo, Ki Young Lee, Jae Kyu Chung; Yeungnam Univ J Med. 1984;1(1):41-48. Published online December 31, 1984; DOI: https://doi.org/10.12701/yujm.1984.1.1.41

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Abstract PDF: The soil bacterium Agrobacterium tumefaciens is a plant pathogen that causes crown gall tumors by infecting the wounded dicotyledonous plants and subsequent integration of bacterial DNA into plant nuclear DNA. Virulent A. tumefaciens strains harbor a large Ti (tumor–inducing) plasmid that carries genes essential for tumorigenesis. In the present study, 13 strains (Malus pumila Mill; A₁₋₃, Populus monilifera; W₁₋₆, Populus tomentiglandlosa; P₁₋₃ and Rosa species; R₁) of Agrobacterium isolated in korean crown gall tumors and plasmids were observed in 6 strains (W₂, W₃, W₆, P₁, P₃ and A₂). The test for crown gall tumor formation was resulted only in ATCC15955 and KW2 strains inoculated into the stem of sun flower and the development was observed for 4 and 6 weeks after inoculation. Above two Ti plasmids (pTi) were purified by cesium chloride-ethidium bromide density gradient centrifugation and digested with restriction enzyme and fragments of pTiATCC15955 and pTiKW₂ observed by EcoR I ; 25&27, Hind III ; 23&21, BamH I ; each 20 and Hpa I ; 12&27. And sizes of pTiATCC15955 and and pTiKW₂ calculated as 200 and 87 kbases. Octopine was isolated from tumor tissue (W₁₋₆ and P₁₋₃) and these strains confirmed as octopine type.

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