• Title/Summary/Keyword: Glycosylated

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A Simple Purification Procedure of Biologically Active Recombinant Human Granulocyte Macrophage Colony Stimulating Factor (hGM-CSF) Secreted in Rice Cell Suspension Culture

  • Sharma Niti;Park Seung Moon;Kwon Tae Ho;Kim Dae Hyuk;Yang Moon Sik
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.6
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    • pp.423-427
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    • 2004
  • A simple purification procedure of bioactive human granulocyte macrophage colony stimulating factor (hGM-CSF) secreted in rice cell suspension culture has previously been described. In this study the protein was purified to apparent homogeneity with an overall yield of $80.1\%$ by ammonium sulfate precipitation and a single chromatographic step involving FPLCanion exchange chromatography. The purified hGM-CSF revealed at least five glycosylated forms ranging from $21.5{\~}29$ kDa, and its biological activity was independent of the glycosylation pattern. This is the first purification report of recombinant hGM-CSF to apparent homogeneity from rice cell suspension cultures.

A Survey of Blood Glucose Testing, Medication, Diet, and Exercise Adherence in Korean Patients with Type2 Diabetes (당뇨병환자의 자가혈당검사, 약물, 식이 및 운동 치료지시이행)

  • Kim Hee-Seung
    • Journal of Korean Academy of Fundamentals of Nursing
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    • v.10 no.2
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    • pp.181-186
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    • 2003
  • Purpose: The purpose of this study was to investigate the adherence to diabetes control recommendations (blood glucose testing, medication, diet, exercise) in patients with type 2 diabetes at home and to analyse the correlation between adherence and blood glucose level. Method: Participants, numbering 214, were recruited from the endocrinology outpatient department of a tertiary care hospital in an urban city. The data were collected by a self report adherence questionnaire. Glycosylated haemoglobin (HbA1c) was determined by the high-performance liquid chromatography technique and fasting blood glucose was analyzed by the glucose oxidase method. Result: Medication adherence was higher than diet or exercise adherence. The frequency of blood glucose testing was lower for middle school graduates than college graduates. Diet adherence was significantly lower for participants who were obese, who did not have a spouse, and who had hyperglycemia. Medication and diet adherence were negatively correlated with HbA1c. Conclusion: A diet education program should be developed for patients with type 2 diabetes who are obese, who do not have a spouse, and who have hyperglycemia.

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A Study on Sexual Function, Sexual Stress, and Quality of Life in Middle Aged Women Patients with Diabetes Mellitus (중년여성 당뇨병 환자의 성기능, 성 스트레스, 삶의 질에 대한 연구)

  • Ahn, Sunyoung;Park, Hyojung
    • Journal of Korean Academy of Fundamentals of Nursing
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    • v.23 no.4
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    • pp.393-401
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    • 2016
  • Purpose: The purpose of this study was to examine the relationships among sexual function, sexual stress, and quality of life in middle aged women patients with diabetes mellitus. Methods: Data were collected through questionnaires distributed to 154 middle aged women patient with diabetes mellitus in one hospital. Results: The mean sexual function score was $12.99{\pm}9.11$. There were statistically significant differences in sexual function according to age, level of education, employment status, time of being diagnosed with diabetes mellitus, number of complications, self-monitoring of blood glucose, menopausal status, and level of glycosylated hemoglobin. The mean sexual stress score was $26.99{\pm}16.88$. The score of quality of life was $79.12{\pm}14.30$. There were statistically significant differences in quality of life according to level of education. Sexual function was negatively correlated with sexual stress (r=-.46 p<.001) and positively correlated with quality of life (r=.32, p<.001), while sexual stress was negatively correlated with quality of life (r=-.36 p<.001). Conclusion: Higher sexual dysfunction in middle aged women patients with diabetes mellitus was correlated with lower sexual stress and improved quality of life, while lower sexual stress was correlated with improved quality of life.

Expression and Purification of Recombinant Human Angiopoictin-2 and Its Analog in Chinese Hamster Ovary Cells

  • Hwang, Su-Jeong;Kim, In-Jun;Go, Gyu-Yeong;Lee, Gyun-Min
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.459-462
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    • 2001
  • Angiopoietin-2 (Ang2) is a naturally occurring antagonist for angiopoietin-l (Angl) and its Tie2 receptor during vasculogenesis, Although angiopoietins have been expressed in several mammalian cell lines, their expression levels are low. Recombinant Chinese hamster ovary (CHO) cell lines expressing a high level of human Ang2 or its analog, human $Ang2_{443}$, with an amino-terminal FLAG-tag were constructed by transfecting the expression vectors into dhfr-deficient CHO cells and subsequent gene amplification in medium containing stepwise increments in methotrexate level. Secreted Ang2 or human $Ang2_{443}$ were purified from the cultured medium using an anti-FLAG- agarose affinity chromatography, The purified Ang2 and $Ang2_{443}$ migrated on SOS-PAGE as a broad band, characteristic of glycosylated protein. Their biological activity in vitro was demonstrated in a serum deprivation-induced apoptosis assay. Ang2 at high concentration, like AngI, can be an apoptosis survival factor for endothelial cells through the activation of the Tie2 receptor.

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Carbohydrate Structure of N- and O-linked Oligosaccharides of Human Erythropoietin Expressed in Chinese Hamster Ovary Cells

  • Lee, Dong-Eok;Ha, Byung-Jhip;Kim, Suk-Joon;Park, Ji-Sook;Yoo, Ree-Ann;Oh, Myung-Suk;Kim, Hyun-Su
    • BMB Reports
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    • v.29 no.3
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    • pp.266-271
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    • 1996
  • A recombinant human erythropoietin (EPO), expressed in Chinese hamster ovary (CHO) cells, is glycosylated at Asn 24, Asn 38, Asn 83, and Ser 126. After release of the N-linked carbohydrate chains by $peptide-N^{4}-(N-acetyl-{\beta}-glucosaminyl)$ asparagine amidase F, the oligosaccharides were analyzed by FACE (Fluorophore-Assisted Carbohydrate Electrophoresis). The O-linked carbohydrate chain was separated by hydrazine, and analyzed by FACE. The monosacccharide composition of recombinant EPO showed man nose, fucose, galactose, N-acetylglucosamine, N-acetylneuraminic acid, and a trace of N-acetylgalactosamine, which are typical monosaccharides in the glycoproteins from the CHO cell. Sequences of N-linked and O-linked oligosaccharides were determined. The structure and composition of oligosaccharides attached to recombinant human EPO, expressed in the CHO cell, are identical to the reported oligosaccharide structure in human EPO isolated from urine.

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An Application of Electrostatic Repulsion Hydrophilic Interaction Chromatography in Phospho- and Glycoproteome Profiling of Epicardial Adipose Tissue in Obesity Mouse

  • Tran, Trang Huyen;Hwang, In-Jae;Park, Jong-Moon;Kim, Jae-Bum;Lee, Hoo-Keun
    • Mass Spectrometry Letters
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    • v.3 no.2
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    • pp.39-42
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    • 2012
  • Phosphorylation and glycosylation are two of the most important and widespread post-translational modifications (PTMs) in an organism. Proteomics analysis of the PTMs has been challenged by low stoichiometry of the modified proteins and suppression effects by high abundance proteins, typically no-functional house-keeping proteins. In this study, a novel method was applied for not only isolating PTM peptides from intact peptides but also concurrently characterizing of glyco- and phosphoproteome using electrostatic repulsion hydrophilic interaction chromatography (ERLIC) packed with silica coated by crosslinked polyethyleneimine. For 2 mg tryptic digest of mouse proteome of epicardial adipose tissue with fat diet, 802 N-glycosylated peptides of 316 glycoproteins and 159 phosphorylated peptides of 75 phosphoproteins were identified using HPLC chip/quadrupole time-of-flight (Q-OF) tandem mass spectrometer.

Association of Clusterin Polymorphisms with Coronary Heart Disease in Koreans

  • Kim, Jun-Young;Kim, Su-Won;Lee, In-Kyu;Yoo, Min
    • Biomedical Science Letters
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    • v.14 no.1
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    • pp.55-58
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    • 2008
  • Clusterin is an 80 kDa heterodimetric glycosylated protein and it plays diverse biological roles in various tissues and organs. Clusterin is reported to be associated with the pathogenesis of coronary artery disease and atherosclerosis. Therefore, we investigated the genotype for the A/G polymorphism at the position 4,183 of clusterin gene in Koreans and compared genotype of patients with control group. 100 patients (Male 63, Female 37), who previously underwent coronary artery stent due to ischemic heart disease and 100 controls (Male 36, Female 64) participated in this study. There was a significant association between 4,183 A/G polymorphism in clusterin gene and coronary artery disease (CAD). The present study shows that clusterin gene A/G polymorphism may be associated with the pathogenesis of CAD. Further studies with larger population may be needed for the development of diagnostic methods at genetic level such as DNA chip.

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Molecular Cloning and Recombinant Expression of the Long Form of Leptin Receptor (Ob-Rb) cDNA as Isolated from Rat Spleen

  • Ju, Sung-Kyu;Park, Jung-Hyun;Na, Shin-Young;You, Kwan-Hee;Kim, Kil-Lyong;Lee, Myung-Kyu
    • BMB Reports
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    • v.34 no.2
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    • pp.156-165
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    • 2001
  • Leptin is a circulating non-glycosylated protein that is mainly produced in adipocytes. Leptin acts in the brain to regulate food intake and energy expenditure. Previously we reported our success in the isolation of a partial cDNA of the long form of the leptin receptor, OB-Rb, from rat spleen, and showed that leptin might also play a role in peripheral immune organs. In the present study, for the first time, the complete coding region of OB-Rb cDNA was cloned from rat splenocytes, and its nucleotide sequence was determined. The cDNA was then further expressed in E. coli and mammalian cells, thereby confirming the functional integrity of this receptor. Prokaryotically overexpressed OB-R protein was then used as an immunizing antigen in BALE/c mice to produce leptin receptor-specific antibodies. By using them, we confirmed the cell surface expression of OB-Rb in transfected CHO cells. It is our belief that the reagents, as produced in this study, will be of great use in further studies of the biological role of rat leptin.

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The Riboflavin Status in Insulin-Dependent Diabetes Mellitus During Growing Period (성장기 인슐린 의존성 당뇨병 환자의 리보플라빈 영양상태)

  • 이정희
    • Journal of Nutrition and Health
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    • v.27 no.7
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    • pp.729-739
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    • 1994
  • Riboflavin status of 17 insulin-dependent diabetic mellitus(IDDM) patients in growing period was evaluated as a function of energy intake and expenditure, biochemical nutritional status and diabetic control indicators. Compared with recommended dietary allowances for Koreans(RDA, 1989), only 35.3% of subjects was at good levels of all nutrients intakes and 52.9% of subjects was below normal level of height and weight. Nutrients consumed below RDA levels were energy(=88.5% of subjects), niacin(64.7%), iron(52.9%) and protein(23.5%) respectively. The riboflavin status was within normal range by urinary riboflavin excretion but 17.6% of subjects was evaluated as showed riboflavin deficiency by erythrocyte glutathione reductase activity coefficient(EGRAC). Correlation between riboflavin intake, urinary riboflavin excretion, EGRA level and diabetic duration were not statistically significant. Correlation analyses indicated that EGRA level was inversely correlated with thiamin, niacin and cabohydrate intake. No significant correlations were found between the EGRA and glycosylated hemoglobin A1(HbA1) (r=-0.464, p=0.129). From this study, it is suggested that IDDM subjects need to maintain balanced diet containing nutrients above RDA for individual activity during growing period. It needs more study whether the current recommended riboflavin allowance is adequate for diabetic patients.

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A Meta-Analysis of Korean Diabetes Patient Studies from 1977 to 2008 (메타분석을 이용한 당뇨중재의 효과분석)

  • Chin, Young-Ran;So, Eun-Sun;Lee, Hyo-Young
    • Journal of Korean Public Health Nursing
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    • v.25 no.1
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    • pp.5-16
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    • 2011
  • Objectives: This study analyzed the effects of diabetes interventions published in journals of nursing, medical and public health, to suggest directions for future research. Methods: A comprehensive search of three databases was undertaken (Korean studies, Riss4U, RICHIS) using research terms such as "diabetes", "nursing", "public health", "intervention" and "program". One hundred fifty four original articles were retrieved and 21 articles were finally analyzed. The analyzed articles satisfied the inclusion criteria and their effect size was calculated. Results: Most studies about diabetes intervention were published form 2001. 2005 (57.1%). The psycho-social variables, 'self-efficacy' and 'self-care' presented good effects on exercise or exercise prescription intervention. In addition, the physiological variables, 'fasting blood sugar' and 'glycosylated glucose' presented good effects on the interventions using four more elements or the interventions held in a public health center. Conclusion: Further research should consider the essential elements that benefit diabetes patients' behaviors or physiological indicators.