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Volume 21(3); September 1997
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Original Articles
Measurement of GAD Antibody in Korean with Diabetes Mellitus.
Hyoung Woo Lee
Korean Diabetes J. 1997;21(3):228-230.   Published online January 1, 2001
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No abstract available.
Measurement of Anti-GAD antibody by EIA and RIA Methods in Korean Diabetic patients: Study for pathogenesis of slowly progressive IDDM.
Han Hyo Lee, Young Goo Shin, Hee Sun Kim, Chang Young Kim, Yon Soo Jeong, Hong Seung Kim, Deok Woo Park, Kap Jun Yoon, Choon Hee Chung
Korean Diabetes J. 1997;21(3):231-242.   Published online January 1, 2001
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background
Sometimes it is difficult to discriminate between IDDM and NIDDM among adults with DM. Some NIDDM patients have autaantibodies and follow the course of IDDM, We call them as slowly progressive IDDM(SPII)DM). Since anti-GAD (Glutamic acid decarboxylase') Ab can be detected both before and for a long pe.riod after the diagnosis of DM it is helpful for the diagnosis of autoimmune diabetes. METHODS: The subjects were 68 diabetic patients who were admitted at Wonju Christian Hospital from May 1994 to Feb 1996. We classified them as IDDM, NIDDM and SPIDDM. We analyzed the following: a studied basic clinical study, oral glucose tolerance test, HLA DR typing, IgM anti-viral Abs, ICA, IAA and nti-GAD Ab. RESULTS: In measurement of anti-GAD Ab, IRMA was more sensitive than EIA. Anti-GAD Ab prevalence was significantly higher in IDDM patients than in NIDDM patients. By IRMA method, Anti-GAD titers showed significant correlation among VELISA, HEXT, IRMA and RSR methods(p<0.001). CONCLUSION: As seen by the results above, the positivity of antiGAD Ab by EIA and RIA method was lower for Korean diabetic patients than for Caucasians. We suggest that the other mechanisms as well as autoimmunity may be involved in the pathogenesis of SPIDDM in Koreans. We need follow-up studies about the clinical and immunogenetic characteristics of these patients.
Distinct Pattern of GAD65 and GAD67 Gene Expression in the Pancreas of NOD Mouse.
In Young Ko, Yup Kang
Korean Diabetes J. 1997;21(3):243-253.   Published online January 1, 2001
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BACKGROUND
Glutamic acid decarboxylase(GAD; EC 4.1.1.15), one of the major B-cell autoantigens in IDDM, is an enzyme which catalyzes the synthesis of major inhibitory neurotransmitter, r-aminobutyric acid (GARA), in the mammalian brain, pancreas and other organs. Two isoforms of GAD, GAD65 and GAD67, have been identified which differ in their intracellular localization. Autoantibodies to GAD have been detected several years before the clinical onset of IDDM, implicating GAD as a leading autoantigen which somehow correlated with the pathogenesis of IDDM. We have determined the characteristics of GAD isoform expression in the pancreas of NOD mouse, an animal model extensively employed in IDDM study, using RT-PCR and Southern blot methods. METHODS: Pancreas was obtained from female NOD mouse(neonate, 4, 8, 12, 16, 20 week-old) and age-matched female ICR mouse. Total cellular RNA was I.solated by acid guanidinium thiocyanate method and employed in the RT-PCR amplification using GAD65- and GAD67-specific primer designed in our laboratory. The PCR product was blotted onto the nylon membrane and subjected to Southern analysis using 32P-ATP labelled hybridization probe. RESULTS: In NOD pancreas, GAD67 was expressed six times higher than GAD65 at neonatal stage. Then, the expression was dramatically decreased from 4 weeks when the pancreatic insulitis begins to occur. After 12 weeks of age, both GAD67 and GAD65 expression was almost undetectable. However, in control ICR mouse, there were no significant differenees between GAD65 and GAD67 expression throughout the ages. And, the expression of both GAD65 and OAD67 was not decreased with ages in contrast to NOD mouse. CONCLUSION: In this experiment, we found that the expression of GAD isoforms in NOD mouse shows distinct pattern in comparison to that of control ICR mouse. The expression of GAD67 was significantly higher than GAD65 in neonatal NOD mouse while, in control ICR mouse, same level of GAD isoforrns expression was observed. This finding clearly suggested the possibility that the expression of GAD isoforms in diabetic NOD mouse is quite distinct and may somehow play a role in the pathogenesis of diabetes although the precise mechanism remains to be unveiled. In addition, our data also supported the hypothesis that expressional pattern, and, if possible, ' the etiophysiological function of GAD isoforms in NOD mouse pancreas may be quite different from that in human pancreas.
Effect of Troglitazone on Glycogen synthase Activity in Human skeletal Muscle Culture from Obese Non-Diabetic and Obese Non-insulin Dependent Diabetes Mellitus.
Leslie Abrams Carter, Theodore Ciaraldi, Robert R Henry, Kyong Soo Park, Hong Kyu Lee
Korean Diabetes J. 1997;21(3):254-261.   Published online January 1, 2001
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BACKGROUND
Skeletal muscle is the principal tissue of insulin resistance in obese and non-insulin dependent diabetic(NIDDM) subjects. Troglitazone, a member of thiazolidinedione class of compounds, has been shown to improve glucose tolerance in insulin resistant state. At the cellular level, troglitazone has been shown to improve insulin action in skeletal muscle, liver and adipose tissue. However, there has been no direct assessment of mechanism of this drug in insulin resistant human skeletal muscle from obese and obese NIDDM subjects. METHODS: To determine the effect of troglitzone on skeletal muscle glycogen synthase(GS) activity in insulin resistant human skeletal muscle, muscle cultures from 7 obese non-diabetic and 8 obese NIDDM subjects were grown for 4 weeks and then fused for 4 days either with or without troglitzone(0~5ug/mL) and harvested for GS activity and protein measurement. GS activity was measured by enzymatic method and protein expression was measured by Western blot using polyclonal antibody specific for C-terminal end of GS protein, RESULTS: Troglitzone increased GS activity(GS activity at 0.1 mM G6P and fractional velocity) dose-dependently in both obese non-diabetic and type II diabetes and the increased GS activity by troglitzone was mostly basal rather than insulin-stimulated. Basal fractional velocity of GS increased 2.8+/-0.7 and 3.7 +/-1.2 fold in obese non-diabetic and type II diabetes respectively. There was no changes in GS total activity and GS protein expression in either group with troglitzone treatment.. CONCLUSION: Troglitazone has effects to improve glycogen synthase activity in skeletal tnuscle of obese and obese NIDDM subjects.
Effect of Glucose on Adherence of Neutrophils to Endothelial Cells.
Seok Man Son, Seok Dong Yoo, In Ju Kim, Yong Ki Kim, Hee Bag Park, Chi Dae Kim, Ki Whan Hong
Korean Diabetes J. 1997;21(3):262-270.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Accelerated atherosclerotic vascular disease is the leading cause of mortality in patients with diabetes mellitus. To clarify the mechanisms that cause macrovascular dysfunction in diabetes, we examined the effect of high glucose on the adhesion of neutrophils to the endothelial cells and release of TNF-a from cultured rabbit aortic endotheIial cells. METHODS: Rabbit aortic endothelial cells in primary culture were prepared by the collagenase digestion method. Cells were incubated for various time upto 24 hours to evaluate TNF-a response to different glucose concentrations(0, 5.5, 11, 22mmol/L). Isolated rabbit neutrophils were incubated with monolayers of rabbit aortic endothelial cells under different glucose condition. RESULTS: After 24 hrs incubation with various concentrations of glucose, neutrophil adherence to high concentration of glucose(11 and 22mM)-treated endothelium was significantly increased(46+/-7 and 64 +/-6%, respectively) compared with adhesion to low concentration of glucose(0 or 5.5mM)-treated endothelium(3l +/-5 and 30+/-3%, respectively), In addition, when TNF-a imrnunoreactivity in the culture medium was measured by enzyme-linked immunoassay after 24 hours of incubation with various concentration of glucose, the secretion of TNF-a from endothelial cells was significantly increased in a concentration-dependent manner upon exposure to high concentration of glucose, CONCLUSION: The results of this study ciemonstrate tht high concentration of glucose stimulates neutrophil adhesion to endothelial cells in association with increased production of TNF-a from endothelial cells. These results suggest that glucose directly causes increased interaction between neutrophil and endothelial cell through a TNF-a-dependent mechaniasm,
Effect of Dietary Polyunsaturated / Saturated Fatty Acid on Membrane Lipid Peroxidation of Red Blood Cells and Hepatic Intracellular Organelles in Streptozotocin Induced Diabetec Rats.
Hyung Joon Yoo
Korean Diabetes J. 1997;21(3):271-279.   Published online January 1, 2001
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BACKGROUND
Lipid peroxidation in tissues and in tissue fractions is a degradative free-radical process that primarily involves polyunsaturated fatty acids; it has been implicated as a major contribution to many types of tissue damage, especially in diabetes mellitus. This study was designed to investigate the effect of P/S dietary composition on the lipid peroxidation of biomembranes in streptozotocin-induced diabetic rats. METHODS: Diabetic Sprague Dawley rats weighing about 200g were randomly divided into 3 groups: Group L(n=8) fed with P/S ratio 0.4, Group M(n=8) fed with P/S ratio 1.2, and Group H(n=8) fed with P/S ratio 2.0. Diabetes was induced by daily intraperitoneal injection of streptozotocin(20mg/kg) for 5 days. After feeding for 6 weeks, plasma cholesterol, HDL-cholesterol, RBC membrane peroxidation and hepatic mitochondrial lipid peroxidation were measured. RESULTS: Total cholesterol(mmol/L) was decreased and HDL-cholesterol/total cholesterol % ratio was increased by increment of P/S ratio in a doseindependent manner(L 3.4+/-0.32 and 8.7; M 2.5+/- 0.29 and 12.6; H 2.5+/-0.29 and 12.3). RBC membrane lipid peroxidation(nmol/mI. packed RBC) was higher in H(3.21+/-0.20) than that in L(2.02+/-0.19) or M(2.40+/-0.21) (p<0.05). Hepatic mitoehondrial lipid peroxidation (nmol/g protein) was lower in L(7.5+/- 1.25) than that in M(11.7+1.57) or H(14.0+2.04) (p<0,05). CONCLUSION: P/S dietary increment increased the lipid peroxidation of biomembranes(RBC membrane an4 hepatic mitochondrial membrane) in streptozotocin-induced diabetic rats.
Lipopolysaccharide-Induced Changes in Vascular Reactivity of Diabetic Rat Aorta.
Ye Kyung Seo, Sang Won Chung, Jik Hwa Nam, Byoung Ho Sin, Dong Hee Kim, Jung Guk Kim, Sung Woo Ha, Bo Wan Kim
Korean Diabetes J. 1997;21(3):280-288.   Published online January 1, 2001
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Backgound: Hemodynamic deteriorations in diabetes mellitus may be mediated by increased contractile response to catecholamines and/or by decreased relaxative response to vasodilators such as acetylcholine(Ach). Decrease in peripheral vascular reactivity to vasoconstrictor was known to be an ominous sign that happens during sepsis or after injection of bacterial lipopolysaccharides(LPS). In this study, we compared the effects of LPS on function of diabetic rat aorta with impaired vascular reactivity with those of control rat aorta. METHODS: Contractile responses to cumulative concentrations(10'M to 3X10'M) of norepinephrine (NE) were measured in aorta isolated ftom the control and 4 to 5-week streptozotocin-induced diabetic rat at 6 hours after LPS treatment to compare with contractile responses of untreated group. We measured relaxative responses to cumulative concentrations(10'M to 10M) of Ach and nitroprusside (NTP) in these aortas contracted submaximally by NE. RESULTS: Diabetic rat aortas showed significantly more impairment in relaxative responses to Ach than control rat aortas before LPS treatment(p0.05 = 0.0l). LPS treatment in those diabetic rat aortas decreased contractile responses to NE by 26.6%(p < 0.01); the changes were sirnilar to those of control (30.9%, p0.01). Relaxative responses to Ach were also significantly decreased by 25.0%(p 0.01) after L.PS treatment; the changes were similar to those of control(34.1%, p0.01). However relaxative responses to NTP were not changed in control and d.iabetic rat aortas by LPS treatment. CONCLUSION: These results suggest that diabetes may induce impairment in endothelium-dependent vascular relaxation and there rnay be no difference of L,P,S-induced effects on hemodynamic deterioration between 4 to 5-week diabetic and control rats.
Prevalence of ICA and anti-GAD, HLA DRB1 / DQA1 / DQB1 Polymorphism in Korean IDDM Patients.
Yong Soo Park, Jin Ho Shin, Jin Bae Kim, Woong Hwan Choi, You Hern Ahn, Tae Wha Kim, Mok Hyun Kim, Sei Won Yang, Seung Duck Hwang, Hee Bal Rhee
Korean Diabetes J. 1997;21(3):289-299.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Although the HLA class II genes are cleaily associated with insulin-dependent diabetes mellitus(IDDM) in all ethnic. Groups, considerable variation in the associated genotypes is observed among the different ethnic groups. Mathods: In order to estimate what degree genetic and environmental determinants influence the true incidence of IDDM, ICA by imrnunohistochemistry, anti-GAD prevalence by radioimmunoprecipitation and HLA-DRB1, DQAl, and DQB1 polymorphisms after PCR amplification of genomic DNA were analyzed in 131 cases of IDDM, whose age at diagnosis were less than 15. RESULTS: 56% of them(73/131) were anti-GAD positive. 43% IDDM(56/131) were ICA positive. HLA DR3 and DR9 were susceptibility markers, whereas DR2 and DR5 were protective markers. DR3/4,, DR3/9, and DR3/X(X: other than 3, 4) were susceptible genotypes. HLA DQA1*0301 allele was increased, and DQB1*0301 and DQB1*0602 were decreased in IDDM. Not only HLA DQA1 Arg, but also DQBl non-Asp were found to be independent marker for IDDM, but their strength of association was weak. The highest prevalence of anti-GAD was observed in thosc homozygous for DR4(87.5%), exceeding that(47.8%) in those without this allele, and those with one DR4(63.5), whereas the highest prevealence of ICA was found in those homozygous for DR3(10G%), exceeding that in those with one DR3(64.3%) or in those without this allele(36.7%). There was a significant difference in numbers of HLA DQ susceptibility heterodimers in anti-GAD positive or negative patients. Conelusion: The prevalence of islet-specific auto-antibodies were present at comparable sensitivity and specificity in Korean IDDM patients. We could also assess that the immunoenetic markers for IDDM among Caucasians likewise confer disease susceptibility among Koreans. However, different HLA susceptibility alleles and a lower strength of association with known susceptibility markers, presumably because of differences in the genetic make-up of the population or in linkage disequilibrium patterns compared with other racial groups.
Increased Serum 8-hydroxy-Guanine Levels in Diabetic Patients.
Byung Sool Moon, Yun Yong Lee, Chan Soo Shin, Kyong Soo Park, Seong Yeon Kim, Hong Kyu Lee, Su Jin Park, Myung Hee Chung
Korean Diabetes J. 1997;21(3):300-307.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Production of reactive oxygen species(ROS) increased in diabetic patients and oxidative damage may contribute to the development of diabetic complications. 8-OH-deoxyGuanosine (oh8dG) and 8-OH-Guanine(ohSGua) are known as excellent markers of the oxidative darnage to DNA. This study was performed to investigate whether serum 8-OH-guanine increased in diabetic patients and whether the glycemic control(HbAlc) is related to the levels of serum 8-OH-guanine. METHOD: In this study, 28 patients with diabetes mellitus was studied, We also included 27 nondiabetic healthy controls whose age, sex, and BMI were matehed to the diabetic patients. Serum 8-OH-Guanine was assayed by high performance liquid chromatography after antibody-based purification with monoclonal antibodies to S-OH-Guanine. RESULTS: The levels of serum 8-OH-Guanine was significantly higher in diabetic patients than in normal controls(4.02+/-3.77 pmol/mL vs. 0.89+/-0.63 pmol/ mL, p<0.01). Serum 8-OH-Guanine concentration was not related to age, HbAlc, duration of diabetes, creatinine clearance, total cholesterol, triglyceride, and HDL-cholesterol. Conclusions: We found a significant increase in serum 8-OH-guanine levels from diabetic patients compared with their respective controls. These results suggest that diabetic patients have significantly increased oxidatively damaged DNA. The factors regulating the oxidative damage to DNA should be further investigated.
Clinical and Coronary Angiographic Findings of Coronary Artery Disease in Patients with Noninsulin Dependent Diabetes Mellitus.
Bon Jeong Ku, Bong Soo An, Jin Ok Jeong, In Whan Seong, Eun Seok Jeon, Min Ho Shong, Heung Kyu Ro, Young Kun Kim
Korean Diabetes J. 1997;21(3):308-313.   Published online January 1, 2001
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BACKGROUND
Diabetes mellitus has a major impact on cardiac morbidity and mortality. We analyzed the clinical characteristics and coronary angiographic findings in patients with a coronary artery disease and with noninsulin dependent diabetes mellitus. METHODS: We retrospectively reviewed the clinical and coronary angiographic findings in 74 patients with noninsulin dependent cliabetes mellitus and in 328 non-diabetic patients who had a coronary insufficiency. RESULTS: Significant coronary occlusions and multiple coronary artery diseases were more common in diabetics than in non-diabetics. The patients with noninsulin dependent diabetes mellitus has several limiting factors to perform the successful percutaneous coronary angioplasty, it could be a possible factor for rapid deterioration of coronary insufficiency in patients with diabetes mellitus. Smoking and hypertension significantly increased the incidence of signiftcant coronary occlusions whereas obesity and hypercholesterolemia did not any significant influence on incidence of the significant coronary occlusions. The longer duration of diabetes rnellitus associated with higher incidence of significant coronary occlusions and multiple coronary diseases. CONCLUSION: Significant coronary occlusions and multiple coronary artery diseases were occurred more common in diabetics than in non-diabetics. Coronary angiography should be performed routinely in noninsulin dependent diabetes.
Case Report
A Case of Diabetic Muscle Infarction in a Patient with Insulin Dependent Diabetes Mellitus.
Joon Ho Jang, Jae Hyun Nam, Woong Chul Kang, Jung Il Jung, Suk Ho Kwon, Yong Suck Yoon, Bong Soo Cha, Young Joon Won, Young Duk Song, Sung Kil Lim, Kyung Rae Kim, Hyun Chul Lee, Kap Bum Huh
Korean Diabetes J. 1997;21(3):314-320.   Published online January 1, 2001
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AbstractAbstract PDF
Diabetic muscle infarction(DMI) is an uncommonly reported complicatian of diabetes mellitus, DMI tends to occur in younger, poorly controlled diabetic patients with other end organ complications(retinopathy, nephropathy, neuropathy). The typical feature of DMI is abrupt onset of thigh pain, tenderness, and swelling, over a period of days, and a firm mass develops. There are no associated systemic symptoms or signs indicative of infection and no skin discoloration suggestive of cellulitis or thrombophlebitis. The patient was diagnosed as DMI with the findings of ultrasonographic, bone scan and magnetic resonance imaging as well as typical clinical and laboratory findings. The painful mass persists for weeks, occasionally with exacerbation of symptoms, and then spontaneously resolves over several montks. Immobilization of the extremity with prolonged bed rest and strict sugar control has had beneficial results. We report a case of diabetic muscle infarction in a 30-year-old woman with insulin dependent diabetes mellitus

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