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β-cell autoantibodies and genetic background associated with insulin dependence in adult-onset diabetes mellitus
Track 2. Clinical Research & Care difference between the treatments. The combination of candesartan and lisinopril reduced BP more than any of the monotherapies and appeared to be the most effective treatment in reducing UACR. Thus the combination of candesartan, and lisinopril, seems a highly effective and well tolerated treatment for reducing BP in hypertensive type 2 diabetic patients with microalbuminuria.
P386 12 Months Post UKPDS and HOT: Impact on BP Targets and Anfihypertensive Drug Prescribing in the Community M.N. MUNRO, R. Larenson, M.D. Feher. Beta Cell Diabetes Centre, Chelsea & Westminster Hospital, London, UK The influence of UKPDS and HOT trials results on community-based prescriptions of antihypertensive drugs in order to achieve lower BP targets for the observed clinical benefit has not been reported. From a single Primary Care Group in Southern England (population 42,000) patients with confirmed type 2 diabetes (n=415) were characterised by current BP recordings and number and class of antihypertensive drug used. From this group, BPs above that achieved in the trials (above 140/80mmHg) were observed in only 40% of subjects, while antihypertensive drug usage was recorded in 54%, of whom 33% were on two or more antihypertensive drugs. When medication usage was stratified according to BP control, 64% of subjects with a systolic BP> 140mmHg were receiving antihypertensive medication and 59% in those with a diastolic BP>80mmHg. Treatment with 2 or more antihypertensives was recorded in 34% in both the higher BP groups. (In comparison, two thirds of UKPDS and HOT patients were similarly treated). Only 3% of patients were on >3 antihypertensive agents. Low usage was recorded for beta-blockers (10%), alpha-blockers (4%) and A-II blockers (2%). Increased use was recorded for calcium channel blockers (18%), diuretics (20%) and ACEI (21%). These data show there has been implementation of evidence based practice in the community. "Newer" compared to "old" antihypertensive treatment appear to be widely used. The number of patients needing antihypertensive treatment will need to be almost doubled if trial-based BP targets are to be reached. In addition, the numbers taking 3 or more antihypertensive drugs will need to be increased nearly 10x if their management reflects UKPDS. The cost of implementing this in the short term is likely to be very substantial in Primary Care.
P387 New Drug Combinations in Diabetic Hypertension; Short Term Effects of Low Dose Moxonidine-ACEI Combination A. COX, A. Zambaninl, C. Mclntosfi, M.D. Feher. Beta Cell Diabetes Centre, Chelsea & Westminster Hospital, London, UK The recent major outcome trials In diabetic hypertension have highlighted the need for combination antihypertensive therapy in order to lower BP and reduce cardiovascular complications. There is little data on the role of moxonidine (a selective imidazoline receptor agonist) in combination antihypertensive treatment in patients with diabetes. In a cross-over study, eight subjects with type 2 diabetes and on conventional dose ACEI for hypertension (mean BP: 176/93, SD:26/6), were assessed three times: (1) after a 2 week run-in phase, with continuation of conventional dose ACEI (2) after 4 weeks low dose moxonidine (200mcg) combination with half usual dose ACEI treatment and (3) after 4 weeks treatment with only conventional dose ACEI. Standardised clinic BP measurements confirmed a mean reduction of 14/12mmHg (A dBP p<0.05 Wilcoxon) with low dose combination therapy and a 14/9mmHg increase (A sBP, dBP p<0.01) after moxonidine withdrawal and reintroduction of the usual dose ACEI. There were no significant changes in weight or fasting glucose or lipid profile. This study has shown the marked BP lowering effects of combining a low dose imldazoline agonist and ACEI, (as well as the effects of monotherapy
$87
withdrawal) and has the potential advantage of minimising drug related side-effects.
1'388 Idiotypic Antibodies in Autoimmunitary Diabetes Mellitus GUSTOVO ACOSTA 1, Maribel Hem~fndez2, Rocio Reyes 3, Martin Herrera4, Carlos Parrao 5, Rao Srinivasa6. J Immunology, Clinica de Inmunodiagntstico, M(xico, City, DE M(xico; 2 Immunology, Clinica de lnmunodiagn6stico, M(xico, City, DE M(xico; s Microbiology, Facultad de Medicina, UNAM, M~xico, City, DE M~xico; 4 Medicina lnterna, Hospital Jutrez de Mdxico, M(xico, City, DE Mdxico; 5 Laboratorio Clfnico, Hospital Judrez de Mtxico, Mdxico, City, DE M(xico; 6 Research and Development, Biomerica, CA, United States of America The regulation of the production of immunoglobulins by antibodies from the same person was postulated by Jerne when he demonstrated secuences of specific aminoacids in the antibodies of each person located in the variable portions of immunoglobulins which are genetically codified and can be antigenic and induce the production of new antibodies against these portions denominated idiotypes and regulate the production of autoantibodies to diminish the inflamatory effects in the pancreatic cells. In the current work the idiotypic induction to regulate the production of (IAA) in 6 patients with diabetes type 1, with a evolution time of 36 months that presents IAA peripheric blood quantified by ELISA and with a substitutive treatmen with human insulin. Individually the gamaglobulins of the 6 patients were purified by the Cohn method it consists of an initial precipitation with ethilic alcohol y then by cromatography affinity using human insulin fixed to Sepharosa 4B. Protocol of specific idiotypic antibodies induction: Daily administration of specific autologous autoantibodies via subcutaneous (0.5 ml) during 40 days. Protocol of inespecific idiotypic antibodies induction: autologous immunoglobulins were purified and administrated in a control group of 40 healthy persons with normal glucose levels and glycosilated hemoglobin at the begining of the study. At the end of the protocol circulanting antibodies weren't detected against human insulin (IAA) by the ELISA method in three patiens, diminishing significantly the dairy doses of insulin, and hemoglobin levels and glucose in fast remained in normal ranges at 60 days. In the control group there weren't variations in the concentration of glucose in fast and glycosilated hemoglobin during 40 days.
1'389 ~8-Cell Autoantibodies and Genetic Background Associated with Insulin Dependence in Adult-Onset Diabetes Mellitns SILVINA N. VALDEZ ~, Ana L. Villanueva l, Anabel Villalba 1, Ruben E Iacono 1, Mauricio P. Sica ', Alejandro C. Cardoso I, Maximiliano Wilda 2, C. Refi 3 Norberto Cedola 3, Gustavo Frechtel 2, Edgardo Poskus i. l Inmunologia, Facultad de Farmacia y Bioqu[mica, UBA e IDEHU (CONICET-UBA), Capital Federal, Buenos Aires, Argentina; 2 Gen(tica y Biolog[a Molecular, Facultad de Farmacia y Bioquimica, UBA, Capital Federal, Buenos Aires, Argentina; 3 Cenexa, Universidad Nacional de La PIata-CONICET, La Plata, Pcia de Buenos Aires, Argentina Immune-mediated Diabetes Mellitus (DM) is early detectable by determination of islet cell autoantibodies (markers). The most useful markers are the autoantibodies to insulin (IAA), antibodies to glutamic acid decarboxylase (GADA) and antibodies to protein tyrosine phosphatase (ICA512A). In this study we determined the presence of immunological and genetic markers in adult-onset diabetic patients. We studied 45 patients treated with insulin after two year from diagnosis ("fast-evolving insulin requiring
Poster Session 1
$88
adults" - IRA(f)), 45 patients with Type 2 DM treated with insulin after 10 years from diagnosis ("slowly-evolving insulin requiring adults "-IRA(s)), 61 patients with conventional (DM2) Type 2 DM and normal controls without personal or familial history of autoimmune disease. Immunological markers were detected by radiobinding assay, and HLA DQB 1 alleles by DNA typing. Results: IAA GADAICA512A 1or moremarkers 2 or moremarkers DQB1Asp57 ./. -4- +/+ IRA(f) 20.0% 35.6% 15.6% IRA(s) 24.4% 8.9% 0.0% DM2 0.0% 3.3% 1.6%
40.0% 33.3% 3.3%
26.7% 0.0% 1.6%
58.8% 41.2% 0~0% 52.4% 42.9% 4.8% 57.1% 35.7% 7.1%
All groups of patients had pancreatic /] cell autoantibodies, indicating that immunological markers are not exclusive of Type 1 DM. As it was previously demonstrated there is an association between insulin requirement and the presence of markers. The IRA(f) patients proved negative for protective alleles. The DQB1 Asp 57-/- phenotype was not significantive prevalent between the studies groups. Analyzing the most frequent susceptibility alleles, DQB 1*0201/0202 and DQB 1.0302, a significative difference was found in IRA(f) (62%) against IRA(s) (43%) and DM2 (43%) patients. Future risk assessment is likely to use mathematical models that incorporate both immunological and genetic markers. The derivation of such models would, however, required prospective study of very large number. In the interim, the present analysis has shown that antibody and genetic testing have the potential to provided and effective means of identifying adult-onset diabetic patients at high risk of insulin requirement.
P390 A New Chimera of Glutamic Acid Decarboxylase 65 Expressed in E. Coli (Trx-M161TGAD65) Useful for the Measurement of GAD65 Antibodies in Diabetes Mellitus JAVIER SANTOS i, Erika A. Anton 1, C.M. Buslje l, Silvina N. Valdez 2, Ana L. Villanueva 2, Mauricio Sica 2, Ruben F. Iacono 3, P. Maffia I , Mario R. Ermacora I, Edgardo Poskus 3 / Ciencia y Tecnologia, Universidad Nacional de Quilmes, Bernal, Pcia de Buenos Aires, Argentina; 2 lnmunologia, Facultad de Farmacia y Bioqufmica, Buenos Aires, Argentina; 3 ldehu, Conicet, Buenos Aires, Argentina Measurement of GAD65 antibodies (GADA) is widely used for the early detection of individuals at risk of developing type 1 diabetes. The best available tests for GADA use properly folded recombinant GAD65 as the antigen. Therefore, there exists a strong interest in a facile and economically sound expression and purification procedure for this protein. Wild-type GAD65 cannot be expressed in E. coli in appropriate yields and a correct conformation. However, we were able to produce in this host a protein fusion thiroredoxin-GAD65 that folded properly with reasonable yield (Papouchado et al. 1997, Eur. J. Biochem. 246, 350-359). Now, we report the construction of a new improved GAD65 hybrid gene that was prepared by joining engineered eDNA obtained from human and rat tissues and subsequent genetic engineering. Despite the numerous changes introduced in the DNA, the new gene codes for human GAD65 with a single amino-acid substitution: Met 161/EThr. This change avoids the co expression of a 48-kDa by-product from an internal translation site that disturbed the purification of our former construction. Also, a proteolytic by-product that co purifies with Trx-M161T GAD65 was identified. The new GAD65 variant was expressed and easily purified, yielding an antigen that performed equally or better than wild-type GAD65 in the reference radiobinding assay for GADA. The procedure provides an inexpensive source of large amounts of fully active and immunochemically competent GAD65
t'391 Association of Acetylator Phenotype with Type 2 Diabeties in a Bangladeshi Population R.B. ZAID, R.A. Bethe, S. Islam, S. Neelotpol, M. Nargis, K. Ahsan, A.K. Azad Khan, L. Ali. Research Division, BIRDEM, Dhaka, Bangladesh Type 2 diabetes mellitus has a strong genetic component and thus genetic markers are widely studied to clarify the transmission of the disease. The acetylator phenotype (AP) has been related to the risk of developing Type 2 diabetes mellitus as well as its complications. The acetylator system in the liver determines the rate of acetylafion of drugs including isoniazid and some sulphonamides, and fast and slow acetylators are found to have a bimodal distribution in individual ethnic groups. Several studies suggested an excess of rapid acetylators among Type 2 diabetes mellitus. This may be explained by an increased risk to develop Type 2 diabetes mellitus among subjects with the rapid acetylator capacity. The aim of the study was to define the possible association of AP in patients with type 2 diabetic. Four hundred forty eight type 2 diabetes and 75 control subjects of Bangladesh origin received oral sulphadimidine in dosages of 250 mg in cases of body weight less than 50 kg, 500 mg in cases of 51-80 kg and 750 mg in cases of more than 81 kg respectively. Serum concentrations of free and total sulphadimidine were measured by colorimetric techniques and phenotypes were calculated by the method of Evans. Total 81.9% of the diabetic subjects were fast AP in contrast to 66.7% in control subjects. There was statistically significant difference between diabetic and nondiabetic subjects regarding the distribution of acetylator phenotype (P=0.005). The results showed clear bimodal distribution in both diabetic and nondiabetic cases. The results suggest that AP is probably associated with diabetes in Bangladeshi population
I'392 PAF Acethylhydrolase (PAFah) Gene Polymorphism Is Not Associated with Diabetic Nephropathy in Type II Diabetic Patients S. NEUGEBAUER, T. Baba, T. Watanabe. Third Department oflnternal Medicine, Fukushima Medical University, Fukushima, Japan
Background and Aims: PAF acethylhydmlase deficiency due to a G to T transition at nucleotide 994 (exon 9) of the PAFah gene occurs in 4% of the Japanese population, resulting in an increase of PAF in the blood. Increased vascular permeability, inflammation, allergic asthma or thrombosis were often observed in patients with PAFah deficiency. According to experimental studies an increase of PAF in the kidney may cause a reduction of GFR, renal blood flow, and water and electrolyte excretion, mesangium proliferation and albuminuria. A mutation of PAFah may consequently aggravate the progression of diabetic nephropathy. Therefore, we investigated the association of the PAFah gene polymorphism with diabetic nephropathy in Japanese Type lI diabetic (non-insulin-dependent) patients. Materials and Methods: Genotyping was performed by the restriction fragment length polymorphism method using DNA from leucocytes of 43 Type II diabetic patients with overt diabetic nephropathy (persistent dip stick-positive proteinuria or end-stage renal failure), 41 diabetic patients without nephropathy, and 82 healthy volunteers. Differences in genotype distribution and allele frequencies of the PAFah gene among the groups was assessed by chi-square test and Fisher's exact test. Results: There was a trend to higher prevalence of homocygotes for the T allele in diabetic patients with nephropathy (6.98%) versus diabetic patients without nephropathy (0%) (3.7% in controls). However, this trend did not reach a significant level (p = 0.0850). The allele frequency (carrier) of the T allele in diabetic patients with nephropathy was 13.95% (21%), versus 13.4% (26.8%) in diabetic patients without nephropathy and 15.2% (26.8%) in the normal controls. There was no significant difference in allele frequency and genotype distribution among the study groups (p = 0.9191 and p = 0.0965 respectively).
P386 12 Months Post UKPDS and HOT: Impact on BP Targets and Anfihypertensive Drug Prescribing in the Community M.N. MUNRO, R. Larenson, M.D. Feher. Beta Cell Diabetes Centre, Chelsea & Westminster Hospital, London, UK The influence of UKPDS and HOT trials results on community-based prescriptions of antihypertensive drugs in order to achieve lower BP targets for the observed clinical benefit has not been reported. From a single Primary Care Group in Southern England (population 42,000) patients with confirmed type 2 diabetes (n=415) were characterised by current BP recordings and number and class of antihypertensive drug used. From this group, BPs above that achieved in the trials (above 140/80mmHg) were observed in only 40% of subjects, while antihypertensive drug usage was recorded in 54%, of whom 33% were on two or more antihypertensive drugs. When medication usage was stratified according to BP control, 64% of subjects with a systolic BP> 140mmHg were receiving antihypertensive medication and 59% in those with a diastolic BP>80mmHg. Treatment with 2 or more antihypertensives was recorded in 34% in both the higher BP groups. (In comparison, two thirds of UKPDS and HOT patients were similarly treated). Only 3% of patients were on >3 antihypertensive agents. Low usage was recorded for beta-blockers (10%), alpha-blockers (4%) and A-II blockers (2%). Increased use was recorded for calcium channel blockers (18%), diuretics (20%) and ACEI (21%). These data show there has been implementation of evidence based practice in the community. "Newer" compared to "old" antihypertensive treatment appear to be widely used. The number of patients needing antihypertensive treatment will need to be almost doubled if trial-based BP targets are to be reached. In addition, the numbers taking 3 or more antihypertensive drugs will need to be increased nearly 10x if their management reflects UKPDS. The cost of implementing this in the short term is likely to be very substantial in Primary Care.
P387 New Drug Combinations in Diabetic Hypertension; Short Term Effects of Low Dose Moxonidine-ACEI Combination A. COX, A. Zambaninl, C. Mclntosfi, M.D. Feher. Beta Cell Diabetes Centre, Chelsea & Westminster Hospital, London, UK The recent major outcome trials In diabetic hypertension have highlighted the need for combination antihypertensive therapy in order to lower BP and reduce cardiovascular complications. There is little data on the role of moxonidine (a selective imidazoline receptor agonist) in combination antihypertensive treatment in patients with diabetes. In a cross-over study, eight subjects with type 2 diabetes and on conventional dose ACEI for hypertension (mean BP: 176/93, SD:26/6), were assessed three times: (1) after a 2 week run-in phase, with continuation of conventional dose ACEI (2) after 4 weeks low dose moxonidine (200mcg) combination with half usual dose ACEI treatment and (3) after 4 weeks treatment with only conventional dose ACEI. Standardised clinic BP measurements confirmed a mean reduction of 14/12mmHg (A dBP p<0.05 Wilcoxon) with low dose combination therapy and a 14/9mmHg increase (A sBP, dBP p<0.01) after moxonidine withdrawal and reintroduction of the usual dose ACEI. There were no significant changes in weight or fasting glucose or lipid profile. This study has shown the marked BP lowering effects of combining a low dose imldazoline agonist and ACEI, (as well as the effects of monotherapy
$87
withdrawal) and has the potential advantage of minimising drug related side-effects.
1'388 Idiotypic Antibodies in Autoimmunitary Diabetes Mellitus GUSTOVO ACOSTA 1, Maribel Hem~fndez2, Rocio Reyes 3, Martin Herrera4, Carlos Parrao 5, Rao Srinivasa6. J Immunology, Clinica de Inmunodiagntstico, M(xico, City, DE M(xico; 2 Immunology, Clinica de lnmunodiagn6stico, M(xico, City, DE M(xico; s Microbiology, Facultad de Medicina, UNAM, M~xico, City, DE M~xico; 4 Medicina lnterna, Hospital Jutrez de Mdxico, M(xico, City, DE Mdxico; 5 Laboratorio Clfnico, Hospital Judrez de Mtxico, Mdxico, City, DE M(xico; 6 Research and Development, Biomerica, CA, United States of America The regulation of the production of immunoglobulins by antibodies from the same person was postulated by Jerne when he demonstrated secuences of specific aminoacids in the antibodies of each person located in the variable portions of immunoglobulins which are genetically codified and can be antigenic and induce the production of new antibodies against these portions denominated idiotypes and regulate the production of autoantibodies to diminish the inflamatory effects in the pancreatic cells. In the current work the idiotypic induction to regulate the production of (IAA) in 6 patients with diabetes type 1, with a evolution time of 36 months that presents IAA peripheric blood quantified by ELISA and with a substitutive treatmen with human insulin. Individually the gamaglobulins of the 6 patients were purified by the Cohn method it consists of an initial precipitation with ethilic alcohol y then by cromatography affinity using human insulin fixed to Sepharosa 4B. Protocol of specific idiotypic antibodies induction: Daily administration of specific autologous autoantibodies via subcutaneous (0.5 ml) during 40 days. Protocol of inespecific idiotypic antibodies induction: autologous immunoglobulins were purified and administrated in a control group of 40 healthy persons with normal glucose levels and glycosilated hemoglobin at the begining of the study. At the end of the protocol circulanting antibodies weren't detected against human insulin (IAA) by the ELISA method in three patiens, diminishing significantly the dairy doses of insulin, and hemoglobin levels and glucose in fast remained in normal ranges at 60 days. In the control group there weren't variations in the concentration of glucose in fast and glycosilated hemoglobin during 40 days.
1'389 ~8-Cell Autoantibodies and Genetic Background Associated with Insulin Dependence in Adult-Onset Diabetes Mellitns SILVINA N. VALDEZ ~, Ana L. Villanueva l, Anabel Villalba 1, Ruben E Iacono 1, Mauricio P. Sica ', Alejandro C. Cardoso I, Maximiliano Wilda 2, C. Refi 3 Norberto Cedola 3, Gustavo Frechtel 2, Edgardo Poskus i. l Inmunologia, Facultad de Farmacia y Bioqu[mica, UBA e IDEHU (CONICET-UBA), Capital Federal, Buenos Aires, Argentina; 2 Gen(tica y Biolog[a Molecular, Facultad de Farmacia y Bioquimica, UBA, Capital Federal, Buenos Aires, Argentina; 3 Cenexa, Universidad Nacional de La PIata-CONICET, La Plata, Pcia de Buenos Aires, Argentina Immune-mediated Diabetes Mellitus (DM) is early detectable by determination of islet cell autoantibodies (markers). The most useful markers are the autoantibodies to insulin (IAA), antibodies to glutamic acid decarboxylase (GADA) and antibodies to protein tyrosine phosphatase (ICA512A). In this study we determined the presence of immunological and genetic markers in adult-onset diabetic patients. We studied 45 patients treated with insulin after two year from diagnosis ("fast-evolving insulin requiring
Poster Session 1
$88
adults" - IRA(f)), 45 patients with Type 2 DM treated with insulin after 10 years from diagnosis ("slowly-evolving insulin requiring adults "-IRA(s)), 61 patients with conventional (DM2) Type 2 DM and normal controls without personal or familial history of autoimmune disease. Immunological markers were detected by radiobinding assay, and HLA DQB 1 alleles by DNA typing. Results: IAA GADAICA512A 1or moremarkers 2 or moremarkers DQB1Asp57 ./. -4- +/+ IRA(f) 20.0% 35.6% 15.6% IRA(s) 24.4% 8.9% 0.0% DM2 0.0% 3.3% 1.6%
40.0% 33.3% 3.3%
26.7% 0.0% 1.6%
58.8% 41.2% 0~0% 52.4% 42.9% 4.8% 57.1% 35.7% 7.1%
All groups of patients had pancreatic /] cell autoantibodies, indicating that immunological markers are not exclusive of Type 1 DM. As it was previously demonstrated there is an association between insulin requirement and the presence of markers. The IRA(f) patients proved negative for protective alleles. The DQB1 Asp 57-/- phenotype was not significantive prevalent between the studies groups. Analyzing the most frequent susceptibility alleles, DQB 1*0201/0202 and DQB 1.0302, a significative difference was found in IRA(f) (62%) against IRA(s) (43%) and DM2 (43%) patients. Future risk assessment is likely to use mathematical models that incorporate both immunological and genetic markers. The derivation of such models would, however, required prospective study of very large number. In the interim, the present analysis has shown that antibody and genetic testing have the potential to provided and effective means of identifying adult-onset diabetic patients at high risk of insulin requirement.
P390 A New Chimera of Glutamic Acid Decarboxylase 65 Expressed in E. Coli (Trx-M161TGAD65) Useful for the Measurement of GAD65 Antibodies in Diabetes Mellitus JAVIER SANTOS i, Erika A. Anton 1, C.M. Buslje l, Silvina N. Valdez 2, Ana L. Villanueva 2, Mauricio Sica 2, Ruben F. Iacono 3, P. Maffia I , Mario R. Ermacora I, Edgardo Poskus 3 / Ciencia y Tecnologia, Universidad Nacional de Quilmes, Bernal, Pcia de Buenos Aires, Argentina; 2 lnmunologia, Facultad de Farmacia y Bioqufmica, Buenos Aires, Argentina; 3 ldehu, Conicet, Buenos Aires, Argentina Measurement of GAD65 antibodies (GADA) is widely used for the early detection of individuals at risk of developing type 1 diabetes. The best available tests for GADA use properly folded recombinant GAD65 as the antigen. Therefore, there exists a strong interest in a facile and economically sound expression and purification procedure for this protein. Wild-type GAD65 cannot be expressed in E. coli in appropriate yields and a correct conformation. However, we were able to produce in this host a protein fusion thiroredoxin-GAD65 that folded properly with reasonable yield (Papouchado et al. 1997, Eur. J. Biochem. 246, 350-359). Now, we report the construction of a new improved GAD65 hybrid gene that was prepared by joining engineered eDNA obtained from human and rat tissues and subsequent genetic engineering. Despite the numerous changes introduced in the DNA, the new gene codes for human GAD65 with a single amino-acid substitution: Met 161/EThr. This change avoids the co expression of a 48-kDa by-product from an internal translation site that disturbed the purification of our former construction. Also, a proteolytic by-product that co purifies with Trx-M161T GAD65 was identified. The new GAD65 variant was expressed and easily purified, yielding an antigen that performed equally or better than wild-type GAD65 in the reference radiobinding assay for GADA. The procedure provides an inexpensive source of large amounts of fully active and immunochemically competent GAD65
t'391 Association of Acetylator Phenotype with Type 2 Diabeties in a Bangladeshi Population R.B. ZAID, R.A. Bethe, S. Islam, S. Neelotpol, M. Nargis, K. Ahsan, A.K. Azad Khan, L. Ali. Research Division, BIRDEM, Dhaka, Bangladesh Type 2 diabetes mellitus has a strong genetic component and thus genetic markers are widely studied to clarify the transmission of the disease. The acetylator phenotype (AP) has been related to the risk of developing Type 2 diabetes mellitus as well as its complications. The acetylator system in the liver determines the rate of acetylafion of drugs including isoniazid and some sulphonamides, and fast and slow acetylators are found to have a bimodal distribution in individual ethnic groups. Several studies suggested an excess of rapid acetylators among Type 2 diabetes mellitus. This may be explained by an increased risk to develop Type 2 diabetes mellitus among subjects with the rapid acetylator capacity. The aim of the study was to define the possible association of AP in patients with type 2 diabetic. Four hundred forty eight type 2 diabetes and 75 control subjects of Bangladesh origin received oral sulphadimidine in dosages of 250 mg in cases of body weight less than 50 kg, 500 mg in cases of 51-80 kg and 750 mg in cases of more than 81 kg respectively. Serum concentrations of free and total sulphadimidine were measured by colorimetric techniques and phenotypes were calculated by the method of Evans. Total 81.9% of the diabetic subjects were fast AP in contrast to 66.7% in control subjects. There was statistically significant difference between diabetic and nondiabetic subjects regarding the distribution of acetylator phenotype (P=0.005). The results showed clear bimodal distribution in both diabetic and nondiabetic cases. The results suggest that AP is probably associated with diabetes in Bangladeshi population
I'392 PAF Acethylhydrolase (PAFah) Gene Polymorphism Is Not Associated with Diabetic Nephropathy in Type II Diabetic Patients S. NEUGEBAUER, T. Baba, T. Watanabe. Third Department oflnternal Medicine, Fukushima Medical University, Fukushima, Japan
Background and Aims: PAF acethylhydmlase deficiency due to a G to T transition at nucleotide 994 (exon 9) of the PAFah gene occurs in 4% of the Japanese population, resulting in an increase of PAF in the blood. Increased vascular permeability, inflammation, allergic asthma or thrombosis were often observed in patients with PAFah deficiency. According to experimental studies an increase of PAF in the kidney may cause a reduction of GFR, renal blood flow, and water and electrolyte excretion, mesangium proliferation and albuminuria. A mutation of PAFah may consequently aggravate the progression of diabetic nephropathy. Therefore, we investigated the association of the PAFah gene polymorphism with diabetic nephropathy in Japanese Type lI diabetic (non-insulin-dependent) patients. Materials and Methods: Genotyping was performed by the restriction fragment length polymorphism method using DNA from leucocytes of 43 Type II diabetic patients with overt diabetic nephropathy (persistent dip stick-positive proteinuria or end-stage renal failure), 41 diabetic patients without nephropathy, and 82 healthy volunteers. Differences in genotype distribution and allele frequencies of the PAFah gene among the groups was assessed by chi-square test and Fisher's exact test. Results: There was a trend to higher prevalence of homocygotes for the T allele in diabetic patients with nephropathy (6.98%) versus diabetic patients without nephropathy (0%) (3.7% in controls). However, this trend did not reach a significant level (p = 0.0850). The allele frequency (carrier) of the T allele in diabetic patients with nephropathy was 13.95% (21%), versus 13.4% (26.8%) in diabetic patients without nephropathy and 15.2% (26.8%) in the normal controls. There was no significant difference in allele frequency and genotype distribution among the study groups (p = 0.9191 and p = 0.0965 respectively).