Molecular mechanisms involved in the etiopathogenesis of malnutrition-modulated diabetes mellitus.

MMDM patients are typically young at onset with low body mass index, require insulin treatment for glycemic control, have insulin resistance, and do not develop ketosis on withdrawal of insulin. WHO's revised classification in 1999, based on the etiopathogenesis of the disease, identifies only two categories: type 1 diabetes and type 2 diabetes. MMDM could be considered as type 1b diabetes. Genetic and immunological studies were done on MDDM patients (n = 72) from Cuttack and healthy controls to understand and to justify its inclusion in the category of type 1b diabetes. Antibodies (Abs) to tyrosine pyrophosphatase (IA2-Abs), glutamate decarboxylase 65 (GAD65-Abs), and other minor markers like ICA12 Abs and tissue transglutaminase Abs (TTG-Abs) were studied. HLA-DR and DQ were studied for the genetic markers. Of the MMDM patients 30% were positive for either GAD65 or IA-2 antibodies, and 14% were positive for ICA12 antibodies. All three antibody markers together accounted for 39% of PDDM patients, as some patients were positive for more than one autoantibody. TTG antibodies (specific for Celiac disease) were present in 14/71 (20%) of MMDM patients compared to 3/122 (2%) controls. All four autoantibodies accounted for 53% of PDDM patients, leaving 47% of patients free of known autoantibodies. The autoantibody-negative PDDM patients were analyzed for HLA and MICA markers, showing that DR7-DQ9 and MICA allele 9 are increased in this group compared to healthy controls, which suggests an autoimmune response to an unknown dietary autoantigen. We conclude from our data that an autoimmune mechanism is involved in the etiology of MMDM. In addition, the presence of silent celiac disease seen with MMDM patients, which has not yet been reported, is significant. It is important to note that subclinical celiac disease exists with diabetes mellitus and must be considered in the diagnosis of MMDM.

Clinicoepidemiological and biochemical profile of malnutrition-modulated diabetes mellitus.

Malnitrition-modulated diabetes mellitus ((MMDM) was previously known as protein-deficient diabetes mellitus (PDDM). Its clinical picture is similar to that of type 1 diabetes, but it develops over a background of chronic malnutrition from childhood. In spite of severe hyperglycemia, ketonuria never occurs. MMDM patients are extremely lean and require high doses of insulin-over 2.0 U/kg/day for good glycemic control. Even when optimally controlled, these patients maintain their leanness. Infections of the skin and soft tissues and pulmonary tuberculosis are often seen, whereas micro- and macrovascular complications are rare, even after long-term follow-up. Ultrasonographic evaluation of the abdomen clearly differentiates MMDM from fibrocalculous pancreatic diabetes. Absence of ketonuria and ketosis despite very severe hyperglycemia in emaciated young subjects is the most significant marker of MMDM.

Immunogenetic studies on malnutrition-modulated diabetes mellitus.

Genetic studies of malnutrition-related diabetes are few. We have analyzed the HLA class II gene polymorphism in malnutrition-modulated diabetes mellitus (MMDM), which was previously referred to as protein-deficient diabetes mellitus (PDDM) in the 1985 WHO classification. Insulin-dependent diabetes mellitus (IDDM) is a polygenic disorder with an autoimmune basis for disease development. In addition to HLA, a second susceptibility locus for IDDM has been identified to lie in the major histocompatibility class III region. Both IDDM and MMDM in eastern Indians are associated with DR3-DQ2 but not DR4-DQ8. The presence of autoantibodies to IDDM autoantigens in clinical MMDM either identifies the slow-onset form of IDDM or suggests autoimmunity different from that in IDDM. Our study demonstrates that the presence of GAD65 antibody and DR3-DQ2 positivity in MMDM patients identifies the underlying autoimmune mechanism in the etiology in eastern India. In autoantibody-negative MMDM patients an association with DR7-DQ2 is identified. The date obtained also indicate the possibility that MMDM can coexist with IDDM in these patients and that malnutrition could be one of the reasons for the slower onset in IDDM-prone individuals. The association of DR7-DQ2 suggests that there is a different immunogenetic background to MMDM than to IDDM. MICA is located in the MHC class I region and is expressed by monocytes, keratinocytes, and endothelial cells. Sequence determination of MICA gene identifies trinucleotide repeat (GCT) microsatellite polymorphism in exon 5. Five alleles with 4, 5, 6, and 9 repetitions of GCT or 5 repetitions of GCT with 1 additional nucleotide insertion (GGCT) are identified. The alleles are A4, A5, A5.1, A6, and A9. We studied the association of MICA alleles with IDDM (n = 52) and MMDM (n = 41) patients and healthy controls (n = 73) from Cuttack, eastern India. MICA was typed by PCR amplification, and fragment sizes were determined in an ABI prism DNA sequencer. Allele 9 of MICA is positively and allele 4 negatively associated with MMDM patients compared to controls. Allele 5 is positively associated with IDDM (OR 2.64, P < 0.05) when compared to controls. Our findings suggest that MMDM is immunogenetically different from IDDM in eastern India and that MIC-A is important in the pathogenesis of MMDM patients from Cuttack in eastern India.

Autoantibodies to tissue transglutaminase in patients from eastern India with malnutrition-modulated diabetes mellitus, insulin-dependent diabetes mellitus, and non-insulin-dependent diabetes mellitus.

Antibodies to tyrosine phosphatase (IA2-Ab) and glutamate decarboxylase 65 (GAD65-Ab) are major markers for IDDM in Caucasians. TTG-Ab is specific for celiac disease. Celiac disease is caused by ingestion of the protein gliadin, a component of wheat gluten, and usually resolves on its withdrawal. Ten to twenty percent of celiac disease patients also have IDDM. The aim of the study was to estimate the prevalence of TTG-Ab in MMDM (n = 71), IDDM (n = 74), and NIDDM (n = 216) and 122 controls from Cuttack in eastern India. MMDM patients are typically young at onset with low body mass index, require insulin for glycemic control, have insulin resistance, and do not develop ketosis on withdrawal of insulin. TTG-Ab was evaluated by radioimmunoassay using in vitro translated recombinant human 35S-TTG. In controls, TTG-Ab was present in 3/122 (2%); in MMDM, TTG-Ab was present in 14/71 (20%); 11/74 (15%) IDDM (P < 0.05 vs. controls) and 23/216 (11%) NIDDM (P < 0.05 vs. controls) were also positive for TTG-Ab. We conclude that MMDM, IDDM, and NIDDM patients from Cuttack have a significantly high proportion of TTG-Ab compared to healthy controls. The highest significance is seen with MMDM patients. It is important to note that subclinical celiac disease must be considered in the differential diagnosis of MMDM.

MHC class I chain-related gene a alleles distinguish malnutrition-modulated diabetes, insulin-dependent diabetes, and non-insulin- dependent diabetes mellitus patients from eastern India.

Insulin-dependent diabetes mellitus (IDDM) is a polygenic disorder with an autoimmune basis for disease development. In addition to HLA, a second susceptibility locus for IDDM has been identified to lie in the major histocompatibility class III region. MIC-A is located in the MHC class III region and is expressed by monocytes, keratinocytes, and endothelial cells. Sequence determination of the MIC-A gene identifies trinucleotide repeat (GCT) microsatellite polymorphism in exon 5. Five alleles with 4, 5, 6, and 9 repetitions of GCT or 5 repetitions of GCT with 1 additional nucleotide insertion (GGCT) are identified. The alleles are A4, A5, A5.1, A6, and A9. The aim of our study was to find the association of MIC-A alleles with IDDM, malnutrition-modulated diabetes mellitus (MMDM), and non-insulin-dependent diabetes mellitus (NIDDM) patients. IDDM (n = 52), MMDM (n = 41), NIDDM (n = 212), and healthy controls (n = 73) from Cuttack, in eastern India, were studied. Of the 212 NIDDM patients analyzed, 96 of them were found to be positive for either GAD65 or IA-2 antibodies. Autoantibodies to GAD65 and IA-2 were measured by radioligand binding assay using (35)S-labeled recombinant human GAD65 and IA-2 in an in vitro transcription/translation system. Autoantibody-positive NIDDM patients (n = 96) and adult healthy controls for NIDDM (n = 113) were also compared. These autoantibody-positive NIDDM patients are considered as slow-onset IDDM or latent autoimmune diabetes in adults (LADA) patients. The samples were analyzed for MIC-A by PCR amplification, and fragment sizes were determined in an ABI prism DNA sequencer. The results of the MIC-A typing are: allele 9 of MIC-A is positively associated (OR 3.62; P < 0.001), and allele 4 is negatively associated (OR 0.31; P < 0.05) with MMDM patients compared to controls. Allele 5 is positively associated with IDDM (OR 2.64; P < 0.05) when compared to controls. Allele 5.1 is positively associated in the autoantibody-positive NIDDM patients compared to adult controls. Our findings of a significant increase of allele A9 in MMDM patients compared to healthy controls suggest that MMDM is immunogenetically different from IDDM in eastern India. MIC-A is important in the pathogenesis of MMDM patients from Cuttack. MIC-A alleles distinguish acute-onset IDDM from slow-onset IDDM, indicating that this molecule may be important for delaying the onset of IDDM with the result that these patients are diagnosed clinically as NIDDM.

Association of HLA class II alleles with different subgroups of diabetes mellitus in Eastern India identify different associations with IDDM and malnutrition-related diabetes.

Genetic studies of Malnutrition related diabetes are few. We have analyzed HLA class II gene polymorphism in different types of diabetes mellitus patients from Cuttack in Eastern India. Patients with insulin-dependent diabetes mellitus (IDDM), non-insulin-dependent diabetes mellitus (NIDDM) and malnutrition-related diabetes mellitus (MRDM), which is subdivided into protein-deficient diabetes mellitus (PDDM) and fibrocalculous pancreatic diabetes (FCPD), were studied and their associations with autoantibody markers. IDDM and PDDM were associated with DR3 and DQ2 but not DR4 and DQ8. FCPD was positively associated with DQ9 (A*0201-B*0303). The association of DQ9 with FCPD suggests differences in the genetic background for susceptibility between IDDM and MRDM in the Cuttack population. There is no association seen between HLA-DR-DQ and NIDDM patients from Eastern India. Clinical classification of diabetes into IDDM, NIDDM and MRDM does not identify the underlying pathological mechanisms. Presence of autoantibodies to IDDM autoantigens in clinical MRDM and NIDDM identifies the slow-onset form of IDDM. Due to the absence of autoantibody assays for diagnosis of IDDM in India, slow onset IDDM is not diagnosed and the patients are classified as NIDDM or MRDM. Our study demonstrates that the presence of GAD65 antibody and DR3-DQ2 positivity in MRDM and NIDDM patients in Eastern India would suggest the presence of slow-onset IDDM. Our data would indicate alternatively that MRDM can coexist with IDDM in these patients and malnutrition could be one of the reasons for the slower onset in IDDM-prone individuals.

Lean (underweight) NIDDM - peculiarities and differences in metabolic and hormonal status - a pilot study.

In the present series of 204 patients with NIDDM, 37 were lean and 35 obese. Mean FBG and HbA1C were significantly higher (P<0.02 and <0.01) in the former. Serum lipids such as total cholesterol (Tc) and triglycerides (Tg) were lower (P<0.05) in the lean while HDLc values were similar. Eight lean patients and 6 obese (Mean BMI : 15.7 vs.27.4) having similar age (48.0 vs 47.7 years) and mean duration of diabetes (4.6 vs 4.2 years) were subjected to the study of insulin and C-peptide status as well as beta cell reserve. The mean basal serum insulin (IRI) level was lower in the lean (15.3 vs. 28.9 mu u/ml ; P<0.05) while there was no statistical difference in the basal C-peptide values. Serum samples analysed 2 hours after 75 G of oral glucose and 1 mg I.V. glucagon (Novo) on two consecutive occasions for IRI and C-peptide responses revealed remarkable differences. The rise in IRI was significantly lower (p<0.01) in the lean after oral glucose and glucagon as compared to the obese. But the C-peptide values did not reveal significant difference suggesting similar reserve in beta cell function in both these groups of patients with NIDDM. The disparity between IRI and C-peptide levels observed was most likely due to excess extraction of insulin by the liver in lean-NIDDM, leading to lower peripheral levels. This phenomenon accounts for the occurrence of severe hyperglycemia inspite of good beta cell function in lean NIDDM.

Serum lipid profile in women using levonorgestrel contraceptive implant, Norplant-2.

Cholesterol content of major lipoprotein fractions along with total cholesterol (Tc) and triglyceride (Tg) were measured in 65 samples from 35 women using levonorgestrel contraceptive implant, Norplant-2 from the third month of implantation until the end of second year along with twenty-five healthy controls. There were significant decreases in all the fractional lipid values, Tc and Tg, up to the sixth month followed by a gradual rise of all lipid parameters except HDLc which remained much below the control level. Interestingly, VLDLc exhibited a significant rise at the end of the second year.

PIP: Total cholesterol, high density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL), and very low density lipoprotein (VLDL) were measured in 35 Indian women using the levonorgestrel-containing implant, Norplant, and in 25 controls. 65 blood samples were taken from the 35 Norplant users at 3,6, 12 and 24 months after placing the implants. Assays were done by quantitative fractionation methods. Serum HDL fell significantly at 3 months (p0.02), and further over 6-24 months of use (p0.001), from 40.0 to 31.8 mg/dl. The control value was 49.2. LDL and VLDL fell to a minimum at 6 months, and rose somewhat at 12 and 24 months. Total cholesterol also was depressed to a mean of 128.8 at 6 months, then rose to 148 mg/dl at 24 months. Control total cholesterol was 190.1. Triglyceride levels followed a similar pattern. The ratios of HDL/total cholesterol and VLDL/total cholesterol in this series suggest increased risk of coronary heart disease. In contrast, the fall in total cholesterol to values below 100 mg/dl in some subjects is cause for concern. These data add to the widely discrepant findings of serum lipoproteins in different studies on Norplant users.

Plasma lipids and lipoprotein cholesterol in undernourished diabetic subjects and adults with protein energy malnutrition.

Estimation of triglycerides (Tg), total cholesterol (Tc), HDLs, LDLc, and VLDLc was carried out in 46 undernourished diabetic subjects (UND); 21 untreated and 25 on insulin; 44 well-nourished diabetic subjects (WND); 22 untreated and 22 on insulin; together with 25 patients with protein energy malnutrition (PEM) and 25 healthy controls less than 50 yr of age. Compared with controls, in the untreated diabetic subjects Tg, Tc, LDLc, and VLDLc were significantly higher in both classes, while HDLc was lower only in WND. Among the treated diabetic subjects, Tg were higher in WND, LDLc lower in UND, and VLDLc higher in both. With regard to relative distribution of cholesterol in the untreated patients, HDLc/Tc was lower in WND, but this was not so in UND. HDLc/Tc was higher in treated UND. Between undernourished and well-nourished groups of diabetic subjects in the untreated patients, HDLc was significantly higher and LDLc lower in the former. Both Tc and LDLc were lower in UND on insulin compared with WND. HDLc/Tc was higher and LDLc/Tc lower in both untreated and treated UND. In adults with PEM, mean values of Tg, Tc, and LDLc were much lower than in controls, as well as in both groups of UND. On the other hand, values of HDLc/Tc were higher and LDLc/Tc lower in PEM compared with controls, but this was not so for treated UND. It is evident from the results of this study that the undernourished have lower levels of plasma lipids and a favorable distribution of cholesterol among the lipid fractions from the point of view of vulnerability to development of atherosclerosis.