Non HLA genetic markers association with type-1 diabetes mellitus

The currently available data identified IDDM1 and IDDM2 as 2 susceptibility loci for type 1 diabetes [T1D]. The major histocompatibility complex [MHC]/HLA region referred to as IDDM1 contains several 100 genes known to have a great influence on T1D risk. Within IDDM2, a minisatellite variable number of tandem repeats [VNTR] locus in the insulin gene [INS] promoter region is likely to represent the etiologic polymorphism. The aim of the present work was to study the association between genotypes and susceptibility to T1D among Egyptian diabetic children and their family members. Twenty-five nuclear Egyptian families with 27 children having T1D, aged 3-14 years, their nondiabetic 44 sibs, aged 3-15 years and their parents were included in our study. All studied children were subjected to: detailed history and family pedigree. Thorough clinical examination and anthropometric measurements. Laboratory work up of diabetes including random blood sugar [RBS] and HbA[1]C. Molecular genetics of INS was studied in four steps; nucleic acid purification, amplification, sequencing and haplotyping using flanking single nucleotide polymorphisms [SNPs] as surrogate markers for minisatellite alleles identification. Analysis of variant repeat distribution among Egyptian families combined with flanking haplotypes revealed that all our diabetic children had class I alleles of INS; 9 had class IC+, 9 had class ID+ and 9 had class ID-, while all non-diabetic family members had class III alleles of INS. Therefore the three class I alleles were considered to be equally predisposing to T1D, while class III alleles are dominantly protective. There was significant positive correlations between body mass index [BMI] and both HbA[1]C and AST liver enzyme among diabetic children with class IC+ but not other alleles; indicating that they need close monitoring of their diabetic control and liver functions beside following specific dietary regimens. It can be concluded that all class I alleles [IC+, ID+ and ID-] are equally important susceptibility factors for T1D among Egyptian children, while class III alleles [IIIA and IIIB] are dominantly protective. It is concluded also that our diabetic children with class IC+ are an especially endangered subgroup of diabetics. Genotyping for INS-VNTR alleles is recommended for diabetic children as an important step of diagnostic and follow up regimens and for their non-diabetic family members for family counseling and early identification of potential diabetics. Further studies of INS-VNTR alleles and HLA haplotypes all over Egypt are recommended to define the Egyptian susceptibility loci for T1D and their relations to the clinical and laboratory findings as an important national programs

Identification and supportive intervention programs of the problems of disabled children

According to International Classification of Diseases-Ninth Revision [ICD-9], neurodevelopmental disorders, asthma and physical disabilities are on the top of child disabilities .The aim of our study was to support families with disabled children and teach them how to deal with different difficulties. The present work was conducted on 85 disabled children [53 males and 32 females], aged 1-12 years allocated into 3 groups. Group 1 [neurodevelopmental group]: 45 patients subdivided into cases of cerebral palsy [CP], mental retardation [MR] and epilepsy [15 patients each].Group 2 [bronchial asthma] included 20 patients. Group 3 [physical disability] involved 20 diabetic children. All disabled children were subjected to thorough history taking, family pedigree, clinical examination and diagnostic investigations .The study was conducted in three steps. The first was a preparatory stage which lasted 6 months, an Initial three months of thorough training and evaluation of medical team and paramedical health providers was followed by an in formative and detailed pilot study which was conducted over the next 3 months .The second step lasted for 6 months and involved weekly visits and applying an in formative and standardized questionnaire to obtain their specific disability problems and to do a thorough family training to manage their problems. The third step [next 6 months]; involved bimonthly visits for assessment, reassessment and correction of the specific solutions taught to the parents. The most common problems encountered among cerebral palsy cases were high risk for injuries defective locomotor function, feeding difficulties and chronic constipation [100%, 73.3%, 40% and 33.3% respectively]. In mental retardation subgroup, the most common problems were delayed growth and development, defective family in formation and support for them and delayed socialization [100%, 73.3% and 46.7% in order]. Among cases of epilepsy we found high risk for injuries and inappropriate diets in 100%and inadequate self confidence in 40% of cases. In bronchial asthma group, most problems were related to: dealing with acute exacerbations [100%], indoor air [60%], molds [40%], and dust mites [35%]. While, among diabetic children, problems were related to insulin regimen and technique of injection, blood glucose monitoring, diet regimen and emotional support. The present work confirmed that well trained parents can improve the problems of their disabled children up to a satisfying extent .Feeding difficulties of CP improved in 66.6% and their defective locomotor function improved markedly in 6.7% and partially in 73.3%. Frequency and severity of bronchial asthma improved in 65%. Attitude of diabetics towards disease improved in 71.4%. Poverty, ignorance, non compliance of parents and presence of mixed types of disability were significant obstacles to our training programs. Hence; improvement of the surrounding environment will decrease the frequency and severity of many problems. Treatment of disabled children is a highly demanding task; it is not just a reflex prescription of medications for complications whenever they occur. Family counseling, team work [psychiatrists physiotherapists, pediatricians, etc.], good communications, improved family knowledge, well trained families and early intervention will ameliorate problems of disabled children. It is recommended to do our best to improve the quality of life of disabled children, to properly teach their parents how to deal with disabilities and to conduct research programs all over of Egypt to precisely establish the real problems and the optimal intervention programs for our disabled children

Heterogeneity and clinical relations of INS-VNTR alleles in Egyptian families of children with type i diabetes mellitus

The currently available data identified IDDMI and IDDM2 as 2 susceptibility loci for type I diabetes [T1D]. The major histocompatibility complex [MHC]/HLA region referred to as IDDMI contains several hundred genes known to have a great influence on T1D risk. Within IDDM2, a minisatellite variable number of tandem repeats [VNTR] locus in the insulin gene [INS] promoter region is likely to represent the etiologic polymorphism. Susceptibility effects of the 3 classes of INS, nature of the interaction of the products of HLA and INS-VNTR, the confounding population substructure and ethnicity are still unresolved issues. The aim of the present work was to study the heterogeneity of INS-VNTR alleles among Egyptian children with T1D and their families and to evaluate the clinical findings associated with different alleles. Twenty-four nuclear Egyptian families with 26 children having TID, aged 2-16 years, and their non-diabetic family members [74] as a control group, aged 1-15 years were studied. All studied children were subjected to: Detailed history and family pedigree. Thorough clinical examination and anthropometric measurements .Laboratory work up of DM including random blood sugar [RBS] and HbA[1]C. Molecular genetics of INS was studied in 4 steps; nucleic acid purification, amplification and genotyping, sequencing and haplotyping using flanking single nucleotide polymorphisms [SNPs] as surrogate markers for minisatellite alleles identification. Analysis of variant repeat distribution among Egyptian families combined with flanking haplotypes revealed that all our diabetic children had class I alleles of INS; 9 had class IC+, 9 had class ID+and 8 had class ID-, while all non-diabetic family members had class III alleles of INS [45 had class IIIA and 29 had class IIIB]. Therefore the 3 class I alleles were considered to be equally predisposing to T1D, while the 2 class III alleles are dominantly protective. It can be assumed that IDDM2 may have a multi-locus etiological basis and that racial differences and developmental plasticity with different phenotypes from a single genotype depending on the early environment are reasonable explanations for differences in between various studies. The present work revealed a markedly high familial occurrence of T1D, mild anemia and increased frequency of urinary tract infection [UTI] among all studied diabetic children. The 3 alleles of INS-VNTR had insignificant relations to demographic, clinical and laboratory variables prevailing among Egyptian children with TID .There was significant positive correlations between body mass index [BMI] and both HbA[1]C and AST liver enzyme among diabetic children with class IC+but not other alleles; indicating that they need close monitoring of their diabetic control and liver functions beside following specific dietary regimens. It can be concluded that all class I alleles [IC+, ID+and ID-] are equally important susceptibility factors for T1D among Egyptian children, while class III alleles [lIlA and 1118] are dominantly protective. It is concluded also that our diabetic children with class IC+are an especially endangered subgroup of diabetics. Genotyping for INS-VNTR alleles is recommended for diabetic children as an important step of diagnostic and follow up regimens and for their non-diabetic family members for family counseling and early identification of potential diabetics. Close monitoring and periodic assessment of diabetic control, anemia, urinary tract infection and liver function is recommended for diabetic children especially those with class IC+alleles. Further studies of INS-VNTR alleles and HLA haplotypes all over Egypt are recommended to define the Egyptian susceptibility loci for TID and their relations to the clinical and laboratory findings as an important national project

study of some clinical and biochemical markers of epileptic seizures

The adverse effects of anticonvulsant drugs, duration and expense of therapy and social implications make it essential for accurate discrimination of epileptic from non epileptic seizures [NES]. There is still no single biochemical marker for epileptic seizures and many patients being treated as epileptics are not actually so. Moreover, the coexistence of pseudoseizures with epilepsy is high. Recently few studies had investigated the neuroprotective erythropoietin [EPO] system in the central and peripheral nervous systems. However, the clinical importance of EPO as a specific biochemical marker for epileptic fits is not yet investigated. Sixty children divided into 3 groups were studied. Group I included 20 recently diagnosed epileptics, aged 3.5-15 years. Group II involved 20 children with recent NES, aged 2-14 years. Twenty children suffering fever of unknown origin with lumbar puncture as part of its diagnostic work up, aged 3-15 years represented group III [control group]. All children were suffering no other neurological, hematological or renal diseases. Thorough history, clinical examination and routine investigations, confirmed diagnosis and established exclusion criteria. CT brain, EEG and EMG were done for all patients. Peripheral white blood cells [WBCs], serum creatine kinase [CK] and serum and CSF albumin and erythropoietin [EPO] were measured 24 hours Post-ictally for all patients and on admission of control children. Family history was positive for epilepsy in 20% of epileptic children. Post-ictal symptoms followed more than a half of epileptic seizures and less than a quarter of NES. The most common types of epileptic seizures were generalized tonic-clonic [GTC], generalized tonic [GT], myoclonic then focal seizures. CT brain was normal among most epileptic and all non epileptic patients; with hemorrhage in two epileptics and calcification in only one. EEG showed focal [FEA], generalized [GEA] and multifocal epileptogenic activities [MFEA] among our recent epileptics. Peripheral WBCs, serum CK and CSF levels of EPO showed a significant elevation 24 hours Post-ictally following generalized tonic-clonic epileptic fits and to a lower extent following focal and non epileptic fits. The 3 parameters showed a significant positive correlation with seizure duration. Serum CK levels were markedly elevated [more than 200 U/L] and CSF levels of EPO increased by more than 2 standard deviations in a high percentage of epileptic seizures especially so; GTC seizures, with this marked degree of elevation as a more sensitive factor discriminating epileptic from non epileptic seizures. Post-ictal symptoms, peripheral WBCs, serum CK and CSF levels of EPO are important discriminative factors between epileptic and non epileptic seizures before proceeding to more sophisticated and expensive investigations

study of DNA damage and antioxidant defense in the peripheral leucocytes of diabetic mothers and their newborn infants

Oxidative stress plays a key direct and indirect role in the pathogenesis of several diabetes and pregnancy related complications in both diabetic mothers and their infants. Forty women and their newborn infants divided into two groups were studied. Group I included 20 diabetic women and their newborn infants. Group II consisted of 20 clinically healthy women and their twenty newborns as controls. All involved mothers and newborns were subjected to detailed history, thorough clinical examination, routine laboratory investigations, imaging studies, and specific laboratory investigations including assessment of glycosylated hemoglobin [HbA1C] for diabetic mothers, and erythrocyte antioxidant enzymes [superoxide dismutase, catalase and glutathione peroxidase] and DNA fragmentation assay for mothers and neonates of both groups. It was found that 25% of the diabetic women had diabetic complications. About 1/3 of the newborn infants of diabetic mothers were large for gestational age, 10% of them had major congenital anomalies [cyanotic heart disease and meningomyelocele] and 15% of them died. Levels of erythrocytes glutathione peroxidase and catalase were significantly lower in diabetic mothers and their infants than those in nondiabetic mothers and their infants. DNA damage, mainly in the form of apoptosis was present in diabetic mothers and their infants [60% and 50% respectively]. There was a significant difference between the values of maximal optical density at 200bp and 600bp between both groups. Comparison between diabetic mothers with and without diabetic complications as regards HbA1C, antioxidants and DNA damage showed that erythrocytes catalase was significantly lower in those with complications [means 366 +/- 54 units/g Hb, 426.3 +/- 45.7 units/g Hb respectively] as a possible explanation for complications in this group. There was a significant negative correlation between HbA1C of diabetic mothers [i.e. diabetic control] and glutathione peroxidase level [i.e. antioxidant defense] in their infants. A significant negative correlation was found between DNA damage and erythrocytes antioxidant [Superoxide dismutase in diabetic mothers, and glutathione peroxidase in their infants]. It can be concluded that hyperglycemia causes a significant reduction of antioxidant capacity [reduced catalase and glutathione peroxidase] in the diabetic mothers and their infants compared with controls and this may be the cause of increased DNA damage observed in these individuals which may lead to the development of diabetic complications in the pregnant mothers and congenital anomalies in their infants. It is recommended to maintain a good control of diabetes and combat oxidative stress to lessen diabetic complications in pregnancy and to avoid congenital anomalies

Antiphospholipid antibodies, fasting insulin, and glycosylated haemoglobin in children with type I diabetes in relation to some clinical risk factors

Fifty children divided into 3 groups were studied. Group I included 10 children with recent onset diabetes type I [<6 months]. Their ages ranged from 6-11 years. Group II consisted of 20 children with prolonged diabetes [>5 years] aged 12-15 years. Twenty non-diabetic children aged 6-15 years represented group lll. All studied children were not suffering from any other autoimmune disorder, nor receiving a medication, which is known to affect any of the studied parameters. All children were subjected to detailed history taking, thorough clinical examination and venipuncture while fasting to measure HBA[1c], fasting insulin and antiphospholipid antibodies [APA] [lgG and 1gM]. It was found that duration of diabetes carries on a more significant effect on both SBP and DBP than the diabetes per Se. WHR reached risky levels [>0.8 in girls and>0.9 in boys] in 30% of children with recent onset diabetes, while BMI reached risky levels [>25 kg/m[2]] in only 10% of them. Fifty percent of children with prolonged diabetes [>5 years] had risky high WHR, while only 30%of them had risky high BMI. So, WHR, was found to be a more sensitive risk factor than BMI. Fasting insulin [Fl] levels were significantly lower in all diabetic children [with recent onset and with prolonged disease duration] than in non diabetic children [mean 3.8, 5.2 and 11.45 uU/ml respectively]. F! was found to be significantly affected by APA [lgG] and BP percentiles [P=0.002 and 0.048 in order]. APA [lgG] was significantly higher in 60%of children with recent onset diabetes, and only in 45% of those with diabetes for>5 years [mean 12.39 and 9.72 GPL/ml respectively]. APA [IgM] showed a less marked increase with diabetes, with high levels in only 20% of recent onset diabetics and 5% of those with prolonged diabetes [mean 8.52 and 6.945 MPL/ml respectively]. APA levels [lgG and 1gM] were affected significantly by WHR, BP percentiles and HBA[1c]. APA especially IgG can be added to the diagnostic autoantibodies of type I diabetes and WHR and BP percentiles to the control regimen [along with HBA[1c]] among children with type I diabetes