GST T1, M1, and IRS-1 G972R Genetic Variants Association to Gestational Diabetes Mellitus (GDM) in Egyptian Women: Linkage to Maternal Hyperglycemia.

BACKGROUND: Gestational Diabetes Mellitus (GDM) shares in part the pathogenic mechanisms of multiple genetic interactions. Some of the T2D susceptibility genes are encountered in association with GDM. OBJECTIVE: We aimed to investigate GST T1, M1, and G972R IRS-I gene polymorphisms with the risk of developing GDM. METHODS: In this randomized case-control study, pregnant women with GDM were genotyped by PCR analysis for glutathione s-transferase-T1, M1 variant polymorphisms. RFLP was done for the G972R IRS 1 gene. Their newborns were additionally assayed for the whole of the clinical, laboratory, and genetic aspects. RESULTS: The T allele IRS-1rs1801278 TT genotype was more frequently detected in GDM mothers in comparison to healthy control ones [for TT homozygous variant; OR(CI 95%): 2.05(1.09-3.87, p: 0.025)]. Furthermore, GST T1 null was significantly presented in GDM mothers than those of control mothers [OR (CI95%: 0.29 (0.084-1.02), p:0.04]. Added to the significant correlation of glycemic indices to clinical parameters of infants born to GDM, the M1-null genotype of GST was significantly correlated (p<0.05) to abnormal values of respiratory rates and 1 minute-APGAR score noted for extra NICU care. CONCLUSION: Our results suggested that GST T1null and IRS-1 TT genotypic variants were claimed for GDM development among Egyptian women with a possible impact on their newly born infants.

Genetic Indices Relationship to Hyperglycemia-associated Biomarkers: Consistency with miRNA Expression in Egyptian Children with T1DM

Objective: Micro RNAs (miRNAs) are gaining acceptance as novel biomarkers for the autoimmune disorders. However, miRNA profiles have not been investigated in individuals at risk of or diagnosed with type 1 diabetes mellitus (T1DM). To study the expression pattern of miRNAs in plasma obtained from patients with T1DM and compare with matched healthy controls. Methods: Equal numbers of patients with T1DM (90) and healthy-matched control children (90) were assessed for the expression profile of plasma miRNAs including miRNA-101-5p, miRNA-146-5p, miRNA-21-5p, miRNA-375, miRNA-126, and Let7a-5p using reverse transcriptase polymerase chain reaction methodology and quantitative real-time testing. Results: Analysis showed that miRNA-101, miRNA-21 and miRNA-375 were highly expressed, whereas, miRNA-146-5p, miRNA-126, and miRNA-Let7a-5p showed significantly low levels of expression in T1DM patients compared to controls (p<0.05). In addition, miRNA-101 and miRNA-146 correlated with age at diagnosis of T1DM and disease duration, respectively. Furthermore, multivariate analysis showed that miRNA-126 and Let7a-5p had a significant negative correlation with mean hemoglobin A1c (HbA1c) values. Conclusion: Dysregulation of the six miRNAs analyzed suggested a possible role as biomarkers in T1DM. miRNA-101 was correlated with age at diagnosis while miRNA-146 correlated with disease duration. Two further miRNAs correlated with the existing biomarker, HbA1c.

Genetic Variability of the Paired Box Transcription Factor; PAX8 Gene: Guidance Towards Treatment Strategies in a Cohort of Congenital Hypothyroidism.

The contribution of PAX8 genetic variants to congenital hypothyroidism (CH) is not well understood. We aimed to study the genetic variability of exons 3 and 5 of PAX8 gene among a cohort of children with congenital hypothyroidism in correspondence to their clinical aspect. Blood samples were collected from 117 children (63 girls and 54 boys) with CH and enrolled as cases (Group I). All cases underwent biochemical confirmation with low FT4 and high TSH levels and thyroid gland imaging, along with equal number of matched apparently healthy individuals who served as controls (Group II). Genomic materials for exons 3 and 5 of PAX8 gene were extracted, amplified by PCR, detected by electrophoresis, purified, and sequenced by the Sanger technique through the application of ABI 3730x1 DNA Sequencer. Out of 117 cases, eight different effective PAX8 mutations were detected in exon 3 (G23D, V35I, I34T, Q40P, p.R31C, p.R31H, p.R31A, and p.I47T) in 14 patients with their sonographic findings ranged from normal, hypoplastic to thyroid agenesis. Besides the reported mutations, one novel mutation; R31A was detected in 1 euotopic case. Exon 5 analysis revealed no detected mutations elsewhere. In contrast, all healthy control children showed no mutation and normal sonographic findings. Mutations in exon 3 of PAX8 gene, implies its important role in thyroid development and function, as a first estimate of PA8 mutation rate in Egyptian patients with CH having normal and dysgenetic gland. Using ultrasound is mandatory for diagnosis and guiding the treatment of children with CH.

Correction to: The G178A polymorphic variant of INSL3 may be linked to cryptorchidism among Egyptian pediatric cohort.

In the original publication.

The G178A polymorphic variant of INSL3 may be linked to cryptorchidism among Egyptian pediatric cohort.

Cryptorchidism (CO) is a genital disorder of multifactorial etiology, with serious remote complications. Mutations in insulin-like 3 hormones (INSL3) G/A variant remain a matter of inquiry. We aimed to investigate the association between G178A-INSL3 polymorphism and undescended testis in a cohort of Egyptian children. In this study, a total of 160 children, including 80 cases with primary non-syndromic undescended testes and 80 healthy children with normal external genitalia as controls, both, were analyzed after detailed history, physical examination and imaging for mutations of G178A polymorphism of INSL3 gene by restriction fragment length polymorphism (RFLP) technique. We found most of the undescended testes were inside the inguinal canal mainly on the left side. Genetic analysis revealed that the mutant A allele of G178A INSL3 variant was significantly detected in the patient group with a frequency of 26.2% against 12.5% for control subjects, especially among cases with an evident family history of similar cases as shown by p value = 0.001 and odd's ratio (CI95%) of 0.13 (0.04-0.723). In conclusion, G178A-INSL3 gene polymorphism could be a susceptibility factor for testicular maldescent in Egyptian children. Also, family history of similar cases was considered as significant predictive risk for cryptorchidism, added to the shared genetic links to consanguinity in our locality.

The Impact of Antiepileptic Drugs on Thyroid Function in Children with Epilepsy: New Versus Old.

OBJECTIVES: To investigate the effects of traditional antiepileptic drugs (AEDs) versus newer AEDs on the thyroid hormone profile of children with epilepsy. MATERIALS & METHODS: A total of 80 children with epilepsy were included in this study and were divided into two groups. Group 1 included 40 children with epilepsy on traditional AEDs, and group 2 included 40 children with epilepsy on newer AEDs. Forty healthy children were also included as the control group (group 3). We analyzed the serum levels of free triiodothyronine (FT3), free thyroxine (FT4), and thyroid stimulating hormone (TSH). RESULTS: In epileptic children treated with traditional drugs, there was a significant reduction in the serum level of FT4 and a significant increase in TSH concentration, compared to the control group (P<0.001). Conversely, epileptic children treated with newer AEDs showed no significant changes in the serum concentrations of FT3, FT4, and TSH, compared to the control group. CONCLUSION: Traditional AEDs have more significant effects on thyroid hormone profile, compared to newer AEDs.

Serum Levels of Interleukin-6 and Tumor Necrosis Factor Alpha in Children With Attention-Deficit Hyperactivity Disorder.

BACKGROUND: Attention-deficit hyperactivity disorder (ADHD) is a common disorder in children, but its etiology and pathogenesis are still unclear. AIMS: The aims of this study were to measure the level of serum interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) as markers of immune system involvement in children with ADHD, and to study their correlation with symptoms severity of ADHD. MATERIALS AND METHODS: The study was conducted on 80 children diagnosed as ADHD based on the criteria adapted from the Diagnostic and Statistical Manual of Mental Disorders, 5th Edition. Eighty healthy children of matched age and sex served as a control group. All children enrolled in the study were subjected to history taking, clinical examination, and psychometric tests. Assay for serum IL-6 and TNF-α for all patients and controls was performed using enzyme-linked immunosorbent assay. RESULTS: The mean serum level of IL-6 was 26.11 ± 11.14 and 6.23 ± 2.52 in children with ADHD and controls, respectively. Children with ADHD showed significantly higher serum IL-6 levels than the control group (P = 0.001). Serum IL-6 showed no significant correlation with the intelligence quotient (IQ) or the Abbreviated Conners' Rating Scale scores for parents. However, TNF-α showed no significant differences between the two groups and no significant correlation with the IQ or the Abbreviated Conners' Rating Scale scores for parents. CONCLUSION: Serum IL-6 levels were significantly higher in children with ADHD compared to controls; however, the IL-6 levels did not correlate with ADHD symptoms severity. Increased IL-6 levels may contribute to the etiology of ADHD.

BMI relationship to the onset of puberty: assessment of growth parameters and sexual maturity changes in Egyptian children and adolescents of both sexes.

Background Puberty is the period of human growth and development. To determine the onset of puberty with regards to the effect of higher adiposity, together with growth parameters of the participants at various stages of sexual maturity for both sexes. Methods The study was conducted on 1944 children (8-16) years; 1022 girls (52.6%) and 922 boys (47.4%) were taken at random. Pubertal assessment was done using Tanner staging that assigned breast development in females and pubic and axillary hair in males and females. Testicular volume was recorded using a Prader orchidometer. Height, weight, body mass index (BMI), body mass (BM) fat, body fat percentage, through applying a body impedance analyzer, and others were recorded. Results The mean ages at the onset of puberty for females and males in our study were 10.29 ± 1.1 and 11.34 ± 1.02 years, respectively. Pubic hair (stage PH2) was attained at mean age of 10.72 ± 0.84 and 11.98 ± 1.03 years for females and males, respectively. For axillary hair (stage AH2), the mean age was 12.47 ± 0.68 years for females and 13.8 ± 0.58 years for males. The mean age at menarche was 12.41 ± 0.65 years. In concordance to BM fat and percentage, all pubertal stages started earlier in females with BMI ≥85th percentile comparable to females within average BMI. As for males, no significant relation was noted between mean pubertal ages and BMI values. Conclusions A significant association of mean ages of Tanner stages to excess weight especially in females warranted the increasing awareness about health care, nutritional aspects, and living circumstances.

IL-1ß-31/IL1-RA genetic markers association with idiopathic generalized epilepsy and treatment response in a cohort of Egyptian population.

Background: Neuroinflammation is an important feature of epileptogenesis.Objectives: To investigate the association of Interleukin-1beta-31 (IL-1ß-31) and Interleukin-1 receptor antagonist (IL1-RA) genetic polymorphisms with idiopathic generalized epilepsy and demonstrate their influence on drug resistance in children.Materials and Methods: One hundred children with idiopathic generalized epilepsy were age and gender-matched with apparently healthy controls. Both groups were genotyped for IL-1ß-31, and IL1-RA gene variants, analysis of these single nucleotide polymorphisms (SNPs) was done through restriction digestion of the corresponding polymerase chain reaction (PCR) products by restriction fragment length polymorphism (RFLP) assay.Results: Genotype frequency of rs1143627 TT of IL-1ß-31 and the homozygous IL1RN*I were found to be more prevalent in epileptic patients (p < .05, OR 0.12 and 5.27respectively). Also observed, T allele of IL-1ß-31 and IL1-RAI/I were substantially positively correlated with drug resistance against those who responded well to antiepileptic drugs (AEDs).Conclusions: The significant association with IL-1ß-31T and IL1-RAN*I alleles potentiated their useful role as predictive markers for the development of epilepsy and response to medical therapy.

Serum Neuron-specific Enolase and S100 Calcium-binding Protein B in Pediatric Diabetic Ketoacidosis

Objective: Neuron-specific enolase (NSE) and S100 calcium-binding protein B (S100B) are markers of different neurological disorders. The aim was to investigate the relationship between NSE and S100B serum concentrations and the severity of diabetic ketoacidosis (DKA) in diabetic children. Methods: Eighty children with DKA, 40 with type 1 diabetes mellitus (T1DM) without DKA and 40 healthy controls were enrolled. Severity of DKA was assessed according to blood pH and bicarbonate concentration. Serum NSE and S100B were measured in all participants. In the DKA group serum NSE and S100B were measured at three time points, at admission and at 12 hours and 24 hours after starting treatment. Results: Children with DKA showed significantly higher serum levels of NSE at all time points compared to children with T1DM without DKA and controls (p<0.01), while serum S100B concentrations did not differ between the three cohorts. Children with T1DM but without DKA also had significantly higher serum levels of NSE (p<0.01) compared to healthy controls. Patients with low Glasgow Coma Scale score (GCSS) and those with moderate and severe DKA had significantly higher levels of NSE at all time points (p<0.01 for each) compared to patients with normal GCSS and those with mild DKA. No significant differences were found in serum S100B levels according to the severity of DKA and GCS (p>0.05). Younger age, lower GCSS, higher glucose and HbA1c, lower pH and lower serum bicarbonate were the risk factors associated with elevated NSE. Conclusion: Serum NSE is elevated in all patients with type 1 DM and, in patients with DKA, correlates with severity of DKA. However, serum S100B concentration did not differ between T1DM with or without DKA and healthy controls.

The Potential Implication of FTO rs17817449 Gene Polymorphism on BMI Mediated Risk For Type2 Diabetes Among Obese Egyptian Children And Adolescents.

BACKGROUND: Genetic variations of the FTO gene were associated with obesity and type 2 diabetes determinants in the European population, notably raised blood levels of insulin and glucose. OBJECTIVE: The aim of this study was to test the association of FTOrs17817449 with obesity/BMI and type 2 diabetes risk among obese Egyptian population. MATERIALS AND METHODS: In this case-control study, (PCR-RFLP assay) was used for genotyping FTOrs17817449polymorphism (SNP) in 120 obese children and 120 controls conducted from attendants of genetic & endocrinology Unit and outpatient clinics, Pediatric Department, Faculty of Medicine, Menoufia University Hospitals. In combination with anthropometric measurements of obesity, predisposition to T2D risk was analyzed (fasting insulin, fasting glucose, insulin resistance). RESULTS: Consanguinity was evident in 32.5% of cases. Positive family history of both obesity and T2D was found to be significant statistically (p<0.05). FTO rs17817449G allele was positively associated with WC (Waist Circumference) (Mean ± SD 84.1 ± 9. 3), raised BMI (Body Mass Index) (32.7 ± 3.5), fasting glucose (114.1 ± 12.8mg/dl), fasting insulin (7.2 ± 1.2µU/ml) and insulin resistance (61.1% of cases) (p<0.001). The odds ratio of obesity was 1.75(95%CI 1.02-3.02) for GT and GG genotype. Fasting glucose and fasting insulin showed statistically significant risk for T2D in the obese group. CONCLUSION: Genetic variation in FTOrs17817449(G allele) was definitely associated with raised BMI, BMI z-score and fasting insulin, and lowered QUICKI values, that predicted the risk for type 2 diabetes among obese children harboring the mutant G allele.

The value of serum procalcitonin in acute meningitis in children.

Early diagnosis and initial therapy are important to reduce the complications of bacterial meningitis. We aimed to evaluate the diagnostic value of serum procalcitonin in children with acute meningitis. We included 40 children (4 months-14 years) suspected to have acute meningitis in our study. Based on the clinical scenario, physical examination and complete analysis of cerebrospinal fluid, patients were assigned into two groups: bacterial meningitis group (24 patients) and aseptic meningitis group (16 patients). Twenty-five apparently healthy children of matched age and sex served as a control group. Procalcitonin, C-reactive protein, and leukocyte count were measured initially at the time of admission and again after 72 h. Initially, patients with bacterial meningitis showed statistically significant higher values of serum procalcitonin than both patients with aseptic meningitis and the control groups (p < 0.001). After 72 h of treatment, patients of bacterial meningitis group showed statistically significant lower values of serum procalcitonin than their initial values (P < 0.05). The cutoff point of procalcitonin needed for early diagnosis of bacterial meningitis was >10 ng/ml at the time of admission. However, values of procalcitonin >2 ng/ml had 100% sensitivity. Whereas, the specificity, negative predictive value and positive predictive value of procalcitonin were 63%, 100%, and 67% respectively. Serum Procalcitonin can be used as an early diagnostic marker of acute bacterial meningitis and its differentiation from aseptic meningitis. In acute bacterial meningitis, it can be used to follow the response to antibiotic therapy.

IL-10 polymorphisms and T-cell subsets could affect the clinical presentation and outcome of childhood immune thrombocytopenia in Egyptian population.

The aim is to study IL-10 polymorphisms and IL-10 level and assess their relation to T-cell subsets in childhood immune thrombocytopenia (ITP). In all, 40 (25 acute, 15 chronic) ITP child patients were investigated at time of presentation, compared to 15 healthy, age- and gender-matched controls and followed up for 1 year to determine chronic cases. Studying the effect of IL-10 promoter polymorphism was done by PCR-RFLP, IL-10 level was determined by ELISA, natural killer cells and T-cell subsets were evaluated by flow cytometry. Subjects with IL-10 promoter (1082 AA and 592 AA) genotypes had lower IL-10 levels and had lower CD4%, higher CD8%, lower CD4/CD8 ratio and lower T-reg%. IL-10 polymorphisms had no effect on NK%. IL-10 serum levels and IL-10 promoter polymorphic genotype frequencies are not different between ITP cases and controls; however, in ITP patients, IL-10 promoter (1082 AA and 592 AA) genotypes and associated lower CD4, higher CD8, lower CD4/CD8 ratio is associated with more severe thrombocytopenia at presentation and had a poorer response to first-line treatment. Patients with lower T-reg cells had a higher tendency to develop chronic ITP. IL-10 level and polymorphisms as well as disturbed T-cell subsets percentages are demonstrable effectors of immune dysfunction in ITP and can affect the presentation and outcome of childhood ITP.

Impact of Vitamin D Supplementation on Attention-Deficit Hyperactivity Disorder in Children.

BACKGROUND: The role of nutrients and dietary factors in attention-deficit hyperactivity disorder (ADHD) remains unclear. OBJECTIVES: The primary objective was to evaluate the serum vitamin D level in children with a diagnosis of ADHD. The secondary objective was to detect the effect of vitamin D supplementation on cognitive function in those with vitamin D deficiency. METHODS: A total of 50 children with ADHD and 40 healthy controls were included in the study. We measured the serum level of vitamin D. Patients with vitamin D deficiency were subdivided into 2 groups: one with vitamin D supplementation and the other without vitamin D supplementation. Further assessment and follow-up of children with ADHD was done. The Wisconsin Card Sorting Test, Conners' Parent Rating Scale, and Wechsler Intelligence Scale for Children were performed at baseline and follow-up in all cohorts with an ADHD diagnosis. RESULTS: The diagnosis of vitamin D deficiency was significantly greater in children with ADHD compared with the control group ( P < 0.05). Children with ADHD had significantly ( P = 0.0009) lower values of serum vitamin D (17.23 ± 8.98) than the control group(31.47 ± 14.42). The group receiving vitamin D supplementation demonstrated improvement in cognitive function in the conceptual level, inattention, opposition, hyperactivity, and impulsivity domains. CONCLUSION: Vitamin D supplementation in children with ADHD may improve cognitive function.

Screening for developmental delay in preschool-aged children using parent-completed Ages and Stages Questionnaires: additional insights into child development.

BACKGROUND: Developmental delay is a delay in areas of speech, language, motor, social and cognitive development. Because of the negative impact of intellectual and learning disabilities, early identification of children with developmental and behavioral problems using appropriate screening tests is crucial. OBJECTIVES: Utilization of parent-completed Ages and Stages Questionnaires (ASQs) for detecting the developmental delay in preschool age children and clarification of possible associated risk factors. MATERIALS AND METHODS: This cross-sectional study was conducted on 1012 children aged 24-60 months enrolled from six centers (n=608) and six villages (n=404) located in Menoufia Governorate, Egypt. All children were screened by nine age-based questionnaires in the first stage of assessment. Children whose scores were ≤ cut-off points in one or more of the screened developmental areas were considered to have suspected developmental delay (SDD) and underwent further evaluation in the second stage assessment. RESULTS: Among the 1012 studied children aged 24-60 months, 978 (96.4%) had normal development. SDD had an overall prevalence of 3.4%, with the highest rates of SDD in problem-solving (3%), followed by communication (2.4%), fine motor skills (2.2%) and social-personal domain (1%), with no SDD in gross motor skills. SDD was more commonly observed in boys, with a significant association with both parental education and consanguinity. Problems with learning (32.3%) was the most commonly observed provisional diagnosis, followed by language disorders (29.4%). Children with SDD in more than one area of ASQ skills also had mild to borderline IQ scores. CONCLUSION: The use of of parent-completed ASQs showed an overall prevalence of developmental delay in children aged 24-60 months of3.4%. Male gender, consanguinity and parental education were identified as risk factors for developmental delay. Family counselling about the child's developmental state is needed.

Value of electroencephalographic monitoring in newborns with hypoxic-ischemic encephalopathy treated with hypothermia.

BACKGROUND: The values of electroencephalography (EEG) in neonatal hypoxic-ischemic encephalopathy (HIE) during therapeutic hypothermia (TH) are still uncertain. AIMS: The aim of this study is to detect EEG background, the prevalence of seizures during cooling, and to determine different EEG patterns that can predict brain injury in magnetic resonance imaging (MRI). PATIENTS AND METHODS: Thirty-nine newborns with HIE were subjected to TH. Continuous monitoring by video-EEG was carried out throughout cooling and during rewarming. MRI was done for all newborns after rewarming. The predictive value of EEG background for MRI brain injury was evaluated at 6-h intervals during cooling and rewarming. RESULTS: At all-time intervals, normal EEG was associated with no or mild MRI brain injury. At the beginning of cooling, normal background was more predictive of a favorable MRI outcome than at later time points. After 24 h of monitoring, diffuse burst suppression and depressed patterns had the greatest prognostic value. In most patients, a discontinuous pattern was not associated with poor prognosis. Thirty-one percent developed electrical seizures, and 8% developed status epilepticus. Seizures were subclinical in 42%. There is a significant association between duration of seizure patterns detected on the EEG and severity of brain injury on MRI. CONCLUSIONS: Continuous EEG monitoring in newborns with HIE under cooling has a prognostic value about early MRI brain injury and identifies electrographic seizures, approximately 50% of which are subclinical. Treatment of clinical and subclinical seizure results in a reduction of the total duration of seizure pattern supports the hypothesis that subclinical seizures should be treated.