Dysregulated Apoptosis and Autophagy in Childhood Epilepsy: Correlation to Clinical and Pharmacological Patterns.

OBJECTIVES: We aimed to assess the serum levels of caspase-3 as a marker of apoptosis and microtubule-associated protein 1A/1B-light chain 3 (MAP1-LC3) as an autophagy marker in epileptic children with various clinical and pharmacological types. METHODS: This case-control study was carried out on 90 participants (50 pediatric patients with epilepsy and 40 healthy matched children), the patients were categorized into three groups: Group (A): 25 pharmacosensitive epilepsy, Group (B): 25 pharmacoresistant epilepsy, and Group (C): 40 (age, sex, and body mass index) matched healthy children selected as controls. Serum caspase-3 and MAP1-LC3 were measured in all study groups, using commercially available ELISA kits. RESULTS: Serum caspase-3 was significantly higher among epileptic children, especially in the pharmacoresistant group, cases managed with multiple antiepileptic drugs, and cases with abnormal EEG findings. Conversely, circulating MAP1-LC3 levels showed a significant reduction in epilepsy cases, particularly in pharmacoresistant cases, in cases treated with multiple antiepileptic drugs, and in cases with abnormal EEG data. A significant negative correlation between serum caspase-3 and MAP1-LC3 was found among epileptic children (r = -0.369, p = 0.0083). Serum caspase-3 was a more valid biomarker in helping diagnose childhood epilepsy, while serum MAP1-LC3 was more valid in predicting pharmacoresistant type. CONCLUSION: The study reveals that serum caspase-3 levels were significantly elevated, particularly in pharmacoresistant cases and those managed with multiple drugs. Conversely, MAP1-LC3 levels were significantly reduced in epilepsy cases, suggesting potential involvement of altered apoptosis and autophagy in childhood epilepsy.

Cord Blood Plasma and Placental Mesenchymal Stem Cells-Derived Exosomes Increase Ex Vivo Expansion of Human Cord Blood Hematopoietic Stem Cells While Maintaining Their Stemness.

BACKGROUND: Mesenchymal stem cells (MSCs) have been used for ex vivo expansion of umbilical cord blood (UCB) hematopoietic stem cells (HSCs) to maintain their primitive characters and long-term reconstitution abilities during transplantation. Therapeutic effects of MSCs mainly rely on paracrine mechanisms, including secretion of exosomes (Exos). The objective of this study was to examine the effect of cord blood plasma (CBP)-derived Exos (CBP Exos) and Placental MSCs-derived Exos (MSCs Exos) on the expansion of UCB HSCs to increase their numbers and keep their primitive characteristics. METHODS: CD34+ cells were isolated from UCB, cultured for 10 days, and the expanded HSCs were sub-cultured in semisolid methylcellulose media for primitive colony forming units (CFUs) assay. MSCs were cultured from placental chorionic plates. RESULTS: CBP Exos and MSCs Exos compared with the control group significantly increased the number of total nucleated cells (TNCs), invitro expansion of CD34+ cells, primitive subpopulations of CD34+38+ and CD34+38-Lin- cells (p < 0.001). The expanded cells showed a significantly higher number of total CFUs in the Exos groups (p < 0.01). CONCLUSION: CBP- and placental-derived exosomes are associated with significant ex vivo expansion of UCB HSCs, while maintaining their primitive characters and may eliminate the need for transplantation of an additional unit of UCB.

Serum Levels of Growth-Associated Protein-43 and Neurotrophin-3 in Childhood Epilepsy and Their Relation to Zinc Levels.

BACKGROUND: Epilepsy is one of the most common neurological disorders, and it places a significant economic strain on the healthcare system around the world. Although the exact mechanism of epilepsy has yet to be illustrated, various pathogenic cascades involving neurotransmitters and trace elements have been reported. We aimed to investigate the serum levels of growth-associated protein-43 (GAP-43) and neurotrophin-3 (NT-3) among cohort of Egyptian children with epilepsy and correlate these biomarkers with their zinc levels. METHODS: This case-control study included 50 pediatric patients with epilepsy who were comparable with 50 controls. Neurological assessment and electroencephalogram (EEG) were done to all included children. Biochemical measurements of serum GAP-43 and NT-3 using enzyme linked immunosorbent assays (ELISA), and total antioxidant capacity (TAC) and zinc using colorimetric assays, were performed to all participants. RESULTS: There was significantly frequent positive parental consanguinity among cases with significantly frequent generalized onset seizures (94%) than simple partial seizure (6%). There were significantly lower serum GAP-43 and zinc levels with significantly higher TAC among cases vs. the controls, p˂0.05 for all. There was no significant difference in the serum levels of NT-3 among epileptic children vs. the controls, p = 0.269. Serum Zn was positively correlated with GAP-43 level among epileptic children (r = 0.381, p = 0.006). Serum GAP-43 in diagnosing childhood epilepsy at cut-off point ≤ 0.6 ng/mL showed 78% sensitivity, 62% specificity, positive predictive value (PPV) = 50.6%, negative predictive value (NPP) = 84.9% with AUC = 0.574. CONCLUSION: GAP-43 can be considered a sensitive good negative biomarker in childhood epilepsy which correlated positively with the zinc status.

Biochemical Assessments of Neurotrophin-3 and Zinc Involvement in the Pathophysiology of Pediatric Febrile Seizures : Biochemical Markers in Febrile Seizures.

Febrile seizures (FSs) are a common occurrence in young children and a serious concern in pediatric practice; nevertheless, the causes and mechanisms of FS are still unknown. We hypothesized a relation of neuropeptides such as neurotrophin-3 (NT-3) and growth-associated protein-43 (GAP-43) as well as zinc and the oxidant/antioxidant system with pediatric FS. The study included 100 infants categorized into 50 infants with FS and 50 febrile infants without seizures as controls. Clinical assessments, biochemical assays of NT-3 and GAP-43 using ELISA assay kits, and colorimetric measurements of TAC and Zn were performed to all participants. Overall, significant rises of the values of NT-3 and insignificant increases of GAP-43 were detected in children with FS. At the same time, zinc values and the total antioxidant capacity in serum samples were found to be decreased significantly. In addition, a negative correlation was estimated between NT-3 and zinc levels. Serum NT-3 in diagnosing febrile seizures at cutoff point > 49.62 ng/L showed 100% sensitivity, 46% specificity, positive predictive value (PPV) = 48.1%, and negative predictive value (NPP) = 100% with AUC = 0.678. Significant altered circulating NT-3 and zinc levels in FS may indicate their possible role in the pathogenesis of FS. This may open a way for further research and warrants enlightening of the pathophysiological details of FS.

Analysis of 25-hydroxy cholecalciferol, immunoglobulin E, and vitamin D receptor single nucleotide polymorphisms (Apa1, Taq1, and Bsm1), among sample of Egyptian children with bronchial asthma: A case-control study.

PURPOSE: The current study aimed to assess the serum levels of vitamin D and immunoglobulin E (IgE) among asthmatic Egyptian children and to find out the possible associations of vitamin D receptor (VDR) polymorphisms with bronchial asthma development. METHODS: The study included 100 Egyptian children, 50 asthmatic children who were comparable with 50 age, sex, and body mass index-matched, unrelated healthy controls (HCs) clinical assessments of asthmatic children were done using global initiative of asthma. Pulmonary function tests (forced expiratory volume in 1 second [FEV1], forced vital capacity [FVC], FEV1/FVC ratio) were performed. Enzyme-linked immunosorbent assays of serum vitamin D3 and total IgE were done. VDR-single nucleotide polymorphisms (SNPs) (ApaI, TaqI, and BsmI) detection has performed using polymerase chain reaction through restriction fragment length polymorphism technique. Data analysis was performed using SPSS version 20.0. The studied SNPs were followed the Hardy Weinberg equation. RESULTS: The mean serum level of 25(OH) D3 was significantly lower among asthmatic children (13.46 ng/mL ± 10.50 SD) in comparison to HCs (37.53 ng/mL ± 13. 0.40 SD), P < .05. Vitamin D deficiency was detected in 72% of cases with no significant difference in its level regarding asthma control. There was significantly higher IgE level among asthmatic children (99.83 ku/L ± 233.81 SD) versus HCs (7.52 ku/L ± 3.32 SD), P < .05. Asthmatic children were presented more commonly with TaqI t allele (odds ratio [OR], 2.25; 95% confidence interval [CI], 1.28-3.96; P < .05) and BsmI b allele (OR, 1.83; 95% CI, 1.05-3.21; P < .05). ApaI a allele was not significantly different among patients versus controls (P > .05). TT + Tt and Bb + bb genotypes were significantly higher among cases versus the controls, P < .05 for all. CONCLUSIONS: TaqI and BsmI were associated with risk of bronchial asthma development among Egyptian children. High IgE and Low vitamin D status were frequently occurring among asthmatic children.

Vitamin D receptor rs7975232, rs731236 and rs1544410 single nucleotide polymorphisms, and 25-hydroxyvitamin D levels in Egyptian children with type 1 diabetes mellitus: effect of vitamin D co-therapy.

Purpose: We aimed to examine the possible association role of vitamin D and vitamin D receptor (VDR) single nucleotide polymorphisms (SNPs) in type 1 diabetes mellitus (T1DM) development, glycemic control and complications among a cohort of Egyptian children. Subjects and methods: A prospective case-control study has been conducted on 50 Egyptian children with T1DM who were comparable with 50 controls. Vitamin D and HbA1c were measured. VDR-SNPs [ApaI (rs7975232), TaqI (rs731236) and BsmI (rs1544410)] detection was done by polymerase chain reaction through restriction fragment length polymorphism (PCR-RFLP) technique. Vitamin D supplements were given to the included T1DM children with low vitamin D and reassessments of both HbA1c% and 25(OH)D serum levels were performed in those children three months later. Results: Eighty percent of the included diabetic patients have poor glycemic control. Vitamin D was deficient in 68% and insufficient in 16% of diabetic patients. Significant improvements in both vitamin D and glycemic status among T1DM children, who have low vitamin D and received vitamin D supplementations. There were significantly negative correlations between serum levels of vitamin D with both HbA1c % (r= -0.358, P˂0.05) and daily insulin dose (r=-0.473, P˂0.05). Compared with controls, T1DM children presented more commonly with ApaI a allele (OR: 2.87; 95%CI: 1.39-5.91, P˂0.05) and BsmI b allele (OR: 4.38; 95%CI: 2.30-8.33, P˂0.05). TaqI t allele wasn't significantly differing among patients and controls (P˃0.05). Aa+aa and Bb+bb genotypes were significantly higher among T1DM vs the controls (OR: 3.08;, 95%CI: 1.33-7.15, P˂0.05 and OR: 9.33; 95%CI: 3.61-24.17, P˂0.05respectively). Conclusion: ApaI and BsmI were associated with risk of T1DM development among Egyptian children. Low vitamin D status was frequently occurring among T1DM with significant improvement in the glycemic control of such children when adding vitamin D supplements to the standard insulin therapy.

Validity of biomarkers in screening for neonatal sepsis - A single center -hospital based study.

BACKGROUND: The diagnosis of neonatal sepsis still considered to be a challenge for both clinicians and the laboratory due to the non-specific clinical presentations. The present study aimed to compare and assess the diagnostic & prognostic values of C-reactive protein (CRP), high sensitivity CRP (hsCRP), presepsin, interleukin-6 (IL-6) and procalcitonin (PCT) in neonatal sepsis separately and in combination. METHODS: This hospital-based cross-sectional study has been conducted on 168 neonates recruited from the neonatal intensive care unit (NICU) of Qena University Hospitals, Upper Egypt. Measurements of CRP using latex agglutination test, hsCRP, presepsin, IL6 and PCT assays using commercially available ELISA assay kits were done to all included neonates. RESULTS: There were significantly higher serum levels of CRP among late onset versus early onset sepsis group with significantly higher serum levels of hsCRP and presepsin among early onset compared with the late onset sepsis group (p < 0.05 for all). There were significantly higher hsCRP, presepsin and PCT serum levels in proven versus probable sepsis group (p < 0.05 for all). Significantly higher serum levels of presepsin and PCT were noted among survivors versus non survivors sepsis group (p < 0.05 for all). The cutoff value of the serum level of CRP >6 mg/dl showed lower sensitivity and specificity than that of hsCRP at cutoff >140 ng/ml in diagnosing neonatal sepsis. The cutoff value of presepsin >200 ng/ml showed equal sensitivity and specificity to IL-6 at cutoff >22 pg/ml. The cutoff value of PCT at > 389 pg/ml showed sensitivity and specificity approximate to that of hsCRP. CONCLUSIONS: CRP could be a helpful prognostic marker in late onset neonatal sepsis. hsCRP and PCT have higher diagnostic accuracy in neonatal sepsis in comparison to other studied markers. Both IL-6 and presepsin have equal diagnostic utility in neonatal sepsis, but presepsin could be helpful diagnostic marker in early onset neonatal sepsis.

Myocardial injury induced by scorpion sting envenoming and evidence of oxidative stress in Egyptian children.

In the present study, 45 children in Upper Egypt (less than 16 years old) were admitted to the Pediatric Intensive Care Unit for scorpion envenomation (SE). They were compared with 30 apparently healthy children of matching age and sex as controls. Out of the studied victims, 35 children (78%) showed signs of severe envenomation, while 10 victims (22%) showed signs of mild envenomation. The case fatality was 33%. The serum levels of cardiac markers, cardiac troponin T (cTnT) and I (cTnI), as well as the enzymatic activities of creatine kinase-MB (CPK-MB) and lactate dehydrogenase (LDH) were determined for both victims and controls. In addition, the serum levels of oxidative stress markers, nitric oxide (NO), malondialdehyde (MDA), superoxide dismutase (SOD), reduced glutathione (GSH) and zinc (Zn) were measured. Electrocardiography and echocardiography were done. All the envenomed victims showed significantly higher mean values of cTnT, cTnI, CPK-MB and LDH than control group. These cardiac markers were elevated in severe cases and in non survivors in comparison with mild cases and survivors respectively. Furthermore, the serum levels of NO and MDA were significantly higher while the serum levels of SOD, GSH and Zn were significantly lower in all envenomed victims than the controls (p < 0.05 for all). There were no significant differences in the serum levels of oxidative stress markers among severe and mild cases or between survivors and non survivors victims. There were no significant correlations between the serum levels of cardiac markers and the oxidative stress markers in envenomed victims. In conclusions, oxidative stress occurs in scorpion envenomed children, but does not determine prognosis. Cardiac markers, but not the oxidative stress, remain the most important determining factor for the severity and the outcome of SE.

Circulating Protein Carbonyls, Antioxidant Enzymes and Related Trace Minerals among Preterms with Respiratory Distress Syndrome.

INTRODUCTION: Information about oxidative stress in preterms with Respiratory Distress Syndrome (RDS) is defective, so various researches in this area are required, which may open new roads in understanding the pathogenesis of the disease, hence provide additional helpful therapeutic approaches. AIM: To assess and compare the plasma level of protein carbonyls as a marker for oxidant status and the antioxidant enzymes; Superoxide Dismutase (SOD) and Glutathione Peroxidase (GPx) and the related trace minerals in the form of Copper (Cu), Zinc (Zn) and Selenium (Se) as markers for antioxidant status, in preterms with and without RDS. MATERIALS AND METHODS: A hospital-based case-control study was conducted on fifty-seven preterm neonates (37 preterms with RDS and 20 preterms without RDS) admitted to neonatal intensive care unit of Qena University Hospitals after approval of the University Hospital Ethical Committee. Plasma protein carbonyls assay was done using commercially available ELISA assay kit. Plasma Cu, Zn, Se, erythrocyte SOD and GPx activities assays were done using commercially available colorimetric assay kits. RESULTS: Significant higher plasma levels of protein carbonyls and oxidant/antioxidants ratio (protein carbonyls/{SOD+GPx}) with significant lower plasma levels of Zn, Cu, Se, erythrocyte SOD and GPx activities were found in the preterms with RDS when compared with the preterms without RDS (p<0.001 for all measured markers for both groups). In terms of birth weights and gestational ages, they were negatively correlated with both plasma protein carbonyls and oxidant/antioxidants ratio and positively correlated with plasma copper, zinc, selenium, erythrocyte SOD and GPx activities in a statistically significant manner. Non-significant correlations were found between the measured oxidative stress markers and the severity of RDS. CONCLUSION: Oxidative stress may have a contributory role in the development of RDS among preterms. Lower birth weight and prematurity may increase the susceptibity to oxidative stress among such patients.