Asmaa M. Zahran

Hypoxia Biomarkers, Oxidative Stress, and Circulating Microparticles in Pediatric Patients With Thalassemia in Upper Egypt:

This study aimed to investigate the oxidative stress, hypoxia biomarkers, and circulating microparticles (MPs) in β thalassemia major. The study included 56 children with thalassemia and 46 healthy controls. Hypoxia biomarkers, oxidative stress biomarkers, and total plasma fragmented DNA (fDNA) were detected by the standard methods. The MPs were assessed by flow cytometry. Hypoxia and oxidative stress biomarkers, fDNA, and MPs were higher and total antioxidant capacity (TAC) was lower in patients with thalassemia than the controls. In splenectomized patients and those who had complications, vascular endothelial growth factor (VEGF), malondialdehyde, fDNA, endothelial, platelet, and activated platelet MP levels were higher while, TAC was lower than the nonsplenectomized patients. In conclusion, the increased tissue hypoxia, oxidative stress in β thalassemia, and its relationship with DNA damage and MPs release could explain many complications of thalassemia and may have therapeutic implications. The VEGF could serve as an important indicator for adequacy of blood transfusion in thalassemia.

CD4+ CD25+High Foxp3+ regulatory T cells, B lymphocytes, and T lymphocytes in patients with acute ITP in Assiut Children Hospital.

We aimed to examine the levels of lymphocyte subsets and regulatory T cells in patients with newly diagnosed immune thrombocytopenia (ITP) and their correlation with the course of ITP. The study included 40 pediatric patients with acute ITP and 30 controls. Lymphocytes and regulatory T cells were analyzed by flow cytometry. The percentages of CD19+ and CD8+ cells were significantly increased while that of CD4+ cells and CD4+/CD8+ ratio were significantly decreased. The percentages of CD4+CD25+High and CD4+CD25+High forkhead box protein 3 (Foxp3+) cells and the expression of Foxp3+ in CD4+CD25+High cells were significantly decreased in patients. Age, platelet count, and mean platelet volume (MPV) in patients with brief duration of thrombocytopenia were significantly decreased than in those with prolonged duration. The percentages of CD8+, CD4+CD25+High, and CD4+CD25+High Foxp3+ were significantly increased in patients with brief duration. Age, platelet count and MPV, and CD8+ cells had prognostic significance. CD4+CD25+High Foxp+ T cells may be a helpful prognostic marker in children with acute ITP.

Leukocytes apoptosis and adipocytokines in children with beta thalassemia major.

Abstract β-Thalassemia is a significant public health problem in Egypt. Infectious complications represent the second most common cause of mortality and the major cause of morbidity in β-thalassemia major (BTM). The increased susceptibility of these patients to infectious diseases has been attributed to the abnormalities of the immune system, which is evident by systemic inflammation and immune deficiency. In a case control study, 35 patients with BTM were compared with 30 sex- and age-matched children who served as controls. Serum ferritin, high-sensitive CRP (hsCRP), leptin and adiponectin levels were determined in all subjects. Apoptosis of neutrophils and lymphocytes was measured by the Annexin V-fluoroisothiocyanate binding assay. Serum leptin was significantly lower in patients when compared to controls. In contrast, adiponectin and hsCRP levels were significantly higher in the patients than the controls. Positive correlation was found between adiponectin and hsCRP. BTM patients had significantly higher total leukocytes, neutrophils and lymphocytes compared with controls. BTM children exhibited a significantly increased apoptosis in T-lymphocytes; however, there was no significant difference in the percentage of apoptosis of B-lymphocytes and neutrophils between the patients and the controls. There was a significant negative correlation between serum leptin and the percentage of apoptotic T-lymphocytes. Our BTM patients had a high percentage of apoptotic T-lymphocyte in comparison with controls. In addition, they had disturbed serum levels of adipocytokines and inflammatory markers. These derangements could have a role in the immunological disturbance observed in thalassemic patients.

Dendritic cells in childhood sepsis.

PURPOSE Our aim was to investigate the level and the maturation status of dendritic cells (DCs) in pediatric patients with sepsis and its relation to prognosis. MATERIALS AND METHODS The study included 16 children with sepsis, 24 children with complicated sepsis, and 40 healthy control children. The patients were investigated within 24 hours of intensive care unit admission and after 28 days. Flow cytometric detection of DCs was done. RESULTS Within 24 hours, the levels of both plasmoid DCs and monocytoid DCs and the expression of CD86 and CD83 on DCs were significantly lower in patients than in controls, and the difference was marked in patients with complicated sepsis. The amount of CD86 and CD83 per cell was significantly lower in patients with complicated sepsis. The baseline numbers of monocytoid DCs and plasmoid DCs were higher in the survival patients than in nonsurvival patients. In addition, the expression of CD86 and CD83 on the entire DCs was significantly higher in the survival patients with sepsis. CONCLUSION Sepsis is associated with reduced level of DCs and decreases their maturation. The estimation of DCs number and maturation state may be used as prognostic makers of sepsis.

Does being overweight affect seminal variables in fertile men

The effect of being overweight on seminal variables was assesed in 165 fertile men. Participants were divided into three groups: fertile men with normal body mass index (BMI) (18.5-24.9 kg/m2), fertile overweight men (BMI 25-29.9 kg/m2) and fertile obese men (BMI >30 kg/m2). Medical history was taken, a clinical examination conducted. Semen analysis was undertaken and BMI measured. Seminal reactive oxygen species (ROS) was estimated by chemiluminescent assay, sperm vitality by the hypo-osmotic swelling test and sperm DNA fragmentation by propidium iodide staining with flowcytometry. Fertile obese men had significantly lower sperm concentration, progressive sperm motility and sperm normal morphology, with significantly higher seminal ROS and sperm DNA fragmentation compared with fertile normal-weight men and overweight men (all P < 0.05). BMI was negatively correlated with sperm concentration (r = -0.091; P = 0.014), progressive sperm motility (r = -0.697; P = 0.001), normal sperm morphology (r = -0.510; P = 0.001), sperm vitality (r = -0.586; P = 0.001), but positively correlated with sperm DNA fragmentation percentage (r = 0.799; P = 0.001) and seminal ROS (r = 0.673; P = 0.001). Increased BMI was found to affect semen parameters negatively even in fertile men.

Oxidative Stress, Trace Elements, and Circulating Microparticles in Patients With Gaucher Disease Before and After Enzyme Replacement Therapy

We studied the level of lipid peroxide, nitric oxide (NO), trace elements (TEs), and microparticles (MPs) in Gaucher disease (GD) before and after 1 year of enzyme replacement therapy (ERT). A total of 15 children with GD and 15 healthy controls were enrolled in this study. Serum level of lipid peroxide, NO, and TEs was determined. The MPs were detected by flow cytometry. The level of lipid peroxide was significantly higher in the patients than in the controls even after ERT. Although NO level was normalized in the patients after ERT, zinc and copper were still lower in the patients after ERT. The percentages of various MPs were significantly higher in the patients than in the controls both before and after ERT. There were positive correlations between chitotriosidase and both lipid peroxide and total MPs. Conclusion: The GD is associated with alteration in oxidant and antioxidant status and high level of circulating MPs.

Circulating Endothelial Cells and Endothelial Progenitor Cells in Pediatric Sepsis.

ObjectivesThe aim of the study was to measure the number of circulating endothelial cells (CECs) and circulating endothelial progenitor cells (CEPs) in pediatric patients with sepsis and correlating it with the severity of the disease and its outcome. MethodsThe study included 19 children with sepsis, 26 with complicated sepsis, and 30 healthy controls. The patients were investigated within 48 hours of pediatric intensive care unit admission together with flow cytometric detection of CECs and CEPs. ResultsThe levels of both CECs and CEPs were significantly higher in patient with sepsis and complicated sepsis than the controls. The levels of CECs were higher in patients with complicated sepsis, whereas the levels of CEPs were lower in patients with complicated sepsis. Comparing the survival and nonsurvival septic patients, the levels of CEPs were significantly higher in the survival than in nonsurvival patients, whereas the levels of CECs were significantly lower in the survival than in nonsurvival patients. Serum albumin was higher in survival than in nonsurvival patients. ConclusionsEstimation of CECs and CEPs and their correlation with other parameters such as serum albumen could add important information regarding prognosis in septic pediatric patients.

Effects of royal jelly supplementation on regulatory T cells in children with SLE.

Background and objective To our knowledge, no previous studies have focused on the immunomodulatory effects of fresh royal jelly (RJ) administration on systemic lupus erythematosus (SLE) in humans. Our aim was to study the effect of fresh RJ administration on the disease course in children with SLE with some immunological markers (CD4+ and CD8+ regulatory T cells and T lymphocytes apoptosis). Methods This was an open-label study in which 20 SLE children received 2 g of freshly prepared RJ daily, for 12 weeks. Results The percentages of CD4+ CD25+high FOXP3+cells (CD4+ regulatory T cells) and CD8+CD25+high FOXP3+cells (CD8+ regulatory T cells) were significantly increased after RJ treatment when compared with baseline values. Apoptotic CD4 T lymphocytes were significantly decreased after RJ therapy when compared with baseline values and the control group. Conclusion This is the first human study on the effect of RJ supplementation in children with SLE. Our results showed improvements with 3-month RJ treatment with regard to the clinical severity score and laboratory markers for the disease. At this stage, it is a single study with a small number of patients, and a great deal of additional wide-scale randomized controlled studies are needed to critically validate the efficacy of RJ in SLE.

Regulatory B cells (CD19+ CD38hiCD24hi) in alloimmunized and non-alloimmunized children with β-thalassemia major

BACKGROUND β-Thalassemia major (BTM) is considered the most common hemoglobinopathy in Egypt and is one of the major health problems in our locality. MATERIALS & METHODS We investigated the frequency of B-regulatory cells (CD19(+)CD38(hi)CD24(hi)); (Bregs) among polytransfused alloimmunized and non-alloimmunized children with BTM. The study included 110 polytransfused pediatric patients with β-thalassemia major. Clinical and transfusion records of all studied patients were reviewed. Indirect antiglobulin test was performed to detect the presence of alloantibodies. We used flow cytometry for detection of CD19(+)CD38(hi)CD24(hi) regulatory B cells. RESULTS Alloimmunization was detected in 35.5% of thalassemic patients (39/110). The analysis of our data showed a significantly higher frequency of Bregs (CD19(+)CD38(hi)CD24(hi)) in the peripheral blood of both alloimmunized and non-alloimmunized patients as compared to healthy controls. CONCLUSIONS Our data showed that the frequencies of CD19(+)CD24(hi)CD38(hi) Bregs cells were significantly increased in children with BTM. Our data suggested that Bregs cells could play a role in the clinical course of BTM. The relationship of Bregs to immune disorders in BTM children remains to be determined. Further longitudinal study with a larger sample size is warranted to explore the mechanisms of Breg cells in the disease process in BTM patients.

Splenectomy for patients with β-thalassemia major: long-term outcomes

Background/aim The use of splenectomy for thalassemia major is restricted over concerns of its long-term outcome. The aim of this study was to assess the long-term outcomes of splenectomy for patients with β-thalassemia major. Patients and methods This study included 70 patients with β-thalassemia major. Patients were classified into two groups: 35 patients underwent splenectomy (S group) and 35 patients did not undergo splenectomy (NS group). Patients were assessed by review of medical records, assessment of medical history, and a clinical examination. In addition to complete blood count, liver function tests and serum ferritin were performed. Assessment of lymphocyte populations was carried out by flow cytometry. These investigations were performed at least 2 years after splenectomy in the S group. Results The mean age of the patients who underwent splenectomy was 6.68 ± 2.54 years and the mean postoperative follow-up period was 6.26 ± 3.03 years. Splenectomy improves anemia, but does not reduce iron burden; more patients were found to be on regular iron chelation after splenectomy. Hematocrit and red blood cell indices were significantly increased after splenectomy. Platelet count increased significantly in the S group (644.700 ± 299.400/mm 3 ). There were no significant differences in T-lymphocyte populations between both groups. IgM memory B lymphocytes were lower in the S group compared with the NS group. No overwhelming postsplenectomy infection was reported in this series. Postsplenectomy portal vein thrombosis was reported in one (2.9%) case. Conclusion With long-term follow-up after splenectomy for treatment of thalassemia major, thrombocytosis and the risk of thromboembolic persist. Splenectomy improves anemia, but does not reduce iron burden or the requirement for blood transfusion. Proper preoperative vaccination can reduce the risk of overwhelming postsplenectomy infection.