Adel A. Hagag

Frequency of red cell alloimmunization in patients with sickle cell anemia in an Egyptian referral hospital

INTRODUCTION Sickle cell anemia (SCA) is an important public health issue in Tanta, Egypt. Erythrocyte transfusions may reduce the morbidity of SCA, however, they are associated with numerous risks. Among other risk categories, alloimmunization to red cell antigens may result from transfusions. The objective of this study was to explore the frequency of red cell alloantibodies among SCA patients who received regular transfusions. MATERIALS AND METHODS A total of 42 patients with SCA were included in this study. This work planned to study the presence of alloantibodies to different red cell antigens in multi-transfused SCA patients using the ID card micro-typing system. Clinical and laboratory data were collected and analyzed to find out the frequency, pattern and factors influencing red cell alloimmunization secondary to multiple blood transfusion in these patients. RESULTS Of a total of 42 SCA patients included in the study, 21.4% of patients developed alloantibodies. The most common alloantibodies were anti-K, anti-E and anti-C. The rate of incidence of these alloantibodies was 7.1%, 4.8% and 4.8%, respectively. There was significant association between alloantibody and the rate of transfused blood. The mean age of patients with and without alloimmunization was 12.0 and 6.2 years. CONCLUSIONS Alloimmunization to minor erythrocyte antigens of variable clinical significance is a frequent finding in transfused SCA patients. Regular screening for red cell alloantibodies would provide better management of these patients.

Expression of lung resistance protein and multidrug resistance-related protein (MRP1) in pediatric acute lymphoblastic leukemia

Multidrug resistance (MDR) is a phenomenon by which cells become resistant to unrelated chemotherapeutic agents. The prognostic value that lung resistance protein (LRP) and multidrug resistance-related protein 1 (MRP1) have in the setting of pediatric acute lymphoblastic leukemia (ALL) is controversial. The aim of this study was to investigate the expression of LRP and MRP1 and effect on clinical outcome and prognosis. The mRNA expression of LRP and MRP1 were analyzed in leukemic blasts of 34 pediatric ALL patients. LRP and MRP1 mRNA expression were detected in 41.2% and 35.3%, respectively. Eleven (91.7%) of 12 patients without LRP achieved CR compared with 9 (50.0%) of 18 with LRP expression. Similarly, 11 (100%) of 11 patients without MRP1 expression achieved CR compared with 9 (47.4%) of 19 with MRP1 expression and higher LRP expression rate or MRP1 expression rate was present in patients with relapse than MDR genes negative patients. The expression of either of two genes was associated with poorer 2-year survival. Also, patients expressing both genes had poorer outcomes and had worse 2-year survival. We suggest that MDR expression affects complete remission and survival rates in ALL patients. Thus, diagnosis appears to provide prognostic information for pediatric ALL.

Serum protein S100B and nucleated red blood cell counts as early markers of cerebral damage in neonates with hypoxic ischemic encephalopathy

Hypoxic ischemic encephalopathy (HIE) is a major cause of neonatal mortality and morbidity. Unfortunately, there are no reliable methods to detect brain damage in these patients. The aim of this study is to investigate whether measurement of serum levels of protein S100B and nucleated red blood cells (NRBCs) counts in asphyxiated full-term newborns could be a useful tool for early detection of post asphyxia brain damage. Thirty full term infants with different grades of HIE together with twenty matched controls were enrolled in the study. Serum samples were collected before the lapse of second hour after birth and samples repeated in the second and third days of life for detection of NRBCs count and level of protein S100B. Serum protein S100B and NRBCs counts were significantly increased in HIE group versus control group (P 0.05). Serum protein S100B and NRBCs counts significantly increased in with increasing HIE severity (P < 0.05). Sensitivity and specificity were calculated for protein S100B, which was higher day 3 (96.7% and 95%, respectively). For nucleated red blood cells, sensitivity and specificity were highest at first day (96.6% and 100%, respectively). From the present study it is concluded that serum protein S100B and NRBCs count are useful tools for prediction of brain damage and the expected course of HIE patients.

Protective Role of Silymarin on Hepatic and Renal Toxicity Induced by MTX Based Chemotherapy in Children with Acute Lymphoblastic Leukemia.

Background ALL is the most common childhood malignancy. The children with ALL are treated with methotrexate (MTX) based chemotherapy protocols. MTX causes unpredictable serious hepatic and renal side effects. Silymarin has antioxidant and anti-inflammatory activities and stimulates tissue regeneration. This study aims to evaluate the protective effects of Silymarin on MTX-based chemotherapy-induced Hepatic and renal toxicity in children with ALL. Patients and Methods 80 children with newly diagnosed ALL were enrolled in the study. They were randomly divided into two groups. Group I included 40 children with ages ranging from 4–13 years and the mean age of 6.85± 2.89 years, who received Silymarin 420 mg/day in 3 divided doses for one week after each MTX dose. Group II included 40 children, with ages ranging from 4–12 years and the mean age of 7.30±2.6 years, who received placebo for one week after MTX therapy. For all patients liver functions including serum bilirubin, total proteins, albumin, globulin and albumin-globulin ratio, alkaline phosphatase, ALT and AST, prothrombin time and activity and renal functions including blood urea and serum creatinine, serum cystatin C and urinary N-acetyl-beta-D-glucosaminidase were done to assess hepatic and renal toxicity before and after chemotherapy. Results There were no significant differences between group I and II as regard liver and renal functions before chemotherapy. After chemotherapy, there were significantly higher values of ALT and AST and alkaline phosphatase, and significantly lower Prothrombin activity in group II compared with group I. No significant differences between group I and II were found in total bilirubin, serum protein, and albumin levels. There was significantly lower blood urea, serum creatinine, and cystatin C and urinary N-acetyl-beta-D-glucosaminidase in group I compared with group II. Conclusion Silymarin improved some hepatic and renal functions in children with ALL who received MTX-based chemotherapy protocols. Recommendations: Extensive multicenter studies could be recommended to prove the hepatic and renal protective effects of Silymarin in patients with ALL who received MTX-based chemotherapy protocols.

Study of gonadal hormones in Egyptian female children with sickle cell anemia in correlation with iron overload: Single center study.

OBJECTIVE/BACKGROUND Sickle cell disease is a hereditary hemoglobinopathy characterized by abnormal hemoglobin production, hemolytic anemia, and intermittent occlusion of small blood vessels, leading to tissue ischemia, chronic organ damage, and organ dysfunction including endocrine organs. The aim of this work was to evaluate some gonadal hormones in female children with sickle cell anemia (SCA) in correlation with iron overload. METHODS This study was conducted on 40 female children with SCA with a serum ferritin of > 1000ng/mL, who were attendants at the Hematology Unit, Pediatric Department, Tanta University, Tanta, Egypt in the period from May 2012 to April 2014. Their ages ranged from 11 years to 15years and the mean age value was 12.63±1.36 years (Group I). Forty female children with SCA of matched age with no iron overload served as a control Group (Group II). For all patients in Groups I and II the following were performed/assessed: complete blood count, hemoglobin electrophoresis, serum iron status, serum estrogen, luteinizing hormone (LH), and follicle-stimulating hormone (FSH). RESULTS There were significantly higher serum ferritin and serum iron levels and significantly lower total iron binding capacity, FSH, LH, and estrogen levels in Group I compared with Group II (mean serum ferritin was 2635.1±918.9 in Group I vs. 292.55±107.2 in Group II with a p value of .001; mean serum iron was 196.3±55.6 in Group I vs. 120±16.57 in Group II with a p value of .001 and mean serum total iron binding capacity was 247.3±28.6 in Group I vs. 327.8.7±21.96 in Group II with a p value of .001; mean FSH level was 1.36±0.22mIU/mL in Group I vs. 2.64±0.81mIU/mL in Group II with a p value of .021; mean LH level was 0.11±0.006mIU/mL in Group I vs. 1.78±1.12mIU/mL in Group II with a p value of .003; mean estrogen level was 21.45±10.23pg/mL in Group I vs. 42.36±15.44pg/mL in Group II with a p value of 0.001) with significant negative correlation between serum gonadal hormones and serum ferritin (r=- .835 and p value of .01 for FSH and serum ferritin; r=- .597 and a p value of .01 for LH and serum ferritin; and r=- 0.624 and p value of .01 for estrogen and serum ferritin. CONCLUSION Female patients with SCA with iron overload may have gonadal hormone deficiency with significant negative correlations between gonadal hormones including FSH, LH, estrogen, and serum ferritin. Recommendations include regular iron chelation for prevention of irreversible damage of the ovaries and attaining normal sexual maturation, and regular follow up for females with SCA with assessment of puberty as they are more vulnerable to develop hypogonadism and may require hormonal replacement therapy.

Study of Male Sex Hormone Levels in Male Egyptian Children with Beta-Thalassemia: Correlation with Iron load

BACKGROUND Beta thalassemia is an inherited hemoglobin disorder resulting in chronic hemolytic anemia. RBCs hemolysis and repeated blood transfusions are the major causes of secondary iron overload which leads to deposition of iron in different endocrine glands. Delayed puberty and hypogonadism are the most obvious clinical consequences of iron overload. The aim of this study was to evaluate male sex hormone levels in male children with β- thalassemia major in correlation with iron overload. MATERIAL AND METHODS The present study was conducted on 60 male children with β- thalassemia major with serum ferritin of more than 1000 ng/ml with their age ranging from 11-18 years and mean age value of 14.16±2.48 (Group I) and 60 male children with β- thalassemia major of matched age with no iron overload (Group II). For all children in both groups the following were done: Complete blood count, Hb electrophoresis, serum ferritin, serum iron, TIBC, serum testosterone levels and assessment of testicular volume by ultrasound and Orchidometer. RESULTS Serum iron and ferritin were significantly higher while TIBC, serum testosterone levels and testicular volume were significantly lower in Group I than Group II (Mean serum iron was 221.70 ± 46.76 in group I versus 122.45 ± 14.32 in group II with p value of 0.001, mean serum ferritin was 2595.06 ± 903.43 in group I versus 373.75 ± 6.82 in group II with p value of 0.001, mean serum TIBC was 210.93 ± 18.17 in group I versus 311.40 ± 13.57 in group II with p value of 0.001, mean serum testosterone was 1.01±1.61 in group I versus 2.73±2.66 in group II with p value of 0.006, mean testicular volume was 4.45± 4.92 in group I versus 8.66±7.08 in group II with p value of 0.016). There was significant negative correlation between serum ferritin and serum testosterone and between serum ferritin and testicular volume in studied patients in group I (r = -0.457 and p value = 0.011 for correlation between ferritin and testosterone and r = -0.908 and p value = 0.001 for correlation between ferritin and testicular volume). CONCLUSION Male sex hormone and testicular volume were significantly lower in thalassemic patients with iron overload, significant negative correlation and serum ferritin. RECOMMENDATIONS Regular follow up for thalassemia patients for early detection of iron overload with regular assessment of puberty as thalassemic patients are vulnerable to develop hypogonadism and may require sex hormone replacement therapy.

Study of Serum Levels of Some Oxidative Stress Markers in Children with Helicobacter pylori Infection

BACKGROUND Helicobacter pylori are gram-negative spiral shaped bacteria, with sheathed flagella. H. pylori infection is one of the most common chronic infections in humans. Infection is usually acquired during childhood, and becomes a lifelong infection in most people unless treated. The aim of this study was to evaluate serum levels of oxidative stress indices in children with H. pylori infection. MATERIAL AND METHODS The present study was carried out on 60 children infected with H. pylori including 28 males, 32 females with their age ranging from 7-10 years and mean age value of 8.5 ± 1.65 ( Group I). This study included also 60 children as a control group including 26 males, 34 females with their age ranging from 7-11 and mean age value of 8.99 ± 1.63 (Group II). For all children in groups I the following were done: Diagnosis of H. pylori infection through H. pylori stool antigen testing using enzyme immunoassay kit and gastric antrum mucosal biopsies which were tested for urease activity using Campylobacter like organism test (CLO test). Measurements of serum oxidative stress markers including Superoxide dismutase (SOD), Malondialdhyde, Glutathione, Catalase and Nitric oxide were done in patients and controls. RESULTS Serum nitric oxide and reduced glutathione were significantly lower in patients compared to controls while serum MDA, Serum catalase and Serum SOD were significantly higher in patients compared to controls (nitric oxide was 91.111 ±6.366 in patients versus 107.211±2.121 in controls with p value of 0.001, reduced glutathione in patients was 2.457± 0.081 versus 2.889±0.491 in controls with p value of 0.001, serum MDA in patients was 140.22±5.18 versus 116.22±2.98 in controls with p value of 0.001, catalase was 401.645± 4.344 versus 278.221±71.712 in controls with p value of 0.001 and SOD in patients was 16.936±9.145 versus 5.578±0.231 in controls with p value of 0.001). CONCLUSION H. pylori infection is associated with oxidative stress with significantly lower serum nitric oxide and reduced glutathione and significantly higher serum MDA, catalase and SOD in patients compared to controls. RECOMMENDATIONS Antioxidants may be beneficial adjuvant treatment in H. pylori infection as H. pylori infection is associated with oxidative stress.

Thyroid Function ‘in Egyptian Children with Sickle Cell Anemia in correlation with iron’ load

BACKGROUND Sickle Cell Disease (SCD) is characterized by defective hemoglobin synthesis, hemolytic anemia, frequent thrombosis and chronic organ damage including endocrine organs. AIM To assess thyroid function in children with SCD in correlation and iron load. PATIENTS AND METHOD This study was conducted on 40 children with SCD with iron overload (serum ferritin more than 1000 ng/ml) including 22 males and 18 females with their ages ranging from 11-14 years and mean age value of 11.63±1.36 years and 40 healthy children of matched age and sex as a control group. For all patients; complete blood count, hemoglobin electrophoresis, serum ferritin, serum iron, iron binding capacity and thyroid function including Free Thyroxine (FT4), Free Triiodothyronine (FT3), Thyroid Stimulating Hormone (TSH), Thyroid Peroxidase Antibody (TPOAb) and Thyroglobulin Antibody (TgAb) were done. RESULTS Significantly higher serum ferritin and iron and significantly lower Total Iron Binding Capacity (TIBC) were found in patients compared with controls (mean serum ferritin was 1665.2±1387.65ng/ml in patients versus 192.55±107.2ng/ml in controls with p-value of 0. 007, mean serum iron was 164±83.9 ug/dl in patients versus 89.5±4.5ug/dl in controls with p-value of 0.039, mean TIBC was 238±44.5ug/dl in patients versus 308±11ug/dl in controls with p-value of 0.001). Significantly higher serum TSH and significantly lower Free T3 and Free T4 were found in patients compared with controls with no significant correlation between thyroid hormones and serum ferritin (mean serum TSH was 4.61±1.2 µIU/mL in patients versus 2.11 ± 0.54 µIU /mL in controls with p-value of 0. 045, mean serum FT3 was 2.61 ±1.3 pg/mL versus 3.93±0.47pg/mL in controls with p-value of 0.027, mean serum FT4 was 0.91±0.174 ng/dL versus 1.44± 0.164 ng/dLin controls with p-value of 0.047, r = - 0. 008 and p-value was 0. 973 for correlation between free T4 and serum ferritin, r = -0. 028 and p-value was 0. 9 for correlation between TSH and serum ferritin and r= - 0.259 and p-value was 0.27 for correlation betweenT3 and serum ferritin). There were no significant differences between patients and controls regarding thyroid peroxidase antibody and thyroglobulin antibody (mean serum thyroid peroxidase antibody was 22.45± 4.32 in patients versus 22.45 ± 3.21 in controls with p-value of 0.98 while mean serum thyroglobulin antibody was 12.32 ± 2.65 in patients versus 12.99 ± 2.34 in controls with p-value of 0.76. CONCLUSION Thyroid hormones deficiency may occur in some patients with SCD. RECOMMENDATIONS Regular assessment of thyroid function in children with SCD may be recommended as they are more vulnerable to develop hypothyroidism and may require replacement therapy.

Penile and Testicular Measurements in Male Neonates and Infants: Single Center Egyptian Study

BACKGROUND Universal reference values of penile length, circumferences and testicular volume in newborns and infants are inappropriate to be used in variable ethnic backgrounds. OBJECTIVE The aim of this prospective study was to establish normal reference values for stretched penile length, penile circumference and testicular volume for Egyptian newborn and infants. SUBJECTS AND METHODS This observational cross-sectional study included 1850 healthy male full term newborn and infants applied for routine check-up, aged 0 -24 months, the newborn and infants were recruited from Tanta University Hospital in the period from July 2015 to January 2017. Penile length, penile circumference, testicular volume, weight, length and occipito-frontal circumference were measured. RESULTS The studied infants were divided into five groups. Group I: 1-4 weeks, the mean penile length was 3.51 ± 0.49 cm, penile circumference was 3.95 ± 0.48 cm, and testicular size was (right 1.81 ± 0.44 cm and left 1.67 ± 0.47 cm). Group II: > 1-6 months age, the mean penile length was 3.99 ± 0.46 cm, penile circumference was 4.10 ± 0.38 cm, and testicular size was (right 2.10 ± 0.33 cm and left 2.04 ± 0.27 cm). Group III: >6-12 months age, the mean penile length was 4.45 ± 0.48 cm, penile circumference was 4.21 ± 0.33 cm, and testicular size was (right 2.13 ± 0.33 cm and left 2.06 ± 0.28 cm). Group IV: >12-18 months age, the mean penile length was 4.55 ± 0.54 cm, penile circumference was 4.28 ± 0.32 cm, and testicular size was (right 2.12 ± 0.33 cm and left 2.09 ± 0.32 cm). Group V: >18-24 months age, the mean penile length was 4.89 ± 0.63 cm, penile circumference was 4.45 ± 0.33 cm, and testicular size was (right 2.28 ± 0.45 cm and left 2.25 ± 0.45 cm). There were significant positive correlations between penile length, penile circumference, left and right testicular volumes with each other and also with all other anthropometric measures including: weight, height and head circumference. CONCLUSION AND RECOMMENDATION The age-related values of penile and testicular measurements must be known to be able to determine the abnormal sizes and to monitor treatment of underlying diseases. Our study is a step to achieve accurate reference values of penile and testicular measurements for Egyptian male newborns and infants. Therefore multicenter studies are recommended to establish Egyptian norms.

Prognostic value of Prominin-1 Expression in Egyptian children with Acute Lymphoblastic Leukemia: Two centers Egyptian study

OBJECTIVES Acute lymphoblasstic leukemia (ALL) is the most common childhood malignancy. Prominin-1 is a cell-surface trans-membrane glycoprotein expressed on the stem cell surface and has potential role in diagnostic and prognostic work-up of several stem cell cancers. Aim of this Work: To assess the prognostic value of Prominin-1 expression in Egyptian children with ALL. SUBJECTS AND METHODS This study was conducted on 80 Egyptian children with newly diagnosed ALL and 30 healthy children of matched age and sex as a control group. Patient history, and clinical and laboratory examination results were taken, including complete blood count, serum LDH, bone marrow aspiration with cytochemistry, immunophenotyping, Fluorescent In Situ Hybridization technique for detection of t(9;22) and Flow cytometery for estimation of Prominin-1 expression on blast cells. RESULTS No statistically significant differences were observed between Prominin-1 positive and negative patients regarding age, sex and clinical presentation at diagnosis. No statistically significant differences between Prominin-1 positive and negative patients were observed regarding white blood cells and platelet counts, peripheral blood and bone marrow blast cells percentage while there were significantly higher hemoglobin and LDH levels in Prominin-1 positive patients. There were no significant differences between Prominin-1 positive and negative patients regarding immunophenotyping and t(9;22). There were statistically significant differences in disease outcome between Prominin-1 positive and negative expression with higher rate of relapse and death and lower rate of complete remission in patients with Prominin-1 positive expression (14 cases with Prominin-1 positive relapsed versus 2 cases with Prominin-1 negative, 12 cases with Prominin-1 positive died versus 2 cases with Prominin-1 negative and complete remission occurred in 20 cases with Prominin-1 positive versus 30 cases with Prominin-1 negative) (P =0.017). There was statistically significant difference in disease-free survival (P = 0.0072) and overall survival (P = 0.0424) between ALL patients with Prominin-1 positive and Prominin--1 negative expression. CONCLUSION Prominin-1 is a helpful prognostic marker in patients with ALL; therefore, it should be routinely assessed at diagnosis in ALL patients for better prognostic assessment and should be taken in consideration in designing future therapeutic strategies based on patient-specific risk factors.