Evaluation of the effect of eltrombopag therapy on the platelet collagen receptor glycoprotein VI (GPVI) expression and soluble GPVI levels in young patients with immune thrombocytopenia.

BACKGROUND: Platelet glycoprotein VI (GPVI) receptor is essential for platelet adhesion and aggregation. Eltrombopag is as an effective treatment for chronic immune thrombocytopenia (ITP); yet, its effect on platelet function is not fully characterized. AIM: This prospective study investigated the effect of eltrombopag therapy on platelet function through assessment of GPVI receptor expression and soluble GPVI levels among pediatric patients with persistent or chronic ITP. METHODS: Thirty-six children and adolescents with persistent or chronic ITP were divided equally into two groups either to receive eltrombopag therapy or the standard of care. All patients were followed-up for 12 months with assessment of bleeding score and complete blood count (CBC). Evaluation of GPVI expression using flow cytometry and measurement of its soluble form by ELISA was done at baseline and at 6 months. RESULTS: ITP patients on eltrombopag had significantly lower bleeding score after 6 months of therapy while the quality of life has significantly improved. Platelet count was significantly increased throughout the study. GPVI expression by flow cytometry and soluble GPVI levels were significantly increased after eltrombopag therapy. After 12 months, ITP patients on eltrombopag were able to maintain a good quality of life and low bleeding score. CONCLUSION: Our data suggest that eltrombopag, through its effect on the GPVI receptor expression and its soluble form, might reduce bleeding manifestations and improve the quality of life of chronic and persistent ITP children independent of its effect on the platelet count.

Assessment of modified Salvadora Persica for removal of 134Cs and 152+154Eu radionuclides from aqueous solution.

Biosorption is a simple and economical method utilized to remove hazardous elements from a waste solution. In this study, a low-cost agricultural waste, Salvadora Persica, was modified with iron oxyhydroxide and evaluated as an economic biosorbent to remove cesium and europium radionuclides from their aqueous solutions. The modified biosorbent was characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction (XRD), elemental analysis as well as thermogravmetirc analysis (TGA). The sorption of 134Cs and 152+154Eu radioisotopes was investigated singly in a batch mode as a function of the solution pH, contact time, and the initial concentrations of the studied ions. The kinetic of the removal process was examined and it was found that the reaction obeys a pseudo-first-order model and the intraparticle diffusion is not the sole mechanism dominating the reaction. Temkin and Sips isotherm models provide the best fit for the equilibrium data. In addition, the sorption of cesium and europium ions was a spontaneous and endothermic process as inferred from thermodynamic studies. The reusability for the sorption of cesium and europium ions reveals the feasibility and efficacy of the modified biosorbent.

Erythroferrone Expression in Anemic Rheumatoid Arthritis Patients: Is It Disordered Iron Trafficking or Disease Activity?

PURPOSE: Erythroferrone (ERFE) is well acknowledged for its inhibitory function on hepcidin synthesis in the liver during stress erythropoiesis, thereby ensuring sufficient iron supply to bone marrow erythroblasts. Hepcidin plays an indispensable role in the pathogenesis of anemia of chronic disease (ACD). Thus, ERFE was suggested to protect against ACD in various diseases. Rheumatoid arthritis (RA) is commonly involved with ACD and high hepcidin levels, with a further increase of the latter in active states. The present study is a case-control study that aimed to determine the pattern of ERFE expression in RA patients with concomitant ACD and study its relationship with hepcidin, erythropoietin (EPO) and disease activity. PATIENTS AND METHODS: Fifty-five RA patients with ACD were categorized into active and inactive RA using the disease activity score (DAS28); 15 healthy subjects were included as control subjects. ERFE was measured for patients and control subjects using quantitative real-time polymerase chain reaction, in addition to testing for CBC, ESR, CRP, iron profile parameters and hepcidin. EPO was assessed for patients of both active and inactive RA groups. RESULTS: ERFE and hepcidin showed the highest levels in active RA; ERFE values were similar in control subjects and inactive RA patients, while hepcidin was significantly higher in inactive RA than control subjects. Patients with high ERFE levels had higher RBC, Hct, MCV, hepcidin and EPO levels. Stepwise regression analysis has identified DAS28 and disease duration as the best predictors of ERFE values, whereas ERFE and hepcidin were independent predictors of disease activity. CONCLUSION: We introduce ERFE as a novel marker of RA activity. Although the inhibitory effect of ERFE on hepcidin is not evident, our results still indicate that ERFE may have a beneficial erythropoietic effect in the context of ACD in RA disease activity.

Flow cytometry in detection of Nucleophosmin 1 mutation in acute myeloid leukemia patients: A reproducible tertiary hospital experience.

INTRODUCTION: Nucleophosmin 1 (NPM1) mutation is one of the most frequent gene mutations in adult acute myeloid leukemia (AML), being detected in 35% of all cases and in up to 60% of patients with normal karyotype AML. AML with mutated NPM1 has distinct pathology, immunophenotyping, and confirmed favorable prognostic significance. Hence, AML with mutated NPM1 is a separate entity in the revised 2016 World Health Organization classification. This study aimed to evaluate the use of a reproducible flow cytometry approach in the assay of mutant NPM1 protein in AML patients and to correlate flow cytometric results with the NPM1 gene mutation. METHODS: Eighty-nine newly diagnosed AML patients were evaluated for the expression of mutant NPM1 using flow cytometry and for the presence of NPM1 exon 12 mutations using high-resolution melting polymerase chain reaction (HRM PCR). RESULTS: The NPM1 mutation was found in 35 (39.3%) patients by HRM PCR. These patients showed a significantly higher level of percentage of positive-stained cells (% positive cells) and normalized median fluorescence intensity (MFI) for mutant NPM1 by flow cytometry than the negative mutation group. CONCLUSION: Flow cytometric detection of mutant NPM1 offers a possible tool to indicate NPM1 mutational status.

Combined Immature Platelet Fraction and Schistocyte Count to Differentiate Pregnancy-Associated Thrombotic Thrombocytopenic Purpura from Severe Preeclampsia/Haemolysis, Elevated Liver Enzymes, and Low Platelet Syndrome (SPE/HELLP).

The occurrence of thrombotic microangiopathy (TMA) in pregnancy is an unfortunate emergency condition. Proper diagnosis is mandatory which requires the consideration of two overlapping diagnoses: severe preeclampsia/haemolysis, elevated liver enzymes, and low platelet syndrome (SPE/HELLP) and thrombotic thrombocytopenic purpura (TTP). The long turn-around times of ADAMTS13 testing precludes the timely distinction between the two conditions. We aimed at evaluating schistocyte counts and immature platelet fraction (IPF%), as both increase in TMAs, to discriminate between TTP and SPE/HELLP of pregnancy. IPF% was measured using Sysmex XE-2100 automated hematology analyzer, and schistocyte counts were estimated microscopically as per the International Council for Standardization in Hematology-Schistocyte Working Group guidelines. The study included 30 pregnant patients with SPE/HELLP, 13 pregnant patients with TTP, and 30 women with normal pregnancy. The discrimination between the two patient categories was based on clinical judgment and TTP cases were identified using the PLASMIC score. TTP patients had higher values of IPF% than SPE/HELLP [19.5% (16.9-27.1) vs 13% (9.5-23.25); p < 0.001]; similar results were revealed regarding schistocyte counts [6.5% (3.9-8.6) vs 2.1% (1.6-3.5); p < 0.001]. IPF% and schistocyte counts were able to discriminate between TMA patients and normal pregnant women, and between and SPE/HELLP and TTP patients. Moreover, the discriminatory function of each was improved when the two parameters were used in combination. IPF% analysis should be used in conjunction with manual schistocyte counting in TMA cases to distinguish TTP pregnant patients from patients having SPE/HELLP.

Erythroferrone, the new iron regulator: evaluation of its levels in Egyptian patients with beta thalassemia.

Since iron overload is the commonest cause of morbidity and mortality in ß thalassemia major (ß-TM), it represents one major target in therapeutic management of the disease. The recently discovered erythroid regulator, erythroferrone (ERFE), governed by high levels of erythropoietin, was found to suppress hepcidin expression, thus increasing iron availability for developing erythroid progenitors. We aimed to investigate ERFE levels in Egyptian ß-TM patients as an attempt to understand its role in the prediction of iron overload states. Our study included 70 ß-TM patients, divided into two subgroups according to the degree of iron overload, and 30 sex and age-matched healthy subjects. ERFE gene expression was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR), and serum hepcidin was measured using enzyme-linked immunosorbent assay (ELISA) technique. Both ERFE gene expression levels and transferrin saturation (TS%) values were able to discriminate among cases with different degrees of iron overload, in contrast to hepcidin. TS% was acknowledged as the best predictor of iron overload (AUC 0.893) in comparison with serum hepcidin and ERFE gene levels (AUC 0.807 and 0.677, respectively), and ERFE gene expression was an independent predictor for the estimated TS%. In conclusion, we suggest that using the ERFE gene expression, combined with serum hepcidin estimation, can substantiate the role of estimated TS% as a promising tool in screening for iron overload in ß-TM patients.

Non-invasive Assessment of Significant Dehydration in Infants Using the Inferior Vena Cava to Aortic Ratio: Is it Useful?

OBJECTIVES: The aim of the study was to assess the accuracy of the inferior vena cava to aorta (IVC/Ao) diameter ratio for predicting significant dehydration in infants relative to their percentage weight change and the clinical diagnosis by a physician. METHODS: A prospective observational study was performed on 200 infants presented with acute diarrhea and clinical evidence of significant dehydration whose treatment required intravenous (IV) fluids as determined by the attending physician at the pediatric emergency department of Tanta University Hospital. Weight was recorded at admission before IV fluid treatment and at hospital discharge. The percentage of dehydration was determined using the following formula: (discharge weight - admission weight)/discharge weight × 100%. Patients with a percentage weight change of <5% were considered to be nonsignificantly dehydrated, whereas patients with a percentage weight change >5% were considered significantly dehydrated. The IVC/Ao diameter ratio was measured for all patients before IV fluid rehydration and again at discharge. RESULTS: Only 134 out of 200 dehydrated infants were found to be significantly dehydrated using the gold standard, percentage weight change. Receiver operating characteristics (ROC) curve analysis of the prehydration IVC/Ao ratio showed a sensitivity of 82%, a specificity of 91%, and an accuracy of 87% for predicting significant dehydration in infants at a cut-off point of less than 0.75. In contrast, physician clinical diagnosis showed a sensitivity of 70%, a specificity of 63%, and an accuracy of 73%. CONCLUSIONS: The IVC/Ao diameter ratio can be used as a reliable predictor for diagnosing significant dehydration in infants.

Serum Osteocalcin Levels in Children With Nonalcoholic Fatty Liver Disease.

OBJECTIVES: The aim of the study was to investigate the relationship between osteocalcin and nonalcoholic fatty liver disease (NAFLD) in children with obesity. METHOD: 60 obese children with NAFLD were taken as a patient group and 60 obese children and normal liver with matching age, sex, and body mass index were taken as a control group. Anthropometric measurements, abdominal ultrasonography for diagnosis and grading of NAFLD, and laboratory investigations in the form of liver function tests, lipid profile, fasting serum glucose and insulin, and serum osteocalcin levels were done for all children. Patients with NAFLD were further divided into patients with metabolic syndrome (MS) and patients without MS. RESULTS: Age of NAFLD children was (10.55 ±â€Š2.71), 20 boys and 40 girls, whereas age of children in control group was (10.05 ±â€Š3.51), 24 boys and 36 girls (P > 0.05). Patients with NAFLD showed significant increase in waist and hip circumference, alanine aminotransferase, alkaline phosphatase, total cholesterol, triglycerides, insulin resistance (IR), fasting serum glucose, and insulin, but lower serum osteocalcin level than control group. Serum osteocalcin level is inversely correlated with waist circumference, triglyceride, liver enzymes, fasting serum insulin, fasting serum glucose, IR, and grades of fatty liver. Increase in alanine aminotransferase, total cholesterol, triglycerides, fasting insulin, and IR went with increase in degree of hepatic steatosis. Serum osteocalcin level <44.5 ng/mL is a good predictor for severity of hepatic steatosis with sensitivity and specificity of 80%. CONCLUSIONS: Osteocalcin plays an important role in glucose and lipid metabolism for protection against NAFLD occurrence and progression. Moreover, it could be a useful marker for progression of NAFLD in children with obesity.

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.

New diagnostic biomarker in acute diarrhea due to bacterial infection in children.

BACKGROUND AND OBJECTIVES: Diarrhea is a major cause of morbidity and mortality in children, and diarrhea may be due to infection that is bacterial or non-bacterial. Differentiation between diarrhea from a bacterial or non-bacterial infection is not a simple task, and no single method is present to differentiate between these causes of diarrhea.To evaluate the diagnostic accuracy of soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) and procalcitonin (PCT) in the diagnosis of acute diarrhea due to bacterial infection. PATIENTS AND METHODS: Case control study of forty children with bacterial infection diarrhea diagnosed by stool culture and CRP, 40 children with acute non-bacterial infection diarrhea and 30 age- and sex-matched healthy controls. Stool cultures, serum CRP, PCT and serum sTREM-1 were measured in all children on admission. RESULTS: Children with acute bacterial infection diarrhea had a significant increase in the serum sTREM-1 and PCT levels on admission compared to patients with nonbacterial infection diarrhea and controls (26.3667 ± 16.8184 ng/ml vs 7.2267 ± 6.4174 ng/ml vs 6.7367 ± 5.6479 ng/ml and 39.9933 ± 22.5260 ng/ml vs 1.8533 ± 1.7123 vs 0.2840 ± 0.1208 ng/ml, respectively; P < 0.05). sTREM-1 demonstrated significantly higher sensitivity (93.7%) and specificity (94.3%) in the prediction of bacterial infection as a cause of acute diarrhea in children with an area under the receiver operator characteristic (ROC) curve (95% CI) of 0.94 (0.84-0.99) at a cutoff value of 12.4 ng/ml. CONCLUSIONS: Both serum PCT and sTREM-1 are valuable in the early diagnosis of acute bacterial infection-induced diarrhea in children, and there was markedly higher diagnostic discriminatory power for sTREM-1.

Immune Status 'in Children with Beta Thalassemia' in Correlation 'with Iron Overload': Single Center Egyptian Study.

BACKGROUND: 'Beta thalassemia is inherited hemoglobin disorder resulting in chronic hemolytic anemia that requires lifelong transfusion therapy'. 'Repeated blood transfusions and RBCs hemolysis are the main causes of iron overload', which in addition to immune abnormalities, are common predisposing factors to infections in patients with thalassemia. The Aim of this Work: The aim of this work was to study immune status including T lymphocyte subsets and serum immunoglobulin levels 'in children with beta- thalassemia in correlation with iron overload'. PATIENTS AND METHODS: The present 'study was conducted on 40 children with beta thalassemia major under follow up at Hematology Unit, Pediatric Department, Tanta University' 'including 24 males and 16 females with mean' age value of 9. 22 ± 3.9 years and 20 'healthy children of matched age and sex as a control group'. All children included in the study were subjected to; 'complete blood count, Hb electrophoresis, serum iron status', T cell subsets including CD3, CD4 and CD8 and serum immunoglobulin levels including IgM, IgA and IgG. RESULTS: 'Pallor and jaundice were the most common presenting' clinical manifestations. Infective episodes 'were significantly higher in patients' compared with controls. There were significantly lower Hb, MCV and MCH levels and significantly higher WBCs and platelets counts, reticulocytes and lymphocytes percentage in patients than controls and no significant differences in MCHC between patients and controls. Serum ferritin and iron were 'significantly higher but TIBC was significantly lower in' patients than controls. CD3, CD4 and IgM were significantly lower but CD8, IgG, and IgA 'were significantly higher in patients than controls' with negative correlation between CD3, CD4, IgM and ferritin and positive correlation between CD8, IgG, IgA and ferritin. CONCLUSION: Iron overload can affect humeral and cell mediated immunity in patients with beta thalassemia with reduction of IgM, CD3 and CD4 and elevation of CD8, IgG, and IgA. RECOMMENDATIONS: Regular follow up of patients with beta thalassemia for detection of iron overload as it affects humeral and cell mediated immunity.

BLOCKADE OF PGE2, PGD2 RECEPTORS CONFERS PROTECTION AGAINST PREPATENT SCHISTOSOMIASIS MANSONI IN MICE.

Schistosomiasis is a chronic disease with considerable social impact. Despite the availability of affordable chemotherapy, drug treatment has not significantly reduced the overall number of disease cases. Among other mechanisms, the parasite produces PGE2 and PGD2 to evade host immune defenses. To investigate the role of PGE2 and PGD2 in schistosomiasis, we evaluated the effects of L-161,982, Ah6809 (PGE2 receptor antagonists alone of combined with each other) and MK-0524 (PGD2 receptor antagonist) during prepatent Schistosoma mansoni infection. Drugs were administered intraperitoneally an hour before and 24 hours after infection of C57BL/6 mice with 100 Schistosoma mansoni cercariae. L-161,982, Ah6809, their combination and MK-0524 caused partial protection against pre-patent S. mansoni infection which was mediated by biasing the immune response towards Th1 phenotype. These results showed that blockade of PGE2 and PGD2 receptors confers partial protection against pre-patent S. mansoni infection in mice and that they may be useful as adjunctive therapy to current anti-schistosomal drugs or vaccines.