Rania A. Zayed

Detection of HCV RNA in the peripheral blood mononuclear cells of serum HCV RNA-negative Egyptian patients under interferon treatment.

Introduction:Despite recent success after the introduction of combination therapy with interferon (IFN)-&agr; and ribavirin, approximately 60% of patients with hepatitis C virus (HCV) genotype 4 fail to respond. Resistance to antiviral therapy remains a serious problem in the management of chronic hepatitis C. In most patients, HCV RNA could be detected in peripheral blood mononuclear cell (PBMC). Objective:The authors aimed to investigate the predictive value of HCV RNA in PBMC of patients with chronic hepatitis C after IFN treatment, which may act as the source of HCV reinfection of hepatic cells. Methods:Seventy patients with chronic hepatitis C were treated with IFN plus ribavirin for 48 weeks; they all achieved clearance of HCV RNA from serum. At the end of treatment, PBMC and serum were examined by real-time polymerase chain reaction for detection of HCV RNA. Six months later, HCV RNA in serum was monitored to detect sustained virologic response. Results:Analysis revealed the presence of detectable HCV RNA in the PBMC of 27% of patients despite clearance of serum HCV RNA. During follow-up, 80% of the patients who became serum HCV positive 6 months after the end of treatment had detectable level of HCV RNA in PBMC at the end of treatment. Conclusions:The absence of HCV in the serum of patients by the end of treatment does not exclude future viremia. The patient might still be a source of infection to others. It is strongly encouraged to test for HCV in PBMC to detect lack of response to treatment and persisting infection.

Indoleamine 2,3-dioxygenase and regulatory T cells in acute myeloid leukemia

Background and objectives: The microenvironment of acute myeloid leukemia (AML) is suppressive for immune cells. Regulatory T cells (Tregs) have been recognized to play a role in helping leukemic cells to evade immunesurveillance. The mesenchymal stem cells (MSCs) are essential contributors in immunomodulation of the microenvironment as they can promote differentiation of Tregs via the indoleamine 2,3-dioxygenase (IDO) pathway. The aim of the present work was to evaluate the expression of IDO in bone marrow derived MSCs and to study its correlation to percentage of Tregs. Methods: Thirty-seven adult bone marrow samples were cultured in appropriate culture medium to isolate MSCs. Successful harvest of MSCs was determined by plastic adherence, morphology, and positive expression of CD271 and CD105; negative expression of CD34 and CD45 using flowcytometry. MSCs were examined for IDO expression by immunocytochemistry using anti-IDO monoclonal antibody. CD4+ CD25+ cells (Tregs) were measured in bone marrow samples by flowcytometry. Results: MSCs were successfully isolated from 20 of the 37 bone marrow samples cultured. MSCs showed higher expression of IDO and Tregs percentage was higher in AML patients compared to control subjects (P = 0.002 and P < 0.001, respectively). A positive correlation was found between IDO expression and Tregs percentage (P value = 0.012, r = 0.5). Conclusion: In this study, we revealed an association between high IDO expression in MSCs and elevated levels of Tregs which could have an important role in the pathogenesis of AML, providing immunosuppressive microenvironment.

Fibro-Mark: a panel of laboratory parameters for predicting significant fibrosis in chronic hepatitis C patients

Abstract Background: Fibrosis markers are useful for the prediction of cirrhosis but clinical scores such as King’s score, AST-Platelet ratio index (APRI), Biotechnology research center (BRC), Fibrosis routine test (FRT), Fibro-α score and Fibro-quotient (FibroQ) have limited accuracy for diagnosing significant fibrosis. We hypothesised that new markers (reflecting the balance between hepatic fibrogenesis and fibrolysis) together with other indirect fibrosis markers would together construct a more sensitive and specific score capable of identifying fibrosis than existing scores. Methods: Collagen IV, hyaluronic acid, platelet-derived growth factor (PDGF) and tissue inhibitor of metalloproteinase-1 (TIMP-1) were measured by ELISA, and AST, ALT, platelet count, albumin, total bilirubin, INR and AFP by routine methods in 148 patients with hepatitis C induced liver disease. Stepwise linear discriminant analysis and area under receiver-operating characteristic curves (AUCs) were used to create a predictive score and compare it to others. Results: Patients with significant fibrosis (n = 100, F2–F4) showed 2.08, 2.14, 1.80 and 1.90-fold increase in collagen IV, hyaluronic acid, PDGF and TIMP-1, respectively, over patients with no or mild fibrosis (n = 48, F0/F1)(all p < 0.01). Significant independent predictors of F2–F4 were AFP (AUC 0.79), age (0.76), PDGF (0.74), collagen IV (0.78) and TIMP (0.75), which together formed a five-marker score ‘Fibro-Mark’ for predicting F2–F4. In comparison with other scores, AUC for Fibro-Mark was 0.89, BRC was 0.83, followed by FRT and King’s score (both 0.82), APRI (0.80), Fibro-α (0.70) and finally Fibro Q (0.63). Conclusions: The Fibro-Mark score provides better discrimination in hepatic-fibrosis staging in chronic hepatitis C patients than existing scores.

The association of cytokine genes polymorphisms and susceptibility to aplastic anemia in Egyptian patients

Background and objective: Aplastic anemia (AA) remains a rare disease, with very interesting pathophysiology that is being investigated for years now. The present study aimed to determine the association between cytokine gene polymorphisms (TGF-β1 −509 C/T, TNF-α −308 G/A, IFN-γ +874 A/T) and susceptibility to AA in Egyptian patients. Methods: The study included 80 participants subjected to determination of gene polymorphisms on genomic DNA using polymerase chain reaction-restriction fragment length polymorphism assay. Results: It was found that IFN-γ +874 A/T gene polymorphism is associated with three-fold increased risk of development of AA (odds ratio (OR) 3.116, P = 0.019), while TNF-α −308 G/A gene polymorphism is associated with decreased risk (OR 0.318, P = 0.026). TGF-β1 −509 C/T gene polymorphism showed comparable risk between patients and controls (P = 0.263). Conclusion: IFN-γ +874 A/T gene polymorphism is associated with the etiology of AA in Egyptian patients.

Towards hepatitis C virus elimination: Egyptian experience, achievements and limitations

Worldwide, more than one million people die each year from hepatitis C virus (HCV) related diseases, and over 300 million people are chronically infected with hepatitis B or C. Egypt used to be on the top of the countries with heavy HCV burden. Some countries are making advances in elimination of HCV, yet multiple factors preventing progress; remain for the majority. These factors include lack of global funding sources for treatment, late diagnosis, poor data, and inadequate screening. Treatment of HCV in Egypt has become one of the top national priorities since 2007. Egypt started a national treatment program intending to provide cure for Egyptian HCV-infected patients. Mass HCV treatment program had started using Pegylated interferon and ribavirin between 2007 and 2014. Yet, with the development of highly-effective direct acting antivirals (DAAs) for HCV, elimination of viral hepatitis has become a real possibility. The Egyptian National Committee for the Control of Viral Hepatitis did its best to provide Egyptian HCV patients with DAAs. Egypt adopted a strategy that represents a model of care that could help other countries with high HCV prevalence rate in their battle against HCV. This review covers the effects of HCV management in Egyptian real life settings and the outcome of different treatment protocols. Also, it deals with the current and future strategies for HCV prevention and screening as well as the challenges facing HCV elimination and the prospect of future eradication of HCV.

Role of physical function in predicting short-term treatment outcome in Egyptian acute myeloid leukemia patients: a single center experience

Background Acute myeloid leukemia (AML) is a potentially fatal hematological disease. Along with disease-related factors, patient-related factors, in particular age, are a strong predictor of outcome that influence treatment decisions. Many acute myeloid leukemia risk stratification models have been developed to predict the outcome of intensive chemotherapy. However, these models did not include physical function assessments. Methods This study investigated the impact of several factors, namely the performance status, physical function and age on the short-term outcomes of intensive chemotherapy in a cohort of 50 Egyptian patients with de novo acute myeloid leukemia. Results Complete remission after intensive chemotherapy in these myeloid leukemia patients at Day 28 was 56% and the mortality rate was 12% and 34% at Day 28 and Day 60, respectively. The pretreatment Eastern Cooperative Oncology Group score was significantly correlated with outcomes on Day 28 and Day 60 (p-value = 0.041 and p-value = 0.032, respectively). There were significant correlations between the two-minute walk test and outcomes of therapy on Day 28 and 60 (p-value = 0.032 and p-value = 0.047, respectively) and between grip strength test and outcomes of therapy on Day 28 and 60 (p-value = 0.046 and p-value = 0.047 respectively). Furthermore, there was a significant correlation between chair stand test and outcome of therapy on Day 28 (p-value = 0.023). Conclusion Performance status and physical function assessments were strong predictors of outcome of intensive chemotherapy in acute myeloid leukemia and we recommend the incorporation of these variables in risk stratification models for the personalization of therapy before treating acute myeloid leukemia patients with intensive chemotherapy.

βS globin gene haplotype and the stroke risk among Egyptian children with sickle cell disease

ABSTRACT Background and aim of work: Sickle cell disease (SCD) is an inherited disease of the beta globin gene. The βS globin gene haplotypes are Senegal, Benin, Bantu, Cameroon, Arab-Indian and atypical haplotypes. In SCD, stroke is a life-threatening event in both adults and children. In light of paucity of studies on βS globin gene haplotypes in Egypt, we aimed to determine βS globin gene haplotypes in children with SCD and study their impact on stroke risk. Methods: Fifty-two SCD patients were included in the study, they were 26 males and 26 females with age range from 3 to 18 years old. The PCR-RFLP technique was used for the determination of βS globin gene haplotypes. Transcranial Doppler (TCD) was done to identify patients at risk of stroke. Results: Benin/Benin was the most prevalent haplotype detected in 50% followed by Benin/Bantu in 30.8% of studied patients. TCD study showed that 14/52 (26.9%) patients had abnormally high TCD flow velocities (TCD velocities ≥170 cm/s) and thus considered high stroke risk group, whereas 38/52 (73.1%) patients had TCD flow velocities <170 cm/s and are considered low stroke risk group. Stroke risk was not found to be associated with βS globin gene haplotype (p = .532). Conclusion: This study provides a relevant contribution to our understanding of the anthropological and historical background of the population in Egypt where Benin haplotype is the commonest βS globin gene haplotype and homozygous Benin/Benin is associated with higher stroke risk than other haplotypes.

Extracellular Matrix Proteins Substantiate IL-28B T allele Effect on Histological Outcome of Chronic Hepatitis C

INTRODUCTION AND AIM The correlation between interleukin-28B (IL-28B) polymorphisms and chronic hepatitis C (CHC) progression is debatable. Here, we aimed to evaluate the relation between IL-28B C/T genotypes and the development of cirrhotic liver. Extracellular matrix (ECM) proteins, FibroScan and model for end-stage liver disease (MELD) were used to substantiate the severity of liver disease. MATERIAL AND METHODS IL-28B rs12979860, liver stiffness and ECM proteins were assessed in 272 CHC patients. RESULTS Cirrhosis percentage increased to 10%, 52% and 96% with the increasing number of T alleles (CC, CT and TT, respectively). Also, elevated ECM proteins levels were correlated with the increasing number of T alleles. Interestingly, among cirrhotic patients, liver stiffness, MELD and ECM proteins were significantly (P < 0.0001) higher in patients with TT more than CT genotype. FibroScan, hyaluronic acid, Laminin, Collagen IV and the N-terminal pro-peptide of collagen type III have high accuracy to differentiate liver status in CC from TT genotype. Area under receiver-operating characteristic curve (95% CI) were 1.0 (1.0-1.0), 0.97 (0.96- 1.0), 0.93 (0.85-1.0), 0.98 (0.97-1.0) and 0.93 (0.91-0.97), respectively. CONCLUSION This study suggests that IL-28B T allele affects the natural course of CHC type 4 and also suggests that carriage of the IL-28B C allele protects from unfavorable clinical outcomes in CHC as coexistence of C allele with T allele reduced cirrhosis severity.INTRODUCTION AND AIM The correlation between interleukin-28B (IL-28B) polymorphisms and chronic hepatitis C (CHC) progression is debatable. Here, we aimed to evaluate the relation between IL-28B C/T genotypes and the development of cirrhotic liver. Extracellular matrix (ECM) proteins, FibroScan and model for end-stage liver disease (MELD) were used to substantiate the severity of liver disease. MATERIAL AND METHODS IL-28B rs12979860, liver stiffness and ECM proteins were assessed in 272 CHC patients. RESULTS Cirrhosis percentage increased to 10%, 52% and 96% with the increasing number of T alleles (CC, CT and TT, respectively). Also, elevated ECM proteins levels were correlated with the increasing number of T alleles. Interestingly, among cirrhotic patients, liver stiffness, MELD and ECM proteins were significantly (P < 0.0001) higher in patients with TT more than CT genotype. FibroScan, hyaluronic acid, Laminin, Collagen IV and the N-terminal pro-peptide of collagen type III have high accuracy to differentiate liver status in CC from TT genotype. Area under receiver-operating characteristic curve (95% CI) were 1.0 (1.0-1.0), 0.97 (0.961.0), 0.93 (0.85-1.0), 0.98 (0.97-1.0) and 0.93 (0.91-0.97), respectively. CONCLUSION This study suggests that IL-28B T allele affects the natural course of CHC type 4 and also suggests that carriage of the IL-28B C allele protects from unfavorable clinical outcomes in CHC as coexistence of C allele with T allele reduced cirrhosis severity.

Impact of circulating erythrocyte-derived microparticles on coagulation activation in sickle cell disease

Sickle cell disease (SCD) is characterized by a hypercoagulable state as a result of multiple factors, including chronic hemolysis and the presence of circulating cell-derived microparticles (MPs). The aim of this work was to study the impact of erythrocyte-derived circulating microparticles (glycophorin A; CD235 positive) on coagulation activation and their probable role in contribution to painful crisis in SCD patients. Peripheral blood samples of 25 SCD patients during painful crisis and in steady state were studied for the presence of erythrocyte-derived MPs using flow cytometry. Estimation of D-dimer level, as a marker of coagulation activation, was done using semiquantitative assay. Thirty-six healthy individuals, age- and sex-matched, were included as a control group. Erythrocyte-derived MPs level was significantly higher in SCD patients during painful crisis compared to control group (p = 0.02), but no statistically significant difference was found between erythrocyte-derived MPs level in SCD patients in steady state compared to controls or to SCD patients during painful crisis (p = 0.3 and 0.49, respectively). D-dimer level was higher in SCD patients both during crisis and in steady state compared to controls (p < 0.001). SCD during painful crisis is associated with increased levels of erythrocyte-derived MPs and D-dimer which may contribute to the hypercoagulable state observed in such group of patients.

Tissue factor expression on blood monocytes in patients with hepatitis C virus-induced chronic liver disease

Chronic liver disease (CLD) is a worldwide common pathology characterised by an inflammatory and fibrotic process leading to progressive evolution from chronic hepatitis to cirrhosis. Monocytes play a crucial role in the pathogenesis of inflammation and fibrosis in chronic liver diseases. Activated monocytes increase the expression of tissue factor, a key glycoprotein that participates in haemostatic and inflammatory processes. This study aims to assess the expression of tissue factor on activated peripheral blood monocytes in patients with hepatitis C virus (HCV)-induced CLD in relation to the degree of hepatic insufficiency and haemostatic imbalance. The current study included 60 patients with HCV-induced CLD, categorised after Child–Pugh into four groups: Child A, B, C and C during acute attack of haematemesis, 15 patients each, and 15 healthy subjects were included as normal controls. Immunophenotype characterization was carried out by flow cytometric analysis for identification of monocytes tissue factor expression (CD142) on activated blood monocytes population (CD11b and CD14) in different groups studied. Data demonstrated significant increase (p < 0.05) in the expression of each of CD11b, CD14 and CD142 revealing monocytes activation and increased expression of tissue factor on peripheral blood monocytes in different groups of patients especially cases during acute attack of haematemesis, compared to healthy subjects. Increased monocytes tissue factor expression in patients with HCV may play a key role in the intensification of the inflammatory and immunological processes in conjunction with activation of the coagulation mechanisms. The interaction of all these phenomena may trigger bleeding by perturbing the unstable haemostasis in frail patients with chronic liver disease.