The impact of MCP1-2518A/G and CCR2-V64I genetic polymorphisms in Egyptian sickle cell disease patients.

BACKGROUND: Sickle cell disease (SCD) is an inherited genetic disorders of hemoglobin that causes multisystem morbidity. The pathophysiology of SCD is complex and includes HbS polymerization/sickling, hemolysis, endothelial dysfunction and inflammation. Chemokines are proteins playing an important role in the inflammation process and could be involved the context of pro-inflammatory SCD. Some chemokine polymorphism were found to be associated with clinical complication in SCD. AIM OF THE STUDY: Was to explore the frequency and the possible effect of Monocyte Chemo-attractant Protein 1-2518A/G (MCP1-2518A/G) and Chemokine Receptor 2-V64I (CCR2-V64I) genetic polymorphisms on clinical and laboratory disease-related variables in Egyptian Sickle cell disease patients. PATIENTS AND METHODS: Genotyping of the two genes were performed by PCR-RFLP technique for 80 SCD patients as well as 50 healthy control group. RESULTS: The study revealed that the MCP1-2518 polymorphic genotypes (AG & GG) showed no statistically significant difference in the distribution of the polymorphic genotypes between SCD patients and the controls (p = 0.164). While a significantly higher frequency of the mutant variants CCR2-V64I GA/AA among SCD patients than the control subjects were found (p = 0.032). Regarding the clinic-pathological features, the frequency of recurrent infections, vaso-occlusive crisis, severe vaso-occlusive crisis and number of hospitalization/year were higher in patients harbouring the MCP1-2518A/G and CCR2-V64I polymorphic genotypes than the wild genotype, and gall bladder complications were higher in MCP1-2518 G allele patients, whereas surgical splenectomy were higher in CCR2-V64I A allele patients (p < 0.05). IN CONCLUSION: MCP1-2518A/G and CCR2-V64I genetic polymorphisms may influence the clinical severity of sickle cell disease.

The Influence of Interleukin-2 Gene Polymorphisms on the Risk and Clinical Outcome of Non-Hodgkin Lymphoma.

Polymorphisms in the IL-2 gene are associated with various diseases and cancers including non-Hodgkin lymphoma (NHL). The aim of the study is to assess the impact of IL-2 genetic polymorphisms [- 330 T/G (rs2069762) and + 114 T/G (rs2069763)] on the susceptibility and prognosis of NHL. Sixty patients with NHL as well as 60 age and sex matched healthy control subjects are included in this study. IL-2 genotypes were determined by Polymerase Chain Reaction-Restriction Fragment length Polymorphism assay (PCR-RFLP). Our study revealed that both IL-2 rs2069762 and rs2069763 gene polymorphisms are associated with increased risk of developing NHL; OR = 3.609 (95% CI = 1.527-8.417) and 4.142 (95% CI = 1.637-10.538) respectively. Moreover, the simultaneous presence of both polymorphisms is associated with about 6 fold increased risk of developing NHL. Also, IL-2 rs2069762 and rs2069763 gene polymorphisms increase the risk of unfavorable prognosis with OR = 17.300 (95% CI = 3.392-87.725) and 10.424(95% CI = 1.870-58.413) respectively. These findings suggest that IL-2 (rs2069762) and (rs2069763) gene polymorphisms could be involved in the development of NHL.

CXC Chemokine Receptor Type 5 Gene Polymorphisms in a Cohort of Egyptian Patients with Diffuse Large B-Cell Lymphoma.

BACKGROUND: The chemokine receptor CXCR5 is selectively expressed on B cells; it is involved in lymphocyte homing and the development of normal lymphoid tissue. Its principle ligand is CXCL13 or B lymphocyte chemoattractant. Three polymorphisms in the CXCR5 gene, rs148351692 C/G, rs6421571 C/T, and rs78440425 G/A, have been identified. OBJECTIVE: To assess the genetic polymorphisms of CXCR5 and evaluate their possible contribution to the susceptibility and response to therapy of diffuse large B-cell lymphoma (DLBCL). PATIENTS AND METHODS: Fifty DLBCL (not otherwise specified) patients and 50 control subjects were included in this study. CXCR5 genotypes were determined by PCR-RFLP. RESULTS: Our study revealed that the CXCR5 rs148351692 C/G and rs6421571 C/T gene polymorphisms are associated with an increased risk of developing DLBCL (OR 28.57 [95% CI 8.96-96.56] and 3.45 [1.67-11.83] respectively), while CXCR5 rs78440425 G/A showed no association with the risk of lymphoma. Moreover, the double and triple combined gene polymorphisms are associated with an increased risk of developing DLBCL of approximately 120-fold and 105-fold, respectively. CXCR5 gene polymorphisms had no significant impact on disease outcome or response to therapy. CONCLUSIONS: CXCR5 gene polymorphisms could be considered a potential risk factor for the development of DLBCL.

Influence of esophageal variceal bleeding on iron status in chronic hepatitis C patients.

BACKGROUND: Disorders of serum iron balance are frequently observed in chronic hepatitis C (CHC) patients. Iron overload as well as iron deficiency anemia are common clinical findings in these patients. Variceal bleeding is also a common complication. To date, no study has discussed the influence of esophageal bleeding on iron status in anemic CHC bleeders. OBJECTIVE: Was to study reticulocyte hemoglobin content (CHr) and serum hepcidin levels in anemic CHC and to evaluate the influence of variceal bleeding on patients' iron status. METHODS: Serum hepcidin levels and CHr were assessed in 65 early phase CHC patients (20 nonanemic, 23 anemic nonbleeders, and 22 anemic bleeders), and 20 healthy controls; and were compared with the conventional indices of iron deficiency including mean corpuscular volume, mean corpuscular hemoglobin, red cell distribution width, serum iron, total iron binding capacity, transferrin saturation and ferritin. RESULTS: Hepcidin levels were comparable in patients groups, but were significantly lower in patients than in controls (P = 0.01). Child-Pugh class B patients showed significantly lower hepcidin levels than class A patients. CHr levels were comparable in all groups as well as all iron deficiency indices. Patients with ferritin values or less 100 ng/ml and CHr or less 29 pg/cell or Tfsat or less 16% are more likely to have iron deficiency [odds ratio (OR = 3.93, 95% confidence interval (CI) = 2.54-6.08; OR = 10.50, 95% CI = 1.94-56.55, respectively). CONCLUSION: Esophageal bleeding has an almost no influence on iron status in CHC patients. Serum hepcidin content is influenced by CHC disease rather than by anemia associated with or without esophageal bleeding and it could be used as a marker of early hepatic insufficiency. Assessing CHr content could add a potential utility in the detection of iron deficiency in CHC patients.

TAp73 and ΔNp73 relative expression in Egyptian patients with lymphoid neoplasms.

BACKGROUND AND AIMS: The p73 gene has different isoforms with opposing anti- and pro-apoptotic functions. The pro-apoptotic activities are inhibited by overexpression of the dominant ΔNp73 isoform. The aim of this study was to detect the expression of the TAp73 and ΔNp73 isoforms in Egyptian patients with malignant lymphoid neoplasms. Their expression was analyzed by quantitative RT-PCR. PATIENTS AND METHODS: The study included 30 B-NHL patients, 24 T-NHL patients, 16 ALL patients, 18 CLL patients, 22 patients with reactive lymphoid hyperplasia, and 6 healthy control subjects. RESULTS: ALL and CLL patients expressed both isoforms at higher levels compared to lymphoma patients. Higher expression of TAp73 was found in both B-NHL and T-NHL (around 4-fold and 16-fold, respectively) in comparison to ΔNp73 (2-fold and 14-fold, respectively). In CLL patients both isoforms showed higher expression levels in comparison to normal peripheral blood lymphocytes controls: nearly 27-fold for TAp73 and 233-fold for ΔNp73. All ALL patients showed higher expression of both studied isoforms than controls (9-fold for TAp73 and 386-fold for ΔNp73). The highest ΔNp73 expression along with a higher ΔNp73/TAp73 ratio (67-fold) was found in ALL patients compared with CLL patients (21-fold). CONCLUSIONS: A considerable number of lymphoma patients lacked the expression of either or both isoforms, while all lymphoid leukemia patients expressed both isoforms. The expression pattern differences of p73 isoforms may reflect differences in the biology of these malignancies.

Molecular and serological assessment of parvovirus B-19 infection in Egyptian children with sickle cell disease.

BACKGROUND/PURPOSE: Human parvovirus B-19 (PB-19) is a cause of hemolysis, red blood cell aplasia, and severe conditions in patients with sickle cell anemia, but the molecular mechanisms of the infection are still insufficiently understood. This study aimed to detect PB-19 DNA together with its antibodies in the sera of Egyptian children with sickle cell disease and to assess the contribution of this infection, which causes transient cessation of erythropoiesis, in precipitating severe anemia in some cases. METHODS: One hundred children with sickle cell disease seeking medical advice in the pediatric-hematology clinic were recruited. Sera of the patients were compared with those of 60 healthy children regarding the presence of PB-19 immunoglobulin (Ig)G and IgM as well as detection of its DNA by nested-polymerase chain reaction technique. RESULTS: There were statistically significant differences in the prevalence of PB-19 IgM, IgG, and DNA among patients when compared with controls (p < 0.001, p = 0.001, and p < 0.001 respectively). Acute PB-19 infection detected by positive IgM and DNA was found in 30% of the patients, while chronic PB-19 infection detected by positive IgG and DNA was detected in 24% of the patients. Anemia was worse in children with acute PB-19 infection than in those with chronic infection, while anemia was mild in children with old infection. CONCLUSION: PB-19 infection is detected at high rates among Egyptian children with sickle cell disease and it may result in severe anemia. So, PB-19 must be suspected and screened for in such group of patients.

The clinical relevance and prognostic significance of microsomal epoxide hydrolase gene polymorphisms and their susceptibility to acquired aplastic anemia: an Egyptian study.

BACKGROUND: Microsomal epoxide hydrolase enzyme (mEPHX) is involved in xenobiotics detoxification. Two variants of mEPHX, Tyr113His and His139Arg, have been described. Both may lead to acquired aplastic anemia (AA). OBJECTIVES: Assessing mEPHX genetic polymorphisms and detecting their impact on susceptibility and prognosis in Egyptian AA patients. PARTICIPANTS AND METHODS: mEPHX 113 and 139 genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 100 patients with AA and 100 control subjects. RESULTS: Both mEPHX Tyr113His and His139Arg gene polymorphisms were associated with increased risk of developing AA, and have a significant impact of bad prognosis (p value < 0.01). CONCLUSIONS: These mEPHX gene polymorphisms can be considered as risk factors and predictive molecular markers for prognosis in AA patients.

Impact of Prothrombotic Risk Factors in a Cohort of Egyptian Hemophilia A Patients.

INTRODUCTION: Hemophilias are a group of related bleeding disorders that show an X-linked pattern of inheritance. The clinical phenotype of severe hemophilia may vary markedly among patients as a result of many factors, including genetic prothrombotic risk factors. OBJECTIVES: Our objective was to study the incidence of the most common prothrombotic risk factors for additive effects among Egyptian patients with hemophilia A and their impact on clinical phenotype; annual bleeding frequency and severity of hemophilic arthropathy, as well as the effect of a single variation in these patients. METHODS: This study was carried out in 100 patients with hemophilia A. Genotyping for factor V Leiden (FVL) G1691A, prothrombin G20210A, MTHFR C677T, and A1298C mutations was conducted using a real time-polymerase chain reaction (RT-PCR) assay. RESULTS: Our study revealed mutations in hemophilia patients as follows: prothrombin G20210A (3 %), FVL (14 %), MTHFR C677T (42 %), and A1298C (59 %). Despite a lack of statistical significance when each gene was analysed separately, heterozygosity of prothrombin G20210A or FVL was always associated with either a mild or moderate, but never a severe, clinical presentation. The lowest bleeding frequency (less than once per month) was identified among patients with two heterozygous variants irrespective of the involved genes. In addition, the incidence of hemarthrosis was significantly higher among patients with a wild genotype of the prothrombin gene and FVL, and the average number of affected joints was significantly higher among patients with wild-type prothrombin and FVL genes than among heterozygous patients. CONCLUSION: These prothrombotic mutations have a cumulative effect in amelioration of the severity of bleeding in hemophiliacs. The most prominent effect is that of prothrombin G20210A and FVL, while MTHFR C677A and A1298C gene mutations are less conclusive.

Survivin and cyclin E2 genes expression in a cohort of Egyptian acute leukemia patients: Clinical importance and future prospects.

BACKGROUND: Abnormalities in the control of apoptosis play an important role in leukemogenesis. Survivin is a member of inhibitor of apoptosis proteins family, it prevents apoptosis by blocking caspase activity and play a role in cell proliferation. While, cyclin E2 is one of the cyclins proteins family that controls progression of cell cycle by activation of cyclin dependant-kinase. OBJECTIVE: Was to assess survivin and cyclin E2 genes expression in acute leukemia (AL) patients, and to define their role in the susceptibility of AL, and their correlation with the clinical presentation, laboratory findings, as well as treatment outcome. PATIENTS AND METHODS: This study included 60 de novo AL patients and 40 control subjects to study the expression of survivin and cyclin E2 genes using RT-PCR. RESULTS: Survivin and cyclin E2 genes expression was significantly higher in leukemic patients compared with control subjects (P< 0.001), both genes separately were associated with increased risk of leukemia development and treatment failure (P< 0.01). Moreover, when combining the 2 genes expression, a significant elevation of the risk of leukemia and treatment failure was found (P < 0.01). CONCLUSIONS: Survivin and cyclin E2 genes expression may have clinical relevance and can be considered as molecular risk factors for AL. Also they may be useful as predictive markers for treatment outcome in leukemic patients.

Chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells in vitro.

UNLABELLED: Different therapeutic techniques have been developed for regeneration of articular cartilage injuries, but none has provided an optimal solution to their treatment. Human umbilical cord blood-mesenchymal Stem Cells (HUCB-MSCs) have been considered as promising alternative cell source for cartilage repair. OBJECTIVES: Examining the success rate of MSCs isolation from HUCB as well as chondrogenic differentiation potential of HUCB-MSCs in vitro. MATERIALS AND METHODS: 32 UCB samples were collected, in addition to 5 bone marrow (BM) and 5 peripheral blood (PB) samples, taken as reference controls. Samples were used for mononuclear cells isolation from which MSCs were expanded under complete aseptic conditions, were verified morphologically and through the presence of CD44 and CD105, and absence of CD34. RESULTS: Success rate of UCB-MSCs isolation was (25%), a rate that was lower than those of PB (40%) and BM (80%). Accordingly, certain input parameters have been recommended for successful MSCs isolation from UCB. On selecting samples in which recommended parameters were fulfilled, success rate was increased to 72%. This was together with providing optimal experiment conditions; mainly type of expansion medium, success rate reached 80%. Then, successfully expanded MSCs were subjected to chondrogenic differentiation by culturing in pelleted micromass system in presence of transforming growth factor beta-1 and chondrogenic medium devoid of fetal bovine serum to evaluate their ability to undergo chondrogenesis. Differentiation was verified microscopically using special stains, and proved by reverse transcriptase-polymerase chain reaction for expression of aggrecan and collagen II genes. In conclusion, in vitro differentiation into chondrocytes is possible from HUCB-MSCs.

Expression of IL4 (VNTR intron 3) and IL10 (-627) genes polymorphisms in childhood immune thrombocytopenic purpura.

OBJECTIVE: Immune thrombocytopenic purpura (ITP) is an acquired autoimmune disorder caused by the production of antiplatelet antibodies. These autoantibodies opsonize platelets for splenic clearance, resulting in low levels of circulating platelets. Interleukin 4 (IL4) and interleukin 10 (IL10) are important immunoregulatory cytokines mainly produced by macrophages, monocytes, T cells, B cells, and mast cells. Our study was aimed at detecting the frequency of IL4 (VNTR intron 3) and IL 10 (-627) gene polymorphisms in Egyptian ITP children as genetic markers for ITP risk and clarifying their possible role in the pathogenesis of ITP as well as their correlation with the clinical presentation and laboratory data. METHODS: IL4 (VNTR intron 3) and IL10 (-627) gene polymorphisms were studied in 70 ITP patients and 50 age- and sex-matched healthy controls using polymerase chain reaction-restriction fragment length polymorphism assay (PCR-RFLP). RESULTS: IL4 RP2 and IL10 A alleles were detected more frequently among ITP patients compared to controls. A statistically significant difference was observed in IL10 and IL4 gene polymorphism distribution between acute and chronic ITP patients, with higher A allele and RP2 allele among chronic ITP patients versus acute ITP patients. Combined polymorphisms of IL4 and IL10 genes were associated with greater risk of ITP. CONCLUSION: IL4 and IL10 gene polymorphisms may contribute to susceptibility for ITP in children.

Genetic polymorphism of microsomal epoxide hydrolase enzyme gene in preeclamptic females.

INTRODUCTION: Microsomal epoxide hydrolase enzyme is involved in xenobiotics detoxification. It catalyzes the phase I hydrolysis of epoxides and plays a role in the detoxification processes and in the metabolism of endogenous and exogenous compounds. Preeclampsia, which is one of the most serious complications of pregnancy, may be due to an imbalance between these compounds, such as lipid peroxides and oxygen-free radicals and detoxifying and scavenging substances. Two variants of human epoxide hydrolase enzyme with different enzyme activity have been described; exon 3 polymorphism is associated with lower enzyme activity whereas exon 4 polymorphism is associated with higher activity. The authors tried to investigate the association between these genetic polymorphisms and preeclampsia. METHOD: Thirty preeclamptic females together with 30 normal pregnant females as controls were included in the study. Genotyping for exons 3 and 4 of microsomal epoxide hydrolase enzyme was done by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: There was no statistical significant difference in the distribution of exon 3 genotype between cases and controls (P = 0.4); on the other hand, a highly statistical significant difference was found between cases and controls as regard exon 4 genotype (P = 0.002). CONCLUSION: There may be an association between epoxide hydrolase enzyme polymorphism and the risk of preeclampsia.

Melanoma antigen encoding gene-A3 expression, its clinical significance, and its risk of developing acute myeloid leukemia.

BACKGROUND: Melanoma antigen encoding gene A3 (MAGE-A3) gene, also called cancer/testis (CT) antigen, is a member of MAGE multigene family, which is located on the long arm of the X chromosome, and its expression can be caused by promoter region demethylation. The MAGE-A3 proteins' functions are unknown, but they were found to play a role in cell cycle progression, transcriptional regulation, and drug resistance. The aims of this study were to determine the expression of the MAGE-A3 gene in an Egyptian cohort of de novo acute myeloid leukemia (AML) patients and to define its role in the development of AML and its correlation with clinical presentation, laboratory data, as well as treatment outcome. PATIENTS AND METHODS: This study included 40 de novo AML patients as well as 30 age- and sex-matched normal healthy subjects as a control group. They were all subjected to reverse transcription-polymerase chain reaction assay for the detection of MAGE-A3 gene expression. RESULTS: Our study revealed that 23 AML patients (57.5%) expressed the MAGE-A3 gene, whereas none of the control group subjects (0%) expressed this gene. It was found that the expression of MAGE-A3 gene was associated with an increased risk of AML (odds ratio = 2.763; 95% confidence interval, 1.890-8.041). Regarding treatment outcome, a highly statistical significant difference was found between MAGE-A3-positive and -negative AML patients with P < 0.001, as the MAGE-A3-positive AML patients had a higher incidence of unfavorable treatment outcome, whereas the MAGE-A3-negative patients had a higher incidence of favorable outcome. This clarifies that the MAGE-A3 gene expression was found to have a significant impact on the risk of treatment failure (odds ratio = 3.591; 95% confidence interval, 1.273-10.462). CONCLUSIONS: The expression of MAGE-A3 gene may have a clinical relevance and important role as a risk factor in the development of AML. It may be considered as a prognostic marker and may be useful as a predictive test for treatment outcome in AML.

RAD51 and XRCC3 gene polymorphisms and the risk of developing acute myeloid leukemia.

RAD51 (Rec A homolog of E. coli) is a polymorphic gene and one of the central proteins in homologous recombination-DNA-double-stand breaks (HR-DNA-DSB) repair pathway, which is vital in maintaining genetic stability within a cell. The x-ray repair cross complementing (XRCC3) protein also functions in HR-DNA-DSB repair pathway and directly interacts with and stabilizes RAD51 and the closely related RAD51C. The aim of this study was to determine the prevalence of the RAD51 and XRCC3 repair gene polymorphisms among acute myeloid leukemia (AML) patients and to define their role in development of AML and its correlation with the clinical presentation, laboratory data as well as treatment outcome using polymerase chain reaction-restriction fragment length polymorphism assay in 50 de novo AML patients as well as 30 healthy subjects as a control group. Our study revealed that RAD51 G135C and XRCC3 Thr241Met alleles were associated with increased risk of AML with odds ratio (OR) of 2.833 and 2.909 and 95% confidence interval (CI) of 1.527 to 8.983 and 1.761 to 9.788, respectively. Moreover, when combining the 2 genes polymorphisms, a significant elevation of the risk of AML was found with OR of 3.124 and 95% CI of 1.872 to 11.243. As regards treatment outcome, a highly statistical significant difference was found between XRCC3 genotypes with P value of 0.001, whereas no significant difference was present between RAD51 genotypes with P value of 0.29. This clarifies that XRCC3 gene polymorphisms was found to have a significant impact on the risk of treatment failure with OR of 3.560 and 95% CI of 1.167 to 10.875; however, RAD51 gene polymorphism was not found to have an equivalent effect with OR of 2.813 and 95% CI of 0.933 to 10.828. So XRCC3 gene polymorphism might be considered as a prognostic marker in AML. In conclusion, RAD51 and XRCC3 genes polymorphisms may play an important role in the development of AML.