β-Thalassemia is a heterogeneous disorder caused by mutations that reduce or abolish the synthesis of the β-globin chain. The clinical severity of thalassemia major makes it a priority genetic disease for prevention programs involving population screening of heterozygotes and an optional prenatal diagnosis for carrier couples.
This study aimed to determine the most common β-globin gene mutations in Egypt using a real-time PCR and fluorescently labeled hybridization probes specific for each mutation and to assess the feasibility of introducing this technique in an overall thalassemia prevention program.
The study was carried out on 45 individuals: 37 β-thalassemia carriers [including five amniotic fluid (AF) samples], seven β-thalassemia major cases (including two AF samples), and one normal AF sample. The most common β-thalassemia mutations were characterized by real-time PCR with fluorescently labeled hybridization probes specific for IVSI-110, IVSI-1, IVSI-6, codon 37, and codon 39 in 28/37 (75.7%) carriers.
The most common mutation encountered was IVSI-110 (46%), followed by IVSI-1 (16.2%) and then IVSI-6 (13.5%). Codon 37 and codon 39 were not characterized in any sample. The genotype of the uncharacterized carriers was determined using a less sensitive method (reverse hybridization technique) and a relatively less common set of mutation was characterized as follows: IVSII-1(10.8%), codon 5 (5.4%), IVSII-745 (5.4%), and IVSI-116 (2.7%). The overall number of alleles detected using both techniques was calculated to be 51. The real-time PCR alone, with its assigned probes, detected 38/51(74.5%). Thirteen mutations (13/51=25.5%) remained uncharacterized by this technique (because of the unavailability of the corresponding probes). However, the reverse hybridization technique detected 48/51 alleles (94.1%). However, comparison between both techniques in terms of the shared mutations showed that the real-time PCR detected 38/38 (100%) of these mutations, whereas the reverse hybridization technique detected only 36/38 (94.7%).
Real-time PCR is a very rapid and accurate method for the detection of the β-thalassemia mutation, which may be valuable in cases for which a rapid decision has to be taken. Impediments to prenatal diagnosis as encountered in this study were attributed to refusal of termination of pregnancy by the family for religious/reasons, abortion following amniocentesis, and failure to determine the correct genotype of the AF analyzed.
Myelodysplastic syndromes (MDS) are a group of malignant myeloid hematopoietic disorders. The diagnosis of MDS can be difficult, especially in cases with a low blast count and a normal karyotype. Flow cytometry has been used to distinguish MDS from nonclonal cytopenias. No one single simple flow cytometric parameter has been proposed to be diagnostic of MDS.
The aim of the present study was to evaluate immunophenotypic alterations in typical MDS patients and whether these abnormalities help in the differentiation process between MDS with nonclonal disorder and leukemic patients.
Marrow aspirates from 29 patients, including 13 with MDS, 16 with acute myeloid leukemia, and 18 with nonclonal disorders (normal controls), were examined in this study. Their immunophenotypes were analyzed using flow cytometry. Blasts, nonblast myeloid cells, and monocytes were gated on the basis of CD45 expression and side scatter (SSC).
Comparison among the three groups showed that the granulocytic lineages of MDS showed decreased SSC compared with the controls (P<0.005 and P<0.000, respectively), altered CD45 intensity (P<0.004), decreased CD10-positive granulocytes (P<0.02), and a higher CD56 positive expression in the MDS and leukemic group (P<0.05 and P<0.001, respectively). Also, decreased intensity of CD11b (P<0.03) was observed in the MDS group. The expression rate of CD123+ was significantly higher in MDS patients than that in normal controls (P<0.0001).
Gating of the granulocytic region is a relatively easy method for MDS immunophenotyping. Among the parameters studied, SSC, CD10, CD123, and CD56 were the most useful for differentiating MDS from nonclonal disorders, whereas immunophenotypic changes in MDS appear to be useful for differentiating MDS from nonclonal disorders.
β-Thalassemia major is a very serious blood condition, as affected patients are unable to synthesize enough healthy red blood cells and depend on blood transfusions throughout their life.
The aim of the study was to evaluate the lipid profile in patients with β-thalassemia major.
Fifty patients with β-thalassemia major and 25 healthy controls were included in this study. They were subjected to complete history taking, a thorough clinical examination, and laboratory investigations including complete blood count, liver function test, and assessment of serum ferritin levels and fasting lipid profile including total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglycerides (TGs) levels.
Patients with β-thalassemia major showed significantly lower total cholesterol, HDL-cholesterol, and LDL-cholesterol when compared with controls. Serum TG levels of β-thalassemia major patients were found to be significantly higher than the levels in control individuals. Our results revealed that the lipid profile changed in patients with thalassemia major.
In thalassemic patients, adequate chelation therapy with normalization of serum ferritin level and monitoring of TGs is highly recommended, and they are treated aggressively if the levels are increased. Several interventions including antioxidant therapy and vitamin-lowering and lipid-lowering agents should be used in high-risk patients with β-thalassemia major to decrease the risk of atherosclerosis.
Janus kinases are cytoplasmic tyrosine kinases that mediate signaling from the cytokine receptors to the cell nucleus. Janus kinase 2 mutation (JAK2 V617F) analysis has been endorsed by the WHO for diagnosing polycythemia vera, essential thrombocythemia, and primary myelofibrosis. The aim of this study was to assess JAK2 V617F point mutation in healthy cigarette smokers compared with healthy nonsmokers and to correlate the presence of this mutation with some clinical and laboratory variables.
Group I comprised 34 cigarette smokers who have been smoking 10 or more cigarettes per day, every day of the week, for at least 10 consecutive years. Group II comprised 42 men who were nonsmokers with no history of drug abuse. In addition to routine laboratory investigations, detection of JAK2 V617F point mutation in peripheral blood neutrophils was assessed for all participants.
In this study, we found an increased percentage of JAK2 V617F mutation in cigarette smokers compared with nonsmokers. Further, we found a significant positive correlation between the percentage of JAK2 V617F mutation and age in both groups.
JAK2 V617F mutation has been detected in the healthy population; however, its incidence significantly increases in cigarette smokers. The mechanisms leading to excess JAK2 mutation and the importance of this mutation in smokers are yet to be elucidated and an adequate follow-up of healthy individuals who carry the mutation is recommended.
Serum immunoglobulin (Ig) paraprotein can be detected in a subset of patients with chronic lymphocytic leukemia (CLL) by serum protein electrophoresis and immunofixation electrophoresis. CLL with Ig paraproteinemia had an inferior survival compared with patients with CLL without serum paraprotein.
The present study was carried out on 100 patients with B-CLL (60 men and 40 women) ranging in age from 33 to 75 years. The staging of CLL was performed according to the Binet staging system. Venous blood samples were obtained from B-CLL patients for a complete blood count. Serum was separated for the measurement of lactate dehydrogenase (LDH), β2-microglobulin (β2-MG) levels by ELISA, and Ig paraprotein. Bone marrow aspiration was carried out for all B-CLL cases. Prognostic markers of CD38 and &zgr;-chain-associated protein kinase-70 (ZAP-70) expression were also analyzed.
Twenty-two patients had Ig paraproteinemia of a total of 100 untreated patients with CLL, frequency 22%. There was a highly significant elevation in LDH, β2-MG, CD38, ZAP-70, IgG, and IgM in CLL with monoclonal paraprotein versus CLL without monoclonal paraprotein. There were positive correlations of serum IgG paraprotein and serum IgM paraprotein with advanced Binet stage (P<0.00 and P=0.00), high level of β2-MG (P=0.00 and 0.00) and LDH (P=0.00 and 0.00), CD38 positivity (P=0.00 and 0.02), and ZAP-70 positivity (P=0.00 and 0.01). Also, there was a positive correlation between high serum β2-MG concentration with Binet stage (P=0.00), high level of LDH (P=0.00), CD38 positivity (P=0.00), and ZAP-70 positivity (P=0.02). After a follow-up of 60 months, 15 patients (15%) died. Eight (36.3%) patients with Ig paraproteinemia died during the observation period, whereas among the 78 patients without Ig paraproteinemia, seven (8.9%) died.
β2-MG and Ig paraprotein serve as poor prognostic markers for B-CLL. Patients with CLL with serum Ig paraprotein represent a heterogeneous group with an inferior clinical outcome. Serum Ig paraprotein might be applied for the assessment of prognosis in patients with CLL.
Chronic lymphocytic leukemia (CLL) is an environment-dependent hematologic malignancy where interactions with accessory cells through cytokines and their receptors seem to confer a survival advantage, thus contributing to disease progression. Interleukin-22 (IL-22) is a T-cell-derived cytokine that promotes cell proliferation and survival through interaction with its receptor IL-22RA1, normally absent in normal immune cells, including B and T lymphocytes.
This study aimed to determine the plasma levels of IL-22 and the expression of IL-22RA1 on malignant cells in patients with B-cell CLL (B-CLL), together with their relation to clinical and prognostic characteristics of the disease.
The study was carried out on 62 newly diagnosed B-CLL patients. Twenty-five age-matched and sex-matched healthy individuals served as controls. Patients were diagnosed, according to the International Workshop on CLL guidelines, by cytomophology, immunophenotyping, conventional cytogenetic analysis, and fluorescence in-situ hybridization. Plasma IL-22 levels were measured by an enzyme-linked immunosorbent assay and the expression of IL-22RA1 on leukemic cells was assessed by flow cytometry.
Plasma IL-22 was significantly higher in B-CLL patients (range, undetectable 62.9 pg/ml; median, 6.6) compared with control participants (range, undetectable 6.4 pg/ml; median, undetectable) (P<0.01). IL-22RA1 expression was negative in all normal controls, whereas in B-CLL patients it was positively expressed in 35/62 CLL cases (56%). Taking the median level of IL-22RA1 expression in CLL patients as a cutoff level, overexpression (≥10%) was observed in 32/62 (52%) cases. IL-22RA1 expression correlated significantly positively with plasma levels of IL-22 (rs=0.817; P<0.01). Patients presenting with high CD38 expression had significantly higher plasma IL-22 levels compared with those with low CD38 (undetectable 62.9 pg/ml; median, 19.3 vs. undetectable 50.1 pg/ml; median, 3.1) (P<0.01) as well as overexpression of IL-22RA1. No significant relation could be established between either plasma IL-22 levels or IL-22RA1 expression with other clinical features or prognostic criteria of CLL.
This is the first report to describe the aberrant expression of the IL-22 signaling pathway in B-CLL and to link its overexpression with high CD38 expression, a known poor prognostic marker of the disease.
Idiopathic thrombocytopenic purpura (ITP) in children is usually a self-limiting disorder. It may follow a viral infection or immunization and is caused by an inappropriate response of the immune system. Many viruses, such as parvovirus B19, cytomegalovirus (CMV), Epstein–Barr virus (EBV), and rubella, are implicated in the occurrence of ITP.
The aim of the study was to investigate the occurrence of viral-associated ITP in Egyptian children.
Viral studies included specific immunoglobulin M for rubella, EBV, and CMV. In addition, molecular detection for parvovirus B19 was carried out.
Positive viral markers either by positive serology immunoglobulin M for rubella, CMV, or EBV or by a molecular study for parvovirus B19 were detected in 19 patients (38.8%). The most common viral infection was parvovirus B19 (30.6%), followed by EBV (16.3%), rubella (12.12%), and CMV (10.2%).
We conclude that in a large proportion of children with ITP in our region, an association with markers of acute viral infections similar to those of rubella, EBV, and CMV is present. Moreover, a significant proportion of the children had occult parvovirus B19 viremia. A study of occult viral infections is recommended in children with ITP.
At present, the traditional activated partial thromboplastin time (a-PTT) of 1.5–2.5 times the control value for monitoring unfractionated heparin (UFH) therapy continues to be used in the coagulation laboratory of Alexandria Main University Hospital. This study was conducted for the following reasons: to evaluate the relationship between heparin concentration and a-PTT results using Sysmex CA-1500; to define thea-PTT therapeutic range for our system; and to assess the level of agreement between a-PTT results and those obtained using the anti-Xa assay for monitoring UFH therapy.
A significant positive correlation between a-PTT and anti-Xa assay results was noted (P=0.907, r=0.000, and P=0.098, r=0.000, for therapeutic a-PTT and therapeutic ratio, respectively). Regression analysis was carried out to determine the anti-Xa-derived therapeutic range (a-PTT results that correspond to a plasma heparin concentration of 0.3–0.7 U/ml by anti-Xa assay). The a-PTT therapeutic range was 64.4–107.93 s; the therapeutic range for the a-PTT ratio was 2.13–3.56 and that for the1.5–2.5 control method was 45.27–75.75 s. The agreement between the the a-PTT therapeutic range and the results of the anti-Xa assay was 78%, whereas the agreement between the 1.5–2.5 control method and the anti-Xa assay was 0.097%. Moreover, the potential for over therapeutic levels occurs more frequently with the 1.5–2.5 control method.
Anti-FXa-derived therapeutic range on Sysmex CA-1500 is superior to that obtained using the 1.5–2.5 control method in clinical decision making. Therapeutic ranges for various a-PTT reagent–coagulometer combinations could be provided by reagent manufacturers or central reference laboratories to the institutions that are not equipped to measure anti-Xa or to those for which the access to plasma samples from treated patients is limited.
Over 170 million individuals are infected with hepatitis C virus (HCV) worldwide. The report published by the Egyptian Demographic Health Survey stated that in Egypt, there is an overall anti-HCV antibody prevalence of 14.7% and the number of Egyptians estimated to be chronically infected was 9.8%. Hematological manifestations are among the most common extrahepatic manifestations of HCV infection. Patients with HCV infection can develop peripheral blood cell count abnormalities that are commonly attributed to hypersplenism, antiviral therapy, decreased thrombopoietin levels, and/or autoimmune mechanisms.
Evaluation of bone marrow findings in patients infected with HCV presenting with peripheral blood cytopenias.
This study was carried out on 35 patients with chronic HCV infection presenting with cytopenias. Patients were subjected to history taking, physical examination, and routine laboratory investigations together with bone marrow aspiration and trephine biopsy. Flow cytometry and cytogenetics analysis were carried out in selected patients.
Among the patients studied, B-cell non-Hodgkin lymphoma was the most commonly encountered disorder (31%), followed by idiopathic thrombocytopenic purpura (11%), acute lymphoblastic leukemia, acute myeloid leukemia, myelodysplastic syndrome, and hypersplenism (9% each), and autoimmune hemolytic leukemia (6%), and the least encountered disorders were anemia of chronic disease, aplastic anemia, CLL, myelofibrosis, Evan’s syndrome, and sideroblastic anemia (3% each).
B-cell non-Hodgkin lymphoma was the most frequently detected problem among the patients recruited (31%). There were significant data regarding relation of bone marrow dysplasia and fibrosis with age of patients.