Nitric oxide metabolites and arginase I levels in ß-thalassemic patients: an Egyptian study.

Stored red blood cells become deficient in nitric oxide that limits their ability to transfer oxygen to tissues that need it. The aims of this study are to assess the endogenous nitric oxide metabolites (NOx) and arginase I levels in transfusion-dependent ß-thalassemic patients; to compare these levels in patients transfused with fresh RBCs with patients transfused with old RBCs, ß-thalassemic minor patients, and normal control; and to correlate these levels with some clinical variables. Group I was composed of 23 patients with homozygous ß-thalassemia on hypertransfusion regimen. They were adequately transfused with fresh RBC. Group II was composed of 17 patients with homozygous ß-thalassemia on hypertransfusion regimen. They were adequately transfused with old RBCs. Group III was composed of 30 patients with homozygous ß-thalassemia. They were adequately transfused with fresh RBCs. Group IV was composed of 18 patients with homozygous ß-thalassemia. They were adequately transfused with old RBCs. Both group III and group IV were supposed to be on hypertransfusion regimen, but they did not follow the regimen. Group V was composed of 21 patients of ß-thalassemia minor. Nineteen apparently healthy individuals (HbAA) served as a control group (group VI). In addition to routine laboratory investigations, plasma levels of NOx and serum levels of arginase I were assessed in all subjects. The mean values of plasma NOx were significantly decreased in groups III and IV compared to the other groups. Also, the levels of NOx were significantly decreased in patients who received old RBCs compared to the other groups. There were high serum levels of arginase I in groups III and IV compared to the other groups. There were significant negative correlations between plasma NOx and some hemolytic biochemical markers in groups III and IV. There were significant positive correlations between serum arginase I and some hemolytic biochemical markers in groups III and IV. Also, there was a significant negative correlation between plasma NOx and serum arginase I levels in groups III and IV. In non-adequately transfused patients with ß-thalassemia major, inactivation of NO correlates with hemolytic rate and is associated with the erythrocyte release of cell-free hemoglobin, which consumes NO directly, and the simultaneous release of the arginine-metabolizing enzyme arginase, which limits bioavailability of the NO synthase substrate, arginine, during the process of hemolysis. New treatments aimed at improving arginine and NO bioavailability through arginase inhibition, suppression of hemolytic rate, oral arginine supplementation, predonation testing, and transfusion of fresh RBCs or use of NO donors represent potential therapeutic strategies for this common hemolytic disorder.

Role of urokinase plasminogen activator receptor (CD87) as a prognostic marker in acute myeloid leukemia.

Expression of uPAR (CD87) may play a relevant role in the diagnosis and pathophysiology and prognostic pattern of acute myeloid leukemia. The aims of this study were to find out the prognostic significance of pretreatment detection of CD87 and study the prevalence of CD87 expression and its value as a predictor for survival in AML patients. CD87 expression has been studied on blasts in 110 newly diagnosed AML patients. Immunophenotyping and cytogenetic analysis of these cases were performed. CD87 was positive in 80 (72.7%) cases of patients. The MFI categorized the expression of CD87 into CD87(High) and CD87(Low) expression. Blast cells show that monocytic differentiation has a significantly more CD87(High) expression than CD87(Low) expression. Cytogenetic abnormalities were found in 58.7% of patients with CD87(Low) AML and 41.25% of patients with CD87(High) AML. Cases with CD87(High) expression cells were characterized by a significantly lower survival period especially when co-expressed with CD56, CD34, and/or CD64. There is a negative prognostic influence of the expression of CD87 on the surface of AML blasts, but more tests are necessary to explain the pathophysiological mechanisms behind these findings and to learn about the mechanism that influences the CD87 expression and function.

Incidence of Philadelphia-chromosome in acute myelogenous leukemia and biphenotypic acute leukemia patients: And its role in their outcome.

BACKGROUND: Philadelphia-chromosome positive acute myeloid leukemia (Ph+ AML) is a rare entity and patient prognosis is poor, with short median survival. Biphenotypic acute leukemia (BAL) is a rare disorder that is difficult to diagnose and it displays features of both myeloid and lymphoid lineage. The aim of this study was to highlight the incidence of Philadelphia chromosome and its presence in cases of acute myeloid and biphenotypic leukemia and determine its role in the outcome of these leukemias. SUBJECTS AND METHODS: This study examined 464 subjects with newly diagnosed acute myeloid leukemia: 312 were males and 152 were females. All individuals were subjected to immunophenotyping and conventional karyotyping. FISH was used in failed cases of conventional cytogenetics analysis to quantify disease and to prove positive BCR-ABL fusion gene. RESULTS: the incidence of Ph+ chromosome was found to be higher in BAL (38.4%) than in AML (1.99%). There was statistically significant difference according to the age and the median survival time between the two groups. CONCLUSION: Detection of specific chimeric transcripts in AML and BAL at the time of diagnosis was crucial since it plays an important role for accurate risk stratification and treatment management.

Paraoxonase-1 and oxidative status in common Mediterranean ß-thalassaemia mutations trait, and their relations to atherosclerosis.

AIMS: Investigation of paraoxonase-1 (PON1) activity with oxidative status parameters and the increased susceptibility to atherogenesis in ß-thalassaemia-trait (BTT) subjects. METHODS: Sixty BTT subjects and 20 age- and sex-matched healthy controls were enrolled in the study. Serum PON1, total antioxidant capacity (TAC), malondialdehyde (MDA) and carotid artery intima-media thickness (CIMT) were determined. Qualitative detection of ß-thalassaemia mutations was carried out. RESULTS: Serum PON1 activity and TAC were significantly lower in BTT subjects than in controls (p<0.001), while MDA and CIMT were significantly higher (p<0.001). In BTT subjects, TAC, MDA, and CIMT levels were significantly correlated with serum PON1 (r=0.945, -0.900, 0.940 and -0.922 respectively). Serum TAC and MDA were significantly correlated (r=-0.979). CONCLUSIONS: Oxidative stress is increased, while serum PON1 activity is decreased in BTT subjects. Decrease in PON1 activity is associated with the degree of oxidative stress, anaemia and increase in CIMT. Therefore, BTT subjects may be more prone to development of atherosclerosis.

Prognostic significance of survivin in pediatric acute lymphoblastic leukemia.

Impaired apoptosis is mediated by members of the inhibitor of apoptosis proteins (IAP) family such as survivin. Survivin was described in number of different tumors and found to correlate with poor prognosis in a variety of cancers including hematologic malignancies. The aim of this study was to determine survivin in pediatric ALL and compare it with clinical and hematological findings, response to therapy and outcome. Flowcytometry was used for detection of intracellular survivin and determine its mean fluorescence intensity (MFI) in bone marrow mononuclear cells. Patients were followed up for 28 months after induction therapy. Survivin was detected in 63.3% of the patients BM. In spite of no association of survivin levels with established risk factors (P > 0.05) except with high WBC, there was significant higher level of survivin expression in high risk group patients when patients were stratified into high and standard risk groups. According to response to induction therapy, there was no significant difference, in survivin level between patients who achieved CR, RD and ED. However, patients suffering relapse of the disease, had a significant higher basal level of survivin than patients still in remission. Over expression of survivin is a candidate parameter to determine poor prognosis in ALL patients and it may serve to refine treatment stratification with intensification of therapy in those patients prone to relapse.

The clinical significance of OPG/sRANKL ratio in thalassemia patients suffering from osteopenia or osteoporosis in Egyptian patients.

Osteopenia and osteoporosis are considered major health problems in patients suffering from thalassemia due to increased life expectancy of those patients. Osteoprotegerin (OPG) and receptor activator of NF-kappa-B ligand (RANKL) have been recently implicated in the pathogenesis of various types of osteoporosis. The aim of this study is to evaluate the role of serum OPG/RANKL ratio in patients suffering from thalassemia complicated with osteoporosis. Serum OPG and RANKL were measured in thalassemia patients and 20 healthy control subjects, using ELISA methods. Stastistically, the results demonstrate lower OPG and OPG/RANKL ratio in patients suffering from thalassemia with documented osteopenia or osteoporosis in comparison with control group and patients suffering from thalassemia without osteopenia or osteoporosis. OPG/RANKL ratio could become a promising rapid and cheap screening marker for osteopenia or osteoporosis in patients suffering from thalassemia. Furthermore, OPG may become a therapeutic option in treatment of osteoporosis of various etiologies including thalassemia.

Impact of CD4+CD25high regulatory T-cells and FoxP3 expression in the peripheral blood of patients with systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease characterized by various immunological abnormalities. Regulatory T cells (Tregs) CD4+CD25+ play an important role in maintaining tolerance to self-antigens controlling occurrence of autoimmune diseases. It has been shown that the transcription factor forkhead box P3 (FoxP3) is specifically expressed on CD4+CD25+T cells. FoxP3 has been described as the master control gene for the development and function of Tregs. A decrease in the number of CD4+CD25highFoxP3+ regulatory T cells can play a key role in the loss of tolerance to self antigens. The study was designed to assess expression levels of FoxP3 in peripheral CD4+CD25+ regulatory T cells in patients with SLE and to evaluate the level of some cytokines that are implicated in the extent of the disease activity. The study was carried out on 30 SLE patients, they were 27 females and 3 males, 10 age and sex matched healthy volunteers were studied as a control group. They were divided into two groups: group I: had active disease (12 patients) and group II: had inactive disease (18 patients) according to Systemic Lupus Erythematosus Disease Activity Index. All individual were subjected to CBC, ESR, s.creatinine, RF, CRP, C3, ANA, anti ds-DNA and flowcytometeric assay of CD4+CD25+ (Tregs) and FoxP3 for patients and controls. Quantitative determination of serum interleukin 10 (IL-10) and transforming growth factor-beta1 (TGF-beta1) concentrations in serum samples by ELISA technique. The results revealed a significant decrease of CD4+CD25high cells in peripheral blood in active lupus patients when compared with patients with inactive lupus and those in healthy controls. Intriguingly, the percentage level of FoxP3 on CD4+CD25high cells was significantly decreased in SLE patients with active disease (2.9 +/- 1.05) when compared with those with inactive SLE (3.5 +/- 0.8) and control groups (4.7 +/- 1.2) (P < 0.05). As regard cytokines levels; the level of IL-10 was significantly increased in patients with active and inactive disease (158.8 +/- 50.8, 82.8 +/- 14.08 respectively) when compared with the control group (P < 0.001). While, the level of TGF-beta1 was significantly decreased in patients with active and inactive disease (22.5 +/- 7.03, 29.07 +/- 10.14 respectively), when compared with the control group (P < 0.05). Our data revealed impaired production of Tregs in SLE patients, which may play a reciprocal role with some cytokines to affect the activity of the disease. Tregs cells should be the target to determine the clinical effectiveness of novel therapy to modulate Tregs in vivo besides the conventional treatments.