Plasminogen activator inhibitor-1 (PAI-1) is a key regulator of fibrinolysis; however, the relationship between PAI-1 and the most common diabetic microvascular complication, retinopathy, is unclear.
To examine the association between the 4G/4G polymorphism of the PAI-1 gene with diabetic retinopathy (DR) as well as with the plasma levels of the PAI-1 enzyme among Egyptian patients.
Thirty-three patients who had type 2 diabetes for more than 10 years were compared with 63 patients with proliferative diabetic retinopathy (PDR). Both groups were compared with 48 control individuals. All groups were matched for age and sex. PAI-1 4G/5G genotyping was carried out by a PCR and the PAI-1 levels were measured by enzyme-linked immunosorbent assay testing.
Higher plasma PAI-1 activity was associated with a higher risk of DR. The overall frequency of the 4G allele was 54.54% among type 2 diabetes patients versus 78.79% among PDR patients (P<0.01). Using multivariate logistic regression analyses, patients with PDR had a higher representation of the genotype 4G/4G (P<0.05, odds ratio: 3.15, 95% confidence interval 0.13–0.89) and the 4G/4G patients studied had higher plasma levels of PAI-1 activity.
The PAI-1 gene polymorphism 4G/4G contributed to the genetic susceptibility to DR and a higher PAI-1 plasma level was independently associated with a higher risk of retinopathy among Egyptians.
This study aimed to determine the efficacy of a Nigella sativa (NS) seed supplement on hemodynamics, hemoglobin (Hb) levels, and blood coagulation in patients with type 2 diabetes mellitus.
This study included 94 patients who were divided randomly into three dose groups. Capsules containing NS seeds were administered orally at a dose of 1, 2, and 3 g/day for 12 weeks. In all patients, the hemodynamic markers [systolic blood pressure, diastolic blood pressure, mean arterial pressure, heart rate (HR), and rate pressure product (RPP)], Hb levels, and coagulation markers (prothrombin time, partial thromboplastin time, fibrinogen levels, and platelet count) were determined before treatment and after 4, 8, and 12 weeks.
Patients receiving 1 g/day NS for 12 weeks (group 1) showed nonsignificant changes in the hemodynamic parameters, whereas patients who received a supplement of 2 g/day NS (group 2) showed a significant reduction in the systolic blood pressure, diastolic blood pressure, mean arterial pressure, HR, and rate pressure product, compared with the baseline values. An increase in the NS dose to 3 g/day exerted less effect on the blood pressure and reversed the effect on HR. The Hb levels decreased slightly, but significantly in the three groups. Partial thromboplastin time showed a significant increase in group 2 at the end of the treatment period and a nonsignificant increase in groups 1 and 3 throughout the treatment period. However, the fibrinogen levels increased significantly in groups 2 and 3 compared with the baseline values.
Ingestion of 2 g/day NS seeds for 12 weeks exerted a favorable impact on the hemodynamic parameters. However, NS should be administered with caution in diabetic patients with anemia.
The discrimination between reactive and malignant cell populations in some patients with suspected T-cell lymphoproliferative disorders can be complicated and less straightforward. Often, the dilemma lies in determining whether a population of lymphocytes is reactive or neoplastic. In such cases, T-cell receptor (TCR) gene clonality studies have proved useful as an additional diagnostic tool. The TCRγ gene is a preferred target for TCR gene clonality as it is rearranged at an early stage of T lymphoid development, rearranged in a large percent of T-cell neoplasms, and also because of its relative structural simplicity.
In the present study, we used the BIOMED-2 multiplex primer panel to assess the value of TCRγ gene rearrangement using genescan analysis in 30 patients with suspected T-cell neoplasms and to elucidate its possible role in the diagnosis of such disorders.
TCRγ gene clonality was detected in all 18 patients with suspected T-cell acute lymphoblastic leukemia/lymphoblastic lymphoma. In contrast, it was detected in nine out of 12 patients (75%) with suspected peripheral T-cell lymphoma. It is known that TCRγ monoclonality in peripheral T-cell lymphoma can be detected in patients with more aggressive disease in terms of both clinical presentation and laboratory results. VγfI was the most frequently used Vγ segment in our patients. The sensitivity and specificity of genescan were found to be 0.89 and 0.67, respectively, as compared with histopathology.
It is concluded that TCRγ gene clonality is a very useful diagnostic tool in T-cell neoplasms. As it is relatively simple, it can be used as a preliminary test for clonality assessment, followed by the more complex TCRβ gene rearrangement only in negative cases to improve sensitivity. In addition, it should be used together with heteroduplex and interpreted within the clinical context to improve its specificity.
Connexin 43 (Cx43) inhibits cell proliferation in gap junction-dependent and gap junction-independent mechanisms and contributes to the pathogenesis of many diseases. The aim of this study was to assess Cx43 expression in acute myeloid leukemia (AML) patients with t(8;21) compared with patients without this translocation and correlate this expression with the cyclin-dependent kinase inhibitor P27.
Group I comprised 21 patients with de-novo M2 who were positive for t(8;21). Group II comprised 29 patients with de-novo M2 who were negative for t(8;21). Group III comprised 25 apparently normal individuals, with matched age and sex, which served as the control group. In addition to routine laboratory investigations, detection of Cx43 expression on bone marrow mononuclear cells (MNCs) and P27 in MNC lysate was carried out for all patients.
In this study, we found increased expression of Cx43 on leukemic cells carrying t(8;21) compared with leukemic cells without t(8;21). Compared with normal cells, there was decreased expression of Cx43 on leukemic cells carrying t(8;21). Furthermore, we found significant negative correlation between the count of leukemic cells and Cx43 expression. We found increased levels of P27kip1 protein in MNCs extracted from leukemic patients with positive t(8;21) compared with leukemic patients without this translocation. In addition, we found significant positive correlation between Cx43 expression and P27kip1 protein level.
Our experiments show that Cx43 expression increases in M2 with t(8;21) and may contribute to the growth-arresting effect of leukemogenic AML1–eight-twenty one (ETO) fusion protein, possibly by causing the accumulation of P27kip1 protein. These novel discoveries have shed new light on the mechanisms of AML1–ETO-induced growth arrest and have provided a foundation for exploring additional mutagenic ‘hits’ for AML1–ETO-associated AML. We recommend the use of drugs that improve Cx43 expression in treating M2.
Deletions of the long arm of chromosome 6 (6q) are among the most frequent chromosome aberrations in multiple human tumors, including acute lymphocytic leukemia and non-Hodgkin’s lymphoma (NHL). The accurate characterization of the 6q deletions is of clinical significance in lymphoid malignancies because they are related to the prognosis. The aim of this study was to evaluate the frequency of 6q21 deletion in B-cell chronic lymphoproliferative disorders (BCLPDs) and to evaluate the association of this deletion with other prognostic criteria.
A fluorescence in-situ hybridization probe set on chromosome 6q21 was used to evaluate 6q21 deletion in 63 patients with different BCLPDs, they were 30 patients with chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL), 21 patients with NHL, four patients with Waldenström macroglobulinemia, and eight patients with multiple myeloma.
6q21 deletion was detected in 20% of CLL/SLL (6/30), (40%) of diffuse large B-cell lymphoma (2/6), 33.3% of follicular lymphoma (2/6), and 20% of mantle cell lymphoma (1/5), 25% of multiple myeloma (2/8), and in 50% of Waldenström macroglobulinemia (2/4) patients. None of the two patients with splenic lymphoma or the three patients with unclassifiable B-NHL showed 6q21 deletion. The median percentage of clonal lymphoid cells with deletion (6q21) was 45% (33–88%). CLL/SLL patients with 6q21 deletion had a significantly lower hemoglobin concentration (P=0.006); a significantly higher mean age (P=0.009); percent of peripheral blood prolymphocytes (P=0.001); and level of serum lactate dehydrogenase (P=0.009) compared with those without the deletion. All CLL/SLL patients (100%) with 6q deletion were allocated to stage C and stages III and IV of Binet and Rai staging systems, respectively, whereas only (58.3%) of those without 6q deletion were allocated to the same advanced stages; however, this was not statistically significant. Among NHL patients, 6q deletion was associated with older age (>60 years, P=0.01), lower hemoglobin concentration (P=0.003), higher level of β2-microglobulin (β2M) and lactate dehydrogenase (P=0.01 and 0.00, respectively), and higher incidence of splenomegaly (P=0.003). All NHL patients with 6q deletion were stratified into a high-risk group according to the International Prognostic Index, whereas those without 6q deletion, approximately 44%, were classified as a high-risk group (P=0.02). Staging of NHL patients according to the Ann Arbor system showed no significant difference between patients with 6q deletion and those without.
On the basis of the high frequency of deletion (6q) in BCLPDs, we suggest a pathogenetic role of this deletion in BCLPDs. Our results support previous studies that have shown an impact of 6q deletion on prognosis in subtypes of BCLPDs.
CD200 is a transmembrane protein expressed on multiple cell types (e.g. thymocytes, activated T cells, B cells, dendritic cells, and endothelial cells). It regulates antitumor immunity and its overexpression is associated with poor prognosis in chronic lymphocytic leukemia (CLL). A soluble variant of CD200 (sCD200) is detectable in human serum, and an elevated serum level was shown in patients with CLL.
The aim of the study is to investigate the relationship between the levels of CD200 and clinical staging in patients with CLL.
The study included 50 patients, divided into two groups. The patient group included 30 de-novo patients with CLL (18 men and 12 women), whose age ranged from 49 to 70 years with a mean±SD of 57.82±4.94 years. They were classified according to the modified Rai staging system into three classes: (a) low-risk patients, which included seven cases (stage 0); (b) intermediate-risk patients, which included eight cases (stages I and II); and (c) high-risk patients, which included 15 cases (stages III and IV). The control group included 20 individuals (14 men and six women), whose age ranged from 50 to 72 years with a mean±SD of 58.54±5.1 years. CD200 levels were measured by flow cytometry.
Higher levels of CD200 were observed in the patient group compared with the control group. In addition, there was a significant increase of CD200 expression in the intermediate group of patients compared with low-risk patients, and a significant increase in the CD200 level in the high-risk group compared with intermediate-risk patients.
A positive correlation between the level of CD200 expression and clinical staging system of CLL was observed in our study; thus, measurement of CD200 may have a prognostic role in patients with CLL. In addition, it can be used as a follow-up marker for clinical response to treatment in CLL. CD200 blockade by immunotherapeutic agents may represent a novel approach for clinical treatment of CLL and a number of B-cell-derived neoplasms.
In the Aseer area, in the southern western region of Saudi Arabia, visceral leishmaniasis or Kala-azar is an endemic and serious disease that mainly affects children.
In this study, an anti-leishmania antibody diagnostic test, the indirect hemagglutination (IHA) test, was compared with the gold standard microscopic detection of Leishmania donovani bodies in bone marrow aspiration (LDB/BMA) smears. Factors such as sensitivity, specificity as well as reliability will govern whether health authorities should continue the use of the IHA assay or replace it with more specific and sensitive as well as reliable techniques.
A total of 582 patient samples were included in this study. All were tested by IHA and by microscopic examination for LDB/BMA smears. Results of the tests are statistically analyzed and tabulated.
The comparison showed a poor correlation between the microscopic detection of parasite forms in the gold standard bone marrow smear examination (LDB/BMA) and the serological antibody screening of patient samples by IHA (P⩽0.05). This type of results and correlation poses serious questions relating to the sensitivity, specificity, and ultimately, the reliability of the IHA test in the diagnosis of visceral leishmaniasis.
A better alternative for IHA test might be the introduction of methods that detect the parasite it self or its antigens that can be used in conjunction with microscopic examination for LDB/BMA. In this respect, PCR–ELISA and KATEX agglutination tests represent obvious candidates.
We assessed the cost-effectiveness of postremission high-dose arabinoside (HiDAC)-based chemotherapy in comparison with allogeneic stem-cell transplantation (alloSCT)-based therapy in adult Egyptian patients with acute myeloid leukemia at either an intermediate or a poor cytogenetic risk.
Forty young patients at either an intermediate or a poor cytogenetic risk received postremission intensive therapy (20 HiDAC-based/20 alloSCT-based). We analyzed the overall survival, disease-free survival, and cost effectiveness in both groups.
Patients were followed up over a period of 60 months. The mean overall survival was 36.32 months for the HiDAC-based group, whereas it was 48.8 months in the alloSCT-based group. The mean disease-free survival was 24.8 months in the HiDAC-based group, whereas it was 51.5 months in the alloSCT-based group. The mean total cost in the HiDAC-based group was 51 300 Egyptian pounds (nearly US$9000), whereas it was 88 250 Egyptian pounds (nearly US$15 482) in the alloSCT-based group.
For the postremission therapy in young acute myeloid leukemia patients at either an intermediate or a poor cytogenetic risk, HiDAC-based chemotherapy is less costly and less effective, whereas alloSCT is more costly and more effective in maintaining patients in durable remission.
B-cell chronic lymphocytic leukemia (B-CLL) results from the accumulation of slowly proliferating long-lived CD5-positive B cells with apoptotic defects. The clinical course of CLL is variable and several markers have been proposed to predict the outcome in CLL patients. CLL-upregulated gene 1 (CLLU1), a gene that is exclusively upregulated in CLL cells, was identified and mapped to chromosome 12q22.
The objective of this study was to evaluate the prognostic significance of CLLU1 expression in B-CLL patients and its relationship with other well-established prognostic markers.
Forty newly diagnosed CLL patients were included in the study. The expression levels of CLLU1 were determined by quantitative real-time PCR, CD38 expression was determined by flow cytometry, and the presence of trisomy 12 was evaluated by the fluorescence in-situ hybridization technique. The relationship between CLLU1 expression levels and Binet’s clinical staging of the disease, CD38 expression, and trisomy 12 was evaluated. The response of CLL patients to therapy and progress of the disease were monitored in relation to CLLU1 expression levels.
CLLU1 expression was found to be upregulated 0.4–1120.1-fold in B-CLL cells as compared with normal B cells. CLL patients were segregated into two groups on the basis of high and low CLLU1 expression levels. High expression of CLLU1 was significantly associated with higher absolute lymphocytic count and also higher serum β2 microglobulin and lactate dehydrogenase levels with no significant association detected between CLLU1 expression levels and age of the patients at diagnosis. Significant positive correlations could be detected between high CLLU1 expression levels and both the advanced clinical stages of the disease and the positive expression levels of CD38. High expression of CLLU1 occurred irrespective of trisomy 12 and was associated with failure to achieve complete remission and shorter overall survival in CLL patients.
The present study demonstrated that high CLLU1 expression in B-CLL cells is a significant predictor of disease progression and poor response to therapy. High expression levels of CLLU1 correlate significantly with the known prognostic markers of the disease, mainly the clinical staging and CD38 expression. CLLU1 expression levels are independent of trisomy 12.
The B-cell-activating factor of the tumor necrosis factor family (BAFF) is a homotrimeric type 2 transmembrane protein that also exists in a soluble form. It belongs to the family of tumor necrosis factor ligands and is expressed at the surfaces of myeloid cells and antigen-presenting cells and induces B-lymphocyte proliferation and immunoglobulin secretion.
The aim of this study was to assess the serum BAFF level in patients with idiopathic thrombocytopenic purpura (ITP) and to determine the correlation of its level with response to immunosuppressive therapy (corticosteroids).
The study included 60 participants: 40 patients with newly diagnosed ITP who were followed up for 3 months after immunosuppressive therapy (steroids), divided into responders and nonresponders, and 20 healthy control individuals.
The serum BAFF level was lower in the controls (mean±SD 3.4±2.2 ng/ml) than the ITP patients (responders and nonresponders) before treatment (mean±SD 19.8±2.9 and 20.5±8.8 ng/ml, respectively) with a statistically highly significant difference (P<0.001), but no significant difference between the responders and the nonresponders. After treatment, the BAFF level was still high in the nonresponder group (mean±SD 18±7.6 ng/ml), with a statistically highly significant difference in comparison with the responders and the controls (mean±3.6±1.9 and 3.4±2.2 ng/ml, respectively) (P<0.001). It was found that BAFF decreased markedly among all the participants and in the responder group, with a statistically highly significant difference between them and the nonresponder group. The percentage of change was 84% among responders compared with 10% only among nonresponders. In addition, the BAFF level was inversely correlated with the platelet count.
The serum BAFF level increases in patients with ITP and reverts to normal after treatment in responders but remains high in nonresponders. We recommend further randomized-controlled clinical trials exploring the role of drugs acting on BAFF in the treatment of autoimmune disorders.
The importance of angiogenesis was proved first for solid tumors, and its importance in hematological malignancies was reported later.
The present study aimed to evaluate the prognostic value of the angiogenesis marker platelet-derived growth factor-BB (PDGF-BB) in patients with aggressive non-Hodgkin’s lymphoma (NHL) and identify its significance on treatment response.
The serum level of PDGF-BB was measured in patients with aggressive NHL before and after chemotherapy and compared with that of a healthy control group, and its relationship with certain prognostic parameters was assessed.
Pretreatment serum level of PDGF-BB did not show a significant change as compared with the control group. Its level showed no relation to Ann Arbor stages, International Prognostic Index scores, B symptoms, or extranodal involvement. Chemotherapy led to a considerable increase in serum PDGF-BB level in all NHL patients and in the noncomplete responder subgroup.
From the results mentioned above it could be concluded that PDGF-BB is not vital in the pathogenesis of NHL. Chemotherapy led to elevation of serum PDGF-BB, reflecting activation of angiogenesis, which can be considered one of the mechanisms of conferring chemoresistance to cancer cells or in identification of patients at risk for developing side effects. Future studies are recommended to prove this effect of chemotherapy. If this effect is confirmed, the use of agents blocking the angiogenic pathway of PDGF-BB in combination with chemotherapy may improve the therapeutic effectiveness and sensitize chemoresistant cells.
Our aim is to determine the rate of glutathione S-transferase T1 (GSTT1) null genotypes in acute myeloid leukemia (AML) patients as a risk factor and analyze the prognostic significance of this gene polymorphism.
We genotyped GSTT1 in two groups: the patient group included 30 children with AML who were receiving chemotherapy and the control group included 50 healthy children. PCR amplification was used to assign the GSTT1 genotype for the cases and the controls. The outcomes were compared in the patient group (those with and without GSTT1 genes).
The frequency of GSTT1 null was significantly increased in the AML cases compared with the controls (50 vs. 10%).
The GSTT1 null genotype is a significant risk factor for childhood AML. The frequency of early death was high in GSTT1-negative cases. Patients with the GSTT1-negative genotype had reduced survival compared with those with at least one GSTT1 allele (GSTT1 positive). The frequency of relapse from the end of induction did not show any significant difference in the GSTT1-negative and the GSTT1-positive cases. The GSTT1 genotype might be useful when deciding on appropriate chemotherapy regimens for children with AML.
FoxP3 constitutive expression is necessary for the suppressive function of regulatory T cells (Tregs). The majority of infected patients with hepatitis C virus (HCV) develop only a weak, narrow, or nonpersistent adaptive response to acute infection. The aim of this study was to investigate the frequency of CD4+FoxP3+ Tregs in peripheral blood and liver biopsy tissues in patients with acute and chronic hepatitis C and their potential association with viral load.
The study participants were divided into three groups. Group I comprised 15 recently diagnosed patients with acute hepatitis C. Group II comprised 46 patients with chronic hepatitis C, of whom 21 patients had chronic hepatitis C without hepatic cirrhosis (group IIa) and 25 patients had chronic hepatitis C with hepatic cirrhosis (group IIb). Group III comprised 22 apparently normal individuals and they constituted the control group. Double immunohistochemical analyses were performed on liver biopsy samples for CD4/FoxP3 and CD8/FoxP3. Further, flow cytometric analysis of peripheral blood for CD4, CD8, and FoxP3-positive lymphocytes was carried out.
Most FoxP3+ cells in the peripheral blood and liver tissues were CD4+ cells, and CD8+FoxP3+ cells were very scare. A considerable number of FoxP3+ cells were observed in the portal tracts and fibrous septa, particularly when lymphoid aggregates were present. They were also observed in parenchymatous areas, where they preferentially localized in necrotic areas. In group I patients there were significant positive correlations between CD4+FoxP3+ cells in peripheral blood and alanine aminotransferase, aspartate aminotransferase, total bilirubin, and viral copies. In chronic hepatitis cases, no correlations were found between CD4+FoxP3+ cells in peripheral blood and liver tissues and other laboratory parameters.
FoxP3− Tregs are upregulated in HCV patients, suggesting an important role for Tregs in establishing and/or maintaining HCV persistence. Further studies are needed to examine the role of Tregs in HCV disease pathogenesis and to develop therapeutic approaches to control the balance between Tregs and effector T cells to enhance viral clearance.