Phe354Leu polymorphism of the liver kinase B1 gene as a prognostic factor in adult egyptian patients with acute myeloid leukemia.

Background: The human liver kinase B1 (LKB1) gene is a significant tumor suppressor widely expressed in all fetal and adult tissues. Despite its established role in solid tumors, the biological and clinical implications of LKB1 gene alterations in hematological malignancies have not been sufficiently recognized. Aim: This study aimed to determine the frequency of the LKB1 Phe354Leu polymorphism in adult Egyptian patients with cytogenetically normal AML (CN-AML), evaluate its clinical prognostic significance, and investigate its effect on the therapeutic outcome and patient survival. Methods: Direct sequencing of amplified exon eight of the LKB1 gene was performed to detect the Phe354Leu polymorphism in 72 adult de novo CN-AML patients. Results: The LKB1 Phe354Leu polymorphism was detected in 16.7% of patients and associated with younger age and lower hemoglobin levels (p < 0.001). Patients in the mutated group had significantly higher total leukocytic count and bone marrow blasts (p = 0.001 and p < 0.001, respectively). The most common FAB subtypes in mutated patients were M4 and M2. The relapse rate was significantly higher in the mutated group (p = 0.004). There was a significant association between the FLT3-ITD polymorphism and LKB1 F354L (p < 0.001). The mutated group had shorter overall survival (p = 0.003). In multivariate analysis, the Phe354Leu polymorphism was a significant independent prognostic variable for the overall and disease-free survival of the studied patients (p = 0.049). Conclusion: The LKB1 Phe354Leu polymorphism was diagnosed at younger ages in Egyptian CN-AML patients and represented a poor independent prognostic factor in CN-AML. Patients who carried this polymorphism had shorter overall survival and more frequent relapses. Our findings may provide insight into the design of therapeutic targets, and molecular testing of the LKB1 gene is recommended for proper risk stratification of CN-AML patients.

Prognostic Significance of VEGF and HIF-1 α in Hepatocellular Carcinoma Patients Receiving Sorafenib Versus Metformin Sorafenib Combination.

BACKGROUND: Hepatocellular carcinoma (HCC) is a major health problem. HCC burden has been increasing in Egypt in the past 10 years. Most HCC cases are diagnosed at an advanced stage with limited treatment options. Sorafenib is the standard therapy for advanced HCC, but the effectiveness is not satisfied. Metformin may decrease the risk of HCC development in diabetic patients, reduces tumor invasion, and augments sensitivity to sorafenib; however, safety and efficacy of combined treatment are still unclear. As HCC is characterized by high vascularity, and vascular endothelial growth factor (VEGF) plays an important role in vascularization, many studies questioned if VEGF and HIF-1 α could offer information about HCC response to sorafenib. We conducted this study to assess the benefits from adding metformin to HCC treatment, and appraise the role of VEGF and HIF-1 α in HCC prognosis. METHOD: This was a prospective, randomized study in which 80 advanced measurable patients consecutively treated with sorafenib plus metformin (arm A) or sorafenib alone (arm B), prognostic value of plasma, and tissue levels of VEGF and HIF-1 α were evaluated. RESULTS: We enrolled 61 men and 19 women with a median age of 60 years (range 49-68 years). Fifty-seven patients had Child-Pugh A while 23 had early B, the most common etiology of liver disease was hepatitis C (86%). Sixty percent of patients were diabetic. No significant difference was detected between arm A and arm B regarding response to treatment (p = 0.5), time to disease progression (p = 0.3), or overall survival (p = 0.6). Low VEGF and HIF-1 α plasma levels were significantly associated with better treatment response (p < 0.001 for both), and higher OS (p < 0.001). Patients with high expressions of VEGF and HIF in HCC tissue had significantly poor treatment outcome (p < 0.001, p = 0.03, respectively), and poor OS (p < 0.001, p < 0.001, respectively). CONCLUSIONS: No superior efficacy of adding metformin to sorafenib in HCC treatment. VEGF and HIF-1 α had promising prognostic value in HCC.

Genetic polymorphism of IL-23R influences susceptibility to HCV-related hepatocellular carcinoma.

BACKGROUND: Genetic variations may play an important role in the development of HCC in HCV patients. Variants of IL23R gene were investigated for association with many diseases like chronic inflammatory disorders, RA, inflammatory bowel diseases and the susceptibility to the development of gastric cancer but no data are available concerning the association of IL23R gene (rs11209026) polymorphism with HCC development in HCV patients. Therefore the current study aimed to analyze this polymorphism within the gene to evaluate its contribution to chronic HCV susceptibility and/or HCC development in Egyptian patients. SUBJECTS AND METHODS: One hundred and ninety-two patients with chronic HCV infection were included in this study (92 of them without HCC and 100 of them with HCC). One hundred healthy control subjects with no history of previous liver disease (HBV and HCV infection were negative) were included in the study. The IL23R polymorphism (rs11209026 G>A) were genotyped by real time PCR. RESULTS: We found a significant lower incidence of GA and AA genotype in HCV patients with HCC compared to those without HCC (p=0.026 and 0.040 respectively) and compared to control group (p=0.008 and 0.007 respectively). While, no significant difference between control and HCV patients without HCC groups was found. CONCLUSIONS: Our study suggests that wild type IL-23R GG serves as a risk factor for HCC and supports for the protective role of the rare variant rs11209026 (Arg381Gln) against HCV-related HCC in Egyptian patients.

Serum copper is a simple but valuable prognostic marker in B-cell chronic lymphocytic leukemia.

We investigated the relationship between serum copper and various prognostic factors, time to start treatment, and treatment response in patients with B-cell chronic lymphocytic leukemia (B-CLL) and related disorders. Fifty newly diagnosed CLL patients aged 36-70 years were included. Patients were studied for serum lactate dehydrogenase (LDH), serum copper, direct Coombs' test, serum ß(2) microglobulin (ß(2)M), immunophenotyping for diagnosis of B-CLL, evaluation of CD38 and zeta-associated protein (ZAP-70) expression, and fluorescence in situ hybridization technique for cytogenetic analysis. Fourteen of 50 patients had high serum copper level; they had a significant increase in LDH, serum ß(2)M, incidence of positive Coombs' test, CD38 and ZAP-70, incidence of 17p del, and a decrease in hemoglobin concentration, lymphocyte doubling time and time to start treatment with a lower treatment response rate. No significant difference was found with regard to Rai staging for CLL. These results indicate that serum copper level, a cheap and simple laboratory test, is of great value in CLL patients as it showed a significant association with some important adverse prognostic markers such as increased expression of ZAP-70 and CD38, shorter time to start treatment and poor response to treatment.

Significance of telomere capture in myelodysplastic syndromes.

Addition of telomeres to the ends of broken chromosomes has been observed in many malignant cells through the capture of the ends of other chromosomes as a result of nonreciprocal translocations. In this study, we aimed to evaluate the percentage of nuclei with telomere capture (TC%) as a prognostic marker in myelodysplastic syndromes (MDS) patients. This study included 45 newly diagnosed MDS patients, 36 cases with denovo MDS and 9 cases with therapy-related MDS, and another 35 apparently healthy volunteers as a control group. Telomere capture percentage was investigated with fluorescent in situ hybridization technique using a probe for 15qter. We found that median TC% rate was significantly increased in those with bad cytogenetic abnormalities, patients with blast cells>10% in BM, and patients categorized as high risk according to WHO and IPSS classification; also, there was a significant negative correlation with progression-free survival. Telomere capture serves as a useful marker for the assessment of MDS patient's risk, and also it had a clinical importance for the early detection of disease progression.

Clinical relevance of serum endoglin level in Egyptian hepatocellular carcinoma patients.

BACKGROUND: In hepatocellular carcinoma (HCC), preferential expression of endoglin on endothelial cells of the tumor vasculature versus neoplastic cells has led to the suggestion that it originates from the neovasculature and plays an important role in angiogenesis. Our study aimed to evaluate the role of serum endoglin in the diagnosis of HCC and to correlate it with other studied prognostic markers. METHODS: Sixty hepatocellular carcinoma patients were studied, 60 cirrhotic patients and 45 matched healthy controls. Liver function tests, alpha fetoprotein (alphaFP) and serum endoglin levels were determined. RESULTS: The study revealed a significant increase in alphaFP and endoglin in patients with liver disease compared to controls and in HCC patients compared to cirrhotic ones. There was a positive correlation between both biomarkers and stages of HCC, whereas no significant correlation between endoglin and alphaFP or between both of them and Child Pugh's classification was seen. At a cutoff value of 20 ng/mL, the sensitivity of alphaFP was 75% and the specificity was 90%, in differentiating HCC from cirrhosis. At a 7.5 ng/mL cutoff, the sensitivity of endoglin was 70% and specificity was 65%. On combination of both biomarkers, the sensitivity was increased to 85%. CONCLUSIONS: Serum endoglin is a useful complementary biomarker in the diagnosis of HCC.

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.

HE4 and mesothelin: novel biomarkers of ovarian carcinoma in patients with pelvic masses.

OBJECTIVES: To evaluate the utility of novel serum tumor markers, HE4 and mesothelin either alone or in combination with CA125 in diagnosis and early detection of ovarian carcinoma in patients with pelvic masses. SUBJECTS AND METHODS: Sera were obtained preoperatively from 65 women underwent surgery for a pelvic mass and 25 age- and menopausal status-matched healthy women. All samples were analyzed for levels of CA125, HE4, and mesothelin by serum based immunoassays and patients results were compared to final pathology findings. RESULTS: Of 65 patients with pelvic masses; 41 had histologically diagnosed ovarian cancer, and 24 had benign ovarian diseases. The studied tumor markers were significantly increased in malignant compared to benign cases and healthy subjects, and in benign cases compared to healthy subjects (p<0.001). Based upon Receiver operator characteristic (ROC) curves analysis, HE4 had the highest sensitivity as a single marker in detecting ovarian malignancy (82.9%) and early stage malignancy (76.9%), followed by CA125, then mesothelin. The combination of HE4 and CA125 gave the highest sensitivity in detecting ovarian carcinoma and early stage disease (90.2%, 84.6% respectively). Addition of mesothelin to this combination did not show any improvement in the sensitivity. CONCLUSIONS: As a single marker, HE4 had the highest sensitivity for detecting ovarian carcinoma specially early stage disease. Combined CA125 and HE4 was a more accurate predictor of ovarian malignancy than either alone.

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.