Purpose: To explore the expression and prognostic significance of Hedgehog signaling transcription factor GLI-1 in newly diagnosed acute myeloid leukemia (AML) patients. Methods: Clinical specimens were obtained from 46 recently diagnosed AML patients. Real-time qPCR was used to measure the GLI-1 mRNA expression in bone marrow mononuclear cells.Also, the relationship between GLI-1 mRNA levels and clinical variables and prognostic variables was assessed. Results: GLI-1 was overexpressed in the bone marrow samples of our patients. GLI-1mRNA expression did not differ significantly across different age groups, between both sexes, or between different FAB subtypes (P = 0.882, P = 0.246, and P = 0.890, respectively). GLI-1 expression varied significantly in different risk categories, with the greatest levels observed in 11 patients with poor risk (24.6 versus 22.7) compared to intermediate risk (5.2 versus 3.9; P = 0.006) and favorable risk (4.2 versus 3; P = 0.001). Comparing patients with the wild FLT3 allele to those with the mutant one, GLI-1 gene levels were considerably greater in those with the mutant allele of FLT3.Following induction chemotherapy, the levels of GLI-1 mRNA were significantly higher in 22 patients who did not experience complete remission (CR) diagnosed with de novo non-acute promyelocytic leukemia (APL) compared to 17 patients who did (P = 0.017). Significantly greater levels of expression were observed in each category of the patients with favorable risk; wild FLT3 allele (P = 0.033) and CR failure P = 0.005). Conclusion: GLI-1 overexpression is a risk factor for poor prognosis and could be a novel therapeutic target for AML.

BackgroundChronic lymphocytic leukemia (CLL) is the most common leukemia among adults in the Western world. Autophagy is a self-degradative process that is essential for balancing the energy sources at critical times and in response to stress. BECN1 and phosphoinositide 3-kinase C3 (PIK3C3) genes are crucial for the initiation of autophagy process. SLAMF1 is a costimulatory molecule and microbial sensor involved in genetic pathways that adjust chemotaxis and autophagy.ObjectivesTo study the role of BECN1 and PIK3C3 gene expression and assess SLAMF1 surface expression in CLL and correlate with disease characteristics.Patients and methodsIn this case–control study, 40 newly diagnosed patients with CLL were included as well as 10 age-matched and sex-matched apparently healthy control. RT-PCR was used to measure PIK3C3 and BECN1 genes expression. SLAMF1 surface expression was measured by flow cytometry.ResultsPatients with CLL had lower BECN1 and PIK3C3 gene expressions than the control group (P < 0.001). PIK3C3 showed a statistically significant correlation with CD38 (P = 0.012), whereas BECN1 showed a statistically significant difference with different Binet stages (P = 0.017). Otherwise, there was no significant correlation between expression of both genes and different clinical parameters. SLAMF1 surface expression was statistically lower in the patient group than in the control group (P < 0.001). In addition, it was found to be an independent prognostic marker in CLL, with higher expression being associated with a good prognosis.ConclusionAutophagy genes and SLAMF1 represent promising candidates for future studies with respect to their role in autophagy in CLL, and they may represent targets of treatment.

B-cell-activating factor (BAFF) is a crucial cytokine supporting survival and differentiation of B cells. Dysregulation of BAFF is involved in the pathogenesis of B-cell related autoimmune diseases including immune thrombocytopenia (ITP). The aim of this study was to evaluate the significance of BAFF expression in pediatric ITP patients. Eighty pediatric patients with ITP are subdivided in three groups. Group I included (32 patients) diagnosed with acute ITP less than 3 months, group II (48 patients) diagnosed with persistent ITP (from 3 to 12 months) and chronic ITP (more than 12 months) and group III 20 healthy controls. Complete blood picture, autoimmune profile, antiplatelet antibodies, coagulation profile, bone marrow examination, and RT-PCR were performed to detect the expression for BAF for all participants in this study. BAFF expression levels significantly increased in cases rather than in controls. BAFF Expression Value significantly increased in groups I & II (3.10 ± 1.99&3.29 ± 2.58) compared to controls (0.83 ± 0.45) as p < .001 for both. On the other hand, groups I & II were comparable in BAFF Expression Value (p = .470). BAFF expression increased in ITP patients, implying a function in the disease's pathogenesis.

Background; S100B has been shown to be beneficial as a biomarker in the treatment of adults with mild traumatic brain injury (mTBI). The efficacy of S100B as a biomarker in children, on the other hand, has been a subject of debate. Aim and objectives; was to assess the validity of Protein S 100B in Mild Pediatric Head trauma. Subjects and methods; this was a prospective study, included 160 pediatric patients with mild head trauma presented to Emergency Department. Result; A highly significant correlation between Positive S100B protein and traumatic brain injury with S100B protein value 1554.1±84.0 ng/L. A100 had cutoff value for positive CT Brain finding above 987.5 ng/L, The sensitivity was 81.0%, the specificity was 75%%, the NPV was 86%, the PPV was 68%, and overall accuracy 77%. There was none statistical significant difference regarding severity of brain injury and S100 B (P = 0.225), Conclusion; Serum S100B levels cannot be used to substitute clinical examinations or CT scans in identifying pediatric patients with mild head injuries, but they can be used to identify low-risk kids to avoid excessive radiation exposure.

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.

ObjectiveTo study p53 codon 72 polymorphism in relation to cytogenetic response to imatinib treatment in patients with chronic myeloid leukemia (CML).BackgroundP53 polymorphism involves the substitution of an arginine for a proline at codon position 72. Many studies have investigated a genetic link between this variation and response to treatment in cancer.Patients and methodsThis study was conducted on 54 CML patients presented to the Clinical Oncology Department, Menoufia University during the period from June 2013 to April 2015. They were classified according to their cytogenetic response to imatinib therapy into 40 CML patients, cytogenetic responders to imatinib and 14 CML patients who are cytogenetic nonresponders to imatinib. Patients were genotyped for p53 codon 72 polymorphism using PCR. Follow up of the patients should be done after 3, 6, 9, 12, and 18 months after diagnosis, and was done by complete blood count, conventional cytogenetic, and fluorescence in-situ hybridization.ResultsAge, sex, hematologic, and cytogenetic response to imatinib in CML patients did not differ significantly among p53 codon 72 genotypes (arg/arg, arg/pro, and pro/pro) (P = 0.44, P = 0.45, and P = 0.11, respectively). P53 codon 72 polymorphism did not significantly alter the risk to imatinib cytogenetic unresponsiveness (P = 0.9221).ConclusionIt could be concluded that p53 codon 72 polymorphism is not associated with imatinib unresponsiveness in CML.

MicroRNAs are regulatory RNAs that have been classified as crucial players of cancer initiation and progression. Recent studies have illustrated an association between certain hematological malignancies and dysregulated microRNAs expressions; other studies have focused on the utilization of microRNAs as molecular biomarkers of cancer detection, outcome and response to therapy.

Objective The aim of this study was to evaluate reticulated platelets (RP) as a predicting factor of the outcome in acute coronary syndrome (ACS) patients. Background ACS is due to rupture of an arterial plaque. RPs are newly formed platelets with a greater mass, residual amount of RNA, and an increased prothrombotic potential compared with mature platelets. In this study, the RPs were investigated as participating factors in the pathogenesis of ACS. Patients and methods A total of 50 patients with ACS [33 acute myocardial infarction (AMI) and 17 unstable angina] and 15 clinically healthy individuals as controls were investigated for RPs by means of flowcytometry analysis. Results RPs were highly significantly increased in AMI than in unstable angina and controls. Conclusion High levels of RPs are found in ACS patients and it can be used as a predictor for adverse outcome. Moreover, it can participate in the development of AMI due to its high thrombogenic potential.

Aim of the work: Monocytes are divided into three major subsets based on the expression of the cluster of differentiation CD14 and CD16. The aim of this work was to determine which of the CD16(+) monocyte subpopulations is expanded in rheumatoid arthritis (RA) and its association with disease activity and interleukin-17 (IL17) levels.Patients and methods: Fifty-three RA patients and 20 controls were enrolled in this study. Flow cytometry was performed to detect monocyte subsets and IL17 was measured by ELISA. Disease activity score (DAS28) was assessed.Results: CD14(++)CD16(+) monocyte percentage was significantly higher in long standing RA patients compared with early patients and controls (p < 0.01, p < 0.001 respectively). It was significantly higher in patients with RA disease activity and remission compared with the controls (p < 0.001, p < 0.01 respectively). It was not significantly associated with resistance to disease modifying antirheumatic drugs (DMARDs), C-reactive protein, rheumatoid factor and anti-CCP positivity (p > 0.05). It significantly correlated with IL17 (p < 0.002). CD14(+)CD16(+) monocyte percentage was not significantly correlated with any of the above parameters. IL17 level was significantly higher in patients with early and long standing RA compared to controls (p < 0.01, p < 0.001 respectively). IL17 was higher in RA patients with active disease compared to those in remission and controls (p < 0.01, p < 0.001 respectively). It was higher in RA patients resistant to DMARDs than in responding patients (p < 0.017).Conclusion: CD14(++)CD16(+) monocyte subpopulation was expanded in long standing RA and was correlated with IL17 levels indicating its potential pathogenic importance in RA and may represent an attractive target for future therapeutic interventions. (C) 2015 Egyptian Society of Rheumatic Diseases. Publishing services provided by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Background: Several lines of evidence have shown that inactivation of tumor suppressor genes is closely associated with tumorigenesis in a wide variety of hematological malignancies. The role of downexpression of the RIZ1 gene in hematological malignancy has been reported in many studies, but its levels after receiving induction therapy in these patients have not been evaluated, which was the aim of this study. Objective: The current study aimed to find the relationship between expression of RIZ1 in cases of hematological malignancies before the start of induction therapy and at the end of induction therapy and its possible role as a risk factor in disease progression. Patients and methods: The study included two groups of patients: group I included 26 newly diagnosed patients with hematological malignancies before the start of induction therapy (day 0) and group II included 12 patients at the end of induction therapy (day 28). Group III included 10 individuals with another indication for bone marrow (BM) (such as peripheral blood unexplained cytopenia) who were age and sex matched with the patients and were enrolled in the study as a control group. The following investigations were performed for all the participants: complete blood count, BM sample for morphological examination, immunophenotyping, and cytogenetic analysis. The levels of RIZ1 gene expression were detected by real-time PCR (RT-qPCR) at diagnosis (day 0) and after induction therapy (day 28). Results: The mean levels of RIZ1 gene expression in day 0 patients were significantly lower than those of the control group, and at the end of induction therapy (on day 28), the level was significantly increased to reach close to that in the healthy controls. Furthermore, the levels of the RIZ1 gene were negatively correlated with the blast cell counts in peripheral blood and BM. Conclusion: We conclude that the RIZ1 gene is downexpressed in leukemic patients and its levels increased after induction therapy, indicating its possible role in disease pathogenesis. The relationship between its level and risk factors suggests its role in disease progression. A large-scale study is recommended for the use of the RIZ1 gene in the therapy of hematologic malignancies.

SummaryIntroductionThis study aimed to assess the correlation between the percentage of CD14+ HLA‐DRlow/− immunosuppressive monocytes, plasma arginase 1 level, and disease aggressiveness in patients with B‐cell non‐Hodgkin lymphoma.MethodsForty‐two patients with B‐cell non‐Hodgkin lymphoma and 20 healthy volunteers were enrolled in this study. Peripheral blood CD14+ HLA‐DRlow/– monocytes were detected by Flow cytometry, and their correlation with disease relapse and refractoriness was analyzed.ResultsThe percent of CD14+ HLA‐DRlow/− monocytes was significantly higher in the lymphoma patients than in the healthy controls (control, 9.3 ± 4%; lymphoma, 35.8 ± 20.2%; P < 0.0001), higher in stage III& IV than stage II (stage II, 26.48 ± 17%, n = 26; stage III & IV, 50.8 ± 15.4%, n = 16; P < 0.0001), more in diffuse large cell lymphoma than other pathology types and in relapsed/refractory patients than in patients who achieved remission during follow‐up (relapsed/refractory, n = 18, 45.7 ± 16.7%; remission, n = 16, 21.4 ± 16.2%; P < 0.0001). The arginase I level correlated with increased percent of CD14+ HLA‐DRlow/− monocytes (P < 0.0001).ConclusionIncreased CD14+ monocytes with loss of HLA expression were seen in patients with higher stage disease, more aggressive pathology, and in relapse or refractoriness to treatment. Identifying therapeutic strategies to overcome the suppressive properties of these monocytes could be of value.

OBJECTIVE:Assess the value of baseline interferon-γ-inducible protein-10 (IP-10) levels as a noninvasive maker of liver fibrosis and as a predictor of response to interferon therapy in HCV genotype 4 infected patients.METHODS:Eighty-four HCV genotype 4 infected patients were enrolled in this study. Degrees of liver fibrosis were determined and baseline IP-10 was measured in serum samples collected prior to initiation of treatment using the enzyme-linked immunosorbent assay. Patients were followed up for 1.5 year to assess their response to antiviral therapy.RESULTS:The baseline IP-10 levels were significantly correlated with the degree of fibrosis and had the ability to differentiate between patients with mild, moderate and advanced stages of fibrosis (F0-1: 95.24 ± 33.08 pg/ml, n = 25; F2: 158.70 ± 52.74 pg/ml, n = 37; F3-4: 357.45 ± 162.18 pg/ml, n = 22; P <0.001). Baseline IP-10 levels were significantly lower in patients achieved Early virological response (responders 134.80 ± 60.47 pg/ml, n = 60; non-responders 334.54 ± 168.94 pg/ml, n = 24, P <0.001). Also baseline IP-10 levels were significantly lower in patients who became HCV RNA negative at 24 weeks of therapy (179.52 ± 130.03 pg/ml, n = 78) than non-responders (352.33 ± 132.58 pg/ml, n = 6, P = 0.002). SVR was achieved in 58/68 (85.3%) patients while 10 patients were relapsed. Baseline IP-10 levels differs significantly between patients who achieved SVR at week 24 post therapy and relapsed patients (IP10 level: SVR, 173.52 ± 125.20 pg/ml, n = 58; Relapsed, 216.20 ± 67.72 pg/ml, n = 10, P = 0.021).CONCLUSION:Baseline IP-10 level independently predicts EVR, response at week 24 during therapy and SVR. It also differentiates patients with mild fibrosis from those with moderate and advanced fibrosis.

ObjectiveThe aim of this work is to study neurogenesis using mesenchymal stem cells (MSCs) as a model of stem cells and then follow them as they form neurons.BackgroundMSCs are multipotent adult stem cells present in all tissues. They are present in the bone marrow, and can differentiate in vitro into neurons, glial cells, and myofibroblasts. MSCs have been proposed as sources of stem cells for regeneration of the central nervous system. Thus, one of the goals of regenerative medicine is to ameliorate irreversible destruction of brain tissue and spinal cord by harnessing the power of stem cells to initiate neurogenesis in damaged areas of the brain.Materials and methodsMSCs were cultured from bone marrow aspirate and detected morphologically and by flow cytometric analysis of surface markers CD44 and Oct3/4, then differentiated into neural cells using neural induction media, which consisted of a cocktail of retinoic acid dissolved in DEMSO, recombinant human basic fibroblast growth factor, recombinant human epidermal growth factor, and insulin-like growth factor I, and detected by glial fibrillary acidic protein (GFAP).ResultsThe results of this study showed that MSCs could be isolated from the bone marrow and assumed the typical fibroblastoid morphology and reached 80-90% confluence at about 9 days. They expressed CD44 with a mean ± SD of 81.54 ± 11.58 and CD Oct3/4 with a mean ± SD of 56.12 ± 17.37. MSCs showed positive expression for double expression of CD44-OCT3/4, with a mean ± SD of 54.03 ± 17.42. A highly significant statistical correlation (P < 0.001) was found between age and double expression of CD44-OCT3/4. No statistically significant correlation (P > 0.05) was found between MNCs and double expression of CD44-OCT3/4. MSCs induced with neural induction media show morphological changes consistent with neurogenesis as compared with the symmetric morphologies of the uninduced cells, as shown by an inverted microscope. Induced cells showed positive staining with GFAP whereas uninduced cells showed negative staining.ConclusionMSCs can be isolated successfully from bone marrow aspirate and can be differentiated into GFAP-positive neural cells.

Non-Hodgkin lymphoma (NHL) is one of the most important malignant diseases worldwide. Cytokines may contribute to the clinical and histopathological alterations of the disease, while CD44, the lymphocyte homing receptor, is a putative determinant of lymphoma dissemination. To assess their value in NHL, the levels of TNF-alpha, IL-2 and sCD44 were measured in patients with different stages of NHL and the relation between these levels and tumor burden, presence of B symptoms and other prognostic criteria of the disease was evaluated. Fifty-two patients with NHL before administration of treatment as well as 20 age- and gender-matched controls were enrolled in this study. Clinical and laboratory assessment was done for the studied patients and the levels of TNF-alpha, IL-2 and sCD44 were estimated by enzyme immunoassay. Laboratory assessment included measurement of Creactive protein (CRP), lactate dehydrogenase (LDH) and albumin. LDH and CRP levels were more significantly higher, while albumin level was significantly lower among patients with stage IV as compared to that of patients with stages I/II or III. The levels of TNF-alpha, IL-2 and sCD44 were significantly higher in NHL patients than in controls. The levels of both TNF-alpha and IL-2 were positively correlated with LDH and CRP and negatively correlated with albumin. However, TNF-alpha, but not IL-2, was negatively correlated with hemoglobin (HB). The level of sCD44 was negatively correlated with both albumin and HB and positively correlated with CRP. There were significant positive correlation between the levels of TNF-alpha, IL-2 and sCD44. There was a significant association between the levels of both TNF-alpha and sCD44 and the presence of B symptoms. In conclusion, the occurrence of B symptoms in NHL may be attributed, at least in part, to high level of TNF-alpha. The increased levels of TNF-alpha IL-2 and sCD44 are associated with high tumor burden and poor prognostic criteria and it is suggest that they can be used as prognostic markers in NHL.