Douaa Sayed

Long-term antalgic effects of repetitive transcranial magnetic stimulation of motor cortex and serum beta-endorphin in patients with phantom pain

Abstract Objectives: To assess the long-term analgesic effect of repetitive transcranial stimulation (rTMS) on chronic phantom pain using high frequency stimulation and to measure the serum beta-endorphin level pre- and post-rTMS. Material and methods: The study included 27 patients with unilateral amputation; all patients had chronic phantom pain. The patients were classified into two groups. Seventeen patients received 10 minutes real rTMS over the hand area of motor cortex (20 Hz, 10 second trains, intensity 80% of motor threshold) every day for five consecutive days and 10 patients received sham stimulation. Pain was assessed using a visual analogue scale (VAS) and the Leeds assessment of neuropathic symptoms and signs (LANSS) scale, before and after the first, fifth sessions, one and two months after the last session. Quantitative determination of serum beta-endorphin before and after five sessions was measured. Results: There was no significant difference between true and sham groups in the duration of illness, VAS, LANSS scores and resting motor threshold in upper and lower limb amputation at the base line. VAS and LANS scores of the patients who received real rTMS decreased more over the course of the treatment through the different points of follow-up (after five sessions, one and two months) than those who received sham stimulation. Serum beta-endorphin was increased significantly after real stimulation with no changes in patients received shame. Serum beta-endorphin showed no significant correlation to Hamilton depression, anxiety, VAS and LANS scores in true or sham groups before or after five sessions for rTMS. Conclusion: These results confirm that five daily sessions of rTMS over motor cortex can produce long lasting pain relief in patients with phantom pain and it might be related to an elevation of serum beta-endorphin concentration.

Genetic variability of hepatitis C virus in South Egypt and its possible clinical implication.

Egypt is one of the countries with very high rates of hepatitis C virus (HCV) related morbidity and mortality. However, little is known about geographical and clinical differences in genetic variability of HCV in Egypt. Using direct sequencing and phylogenetic analysis of partial core/E1 and NS5B regions of the HCV genome, HCV genotype/subtype was determined in 129 HCV‐infected patients residing in three governates in south Egypt: Assuit, Sohag, and Qena. According to clinical stage of infection, patients were categorized into four groups: asymptomatic carriers, n = 16; chronic hepatitis C patients, n = 36; liver cirrhosis, n = 54; and hepatocellular carcinoma (HCC), n = 23. Genotype 4a was detected in 80.6%, whereas 1g, 4l, 4n, 4o, 4f, and 4m were identified in 7.7%, 4.7%, 3.9%, 1.6%, 0.8%, and 0.8% of cases, respectively. The prevalence of 4a differed regionally; from 88.5% (in Sohag) to 64% (in Assuit, P = 0.002). Genotypes 4l and 4n had a higher prevalence in Assuit (12.8%, 10.3%) than Sohag (0%, 0%; P ≤ 0.011). Difference in clinical features of determined genotypes/subtypes was observed; more carriers of non‐4a variants (4l and 4n, 4f, or 4m) had chronic hepatitis compared to carriers of 4a (53.3% vs. 23.1%, P = 0.025), while more patients with 4a had liver cirrhosis (45.2% vs. 13.3%, P = 0.023). Two HCV‐4o strains were isolated in this study, both from patients with HCC. In conclusion, geographical diversity of HCV was revealed in this study in southern Egypt. A further case–control study is required to confirm the trends of differential pathogenicity of HCV subtypes, indicated by this study. J. Med. Virol. 81:1015–1023, 2009.

PREVALENCE OF OCCULT HEPATITIS B VIRUS INFECTION IN HEMODIALYSIS PATIENTS FROM EGYPT WITH OR WITHOUT HEPATITIS C VIRUS INFECTION

Background While prevalence of Hepatitis B virus (HBV) in patients with end-stage renal failure (ESRF) who are undergoing dialysis has decreased significantly during the past few decades, it still remains a distinct clinical problem. The immunosuppressive nature of renal disease often leads to chronicity of the HBV infection and an opportunity for nosocomial spread of the infection among dialysis patients. Egypt is among the countries with intermediate endemicity of HBsAg (range, 2%–7%). Large-scale geographic heterogeneity in HBV prevalence has been reported worldwide and HBV prevalence is especially heterogeneous in Egypt. Objectives To assess the prevalence of occult HBV infection (OBI) in hemodialysis patients with or without chronic hepatitis C (HCV) from Minia and Assuit, Upper Egypt, using HBV DNA assays. Patient and Methods Sera from 145 hemodialysis patients with negative HbsAg were investigated for HBV DNA using real-time polymerase chain reaction (RT-PCR). Only serum samples with repeatedly detectable HBV DNA were considered positive. Patients were divided into 2 groups: HCV RNA positive and HCV RNA negative, based on the results of a third generation enzyme linked immunosorbent assay (ELISA) anti-HCV test and HCV RNA PCR. Results HBV DNA was detected in 6 of the 145 patients (4.1%) and HBcAb was detected in 29/145 patients (20%). There were no statistically significant differences in the age, duration of hemodialysis, biochemical parameters, serological markers of HBV, or HBV DNA between patients with and without HCV infection. Conclusions Four percent of the hemodialysis patients had OBI. There was no significant difference in the prevalence of OBI between hemodialysis patients with or without HCV co-infection.

Parathormone – 25(OH)-vitamin D axis and bone status in children and adolescents with type 1 diabetes mellitus

Hamed EA, Abu Faddan NH, Adb Elhafeez HA, Sayed D. Parathormone – 25(OH)‐vitamin D axis and bone status in children and adolescents with type 1 diabetes mellitus.

Monocyte-platelet aggregates and platelet micro-particles in patients with post-hepatitic liver cirrhosis

INTRODUCTION Monocytes are the cells that play a crucial role in the pathogenesis of liver damage and liver cirrhosis (LC), and as platelets, by connecting hemostasis and inflammatory processes, participate in pathogenesis of chronic liver diseases, we aimed to investigate the presence of monocyte-platelet aggregates and platelet micro-particles (PMPs) and their role in LC. PATIENTS AND METHODS The study included 60 patients with post-hepatitic LC and 20 healthy controls. Activated monocytes (CD11b, HLA-DR, CD14, CD16), monocyte-platelet aggregates (CD41/CD14), activated platelets (CD41/CD62) and PMPs were analyzed by flow cytometry. Their relations to the clinical and laboratory data were assessed in the studied group. RESULTS Patients with LC had higher levels of activated platelets, activated monocytes and monocyte-platelet aggregations as compared to healthy controls. PMPs percentage showed no significant differences between patients and controls but significantly increased in both patients with no bleeding and patients with splenomegaly compared to patients without. All studied markers showed no significant differences between patients with thrombocytopenia and those with normal platelet counts and also between patients with different disease stages. Positive correlations between monocyte-platelet aggregates and both activated platelets and monocytes were demonstrated. There were significant negative correlations between PMPs and both age and prothrombin time among patients. CONCLUSIONS The stage of post-hepatitic LC is not the only factor that affects the level of activated platelets, activated monocytes and monocyte-platelet aggregates. PMPs have no influence on thrombocytopenia but may have the potential to influence the progression of clotting activity in LC.

Increased circulating red cell microparticles (RMP) and platelet microparticles (PMP) in immune thrombocytopenic purpura

UNLABELLED It has been suggested that patients with ITP have an increased thrombotic risk compared to the general population and compared to those with other causes of acquired thrombocytopenia. The pro-coagulant role of microparticles in some clinical situations has been reported, yet, very little data is available about microparticles in ITP and their effect. AIM OF THE WORK To assess the levels of red cell microparticles (RMP), platelet microparticles (PMP) and their possible relation to some haemostatic parameters in ITP patients PATIENTS AND METHODS The levels of RMP and PMP in addition to FVIII, FIX, FXI, PC and aPTT were assessed in 29 patients with chronic ITP (8 of them had splenectomy). Ten apparently healthy volunteers served as controls. We compared the levels of the studied parameters in ITP patients with that in controls. Correlations of these parameters with each other and with the platelet count were studied. RESULTS RMP (p=0.0001), PMP (p=0.0001), D- dimer (p=0.048), FVIII (p=0.049), FIX (p=0.0001) and FXI (p=0.0001) were significantly higher in ITP patients compared to controls. aPTT was significantly longer in ITP patients (p=0.0001) but PC showed no significant difference. However, RMP was associated with shorter aPTT. Generally, the coagulation factors were negatively correlated with platelet count in ITP patients. Compared to controls, ITP patients preserved higher levels of RMP and PMP even in those with near-normal platelet count. Splenectomy was associated with lower FIX (p=0.0001) and FXI (p=0.028) and higher RMP (p=0.0001). IN CONCLUSION Chronic ITP was associated with increased levels of RMP and PMP. FVIII, FIX and FXI were increased in ITP patients but showed a negative correlation with platelet count. Splenectomy was associated with increased levels of RMP and lower levels of FIX and F XI. The high level of microparticles in ITP might point towards a prothrombotic tendency.

Immunophenotyping and immunoglobulin heavy chain gene rearrangement analysis in cerebrospinal fluid of pediatric patients with acute lymphoblastic leukemia

The study aimed to assess the diagnostic accuracy of Flow cytometry (FCM) immunophenotyping and IgH gene rearrangements (IGHRs) by real-time PCR in comparison with classic cytology for diagnosing CNS infiltration in pediatric ALL. We concluded that the diagnostic value of FCM and IGHR are two to three times more than that of cytology. Therefore, immunophenotyping by FCM is recommended for routine diagnosis of CSF infiltration. Furthermore, IGHR analysis by real-time PCR appears to be a useful addition in evaluation of CNS infiltration.

Impact of CD34 subsets on engraftment kinetics in allogeneic peripheral blood stem cell transplantation

Summary:Our objective was to evaluate, probably for the first time, the impact of CD34 subsets on engraftment kinetics in allogeneic PBSC transplantation (PBSCT). PBSC graft components were analyzed in 62 cases for the absolute count/kg of total CD34+ and the following subsets: DR− and +, CD71+/−, CD38+/−, CD33+/− and CD61+/−. Time to ANC >0.5 and >1 × 109/l and platelets >20 and >50 × 109/l was reported. The median value for each parameter was used to discriminate rapid from slow engraftment. Four parameters showed significant predictive power of early neutrophil engraftment, namely CD34+/DR− (P=0.002), CD34+/38− (P=0.02), CD34+/CD61− (P=0.04) and total CD34+ cell dose (P=0.04). Four parameters showed significant predictive power of early platelet engraftment, namely CD34+/CD61+ (P=0.02), CD34+/CD38− and total CD34+ cell dose (P=0.04) and CD34+/CD71− (P=0.05). Comparing patients who received >to those who received < the threshold dose(s), only CD34+/CD38− lost its significance for neutrophil engraftment; and only CD34+/CD61+ retained its significance for platelet engraftment (P=0.03); furthermore, the former group required significantly fewer platelet transfusions (P=0.018). We concluded that in allogeneic PBSCT, the best predictor of early neutrophil engraftment is the absolute CD34+/DR− and for early platelet engraftment is the absolute CD34+/CD61+ cell dose.

Silica Nanoparticles Sensitize Human Multiple Myeloma Cells to Snake (Walterinnesia aegyptia) Venom-Induced Apoptosis and Growth Arrest

Background. Multiple myeloma (MM), an almost incurable disease, is the second most common blood cancer. Initial chemotherapeutic treatment could be successful; however, resistance development urges the use of higher toxic doses accompanied by hematopoietic stem cell transplantation. The establishment of more effective treatments that can overcome or circumvent chemoresistance has become a priority. We recently demonstrated that venom extracted from Walterinnesia aegyptia (WEV) either alone or in combination with silica nanoparticles (WEV+NPs) mediated the growth arrest and apoptosis of prostate cancer cells. In the present study, we evaluated the impact of WEV alone and WEV+NP on proliferation and apoptosis of MM cells. Methods. The impacts of WEV alone and WEV+NP were monitored in MM cells from 70 diagnosed patients. The influences of WEV and WEV+NP were assessed with flow cytometry analysis. Results. WEV alone and WEV+NP decreased the viability of MM cells. Using a CFSE proliferation assay, we found that WEV+NP strongly inhibited MM cell proliferation. Furthermore, analysis of the cell cycle using the propidium iodide (PI) staining method indicated that WEV+NP strongly altered the cell cycle of MM cells and enhanced the induction of apoptosis. Conclusions. Our data reveal the biological effects of WEV and WEV+NP on MM cells that enable these compounds to function as effective treatments for MM.

Incidence and characteristics of HBV reactivation in hematological malignant patients in south Egypt

AIM To investigate characteristics of hepatitis B virus (HBV) implicated in HBV reactivation in patients with hematological malignancies receiving immunosuppressive therapy. METHODS Serum samples were collected from 53 patients with hematological malignancies negative for hepatitis B surface antigen (HBsAg) before the start of and throughout the chemotherapy course. HBV reactivation was diagnosed when the HBsAg status changed from negative to positive after the initiation of chemotherapy and/or when HBV DNA was detected by real-time detection polymerase chain reaction (RTD-PCR). For detecting the serological markers of HBV infection, HBsAg as well as antibodies to the core antigen (anti-HBc) and to the surface antigen were measured in the sera by CEIA. Nucleic acids were extracted from sera, and HBV DNA sequences spanning the S gene were amplified by RTD-PCR. The extracted DNA was further subjected to PCR to amplify the complete genome as well as the specific genomic sequences bearing the enhancer II/core promoter/pre-core/core regions (nt 1628-2364). Amplicons were sequenced directly. RESULTS Thirty-five (66%) of the 53 HBsAg-negative patients were found to be negative serologically for anti-HBc, and the remaining 18 (34%) patients were positive for anti-HBc. Five of the 53 (9.4%) patients with hematologic malignancies experienced HBV reactivation. Genotype D1 was detected in all five patients. Four types of mutant strains were detected in the S gene product of HBV strains and were isolated from 3 patients with HBV reactivation: T/S120, L143, and I126. HBV DNA was detected in the pretreatment HBsAg-negative samples in one of the five patients with HBV reactivation. In this patient, sequences encompassing the HBV full genome obtained from sera before the start of chemotherapy and at the time of de novo HBV hepatitis were detected and it showed 100% homology. Furthermore, in the phylogenetic tree, the sequences were clustered together, thereby indicating that this patient developed reactivation from an occult HBV infection. CONCLUSION Past infection with HBV is a risk factor for HBV reactivation in Egypt. Mandatory anti-HBc screening prior to chemotherapy in patients with hematological malignancies is recommended.