Elsayed Mostafa Ali

Performance of two Real-Time RT-PCR assays for quantitation of hepatitis C virus RNA: evaluation on HCV genotypes 1-4.

Accuracy for monitoring of the concentration of hepatitis C virus (HCV) RNA represents a major challenge throughout the management of patients with chronic hepatitis C. To investigate the genotype‐independent efficiency and the accuracy of two real‐time detection reverse transcription‐polymerase chain reaction (RT‐PCR) assays; the Cobas Ampliprep/Cobas TaqMan (CAP/CTM); and the Abbott RealTime HCV (ART), a total of 184 samples with different HCV subtypes were examined; 1b (n = 58), 2a (n = 39), 2b (n = 26), 3a (n = 20), and 4 (n = 41). A robust linear correlation was observed between the two assays applied to genotypes 1b, 2a, 2b, and 3a [the correlation coefficient (R) ranged from 0.99 to 0.98], but not to genotype 4 specimens (R = 0.78). A significant difference in measurements of HCV RNA using CAP/CTM and ART in serum samples with genotypes 1b and 4 was observed (0.72, −0.53 log IU/ml, P < 0.0001, 0.01, respectively). A robust correlation was observed between the HCV core antigen and HCV RNA values by either of the HCV RNA quantitation assays applied to all genotypes with exception of genotype 4, for which R was higher with ART (R = 0.95) than with CAP/CTM (R = 0.80). The lower limit of detection of CAP/CTM and ART were 41.4 and 28.5 IU/ml using the WHO standards, respectively. In conclusion, two RT‐PCR assays had a high efficiency and accuracy for quantitation of HCV RNA of genotypes 2a, 2b, and 3a, but the mean values of HCV RNA differed for genotype 1b and 4. J. Med. Virol. 82:1878–1888, 2010.

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.

The prognostic value of triple negative in stage II/III breast cancer.

Introduction. Breast cancer is no longer seen as a single disease but rather a multifaceted disease composed of distinct biological subtypes with diverse natural history, clinical, pathological, and molecular features. Recent attention has been directed at the molecular classification of breast cancer. Objective. To evaluate the prognostic value of triple-negative subtype in stage II/III breast cancer and to define the role of clinical stage in prognosis of breast cancer. Methods. We used the immunohistochemical technique to divide 255 cases of breast cancer, stages II and III, into four subtypes according to estrogen receptor/progesterone receptor and Her-2 expression. Results. Triple-negative subtype comprised 76.5% of the cases with 12.3% recurrence rate. Luminal A subtype also carried a poor outcome with 16.7% recurrence rate. Conclusion. Triple-negative subtype has the worst overall and disease-free survival in stage II/III breast cancer. Clinical stage is still an independent prognostic factor in the breast cancers of all types.

Preoperative embolization in surgical management of massive thoracic tumors.

Background: The surgical excision of a highly vascular giant tumor may be challenging. The aim of this study was to describe our experience with preoperative percutaneous embolization of massive vascular chest tumors before surgical excision. Methods: From 2009 to 2011, 8 cases of giant vascular thoracic tumor were treated at Assiut University Hospital, Assiut, Egypt, by preoperative embolization of the feeding arteries followed by successful excision after 48 h. Results: The median age of the 8 patients was 39 years. Embolization of their tumors resulted in a reduction of tumor size, and minimal blood loss was observed perioperatively. Perilesional edema and easy differentiation of ischemic tissue facilitated complete surgical removal of the tumors. Conclusions: Preoperative embolization of giant vascular thoracic tumors is useful to decrease perioperative blood loss and facilitate total excision.

Adult Wilms’ tumor: Review of literature

Background. The most common renal tumors in adults is renal cell carcinoma. Wilms’ tumor in subjects older than 16 years is rare; only 3% of Wilms’ tumors are reported in adults, which explain the difficulties in diagnosis and treatment of this tumor entity in this age group. Methods. Patient with stage IV adult nephroblastoma with favorable histology was described, current treatment modalities were discussed, and the literature was reviewed. Results. Nineteen year old female patient is presented with renal mass, abdominal lymphadenopathy, and bilateral pulmonary deposits. Sonar guided biopsy from the renal mass was taken and pathology revealed nephroblastoma. Right nephrectomy was performed and the pathological examination revealed classic histology of nephroblastoma. The case diagnosed as stage IV adult Wilms’ tumor with favorable histology. According to National Wilms’ Tumor Study Group (NWTS-3), multimodal therapy was initiated immediately after surgery. The patient failed to respond to the first line therapy and died due to disease progression. Conclusion. Adult Wilms’ tumor has no specific guidelines and this may lead to improper or incorrect treatment.

Concurrent radiotherapy and chemotherapy for locally advanced squamous cell carcinoma of the head and neck

BackgroundConcurrent chemoradiation is the standard treatment for patients with advanced head and neck squamous cell carcinoma (HNSCC).The present study was carried out to assess the feasibility and efficacy of low-dose gemcitabine as a radiosensitizer when used during radical therapeutic management of patients with locally advanced HNSCC.Patients and methodsFifty-two patients with locally advanced HNSCC (stage III, 50%; stage IVa, 50%) were enrolled during the period from July 2008 to December 2010. All received a course of radiotherapy (70 Gy over 7 weeks) concurrent with weekly infusions of gemcitabine at 50 mg/m2.ResultsAll patients were available for toxicity and response. Severe mucositis (grade 3-4) was observed in 76% of patients. Severe hematological toxicity was uncommon. Xerostomia was the most common late toxicity in 34 patients (65.4%). The rate of complete and partial response rate was 67.3% and 21.1%, respectively, with an overall response rate of 88.45%. Two years progression-free survival and disease-free survival were 46% and 38.46%, respectively.ConclusionUsing low-dose gemcitabine concurrent with radiotherapy maintains high response rate with low systemic toxicity, in spite of severe mucositis in a high percentage of patients.

Characteristics of escape mutations from occult hepatitis B virus infected patients with hematological malignancies in South Egypt.

AIM To investigate the prevalence and virological characteristics of occult hepatitis B virus (HBV) infections in patients with hematological malignancies in South Egypt. METHODS Serum samples were collected from 165 patients with hematological malignancies to monitor titers of HBV DNA, hepatitis B surface antigen (HBsAg), and antibodies to HBV core (anti-HBc) and surface antigens. Serum samples negative for HBsAg and positive for anti-HBc were subjected to nucleic acid extraction and HBV DNA detection by real-time polymerase chain reaction. DNA sequences spanning the S region were analyzed in cases with occult HBV infection. In vitro comparative study of constructed 1.24-fold wild type and S protein mutant HBV genotype D clones was further performed. RESULTS HBV DNA was detected in 23 (42.6%) of 54 patients with hematological malignancies who were HBsAg negative, but anti-HBc positive, suggesting the presence of occult HBV infection. The complete HBV genome was retrieved from 6 occult HBV patients, and P120T and S143L were detected in 3 and 2 cases, respectively. Site directed mutagenesis was done to produce 1.24-fold genotype D clones with amino acid mutations T120 and L143. The in vitro analyses revealed that a lower level of extracellular HBsAg was detected by chemiluminescence enzyme immunoassay (CLEIA) with the clone containing T120 mutation, compared with the wild type or the clone with S143L mutation despite the similar levels of extracellular and intracellular HBsAg detected by Western blot. Southern blot experiments showed that the levels of intracellular HBV DNA were not different between these clones. CONCLUSION Occult HBV infection is common in patients with hematological malignancies and associated with P120T and S143L mutations. 120T mutation impairs the detection of HBsAg by CLEIA.

Surrogate Role of CD85k on Monocytic Lineage Involved Leukemogenesis Biology and Clinical Aspect

Background: Unique receptor involved in leukemogenesis is CD85k; an immuneglobulin receptor for immune tolerance, CD36 is glycoprotein mediates cellular adhesion and metastatic spread, CD14, CD15 considered common monocytic markers. Aims: to investigate CD85k with monocytic lineage involved leukemia (MLIL) markers in leukemia pathogenesis and clinical presentation. Patients and Methods: 47 patients (32 diagnosed acute myeloid leukemia (AML); 15 non-malignant hematological disease as a control), were included, aged from 2 to 80 years, all subjected to peripheral blood (P.Bl) and bone marrow (B.M) examination, immunophenotyping (IPT) using FASC Canto four color flow cytometer (FCM) Becton Dickenson (BD) USA, for CD13, CD33, MPO, HLA-DR, CD34, CD38, CD117, CD14, CD15 and CD36 the Mo Abs supplied by B.D Bioscience, and anti CD85k Mo Abs by Aveda de Coimbra Flamenco, reference No. 1399990130. Results: Frequency of CD85k is 19/32 (59.37%) of AML; 14/14 (M4/M5) 100% positive CD85k, insignificant correlations of CD85k to sex, lymphadenopathy or organomegaly, platelets count and P.Bl blast (P > 0.05), significant to age 50,000 × 109/l, Hb 0.05). Conclusion: Although CD85k is MLIL associated marker, it is not correlated with other MLIL markers with frequency 100% in MLIL and 59.37% in AML, age predisposition is <35 years with no sex variation, significant correlation to progenitor and myeloid markers, it’s a crucial role in leukemogenesis biology, not in clinical presentations, considered good follow up predictor MLIL marker.

Upper Egypt experience in bladder preservation using concurrent chemoradiotherapy

Objective To share our experience in bladder preservation in Upper Egypt, Assiut and Sohag Universities, using different treatment protocols. In Sohag study patients with operable muscle invasive bladder cancer were included and underwent transurethral resection followed by radiochemotherapy (5- fluorouracil and Cisplatin) for bladder preservation. In Assiut study after maximum safe resection of bladder tumor, patients received combined chemo-radiotherapy, 60 Gy of fractionated radiotherapy over 6 weeks, with Cisplatin and Gemcitabine. Results In Sohag study the age of patients ranged from 35–72ys with Median 56 years, 24 patients were male (80%) and 6 patients were female (20%). In Assiut study the mean of age was 57.30 years, median 58.5 years with peak incidence in 7th decade (9 cases) then in 6th decade 7 cases (23.33%). Performance status was represented as following, 23 patients (76.6%) were scale 1 and seven patients (23.3%) were scale 2. In Assiut study, 90% of patients were disease free at the time of cystoscopic reevaluation. Of concern is that within 18 months of follow up in Assiut study, 7 of 27 (74%) complete responding patients have had local recurrence and 66.7% of all cases. The recurrence free survival in Sohag study at the median follow up (17 months) was 84% and at the end of follow up (30 months) was 70%. The overall survival at the median follow up was 95%, and at the end of follow up was 84%. The disease free survival in Assiut study was 66.7% and the overall Survival in Assiut study was 76.7. Conclusion Three significant prognostic factors were detected for overall survival, performance status, tumor size and residual of tumor and two significant prognostic factors were detected for disease free survival, tumor size and residual of tumor in Assiut study. And it was nearly similar to that reported by Sohag study as they found the completeness of TUR and early stage of the tumor had the strongest impact in response to treatment.