Identification of the possible therapeutic targets in the insulin-like growth factor 1 receptor pathway in a cohort of Egyptian hepatocellular carcinoma complicating chronic hepatitis C type 4.

BACKGROUND: Molecular targeted drugs are the first line of treatment of advanced hepatocellular carcinoma (HCC) due to its chemo- and radioresistant nature. HCC has several well-documented etiologic factors that drive hepatocarcinogenesis through different molecular pathways. Currently, hepatitis C virus (HCV) is a leading cause of HCC. Therefore, we included a unified cohort of HCV genotype 4-related HCCs to study the expression levels of genes involved in the insulin-like growth factor 1 receptor (IGF1R) pathway, which is known to be involved in all aspects of cancer growth and progression. AIM: Determine the gene expression patterns of IGF1R pathway genes in a cohort of Egyptian HCV-related HCCs. Correlate them with different patient/tumor characteristics. Determine the activity status of involved pathways. METHODS: Total ribonucleic acid (RNA) was extracted from 32 formalin-fixed paraffin-embedded tissues of human HCV-related HCCs and 6 healthy liver donors as controls. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) using RT2 Profiler PCR Array for Human Insulin Signaling Pathway was done to determine significantly up- and downregulated genes with identification of most frequently coregulated genes, followed by correlation of gene expression with different patient/tumor characteristics. Finally, canonical pathway analysis was performed using the Ingenuity Pathway Analysis software. RESULTS: Six genes - AEBP1, AKT2, C-FOS, PIK3R1, PRKCI, SHC1 - were significantly overexpressed. Thirteen genes - ADRB3, CEBPA, DUSP14, ERCC1, FRS3, IGF2, INS, IRS1, JUN, MTOR, PIK3R2, PPP1CA, RPS6KA1 - were significantly underexpressed. Several differentially expressed genes were related to different tumor/patient characteristics. Nitric oxide and reactive oxygen species production pathway was significantly activated in the present cohort, while the growth hormone signaling pathway was inactive. CONCLUSIONS: The gene expression patterns identified in this study may serve as possible therapeutic targets in HCV-related HCCs. The most frequently coregulated genes may serve to guide combined molecular targeted therapies. The IGF1R pathway showed evidence of inactivity in the present cohort of HCV-related HCCs, so targeting this pathway in therapy may not be effective.

Gallbladder Papilloma in a Child Unmasking Metachromatic Leukodystrophy: A Case Report With Review of Literature.

Background: Metachromatic leukodystrophy (MLD) is a lipid storage disease characterized the accumulation of sulfatides in different viscera including the gallbladder. Case report: A 2-year-old girl had upper right quadrant lesion that was preoperatively thought to be a biliary cystadenoma. Histologically, the gallbladder lesion was a tubulo-villous papilloma with multiple foci of papillary mucosal hyperplasia. Many storage histiocytes containing metachromatic granules, characteristic of MLD, were present in the tips of the papillae. MLD was later confirmed by enzyme studies. Conclusion: Gallbladder papilloma can be the presenting feature of MLD.

Cervical intraepithelial lesions in females attending Women's Health Clinics in Alexandria, Egypt.

BACKGROUND: Data from Egyptian studies provide widely varying estimates on the prevalence of preinvasive cervical lesions. The aim of this study was to estimate the rate of cervical intraepithelial neoplasia (CIN) in Egyptian women living in Alexandria to clarify the need for implementing a national organized screening program and a vaccination program in our community. MATERIALS AND METHODS: The study was conducted over a 6 years period and covered the different socioeconomic levels to have a representative sample for women living in Alexandria. All women included did not have any cervical disorder related complaints. Conventional Pap smears were obtained and diagnosed using the Bethesda system. Women with abnormal Pap smears were managed according to the 2006 consensus guidelines within the available facilities. Persistent abnormal cytological results were referred for colposcopic biopsy. Histological results were grouped into: Reactive changes, CIN 1, CIN 2/CIN 3 and adenocarcinoma in-situ (AIS). RESULTS: Out of the 6173 smears included in the study 6072 (98.36%) were normal and only 101 (1.63%) were abnormal. After colposcopic biopsies, 0.08% had CIN 1, 0.03% had CIN 2, 3 and 0.01% had AIS. CONCLUSION: We concluded that cervical cancer screening programs, although life-saving for a number of women, are not a sufficiently high priority in our community. Money for national health screening programs should preferably be directed more towards recruiting women for breast cancer screening, since breast cancer accounts for about 33% of all female cancers in Egypt ranking number one, while cervical cancer ranks number 13.

Lymphadenopathy in a Series of Egyptian Pediatric Patients and the Role of Pathology in the Diagnostic Workup.

Assessment of lymphadenopathy in children represents a diagnostic challenge because of the extensive differential diagnoses including reactive and malignant conditions. Knowledge of the etiologic pattern of lymphadenopathy in a given geographical region is essential for making a confident diagnosis or suspecting a disease. Hence, the present study was carried out to identify different etiologies of lymphadenopathy in children in our region, and assess parameters commonly associated with malignancy, with an emphasis on the role of pathology in the diagnostic workup. One hundred and twenty patients aged one month to 18 years were included in the study. They were sorted into neoplastic and non-neoplastic (Infectious and non-infectious). In only 56 patients, biopsy, whether fine needle aspiration cytology (FNAC), core needle or excision biopsy, was essential to reach the final diagnosis. Sensitivity of FNAC in the differentiation between neoplastic and non-neoplastic lymphadenopathy was 92.3%, specificity 90.0%, with a diagnostic accuracy of 91.3%. We concluded that malignancy should be suspected in the following conditions: presence of abdominal or multiple symptoms, symptoms duration of 1-6 months, generalized lymphadenopathy, multiple groups of lymph node (LN) involved, LN size > 2 cm, amalgamated, hard, fixed and non-tender LNs, certain abnormal CBC findings, blast cells in blood film and elevated LDH level. In such cases, LN biopsy is highly recommended. A final diagnosis was achieved after integrating information from history and clinical findings with those of the laboratory, radiological, pathological and microbiological findings. Accordingly, an algorithm for primary diagnostic evaluation of children with lymphadenopathy is suggested.

Prostate-specific membrane antigen (PSMA) immunoexpression in the neovasculature of colorectal carcinoma in Egyptian patients.

In Egypt colorectal carcinoma (CRC) is the most common type of malignancy of the digestive system. Selectively inhibiting neoangiogenesis by targeting tumor-associated blood vessels is an important therapeutic strategy. Prostate specific membrane antigen (PSMA) is expressed in the tumor-associated neovasculature of most solid cancers making it an interesting therapeutic target. We thought to study the expression of PSMA in a series of CRCs in order to test for its possible use as a target for antiangiogenic cancer therapy in Egyptian patients. One hundred CRC cases were retrieved. Representative sections from each tumor were subjected to immunohistochemistry using PSMA antibodies and CD31 antibodies as reference marker. Accordingly vascular endothelial cell immunoreactivity was semiquantitatively scored. PSMA immunostaining was positive in the neovasculature of 75% of tumors. A statistically significant relation was found between PSMA immunostaining and distant metastasis as well as vascular invasion. The present findings strengthen the evidence on the potential usefulness of PSMA as a therapeutic vascular target. This study is the first to demonstrate a positive relation between PSMA expression in CRC and distant metastasis as well vascular invasion, suggesting that PSMA may play a significant role in vascular invasion and subsequent metastasis.

Should fine-needle aspiration cytology be the first choice diagnostic modality for assessment of all nonpalpable breast lesions? The experience of a breast cancer screening center in Alexandria, Egypt.

Fine-needle aspiration cytology (FNAC) of breast masses has been replaced by ultrasound-guided core-needle biopsy (USG-CNB) in many countries. However, in Egypt, breast FNAC continues to play the major role in diagnosing breast masses. In this prospective study, we evaluated the efficacy of USG-FNACs performed at a breast cancer screening center by comparing the FNAC results with the corresponding definitive histological examination outcome. We also investigated the role that CNB can play as a complementary diagnostic tool for FNAC in selected cases. A total of 229 consecutive nonpalpable breast masses were included in this study. Each FNAC was placed into one of four categories: 3.5% nondiagnostic, 13.5% benign, 12.3% atypical/suspicious (indeterminate), and 70.7% malignant. The overall diagnostic accuracy was 98.9%, with a specificity and sensitivity of 99.3 and 96.7%, respectively. The overall positive predictive values and negative predictive values were 99.3 and 96.7%, respectively. Only 37 masses (16%) were converted to CNB, with the indeterminate cytology being the most common cause (54%) for this conversion. Two cases demonstrating the superior benefit of FNAC over CNB are illustrated. Although we started the study by reserving CNB as a first choice to assess microcalcifications without architectural distortion, we ended the study by deciding to perform combined FNAC and CNB for this type of lesions. In conclusion, aiming to maximize the preoperative diagnosis of cancer, it would be cost efficient and time saving to use FNAC as a first-line investigation to benefit from the wealth of cytological information yielded, followed by CNB in selected cases.

Lymphatic vessel density as prognostic factor in breast carcinoma: relation to clinicopathologic parameters.

Angiogenesis and lymphangiogenesis are essential for breast cancer growth and progression. This study aimed at investigating lymphatic microvessel density (LVD) and microvessel density (MVD) as prognostic markers in breast carcinoma. Forty breast carcinomas were immunostained for D2-40, CD31 and VEGF. Median lymphatic and blood microvessel densities, as well as VEGF expression, were related to each other and to clinicopathologic parameters including lymph node (LN) status. The efficacy of haematoxylin and eosin (H&E) in detecting lymphatic vessel invasion (LVI) compared to D2-40 immunostaining was also investigated. D2-40 stained normal lymphatic endothelium and myoepithelial cells, but with different staining patterns. D2-40 LVD related significantly to CD31 counts (r=0.470; p=0.002), and LN metastasis (Mann-Whitney U=101.500; p=0.043); however, it did not relate to age, tumor grade, tumor size or LVI. D2-40 identified LVI in 3 more cases (7.5%) than those detected by H&E. VEGF was expressed in 85%of cases, and was significantly related to CD31 and D2-40 counts (p=0.033 and 0.007, respectively). In conclusion, D2-40 LVD showed a significant association with LN metastasis, and can be considered to segregate patients with positive from those with negative LNs. D2-40 enhances the detection of LVI relative to H&E staining reflecting a potential for lymphatic metastatic spread and possible poor prognosis.

Evaluation of the Novel Monoclonal Antibody Against DOG1 as a Diagnostic Marker for Gastrointestinal Stromal Tumors.

BACKGROUND AND OBJECTIVES: Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract. With the growing effectiveness and availability of first and second-generation tyrosine kinase inhibitor (TKI) drugs, the accurate diagnosis of GIST has become imperative. The problem is that some GISTs with KIT or Alpha-type platelet-derived growth factor receptor (PDGFRA) mutations may have low KIT expression by immunohistochemistry yet will still benefit from TKI drugs. Molecular analysis is a costly and laborious process. Therefore the emergence of a new sensitive immunohistochemical marker for GISTs would be ideal. Recently antibodies against "Discovered on GIST-1" (DOG1) have been generated. The aim of this study was to evaluate the monoclonal DOG1.1 antibody as a diagnostic marker for GISTs and to compare immunohistochemical staining and diagnostic efficacy of DOG1.1 with that of KIT in GISTs. MATERIALS AND METHODS: Forty seven paraffinembedded GISTs were immunostained with both KIT and DOG1.1 antibodies. Immunoreactivity was graded semiquantitatively from 0 to 4. Some other mesenchymal tumors were included in the study and stained for both markers to test for their specificity. RESULTS: Out of the 47 GISTs, 44 were immunoreactive for both KIT and DOG1.1 antibodies (93.62%). Two cases (4.25%) were KIT-positive DOG-negative and the remaining case was DOG-positive KIT-negative (2.13%). A statistically significant concordance was found between KIT and DOG1.1 immunoreactivity (p=0.004), with moderate agreement between immunostaining scores (kappa =0.379). As regards tumor site, a statistically significant association was found between high DOG1.1 scores and gastric GIST (p=0.008). High KIT and DOG1.1 immunostaining scores were significantly associated with highrisk tumors (p=0.002 and p=0.002 respectively). DOG1.1 immunoreactivity was focal in more than half of the cases. The overall diagnostic accuracy of DOG1.1 was 96.5%, with a specificity and sensitivity of 100% and 95.7%, respectively. The overall positive predictive values and negative predictive values were 100% and 84.6%, respectively. As for KIT, The overall diagnostic accuracy was 94.8%, with a specificity and sensitivity of 81.8% and 97.8%, respectively. The overall positive predictive values and negative predictive values were 95.8% and 90%, respectively. CONCLUSIONS: This study has demonstrated that DOG1.1 is a more specific marker for GIST than KIT, yet its sensitivity is a little inferior to that of KIT owing possibly to the focal staining of DOG1 antibody in many cases. In conclusion, in our institution, in which cost effective health care is a priority, we recommend using DOG1 as the first choice antibody for the diagnosis of GIST as it is a more specific marker. If it is negative, then KIT is tried. If this latter is also negative, then several other antibodies should be tested. And owing to the potential therapeutic significance of GIST diagnosis, if still the results of immunohistochemistry are ambiguous, mutational analysis should be considered to confirm the diagnosis. KEY WORDS: GIST - DOG1 - KIT - Immunohistochemistry.

Osteopontin and C-reactive protein in Egyptian patients affected with tuberculous and malignant pleural effusion.

BACKGROUND: The first step in the evaluation of patients with pleural effusion is to determine whether the effusion is a transudate or an exudate. Osteopontin (OPN) is a pleiotropic integrin-binding protein with many functions. We assessed pleural effusion and serum concentrations of OPN and C-reactive protein (CRP) in patients with different types of pleural effusions. METHODS: The current study comprised three groups: 20 patients with transudative effusion, 30 patients with malignant effusion and 30 patients with tuberculous effusion. OPN was analysed using a commercially available enzyme-linked immunosorbent assay kit. RESULTS: OPN effusion values were significantly higher in exudates (both malignant and tuberculous effusion cases) compared with transudative effusion. Also when compared separately, patients with tuberculous effusion and those with malignant effusion had a significantly higher fluid and OPN effusion/serum ratio than those with transudative effusion. Patients with tuberculous effusion had a significantly higher serum CRP effusion and effusion/serum ratio of CRP than those with malignant or transudative effusion. CONCLUSION: OPN is significantly increased in exudative effusion compared with transudative ones. However, serum OPN and effusion/serum OPN ratio were not significantly different in patients with malignant from those with tuberculous effusions. The lack of difference in serum OPN and effusion/serum OPN ratio between patients with malignant and those with tuberculous effusion may be attributed to the heterogeneity of the malignant effusion group. Receiver-operating characteristic (ROC) curve analysis has shown that effusion/serum CRP ratio outperformed effusion/serum OPN ratio as a diagnostic marker for tuberculous pleural effusion.