Promoter Hypermethylation of Tumor suppressor Genes correlates with Tumor grade and Invasiveness in patients with Urothelial Bladder Cancer

Purpose: To investigate the promoter methylation status at selected loci which encode for key proteins involved in apoptosis, DNA repair, cell cycle control and progression in urothelial cell carcinoma of bladder and compare the findings from tissue samples with that of plasma. Methods: Total genomic DNA was isolated from 43 non-muscle invasive (low grade) and 33 muscle invasive (high grade) urothelial bladder cancer samples along with 10 control cases of normal bladder mucosa. Promoter methylation status was investigated for RASSF1A, APC, MGMT, CDKN2A and CDKN2B genes using real-time methylation-specific PCR with SYBR® green. Plasma samples from 16 patients with muscle invasive high grade bladder cancer were also subjected to similar analyses. Results: Promoter hypermethylation was frequently observed in RASSF1A, APC and MGMT gene promoters (p-value \u3c 0.001). The methylation was more prominent in the muscle invasive high grade bladder cancer when compared to non-muscle invasive low grade group (p-value \u3c 0.001) and normal bladder mucosa (p-value \u3c 0.05). The RNA expression of RASSF1A, APC and MGMT was also found to be decreased in the muscle-invasive high grade bladder cancer when compared to the non muscle invasive low grade group (p-value \u3c 0.05). RASSF1A, MGMT and CDKN2A showed comparable results when data from 16 plasma samples was compared to the corresponding tissue samples. Conclusion: Our results suggest that epigenetic silencing of RASSF1A, APC and MGMT genes is strongly associated with invasive high grade urothelial bladder cancer. Thus, status of promoter methylation has the potential to serve as valuable tool for assessing aggressiveness of urothelial cell carcinoma of bladder

Clinical, pathological and molecular factors predicting Axillary Node involvement in primary Breast Cancer in Pakistani women

Background: Axillary lymph node involvement in primary breast cancer is one of its most important prognostic features. Thus any factors that may predict axillary lymph node involvement in this setting could be potentially helpful in treatment planning and other interventions. Objective: The objective of this study was to evaluate clinical, pathological and immuno-histochemical markers in univariate and multivariate analysis, which may be helpful predictors of axillary lymph node involvement in breast cancer. Method: A retrospective analysis of 555 cases. Of these 58% had axillary nodal positivity and 42% were negative. Conclusion: Factors of no significance included patient’s age, height, weight, age of first pregnancy, parity, marital status, menopausal status, family history of breast cancer, side of tumor. In univariate analysis the age of menarche, duration of symptoms, tumor size, site in outer quadrant, S phase and skin and nipple involvement all predicted axillary nodal involvement. The length of breast-feeding, increased intraductal component and increased PCNA were inversely proportional to nodal involvement. In multiple regression analysis however only size of the tumor, involvement of the skin and nipple and disease in the outer quadrant of breast were the factors, which assumed significanc

Combined Toxicity and Bioconcentration of Fluoride and Arsenic in African Catfish Clarias gariepinus (Burchell, 1822)

Laboratory experiments were performed to examine the combined toxic effects of two important aquatic contaminants viz., arsenic and fluoride on African catfish, Clarias gariepinus. Additionally, the bio concentration factors (BCFs) of the two contaminants in tissues and blood of catfish were also determined. The LC50 for sodium fluoride and arsenic trioxide were determined to be 619.3 mg L-1, 30.3 mg L-1, respectively. Erratic swimming movements with hyperactivity, loss of equilibrium, augmented air gulping and decreased food consumption were observed in the experimental groups. In co-exposure groups of arsenic and fluoride, the concentration of fluoride in fish tissues increased with increasing water fluoride concentration in the test aquaria with significant differences (

Chemical characterization of atmospheric particulate matter in Delhi, India, part II: Source apportionment studies using PMF 3.0

World Bank reports Delhi as a second most polluted megacity in the world for particulates pollution. In Delhi, PM10 (d ≤ 10 μm) aerosol samples were monitored throughout 2008 and their characterization for major chemical elements (Na, Mg, Al, Si, P, S, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, As, Br, Sr, Ba, Pb, Cd, Sn and Sb) and ions (Cl-, NO3-, SO42-, Na+, NH4+, K+, Mg2+ and Ca2+) have been documented in an earlier study. To resolve complexity in source apportionment for chemical constituents in PM10, UNMIX 6.0 and Positive Matrix Factorization (PMF 3.0) models are applied. Four factors were derived to explain routine sources of PM10 (crustal origin, road-traffic and secondary aerosols). Factor-1, designated as road-traffic source, has been determined by temporal correlation among Pb, Cu, Zn, Ni and V with strong correlation between Pb and Zn. This source factor-1 has shown more than 60% contribution to receptor site. Factor-2, referred as crustal origin due to strong inter-relationship among Si, Fe, Al, Ca and Mg, has also shown to be significant contribution to similar species in receptor matrix. Factor-3 ( NH4+, NO3-) has been differentiated due to contribution of secondary aerosols in the receptor region. This factor-3 has indicated major fraction of these ionic species for their uniform percentage variability, where mean values have been projected close to 75th percentile. Surprisingly, source factor-4 has explained the specific chloride source in the region with major contribution of 86%. For policymakers, results presented would serve as benchmark of source apportionments in Delhi

Epidermal Growth Factor Receptor (EGFR) as a Prognostic Marker: an Immunohistochemical Study on 315 Consecutive Breast Carcinoma Patients

Objective: To assess the independent and interdependent prognostic value of epidermal growth factor receptor (EGFR) in carcinoma of breast in female population. The Type I family of growth factor receptors includes epidermal growth factor receptor (EGFR also known as EGFRI). Methods: The expression of EGFR protein was analysed immunohistochemically on 315 tumour specimens of infiltrating ductal carcinoma of breast. These patients also had axillary lymph nodes sampling. Results: Overexpression and/or amplification of EGFR was observed in 70 (22.00%) tumours. Eleven (16%) were grade I, 43 (61%) grade II and 16 (23%) grade Ill tumours. Axillary lymph node metastasis had significant correlation with intensified positivity of EGFR (p\u3c 0.05). Significant number of EGFR positive patients developed local recurrence and distant metastases to brain, liver and bone (p\u3c 0.05). EGFR positivity showed significant correlation with the disease free and overall survival (p\u3c 0.05). At a median follow-up of48 (4 years) months in EGFR positive patients, the overall survival was 3.39 years and disease free survival was 2.86 years. EGFR negative tumour patients showed a better survival. In this group the overall survival was 4.62 years and the disease free survival was 4 years. Conclusion: EGFR analysis can be a useful indicator for the selection of patients who are at the high risk, for hormonal therapy decisions and can be useful as a target for new treatment modalities

Flow cytometric analysis of childhood leukemias

Objective: To collect demographic data for childhood (less than 15 years) leukemias in Karachi, describe the accuracy of the cell surface markers routinely used in the flow cytometric analysis of leukemic cells and arrive at an ideal panel of antibodies for analyzing leukemic samiples. Materials and Methods: Data from 62 consecutive cases of childhood leukemias referred to the Department of Pathology, Aga Khan University Hospital. (AKUH) between January 1995 and December 1998 was analyzed using Epi Info Version 6. Flow cytometry on all samples was performed using standard protocols. Results: The mean age of patients was 8.2 years and 49 (79%) were males. Fifty (81%) had acute lymphoblastic leukemias of which 50% were CD1O positive and 24% CD10 negative Pre-B cell leukemias. Among all Pre B cell All 98% were positive for CD19, 96% for CD22, 89% for HLA-DR and 67% for CD10. Of the 10 AML cases, 100% were positive for CD33, 90% for CD13, 80% for CD19 and 70% for HLA-DR. Conclusion: The mean age in this study population was significantly higher and percentage of CD10 positive Pre-B All is lower than that in the West. Both these factors might be responsible for the poorer prognosis of these patients. It is not possible to specify a minimum or maximum panel of antibodies that should be used for phenotyping all cases of childhood leukemias. A certain degree or redundancy is essential in any panel of antibodies used for flow cytometry of leukemias

Histological pattern of central nervous system neoplasms

Objective: To see in a large series of patients the histological pattern of CNS neoplasms and to provide a comprehensive data about the frequency of CNS neoplasms in Pakistani patients and to give a clearer idea about their prevalence. Method: Retrospective study of all consecutive cases of CNS neoplasms diagnosed at The Aga Khan University Hospital (AKUH) between 1st January 1994 and 31st December, 1999. Setting: The section of Histopathology, AKUH, Karachi. Observation: Our series included a large number of cases i.e., 1110 . There are very few local studies on CNS tumours and none so large . Our data was correlated with published Western and Pakistani data . Glial tumours were the commonest primary CNS neoplasms followed by meningiomas . Among glial tumours , astrocytomas comprised the largest group and the majority were high grade (Ill and IV ) astrocytomas. Percentages of anaplastic oligodendrogliomas and ependymomas were also significant. Male to female ratio was 1.6:1. However, meningiomas were more common in females. Almost half of the CNS tumours in patients 15 years or younger were in the posterior cranial fossa . Metastatic tumours were much lower as compared to the West. Conclusion: Except for the high percentages of anaplastic oligodendrogliomas and ependymomas and the low percentage of metastatic tumours, most of our findings roughly correspond with the published data

Immunohistochemical Estrogen receptor determination in human Breast carcinoma: correlation with histologic differentiation and age of the patients

An immunohistochemical assay for the measurement of estrogen receptor (ER) has been evaluated on 290 consecutive human breast biopsy and mastectomy specimens in the year 1992 at The Aga Khan University Hospital laboratories. Immunohistochemical localization of estrogen receptor on frozen/paraffin section was scored in a semi-quantitative fashion incorporating both the intensity and the distribution of specific staining. Histologic grading of the tumour was performed according to Bloom’s method. In this study, 21% of the tumours were estrogen receptor negative, 15% were weak positive, 25% intermediate positive and 39% strong positive. Fifty percent of the well differentiated tumours showed strong ER positivity against 27% of the poorly differentiated tumours. Seventy eight percent of all negative estrogen receptors were in patients younger than 50 years of age (pre-menopausal group), while 52% of strong estrogen receptor positivity was observed in patients older than 50 years (post- menopausal). This study demonstrates the value of immunohistochemical method to determine the ER status in patients with advanced breast cancer

Diagnostic evaluation of fine Needle aspiration cytology in the management of palpable Breast lesions

A total of 113 fine needle aspirates of the breast masses were evaluated in which the subsequent biopsy or mastectomy specimen were also available for histological examination. The age ranged from 16 to 80 years with a mean of 42 years. In benign conditions the mean age was 34.7 years while in malignant cases it was 48 years. The cytological diagnoses were compared with the histological results which revealed that the specificity and sensitivity of fine needle aspiration cytology in the palpable breast lesions was 86.1% and 89.2% respectively with a positive predictive value of 93% and efficiency of 88.2%. Similar statistics from other series in which the cytological results of breast lesions were compared with histological results, revealed almost same results which suggest that fine needle aspiration cytology is an effective and accurate technique for the diagnosis and management of palpable breast lumps

Transitional cell carcinomas of the urinary bladder. A histopathological study

Objective: To determine the histological grading and muscle invasion in Transitional Cell Carcinomas of the Urinary Bladder; and to evaluate whether any correlation exists between tumour grade and muscle invasion. Method: A Retrospective study of all consecutive cases of Transitional Cell Carcinomas of the Urinary Bladder diagnosed at Aga Khan University Hospital between 1st Jan 1997 and 31st Dec 2000. Results: This study included 495 cases of Transitional Cell Carcinoma of the Urinary Bladder. M: F ratio was 4:1. Ages of patients ranged from 30 years to 87 years. Mean age in males was 59.1 years and in females 58.8 years. Forty four percent cases were Grade II, and 29.5% Grade Ill. None of the Grade I lesions were invasive, 10% Grade II tumours, 61 .41% of Grade Ill and all Grade IV tumours were invasive. Conclusion: There is a definite correlation between advancing tumour grade and muscle invasion