Cytological and histopathological correlation of thyroid lesions.

OBJECTIVE: To determine accuracy of cytological diagnosis in comparison with the corresponding histopathological diagnosis of thyroid lesions. METHODS: The retrospective study was conducted at the Shaukat Khanum Memorial Cancer Hospital, Lahore, Pakistan, and comprised data from January to December 2017 of all in-patient cases of thyroid cytology with their histopathological diagnosis. Both Haematoxylin and Eosin stain slides and cytological smears were reviewed. True negative, true positive, false negative and false positive cases were marked using the criteria defined in Table-1. RESULTS: Of the total 36 cases, 5(13.9%) were non-diagnostic or unsatisfactory for cytological assessment. Cytological diagnosis achieved sensitivity of 82.3%, specificity 64.3%, positive predictive value 73.6%, negative predictive value 75%, false positive rate 35.7% and false negative rate 17.6%. The diagnostic accuracy of cytological diagnosis was 63.9%. CONCLUSIONS: There was significant cytological and histopathological concordance of thyroid lesions.

Prognostic Significance of Percentage Necrosis in Clear Cell Renal Cell Carcinoma.

OBJECTIVES: The consensus conference of the International Society of Urological Pathology (ISUP), held in 2012, made recommendations regarding prognostic parameters of renal tumors. There was a strong consensus that tumor morphotype, pathologic tumor stage, and tumor grade are prognostic indicators of poor outcome. It was also agreed upon that prognostic significance of tumor necrosis is in evolution, and both microscopic and macroscopic tumor necrosis should be documented in percentages. The aim of our study was to explore the impact of tumor necrosis on metastasis-free survival in clear cell renal carcinomas (ccRCCs) in Pakistani patients. METHODS: We retrieved 318 consecutive in-house cases of ccRCC resections from 2014 to 2020 through hospital archives. Histologic slide review was done for assessment of tumor necrosis, tumor stage, and World Health Organization/ISUP grade. The follow-up data to assess metastasis-free survival were available in hospital archives. RESULTS: In multivariable analysis performed by logistic regression model, tumor necrosis was an independent poor prognostic indicator (P = .0001): group 1 (reference group), 0% necrosis; group 2, 1% to 10% necrosis (adjusted odds ratio [AOR], 8.71; 95% confidence interval [CI], 3.62-20.98); and group 3, more than 10% necrosis (AOR, 9.48; 95% CI, 3.99-22.725). CONCLUSIONS: Tumor necrosis is an independent predictor of poor outcome in ccRCCs.

Morphological Spectrum and Survival Analysis of Diffuse Midline Glioma With H3K27M Mutation.

Background Diffuse midline gliomas with the H3K27M mutation are now recognized as separate entities due to their unique molecular signature, clinical features, and adverse outcome. Objective To determine the morphological spectrum and survival rate of diffuse midline gliomas with H3K27M mutation. Material and methods This retrospective study was conducted between January 2015 and January 2021 at Shaukat Khanum Memorial Cancer Hospital and Research Centre. Medical records of 28 cases of H3K27M-mutated midline gliomas were retrieved. Case slides were reviewed and the pertinent histological spectrum was evaluated. Results The mean age of patients was 24.36 ± 14.06 years. There were 21 (75%) males and 7 (25%) females. Biopsy was performed in 22 (78.6%), total resection in 1 (3.6%) while subtotal resection was done in 5 (17.9%) cases. Histologically, a spectrum of morphologies was noted with pilocytic astrocytoma (WHO grade 1) at one end and glioblastoma (WHO grade IV) at the other end. Immunohistochemically, all 28 cases were positive for Histone 3 immunohistochemistry. ATRX was performed in 7 (25.0%) cases with loss of ATRX expression in 3 (10.7%) and retained expression in 4 (14.3%) cases. Ki67 was <5% in 6 (21.4%), 5-10% in 1 (3.6%), 11-15% in 1 (3.6%), 16-20% in 3 (10.7%), 21-25% in 4 (14.3%), and 26-30% in 2 (7.1%) cases. The mean survival was 8.00 ± 9.39 months. Out of 28 patients, 15 (62.5%) patients died of disease. Conclusion Diffuse midline gliomas with H3K27M mutation is an aggressive entity with a broad morphological spectrum.

Recurrent fusions in PLAGL1 define a distinct subset of pediatric-type supratentorial neuroepithelial tumors.

Ependymomas encompass a heterogeneous group of central nervous system (CNS) neoplasms that occur along the entire neuroaxis. In recent years, extensive (epi-)genomic profiling efforts have identified several molecular groups of ependymoma that are characterized by distinct molecular alterations and/or patterns. Based on unsupervised visualization of a large cohort of genome-wide DNA methylation data, we identified a highly distinct group of pediatric-type tumors (n = 40) forming a cluster separate from all established CNS tumor types, of which a high proportion were histopathologically diagnosed as ependymoma. RNA sequencing revealed recurrent fusions involving the pleomorphic adenoma gene-like 1 (PLAGL1) gene in 19 of 20 of the samples analyzed, with the most common fusion being EWSR1:PLAGL1 (n = 13). Five tumors showed a PLAGL1:FOXO1 fusion and one a PLAGL1:EP300 fusion. High transcript levels of PLAGL1 were noted in these tumors, with concurrent overexpression of the imprinted genes H19 and IGF2, which are regulated by PLAGL1. Histopathological review of cases with sufficient material (n = 16) demonstrated a broad morphological spectrum of tumors with predominant ependymoma-like features. Immunohistochemically, tumors were GFAP positive and OLIG2- and SOX10 negative. In 3/16 of the cases, a dot-like positivity for EMA was detected. All tumors in our series were located in the supratentorial compartment. Median age of the patients at the time of diagnosis was 6.2 years. Median progression-free survival was 35 months (for 11 patients with data available). In summary, our findings suggest the existence of a novel group of supratentorial neuroepithelial tumors that are characterized by recurrent PLAGL1 fusions and enriched for pediatric patients.

Secretory Carcinoma Of Salivary Gland: A Clincopathological Analysis.

BACKGROUND: Secretory carcinoma of the salivary gland (SC) is a new entity that harbours a specific ETV6 gene rearrangement. The clinical behaviour of this tumour is not well-known as it is a relatively new entity but it is generally considered as a tumour of low malignant potential. The objective of the study was to find out the frequency of ETV6 translocation in cases diagnosed based on histology and immunohistochemistry, to study morphological features and immunohistochemical findings of our cases and to determine the survival and disease-free status of our patients. METHODS: Twenty-five diagnosed cases of SC were retrieved from the archives of SKMCH and RC. Diagnosis was made primarily based on morphology and immunohistochemistry. Immunohistochemistry includes S100, p63, mammaglobin, DOG 1, GCDFP-15, TTF-1, GATA3, SMA, AMA, and AR. The diagnosis was further confirmed by molecular testing, i.e., Fluorescence in situ hybridization (FISH) studies to observe specific ETV6 gene break. Follow up of the patients was done by developing a questionnaire. Statistical analysis of the data was done using SPSS-23.0. RESULTS: The mean age of diagnosis was 41±17.4 and the male to female ratio was 1.5:15. The mean size of the tumour was 45.48±27.35. The most common site of the tumour was parotid gland (60%). On morphology, SC showed a wide range of morphological patterns, most common being the tubular, microcystic, intraductal, and papillary. Immunohistochemical stains mammaglobin (22/22), GCDFP-15(15/15) and GATA3 (10/10) showed 100% positive result. However, all cases were negative for p63 (0/18) and DOG 1(0/11). ETV6 break was seen in 17/17 cases (100%). The mean disease-free survival was 75 months and the overall survival was 51.90±2.80 months. CONCLUSIONS: This study highlights the presence of specific molecular alteration in all cases, which were diagnosed based on morphology and immunohistochemistry.

Androgen Receptor Expression In Endometrial Carcinoma And Its Correlation With Estrogen Receptor And Progesterone Receptor And Clinicopathological Findings.

BACKGROUND: The objective of the study is to analyze the expression of androgen, estrogen and progesterone receptor in different types of endometrial carcinomas and to correlate the androgen receptor expression with estrogen and progesterone receptor and the clinicopathological parameters like lymphovascular invasion, grade of the tumour, size of tumour and extent of myometrial invasion.. METHODS: It is a cross-sectional analytical study design with a simple random sample of a total of 54 cases of different types of endometrial carcinomas from the year 2017. Immunohistochemical stains androgen receptor, estrogen receptor, and Progesterone receptor were applied in all the cases. The Pearson Chi-square test of independence was applied to measure association and P-value is calculated to check the significance of the results. RESULTS: Androgen receptor expression was observed in 73% of low-grade endometrioid carcinomas, 62.5% of high-grade endometrioid carcinomas, 62% of serous, 20% of clear cell and 18% of carcinosarcomas, respectively. Androgen positive tumours were also positive for estrogen and progesterone in most of the cases, except 3 serous carcinomas and one low-grade endometrioid carcinoma. However, no significant relation was observed between androgen expression and prognostic parameters like the lymphovascular invasion, size of the tumour and myometrial invasion. CONCLUSIONS: Maximum expression of androgen receptor was observed in endometrioid and serous carcinomas, while carcinosarcomas and clear cell carcinomas showed minimum expression with no significant correlation between androgen receptor expression and clinicopathological parameters.

The Hot Pleura: Isolated Pleural Metastases From Renal Cell Carcinoma.

Renal cell carcinoma (RCC) is the most common solid renal malignancy that metastasizes typically to lungs, bones, liver, adrenals, and lymph nodes. Isolated pleural metastases of RCC are extremely rare, with only few cases reported in the literature. We report one such case of a 60-year-old woman, a known case of RCC of the left kidney who underwent nephrectomy, and on follow-up, PET/CT scan revealed diffuse thick nodular hypermetabolic left pleural thickening, which was later biopsied and turned out to be pleural metastases from RCC. No other site of metastases was found.

Lhermitte-duclos disease (dysplastic cerebellar gangliocytoma): A case report.

Lhermitte-Duclos disease (LDD) is a relatively uncommon condition of the cerebellum. It is generally characterised as a hamartomatous lesion of posterior fossa and is common in the third and fourth decades of life. According to the World Health Organisation, it is classified as a grade I tumour with potential for recurrence. Otherwise, this disease is generally associated with good prognosis. Malignant transformation of LDD has not yet been reported. However, genetic counselling of the patient is recommended with active surveillance. Since LDD is believed to be a pathognomonic feature of Cowden syndrome, which is a multi-system autosomal dominant hereditary disorder characterised by multiple hamartomas and an elevated risk of benign and malignant neoplasms, we decided to report this important entity considering its rarity and high clinical significance.

Diagnostic Utility of BCOR Antibody in Clear Cell Sarcomas of Kidney.

Purpose. Clear cell sarcoma of the kidney (CCSK) is an uncommon malignant renal tumor. It is the second most common renal pediatric renal malignancy after Wilms tumor. It exhibits a heterogeneous morphology, with overlapping features with its close differentials, which results in diagnostic challenges. There was no specific immunohistochemical marker in the past, to help in this regard. BCOR antibody has recently been suggested to be helpful in the differential diagnosis. We aim to study the utility of the BCOR antibody in the diagnosis of CCSK. Methods. We selected a total of 27 cases of CCSK (n = 12), Wilms tumor (n = 12), and congenital mesoblastic nephroma (n = 3). All cases were evaluated for the extent and intensity of nuclear labeling for BCOR antibody by immunohistochemistry (IHC). Results. We found that BCOR IHC was positive in 11 out of 12 cases with diffuse and strong staining in 8 of the cases. None of the cases of Wilms tumor and congenital mesoblastic nephroma were positive. Only 2 cases of Wilms tumor showed minimal and weak staining in <5% of cells. Conclusion. Diffuse and strong nuclear staining for the BCOR antibody is highly specific for CCSK among common pediatric renal malignancies. Our study confirms that BCOR IHC is a good IHC marker for the diagnosis of CCSK.

Absence of the TRIP13 c.1060C>T Mutation in Wilms Tumor Patients From Pakistan.

BACKGROUND AND AIM: Wilms tumor (WT) is the most common childhood malignant renal tumor. Germline mutations in several WT predisposition genes have been identified. However, the fundamental cause of most WT patients remains unexplained. Recently, a founder mutation, c.1060C>T (p. Arg254X) in a mitotic spindle checkpoint gene, TRIP13, was reported in 5 unrelated children with WT from the United Kingdom, of Pakistani descent from Azad Kashmir region. This observation suggests other children with WT in Pakistan may also harbor this mutation. We conducted the first study to assess the contribution of TRIP13 c.1060C>T mutation to WT in Pakistan. MATERIALS AND METHODS: Constitutional genomic DNA from 68 Pakistani individuals including unrelated WT cases (n=26) and one (n=10) or both (n=32) of their parent(s) were screened for the TRIP13 c.1060C>T mutation using DNA sequence analysis. We also included positive controls in the analyses. RESULTS: The median age of WT diagnosis was 3.0 years (range, 0.75 to 10). The TRIP13 c.1060C>T mutation was not found in any WT patient (n=26) or their parents (n=42). Twenty-four patients (92.4%) presented with unilateral tumor and 2 patients (7.7%) were diagnosed with synchronous bilateral WT. Thirteen patients (50%) reported parental consanguinity. Thirteen patients (50.0%) belonged to the Punjabi ethnicity and 1 patient (3.8%) had a Kashmiri background. Four patients (16.7%) reported a family history of WT or other malignancies. The predominant histologic subtype was stromal (46.2%). The majority of patients presented with >5 cm of tumor size (81%). None of the patients had a personal or family history of congenital anomalies, or associated genetic syndromes. CONCLUSIONS: Our findings suggest that TRIP13 c.1060C>T mutation may be infrequent in Pakistani WT cases. Further evaluation of this mutation in a large number of WT patients of Kashmiri heritage and various ethnic backgrounds from Pakistan is warranted.

Structural weakening of the colonic mucus barrier is an early event in ulcerative colitis pathogenesis.

OBJECTIVE: The colonic inner mucus layer protects us from pathogens and commensal-induced inflammation, and has been shown to be defective in active UC. The aim of this study was to determine the underlying compositional alterations, their molecular background and potential contribution to UC pathogenesis. DESIGN: In this single-centre case-control study, sigmoid colon biopsies were obtained from patients with UC with ongoing inflammation (n=36) or in remission (n=28), and from 47 patients without colonic disease. Mucus samples were collected from biopsies ex vivo, and their protein composition analysed by nanoliquid chromatography-tandem mass spectrometry. Mucus penetrability and goblet cell responses to microbial stimulus were assessed in a subset of patients. RESULTS: The core mucus proteome was found to consist of a small set of 29 secreted/transmembrane proteins. In active UC, major structural mucus components including the mucin MUC2 (p<0.0001) were reduced, also in non-inflamed segments. Active UC was associated with decreased numbers of sentinel goblet cells and attenuation of the goblet cell secretory response to microbial challenge. Abnormal penetrability of the inner mucus layer was observed in a subset of patients with UC (12/40; 30%). Proteomic alterations in penetrable mucus samples included a reduction of the SLC26A3 apical membrane anion exchanger, which supplies bicarbonate required for colonic mucin barrier formation. CONCLUSION: Core mucus structural components were reduced in active UC. These alterations were associated with attenuation of the goblet cell secretory response to microbial challenge, but occurred independent of local inflammation. Thus, mucus abnormalities are likely to contribute to UC pathogenesis.

Structure-based pharmacophore models to probe anticancer activity of inhibitors of protein kinase B-beta (PKB ß).

Protein kinase B-beta (PKBß/Akt2) is a non-receptor kinase that has attracted a great deal of attention as a promising cancer therapy drug target. In mammalian cells, hyperactivation of Akt2 exclusively facilitates the survival of solid tumors by interfering with cell cycle progression. This definite function of Akt2 in tumor survival/maintenance provides the basis for the development of its antagonists with the aim of desensitizing cell proliferation. In order to find novel and potent Akt2 inhibitors, structure-based pharmacophore models have been developed and validated by the test set prediction. The final pharmacophore model was used for hits identification using public chemical databases. The hits were further prioritized using drug-like filters which revealed 14 potential hit compounds having novel chemical scaffolds. Our results elucidate the importance of three hydrogen bond acceptors (A), one hydrogen bond donor (D), one hydrophobic group (H), and one positive ionic charge (P) toward inhibition of the Ak2. One of our selected hits showed 68% cell apoptosis at 8 µg/ml concentration. We proposed various chemical scaffolds including benzamide, carboxamide, and methyl benzimidazole targeting Akt2 and thus may act as potential leads for the further development of new anticancer agents.

Diagnostic accuracy of glypican-3 in differentiating hepatocellular carcinoma from metastatic liver tumours.

OBJECTIVE: To determine the diagnostic accuracy of Glypican-3 in differentiating hepatocellular carcinoma from metastatic liver tumours while taking histopathology as the gold standard.. METHODS: The cross-sectional study was conducted at Shaukat Khanum Memorial Cancer Hospital, Lahore, Pakistan, from January 1 to June 30, 2016, and comprised cases of malignant liver tumours. Samples were collected from the pathology department. Glypican-3was applied on them. Tumours were classified as positive when they showed >5% positivity and negative when showing <5% positivity. Frequencies and percentages of cases showing GPC3 positivity and negativity along with frequency and percentages of hepatocellular carcinoma and metastatic tumours were calculated. RESULTS: Of the 240 patients, 143(59.58%) were males and 97 (40.42%) were females. Overall mean age was 54.65 ± 13.46 years. On histopathology, 134 cases were hepatocellular carcinoma (55.83%) and 106 (44. 17%) cases turned out to be metastatic carcinoma. Glypican-3staining was positive in 116 (48.33%) cases and negative in 124(51.67%). Sensitivity was 82%, Specificity 94.33%, positive predictive value 94.82% and negative predictive value 80.64%. Diagnostic accuracy was 87.5%. CONCLUSIONS: Glypican-3 was found to be a highly sensitive and specific Immunohistochemistry stain distinguishing hepatocellular carcinoma from the clear majority of metastatic carcinomas of the liver.

Granulomatous Interstitial Nephritis: A Rare Cause of Acute Kidney Injury.

Acute granulomatous interstitial nephritis (GIN) is a rare cause of acute kidney injury (AKI) but treatable. It is present in 0.5 to 0.9% of native renal biopsies. Treatment with moderate dosage of steroids is associated with good prognosis. We herein review a case of a 48-year lady having AKI following drug-induced damage [non-steroidal anti-inflammatory drugs (NSAIDs) and antibiotics]. Her renal functions did not improve after one month of drug withdrawal, so renal biopsy was done that showed acute GIN. She was treated with intravenous then oral steroids that completely resolved AKI.

Fluorescence in Situ Hybridization (FISH) for Differential Diagnosis of Soft Tissue Sarcomas

Introduction:Soft tissue sarcomas are rare tumors comprising 1 percent of solid malignancies. The latest edition of WHO soft tissue pathology lists 94 benign and malignant soft tissue tumors. Many of these show a large degree of morphological overlap. Immunohistochemistry has been shown to be reliable in many cases for differential diagnosis of lesions, although cytogenetic tests are considered the gold standard for many entities.Fluorescence in-situ hybridization (FISH) is a cytogenetic technique that uses fluorescent probes that bind to only those parts of the chromosome which have a high degree of sequence complementarity. Many soft tissue tumors show recurrent genetic mutations that are now being used as diagnostic markers. Knowledge of the molecular identity allows prediction of behavior, prognosis and treatment response. Objective:The aim of this study was to identify genetic mutations in soft tissue sarcomas using FISH testing and to assess correlations with histological diagnosis. Material and methods:A total of 25 cases of different soft tissue sarcomas diagnosed on histology with the help of immunohistochemical staining and for which FISH studies were requested were included in this study. Three pathologists with a special interest in soft tissue sarcomas reviewed the cases. FISH tests for EWS, the X:18 translocation, FOXO1 and MDM2 were respectively applied for 8 cases of Ewing sarcoma, 8 cases of synovial sarcoma, 2 cases of rhabdomyosarcoma and 7 cases of dedifferentiated liposarcoma and atypical lipomatous tumors/well differentiated liposarcomas. Results:EWS gene fusion was detected in 7 out of 8 cases of Ewing sarcoma and the X:18 translocation was positive in 3 of the 8 cases of synovial sarcoma. FOXO1 was not detected in either of the two rhabdomyosarcomas. MDM2 by FISH was detected in only one out of 5 cases of atypical lipomatous tumors and 1 out of 2 dedifferentiated liposarcomas. Conclusion: FISH is a useful adjunct in the diagnostic assessment of different types of soft tissue sarcomas. It is easy to set up, is relatively inexpensive and has the ability to diagnose sarcomas with great accuracy, especially in cases which can not be accurately classified even after thorough histological and immunohistochemical evaluation. It may play a very important role in the accurate diagnosis and correct management of patients.

Pharmacoinformatic Approaches to Design Novel Inhibitors of Protein Kinase B Pathways in Cancer.

BACKGROUND: Protein kinase B (PKB/Akt) belongs to the AGC superfamily of related serine/ threonine kinases with three structurally homologous mammalian isoforms, Akt1 (PKBα), Akt2 (PKBß), and Akt3 (PKBγ). Besides sharing a similar structural topology, the difference in their physiological functions and tissue distribution makes Akt a cardinal node in diverse signaling pathways involving cell growth, survival, and proliferation. Various immunohistochemical studies have reported that the constitutive hyperactivation of Akt signaling is responsible for several types of human cancer, poor prognosis, as well as chemotherapeutic and radiotherapeutic resistance. Thus, inhibition of Akt activation represents a promising concept to induce cell apoptosis in various cancers and evade chemotherapeutic resistance. However, development of potent and selective inhibitors of Akt kinases as suitable antagonists remained gloomy and thus, only handful of compounds were selected for the clinical investigation but none of them could reach the market for routine clinical usage to circumvent cell proliferation and resistance to chemotherapeutic agents in cancer. Recent reports on achieving isoform selectivity by designing inhibitors against PH domain of Akt, together with availability of crystal structures of the PH domain of Akt1, open the possibility of structurebased design. METHODS: In this article, various biological regulatory networks by which Akt and its substrates regulate cell growth and survival and several SAR and QSAR strategies in combination with molecular docking studies on selective inhibitors of Akt subtypes have been highlighted to further probe the selectivity of ligand-Akt subtypes interactions. RESULTS: Structure-based drug design studies revealed that the interactions of structurally diverse compounds with Glu121, Ala123, Asn171, Asp184, Glu228 and Ala230 amino acid residues in CAT domain and Arg23, Arg25, Lys30, Asn54 and Arg86 amino acid residues within PH domain play an important role in attaining significant inhibitory potency. CONCLUSION: Isoform selective inhibition of Akt might have clinical significance and thus, should be taken into account in future investigations. Moreover, an up to date isoform selective chemical data is required to further validate already reported isoform selective binding hypothesis.

A 2D-QSAR and Grid-Independent Molecular Descriptor (GRIND) Analysis of Quinoline-Type Inhibitors of Akt2: Exploration of the Binding Mode in the Pleckstrin Homology (PH) Domain.

Protein kinase B-ß (PKBß/Akt2) is a serine/threonine-specific protein kinase that has emerged as one of the most important regulators of cell growth, differentiation, and division. Upregulation of Akt2 in various human carcinomas, including ovarian, breast, and pancreatic, is a well-known tumorigenesis phenomenon. Early on, the concept of the simultaneous administration of anticancer drugs with inhibitors of Akt2 was advocated to overcome cell proliferation in the chemotherapeutic treatment of cancer. However, clinical studies have not lived up to the high expectations, and several phase II and phase III clinical studies have been terminated prematurely because of severe side effects related to the non-selective isomeric inhibition of Akt2. The notion that the sequence identity of pleckstrin homology (PH) domains within Akt-isoforms is less than 30% might indicate the possibility of the development of selective antagonists against the Akt2 PH domain. Therefore, in this study, various in silico tools were utilized to explore the hypothesis that quinoline-type inhibitors bind in the Akt2 PH domain. A Grid-Independent Molecular Descriptor (GRIND) analysis indicated that two hydrogen bond acceptors, two hydrogen bond donors and one hydrophobic feature at a certain distance from each other were important for the selective inhibition of Akt2. Our docking results delineated the importance of Lys30 as an anchor point for mapping the distances of important amino acid residues in the binding pocket, including Lys14, Glu17, Arg25, Asn53, Asn54 and Arg86. The binding regions identified complement the GRIND-based pharmacophoric features.

Right-Sided and Left-Sided Colon Cancers are Two Distinct Disease Entities: an Analysis of 200 Cases in Pakistan.

BACKGROUND: There is growing evidence that there are differences in histological and genetic characteristics along with clinical behavior between right- and left-sided colon carcinomas. We have compared various parameters of the two types and assessed associations of the results with prognosis in patients in Pakistan. MATERIALS AND METHODS: We reviewed 200 cases from our institutional database; 100 cases of right-sided and 100 cases of left-sided colon cancer. Parameters including age, gender, TNM stage, histological features and clinical outcome were analyzed. RESULTS: The patients with right-sided colon cancer were significantly older as compared to their counterparts with left-sided cancer. They presented with a lumbar mass rather than symptoms of obstruction and perforation as seen in left-sided colon cancers, and the histology showed higher percentage of poorly differentiated tumors with advanced pT stage. Moreover, Crohn's-like reactions, intra tumoral lymphocyte responses and other poor prognostic factors like lymph vascular invasion and perineural invasion were more common in right-sided cancers. CONCLUSIONS: We found that right- and left-sided colon cancers are different from each other in terms of clinical presentation, histology and clinical behavior. Right-sided colon cancers are more aggressive and are associated with poorer clinical outcome as compared to left sided colon cancers in our population.

Genotypes of hepatitis C virus in relapsed and non-respondent patients and their response to anti-viral therapy in district Mardan, Khyber Pakhtunkhawa, Pakistan.

Hepatitis C is a blood-borne infectious disease of liver, caused by a small enveloped, positive-single stranded RNA virus, called the hepatitis C virus (HCV). HCV belongs to the Flaviviridae family and has 6 genotypes and more than 100 subtypes. It is estimated that 185 million people are infected with HCV worldwide and 5% of these are in Pakistan. The study was designed to evaluate different genotypes of HCV circulating in District Mardan and to know about the behavior of these genotypes to different anti-viral regimes. In this study 3,800 patients were exposed to interferon alfa-2a plus Ribavirin treatment for 6-months and subjected to real-time PCR to check the viral response. Among these 3,677 (97%) patients showed no detectable HCV RNA while 123 (3%) patients (non-responders) remained positive for HCV RNA. Genotypes of their analyzed showed that most of them belonged to the 3a genotype. Non-responders (123) and relapsed (5) patients were subjected to PEG-interferon and Ribavirin therapy for next 6 months, which resulted into elimination of HCV RNA from 110 patients. The genotypes of the persisting resistant samples to anti-viral treatment were 3b, 2a, 1a and 1b. Furthermore, viral RNA from 6 patients remained un-typed while 4 patients showed mixed infections. HCV was found more resistant to antiviral therapy in females as compared to mals. The age group 36-45 in both females and males was found most affected by infection. In general 3a is the most prevalent genotype circulating in district Mardan and the best anti-viral therapy is PEG-interferon plus Ribavirin but it is common practice that due to the high cost patients receive interferon alfa-2a plus Ribavirin with consequent resistance in 3% patients given this treatment regime.

Loss of E-cadherin expression is not a prerequisite for c-erbB2-induced epithelial-mesenchymal transition.

Recent research into the mechanisms of tumour cell invasiveness has highlighted the parallels between carcinogenesis and epithelial-mesenchymal transition (EMT), originally described as a developmental transdifferentiation program but also implicated in fibrosis and cancer. In a model system for mammary carcinogenesis, we previously observed that induced signalling from a homodimer of the c-erbB2 (HER2) receptor tyrosine kinase in an initially non-malignant mammary cell line caused EMT where i) cell scattering occurred before downregulation of the cell-cell adhesion molecule E-cadherin and ii) the progress of EMT was dramatically delayed when cells were grown at high density. Here, we have further analysed these phenomena. Ectopic expression of E-cadherin concomitant with c-erbB2 signalling was unable to impede the progression of EMT, suggesting that E-cadherin downregulation is not required for EMT. Furthermore, fibroblast-like cells isolated after EMT induced in the presence or absence of ectopic E-cadherin expression showed highly similar morphology and vimentin expression. E-cadherin expressed in these fibroblastic cells had a subcellular localisation similar to that found in epithelial cells, but it exhibited a much weaker attachment to the cytoskeleton, suggesting cytoskeletal rearrangements as an important mechanism in EMT-associated cell scattering. We also investigated whether density-dependent inhibition of EMT is mediated by E-cadherin as a sensor for cell-cell contact, by expressing dominant-negative E-cadherin. While expression of this mutant weakened cell-cell adhesion, it failed to facilitate EMT at high cell densities. These results indicate that loss of E-cadherin expression is a consequence rather than a cause of c-erbB2-induced EMT and that density­dependent inhibition of EMT is not mediated by E-cadherin signalling.