Comparative study of immunohistochemical analysis in tissues and molecular detection in exfoliated tumour cells in urine of human telomerase reverse transcriptase [hTERT] in urinary bladder cancer in Egyptians

Bladder cancer ranks the third most common malignancy in Egypt following to breast cancer and leukemia. Telomerase plays important roles in cancer development and promotion. Its activity is present in most human malignant tumor cells. Its activity was also detected in voided urine and in bladder washes of patients with bladder cancer, making it a potential marker for non invasive detection of bladder cancer in urine. The present study aims to correlate the expression of the hTERTmRNA in exfoliated tumour cells in urine to the in situ expression of hTERT protein in the corresponding tumour specimens and to evaluate the relationship to hTERT expression and the clinicopathologic tumour characteristics. The study comprised twenty three bladder cancer cases [22 urothelial and one squamous cell carcinomas]. Semiquantitative immunohistochemical detection of hTERT expression was evaluated using combined score evaluated by two examiners which included scoring of intensity and percentage of positivity Semiquantitative expression of hTERTmRNA relative to housekeeping gene GAPDH mRNA was evaluated from RNA extracted from exfoliated cells in urine. Expression of hTERT by immunohistochemistry [IHC] and RT-PCR was detected in 100% of the bladder cancer series. Both methods were significantly correlated [p=0.004]. There was no correlation detected between hTERT expression by both methods and clinicopathologic characteristics of the tumours represented by stage and grade. The high concordance between the semiquantitative expression of hTERT protein by IHC in tumour sections and hTERTmRNA in exfoliated tumour cells validate the potential use of hTERT as a diagnostic non invasive marker for diagnosis of bladder cancer in high risk Egyptian patients and in the follow up following cystoscopic resection of superficial tumours. To the best of our knowledge this is the first study which compares hTERT in situ expression in bladder tumours and hTERT mRNA in exfoliated tumour cells in urine

Immunohistochemical Detection of HCV in nerves and muscles of patients with HCV associated Peripheral Neuropathy and Myositis

Background: Chronic hepatitis C Virus [HCV] infection may be associated with numerous extrahepatic manifestations, such as mixed cryoglobulinaemia, membranoproliferative glomerulonephritis, sicca syndrome. Cryoglobulinaemia [CG] is a condition characterized by the presence of serum proteins that reversibly precipitate in the cold. The objective of the present work was to study the histopathological changes in neuromuscular biopsies in patients with HCV associated peripheral neuropathy, or myopathy; with and without cryoglobulinemia, and to assess the presence of HCV in nerve and muscle tissues of those patients which might clarify some pathogenetic mechanisms for neuropathy, and myopathy associated with HCV

Methods: The study was conducted on 17 cases of HCV infected patients with peripheral neuropathy and myositis. All patients were subjected to thorough laboratory investigations, neurological examination, electrophysiologic studies including nerve conduction, and needle EMG studies

Results: Histopathological examination of nerve biopsies showed features of vascultis in 2/10 cases, interstitial inflammatory infiltrates in 5/10. Muscle biopsies showed intense inflammatory reaction, degenerative changes in the muscles of 3/10 cases diagnosed as myositis. Immunohistochemical results, showed in nerve biopsies, 7/10 cases with positive reaction for HCV with nuclear and perinuclear staining.. Two patients showed positive reaction in the epineural, and endoneural blood vessels and a negative reaction in nerve bundles, while in five patients, reaction was only positive in the nerve bundles. In muscle biopsies, 7/10 cases showed positive reaction for HCV in the nuclei of the muscle fibers, including the cases presented with myositis

Conclusion: The presence of HCV particles in nerve and muscle biopsies of patients with peripheral neuropathy suggests a virus triggered immune mediated mechanism

Immunohistochemical detection of HCV in nerves and muscles of patients with HCV associated peripheral neuropathy and myositis

Chronic hepatitis C Virus [HCV] infection may be associated with numerous extrahepatic manifestations, such as mixed cryoglobulinaemia, membranoproliferative glomerulonephritis, sicca syndrome, Cryoglobulinaemia [CG] is a condition characterized by the presence of serum proteins that reversibiy precipitate in the cold. The objective of the present work was to study the histopathological changes in neuromuscular biopsies in patients with HCV associated peripheral neuropathy, or myopathy; with and without cryoglobulinemia, and to assess the presence of HCV in nerve and muscle tissues of those patients which might clarify some pathogenetic mechanisms for neuropathy, and myopathy associated with HCV. The study was conducted on 17 cases of HCV infected patients with peripheral neuropathy and myositis. All patients were subjected to thorough laboratory investigations, neurological examination, electrophysiologic studies including nerve conduction, and needle EMG studies. Histopathological examination of nerve biopsies showed features of vascultis in 2/10 cases, interstitial inflammatory infiltrates in 5/10. Muscle biopsies showed intense inflammatory reaction, degenerative changes in the muscles of 3/10 cases diagnosed as myositis. Immunohistochemical results, showed in nerve biopsies, 7/10 cases with positive reaction for HCV with nuclear and perinuclear staining. Two patients showed positive reaction in the epineural, and endoneural blood vessels and a negative reaction in nerve bundles, while in five patients, reaction was only positive in the nerve bundles. In muscle biopsies, 7/10 cases showed positive reaction for HCV in the nuclei of the muscle fibers, including the cases presented with myositis. The presence of HCV particles in nerve and muscle biopsies of patients with peripheral neuropathy suggests a virus triggered immune mediated mechanism

Evaluation of E- cadherin and telomerase expression in urothelial carcinoma of the urinary bladder

E-cadherin is a transmembrane glycoprotein involved in intercellular adhesion. A loss or reduction in e-cadherin expression has been linked to the invasive phenotype of a wide variety of human neoplasms. including bladder tumors. Human telomerase reverse transcriptase [hTERT] is a catalytic subunit of telomerase and is a potentially useful diagnostic marker for cancers in many neoplasms, increased telomerase activity is associated with poor clinical outcomes. The objective of the present study was to evaluate the immunohistochemical expression and the potential prognostic value of E-cadherin and telomerase catalytic subunit [hTERT]in urothelial carcinoma of the urinary bladder by comparing them to other prognostic parameters as tumor grade, depth of invasion, and stage. The study comprised 54 patients with bladder tumors who underwent TUR or cystectomy. Formalin fixed paraffin sections from the tumors were processed with a hot, citric acid antigen retrieval method, followed by immunostaining using a monoclonal antibody E-cadherin [NCL-E-cad] Novocastra, mouse monoclonal antibody NCL-L-hTERT [Novovastra]and peroxidase detection system [Novocastra]. Showed loss of normal membrane E-cadherin immunoreactivity in 49 [90.7%] patients. Abnormal expression of E-cadherin was associated with muscle invasive disease [p = 0.0002] and high grade tumors [p = 0.0002].Telomerase [hTERT] was detected in 5 1/54 [94.4%] of urothelial carcinoma, and negative in all normal urothelium adjacent to the neoplasm in cystectomy specimens. No association was found between telomerase and other prognostic factors. In bladder cancer altered E-cadheri n expression is associated with the degree of invasiveness, and tumor grade. However, Telomerase [hTERT] detection by immunostaining is a highly sensitive diagnostic marker for urothelial malignancy but its value as a prognostic marker needs further assessment

Value of DNA image cytometry, and cell proliferation by pCNA index in bone marrow biopsies infiltrated by non-Hodgkin's lymphoma

The assessment of DNA ploidy, and proliferative activity by image analysis [IA], was carried out on 17 bone marrow trephine biopsies [BMTBs], from patients with Non-Hodgkin's Lymphoma [NHL]. Comparative estimation of proliferative activity, was performed using immunohistochemical staining of proliferating cell nuclear antigen [PCNA]. In the twelve bone marrow biopsies, infiltrated by NHLs, aneuploidy was detected in 11[91.6%] cases, S Phase Fraction [SPF], ranged from 15.6-32.5, PCNA labeling index, ranged from 4% to 70% with highest score in intermediate grade NHL mixed small and large cell type. A significant correlation was found between SPF and tumor cell burden [TCB], nuclear surface area and nuclear area variance. However no correlation was found between SPF and PCNA index. For bone marrow biopsies, image analysis has advantages over flowcytometry, including better sampling by histologic correlation, the ability to use limited material, and the supplementary information of prognostic indicators, assessed on comparable paraffin blocks. DNA image analysis and PCNA index by immunostaining are complementary methods in assessment of proliferative activity of NHL. However each technique has its advantages and limitations

Ultrastructure study of experimental cutaneous leishmaniasis before and after dapsone intake

Leishmania strain was isolated from a human case of cutaneous leishmaniasis and inoculated into experimental animals. Group of infected animals were given dapsone at a dose of 25 mg/kg for 3 weeks. Results showed that infected animals suffered from autoamputation of the inoculated foot pad. On the other hand, those receiving dapsone showed complete clinical cure. Transmission electron microscope of both groups revealed the fine structure of Leishmania amastigotes. Those given dapsone showed considerable reversible changes, which did not affect parasitic virulence. The latter has been demonstrated by their ability to infect experimental animals leading to pathological lesion. It should be pointed out that patients treated with this drug and showed marked clinical response must be very carefully examined for fear of residual parasites, which may be the cause of relapse later on

value of immunohistochemical detection of p53 protein in endometrial hyperplasia and carcinoma as a diagnostic and prognostic marker

50 cases of formalin-fixed paraffin embedded tissue sections of endometrial lesions representing 21 endometrial hyperplasia [7 simple, 9 complex and 5 complex atypical] and 20 endometrial carcinomas were studied for the detection of P53 protein expression using the ABC immunohistochemical method. 9 out of the 29 cases [30.1%] of endometrial carcinomas showed positive expression for the P53 protein. On the other h and, P53 protein could not be detected in all the cases of endometrial hyperplasia. No statistically significant relation was found between P53 protein expression and the patient's age or the tumor grade. These findings may indicate that P53 mutation is late event in endometrial tumorigenesis