Immunohistochemical Expression of P53; Ebv-Lmpi and Cyclin D1 Proteins in Oesophageal Carcinoma: A Pilot Retrospective Study of 26 Cases

Introduction: The development of oesophageal carcinoma has been attributed to various environmental factors and its incidence varies regionally. The development of this disease is known to occur in recognized histological stages from normal through dysplasia to the malignant stage. Like other cancers; the diagnosis of oesophageal cancer in its premalignant stage would improve the survival. The diagnosis of this cancer on cytomorphology alone is usually done in the late stage of the disease. To be able to diagnose this disease in its early stage; specific tumour markers must be found. The objective of this study was to evaluate p53 tumour suppressor gene protein expression; Epstein-Barr virus latent membrane protein expression and cyclin DI cell cyase protein expression in malignant and normal oesophageal tissues to see whether any variation in their expression in these tissues could be of diagnostic or prognostic value. Methods: 26 archival formalin-fixed paraffin wax embedded tissue blocks of oesphageal carcinoma and 6 of normal oesophagus obtained by endoscopy were studied. 5?m thick tissue sections were cut onto poly-L-Lysine coated microscope slides and dried at 600C for 60 minutes. p53 gene protein expression; EBV-LMPI protein expression and cyclin DI expression were studied immunohisto chemically in these tissue sections. Sections were dewaxed and hydrated to Tris-buffered saline; pH 7.6. Appropriately diluted primary antibodies to p53; EBV-LMPI and Cyclin DI were applied to different sections and incubated overnight at 40OC in a humidity chamber. Sequential applications of other reagents in a three-stage peroxidase antiperoxidase method were applied for chromogen immunoreaction for light microscope visualization. The sections from normal oesophageal tissues were processed with carcinomatous tissues.Results: p53 gene protein was overexpressed in 17 of 26 cases of carcinoma; EBV-LMPI was expressed in 12 of 26 cases of carcinoma; cyclin DI protein was expressed in 14 of the 26 cases of carcinoma; 10 cases of p53 expression were also associated with EBV-LMPI protein expression; 7 cases of p53 protein overexpression did not express EBV-LMPI; 2 cases of EBV-LMPI protein expression did not express p53 protein and 7 cases did not express both p53 and EBV-LMPI proteins. Conclusion: Overexpression of p53 tumour suppressor gene protein in tumour cells of 17 of 26 cases of oesophageal carcinoma while no such expression was demonstrated in normal oesophagus; may have diagnostic and prognostic value. EBV-LMPI expression in tumour cells of 10 of 26 cases of oesophageal carcinoma may also be of value in diagnosis and pathogenesis. Cyclin DI was overexpressed in 14 of the 26 cases and may have diagnostic and prognostic value

Diagnostic utility of fine-needle aspiration cytology in the management of liver disease in a district hospital.

BACKGROUND: Fine needle aspiration (FNA) cytology is a cost effective technique of obtaining cellular specimens for diagnoses. It has many advantages over large needle core biopsies that makes it quite suitable for outpatient department and institutions without facilities for histopathology diagnosis. Any site of the body can be sampled with FNA technique. OBJECTIVE: To assess the diagnostic utility of fine needle aspiration cytology technique in management of patients with liver disease in resource limited settings. DESIGN: Prospective cross sectional study. SETTING: Patient evaluation and FNA aspiration were done in Murang'a and Machakos district hospitals and specimen staining and microscopy were done in Centre for Clinical Research, Kenya Medical Research Institute. SUBJECTS: Patients who were suspected to have liver disease were referred to the medical outpatient clinics in Murang'a and Machakos district hospitals for evaluation. In addition those on the medical wards in the two hospitals who were suspected to have liver disease were also evaluated. INTERVENTIONS: The patients with liver disease were identified after physical and abdominal ultrasound examination. Those with hepatomegaly were selected for the study. Liver fine needle aspiration was done using 21 gauge fine needle attached to a 10 ml syringe. Focal lesions were targeted as much as possible. Thin smears of specimens obtained were spread onto microscope slides which were then dropped into 95% ethanol and left for 30 min in order to fix the material. The slides were then removed, dried and transported to KEMRI where they were stained with Papanicolaou stain then evaluated on the light microscope. RESULTS: One hundred and twenty patients with suspected liver disease were evaluated in the two hospitals; 70 in Machakos and 50 in Murang'a. Fifty patients were identified to have liver disease and fine needle aspiration done aseptically. Twenty one (42%) of the 50 patients had malignant cells; 15(30%) of the 50 patients had necrotic material aspirates 2 (4%) shoved fatty changes later showed to be liver cirrhosis; 11 (22%) showed normal hepatocytes. CONCLUSION: The results support the diagnostic utility of FNA cytology in the management of liver disease, especially in rural setting.

Pneumocystis carinii pneumonia in HIV/AIDS patients at an urban district hospital in Kenya.

BACKGROUND: Pneumocystis carinii pneumonia has generally been regarded to be an uncommon opportunistic infection in HIV infected individuals in sub-Saharan Africa. The reason for this has not been clear but postulates included a lack of suitable pathogenic types in the African environment, diagnostic difficulties and the more commonly held belief that African HIV infected individuals were dying early from common non-opportunistic pathogens before severe degrees of immunosuppression occured. Recently a trend has emerged at the Mbagathi district hospital whereby an increasing number of HIV infected patients are empirically treated for Pneumocystis carinii pneumonia (PCP) based on clinical and radiological features. OBJECTIVE: To determine the prevalence of PCP and clinical outcomes of HIV infected patients presenting at the Mbagathi District Hospital, Nairobi with the presumptive diagnosis of PCP. SETTING: Mbagathi District Hospital, a 169-bed public hospital in Nairobi, Kenya. METHODS: Patients presenting with a sub-acute onset of cough and dyspnoea were eligible for the study if they were found to have bilateral pulmonary shadows and had negative sputum smears for AFBS. Consenting patients who had no contraindication to fiberoptic bronchoscopy had a clinical evaluation which was followed with a fiberoptic bronchoscopy procedure where bronchoalveolar lavage fluid (BALF) was obtained. BALF was examined for cysts of P. carinii using toluidine blue stain and immunofluorescent antibody test (IFAT). BALF was also processed for fungi, bacteria and mycobacteria using routine procedures. Standard treatment with high dose cotrimoxazole was offered to all patients who were then followed up until discharge from hospital or death whichever came first. RESULTS: Between June 1999 and August 2000 a total of 63 patients were referred for bronchoscopy. Of these four declined to undergo the fiberoptic bronchoscopy procedure, four died before the procedure could be done, one was judged too sick to undergo the procedure and three had been on cotrimoxazole for longer than five days. Thus 51 patients underwent bronchoscopy. Pneumocystis carinii stain was positive in 19 (37.2%) while death occured in 16 (31.4%) of the 51 patients. There were more deaths in those without PCP but this difference was not statistically significant (odds ratio 0.68 (95% CI 0.35-1.32; P=0.2). CONCLUSION: PCP was found to be common in HIV infected patients presenting with clinical and radiological features of the disease. The mortality rate for patients with a presumptive diagnosis of PCP is high. This study suggests that cotrimoxazole preventive therapy may be a useful intervention in symptomatic HIV infected patients in Kenya for the prevention of PCP and may avert deaths from this disease.