Evaluation of the diagnostic value of serum and tissue apoptotoic cytokeratin- 18 in patients with chronic hepatitis C

Hepatitis C virus [HCV] is considered the most common aetiology of chronic liver disease [CLD] in Egypt. The disease severity ranges from mild illness to cirrhosis and hepatocellular carcinoma. A role for apoptosis in liver damage caused by HCV chronic infection has been suggested. Cytokeratin 18 [CK-18] is the major intermediate filament protein in the liver and is a known caspase substrate in hepatocyte apoptosis. Therefore, we analysed the serum and tissue levels of CK-18 in patients with chronic HCV infection to evaluate its role in hepatocyte apoptosis. We also correlated CK-18 expression with the severity of hepatic pathology. This study examined 80 Egyptian patients with liver disease. There were 69 patients with chronic hepatitis C and 11 patients with hepatitis C-induced cirrhotic changes. Fifteen healthy controls were also included in the study. The levels of CK-18 fragment were quantified in paired serum and liver biopsy samples. The serum and tissue CK-18 levels were reduced in chronic HCV patients compared to early cirrhosis patients. This result indicates that serum levels of CK-18 and the hepatic expression of CK-18 might play an important role in disease progression. The serum and tissue levels of CK-18 were significantly increased and directly correlated with inflammation severity, stage of fibrosis, and ALT levels in the chronic HCV group and the cirrhotic liver group. There was no significant difference in viral load between patient cohorts. The serum level and the hepatic expression of CK-18 are related to disease activity and are directly correlated with METAVIR scoring. This result suggests that serum CK-18 levels may be useful for monitoring disease activity in chronic HCV and liver cirrhosis patients

In vitro differentiation of mice bone marrow cells into hepatocyte-lik cells

Recent publications showed that highly selected hone marrow derived stem cells can differentiate into hepatocyte like cells. Bone marrow, considered as a rich source of adult stem cells, contains not only hematopoietic and endothelial stem cells, but also precursors of other tissues of mesenchymal origin. The different growth factors used in the culture media are critical factors directly affecting the hepatocytic differentiation and expansion process. Differentiation of adult bone marrow stem cells [BMC] into hepatocyte-like cells is commonly performed by continuous exposure to a cytokines-cocklail. Here, we investigated whether the in vitro differentiation efficacy can be enhanced by sequential addition of liver-specific factors in a time-dependent order that closely resembles their secretion pattern during in vivo liver embryogenesis. Fibroblast growth factors [FGFa and b], Leukeamia inhibitory factor [L1F] Stem cell factor [SCF], Hepatocyte growth factor [HGF] and Oncostatin M [OSM] were used as differentiation factors. Mice bone marrow cells were cultured for one week in a primary culture media and the adherent cells were then subcultured in the directed differentiation media including the above mentioned growth factors added either as a cocktail or in a sequential manner for comparison. In the course of cell differentiation, cell morphology was observed, and the expression of albumin by the transdif Terentiated cells was examined by immunofluorescence and Western blot analysis. Some epithelial-like cells or polygonal hepatocyte like cells appeared in the directed differentiation medium within 12 days, and the number and size of colonies of epithehial-like cells or polygonal cells increased in the course of the cell directed differentiation. The sequential addition of growth factors setup featured relatively earlier and more intense differentiation to hepatocyte like cells. By immunofluoresence analysis albumin expressions appeared, lasted and increased throughout the later directed differentiation. Albumin was found in the cell membrane and scattered in the cytoplasm by immunofluorescent staining. On day 21, the ratio of albumin-positive cells was 85% in the sequentially exposed cells in contrast to the cocktail treatment where the ratio of albumin expressing cells was 65%. We propose that the addition of the hepatocyte growth factor to the culture media is the pivotal step in the trans-differentiation protocol. Moreover the sequential induction of the differentiation process, analogous to in vivo liver development, is crucial for in vitro differentiation of adult mice BMC into functional hepatocyte-like cells

SEN virus infection in Egyptian chronic hepatitis C and haemodialysis patients

SEN virus [SENV] has been tentatively linked to transfusion-associated non A-E hepatitis. The aim of the present study was to determine the prevalence of SENV among Egyptian patients with HCV-related chronic liver disease [CLD] and haemodialysis [HD] patients and to assess the clinical effect of SENV infection on coexistent hepatitis C either in the severity or the probability of developing hepatocellular carcinoma [HCC]. Polymerase chain reaction [PCR] was used to detect SENV-D and SENV-H DNA in serum samples of 74 HCV-related CLD patients, 45 uraemic patients on maintenance HD and 28 healthy controls. SENV DNA was detected in 13.5%, 11.1%, and 7.1% of CLD, HD patients and healthy controls respectively with no significant differences between patients and control group. No statistically significant differences were demonstrated between SENV infected and non infected CLD or haemodialysis patients regarding the clinical and biochemical parameters. SENV infection was significantly higher in CLD patients with HCC [33.3%] than without [8.5%] [p<0.05]. In conclusion, SENV does not seem to be a common infection in Egyptian patients. It has no apparent influence on the severity of co-existent HCV related CLD but it could be a risk factor for developing HCC in these patients. Further studies are needed to define the aetiopathogenic role of SENV infection in HCC development

Epidemiologic approach for early detection and control of renal and urinary tract diseases in rural populations

The dipstick testing, microscopic examination of urine and urine cytology were performed for inhabitants from two rural villages [El Shobak El Sharki, V.I and El Katta, V.2] in Giza G. The proliferating cell nuclear antigen [PCNA] and Schistosoma haematobium antigen were done by immuno-histochemical stain to confirm diagnosis. Also, they were subjected to medical questionnaire, clinical examination, ultra-sonography of kidneys and urinary tract. The results showed that V.2 had higher percentage of haematuria, proteinuria, glucosuria and lower urinary tract infection than V.I. Crystaluria was higher in V.I. Sensitivity of dipstick testing compared to microscopic examination was 26.6%, and specificity was 78.7%. Lower urinary tract infection cytologically detected was 44.2% sensitivity and 62.5% specificity compared to pyuria detected by microscopic examination of urine. Among those suffering variable urinary abnormalities, schistosome antigen was not detected in any fixed urine samples in comparison to corresponding confirmed positive controls. Urine cytology detected urinary tract infection, Crystaluria, dysplasia and atypia, squamous metaplasia and transitional cell carcinoma [TCC]. PCNA positivity was found in TCC [100%], dysplasia [50%] and squamous metaplasia [28.6%]. So, microscopic examination of urine proved valuable for tract abnormalities as pyuria, haematuria and crystaluria. Also, urine cytology is a must for malignancy of urinary tract especially in adult males

Autolytic changes in blood cells of albino rats a post-mortem morphologic study

The morphology of various types of cells in the peripheral blood of albinorats cadavers was investigated. The material comprised 32 adult male albinorats from which ante-mortem blood samples were compared to postmortem samplestaken every hour up to 6 hours after death. The rat corpses were eitherexposed to the environmental temperature [31 +/- 3C], or kept at +10C in anincubator. The red cell count, and the total white cell and platelet countsdecreased significantly during the first few hours postmortem in theenvironmentally-exposed group of corpses. Red cells were quite rapidlytransformed from the normal rounded discoid configuration to acquireinvaginations, crenations and later on, spheroidal configuration and membraneabsorption. Rapid deterioration of the staining properties, and markedmorphological changes in many leucocytes occurred quite rapidly after death. Lymphocytes seemed to be the most resistant to the effect of autolysis. Thepresent morphological observations and the quantitative results suggested thatvarious cellular elements of the blood seem to be quite resistant to autolyticeffects in the early postmortem period, and many cells apparently retain theirnormal morphology for longer periods of time in the blood of rat cadavers keptat reduced temperature