Objective: to study the activity of proapoptotic signal protein caspase-3 for determination of peculiarities of apoptosis regulation under liver chronic diseases.Subjects and methods. The immunohistochemical analysis of caspase-3 activity in 5 liver biopsies of the patients with mono infection of chronic hepatitis B and 5 liver biopsies of the patients with mixed infection of tuberculosis, chronic hepatitis C and human immunodeficiency virus was fulfilled. Morphological and morphometric analysis of serial microphotographs was performed using an image analysis system (microscope Leica DM 2500, digital camera Leica DFC320 R2 and a computer).Results. The activity of caspase-3 as dark brown granularity was revealed in all tis-sue components of liver (hepatocytes, epithelium of bile ducts, endotheliocytes, Kupffer cells of sinusoids, in compositions of lymphohistiocyte infiltrations). The maximal activity was discovered in hepatocytes nuclei. The expression of caspase-3 was significantly higher in liver biopsies of the patients with mixed infection. It is typical that the immunoreactive hepatocytes had not any morphological marks of apoptosis.Conclusion. The caspase-3 expression of proapoptotic signal protein caspase-3 may serve as an early marker of liver damage including the possibilities of apoptosis development.

The aim of this work was to perform a statistical analysis of quantitative changes of the cell population structure of the liver biopsy specimens in patients with chronic hepatitis B. The data from computer microscopy of 707 serial section micrographs of liver biopsies were analyzed. The investigation showed an existence of statistically significant differences in cell populations of portal zones from populations of medium and central zones. The dependences between some characteristics of cell populations and blood indexes were discovered in terms of the dependence between different range evaluations.

Objective : to study the activity of proapoptotic signal protein caspase-3 for determination of peculiarities of apoptosis regulation under liver chronic diseases. Subjects and methods . The immunohistochemical analysis of caspase-3 activity in 5 liver biopsies of the patients with mono infection of chronic hepatitis B and 5 liver biopsies of the patients with mixed infection of tuberculosis, chronic hepatitis C and human immunodeficiency virus was fulfilled. Morphological and morphometric analysis of serial microphotographs was performed using an image analysis system (microscope Leica DM 2500, digital camera Leica DFC320 R2 and a computer). Results. The activity of caspase-3 as dark brown granularity was revealed in all tis-sue components of liver (hepatocytes, epithelium of bile ducts, endotheliocytes, Kupffer cells of sinusoids, in compositions of lymphohistiocyte infiltrations). The maximal activity was discovered in hepatocytes nuclei. The expression of caspase-3 was significantly higher in liver biopsies of the patients with mixed infection. It is typical that the immunoreactive hepatocytes had not any morphological marks of apoptosis. Conclusion. The caspase-3 expression of proapoptotic signal protein caspase-3 may serve as an early marker of liver damage including the possibilities of apoptosis development.

Background: Apoptosis is a genetically programmed form of a cell death that plays a major role in pathogenesis of chronic liver diseases. As Caspase-3 activation is required to produce apoptotic chromatin condensation and DNA fragmentation it is used in the study of apoptosis. Subjects and methods: Liver biopsies of patients with heroin abuse and coinfection of tuberculosis, chronic hepatitis C, human immunodeficiency virus (TB, HCV, HIV) were investigated. For comparison liver biopsies of patients with chronic hepatitis B (HBV) were used. All biopsies were performed according to routine medical program, using standard Mengini procedure. Material was fixed in formalin, embedded in paraffin and cut 7 micrometers in thickness. For immunohistochemical detection endogenous peroxidase activity was blocked with 3% hydrogen peroxide and the sections were stained using Immunohistochemistry kit according to the manufacture’s guidelines. Cleaved Caspase-3 (Asp175) served as primary antibody. Biotinylated secondary antibody and streptavidin-conjugated peroxidase were used for detection using DAB as chromogen. Nuclei were counter stained with Harris Hematoxylin. Negative controls did not contain primary antibodies. In addition to the immunohistochemical study of Caspase-3 numerical score was established for each biopsy specimen both for grading of necroinflammatory activity (Knodell et al.) and stage of fibrosis (French METAVIR). Results: Our investigation showed that positive reaction of Caspase-3 strongly varies in different liver biopsies; activation is revealed in nuclei and cytoplasm of hepatocytes and some Kupffer cells as well as in endotheliocytes of sinusoids. In portal tracts the activation occurs in some endotheliocytes of interlobular portal veins, in cells of lymphohistiocyte infiltrates. In liver biopsies of patients with coinfection the Caspase-3 expression was noticed in 30-50% of nuclei, whereas in liver biopsies of patients with HBV mono infection the index did not exceed 10%. Index of histological activity according to Knodell of patients with coinfection reached 12-15 balls, fibrosis stage according to METAVIR – F2; in patients with HBV these indexes were respectively 4 balls and F-1. Conclusions: Our study demonstrated the expression of Caspase-3 was significantly higher in cases of chronic hepatitis with more severe histological necroinflammatory activity. Especially high activation of Caspase-3 was in liver biopsies of patients with coinfection and heroin abuse. Our results suggest high level of apoptosis in liver biopsies of such types of patients.

The aim of our study was the comparison of the synthetic glucocorticoid dexamethasone (DEXA) influence on T-lymphocytes in central (thymus) and peripheral (spleen, lymphatic nodes) immune organs. For that reason therapeutic doses of DEXA were used followed by histological, histochemical (TUNEL) as well as computerized histomorphometrical investigations. In the study 36 young adult Wistar rats performed. 1–7 days after 3 days injection of DEXA (total dose 1,2 mg/rat i.p.) the material was taken for futher investigations. First days after DEXA administration in therapeutic doses the number of apoptotic cells was considerably increased in the cortical part of thymus. No significant changes were in rest of thymus as well as in peripheral immune organs. 7 days after DEXA-induced injury the number of apoptotic cells had decreased almost to the normal level. Our investigations conclude that the most sensitive for the dexamethasone-induced T-lymphocyte apoptosis is cortex of thymus while the changes in medullary area of thymus and peripheral immune organs – spleen and perithymic lymphatic nodes – are less significant. Week after drug cessation the apoptotic changes are almost at normal level in both types of lymphoid organs.

The cell population analysis of liver biopsies from the patients with both chronic hepatitis, chronic viral hepatitis C (HCV) and chronic viral hepatitis B (HBV) included the comparative evaluation of the specific part of non-parenchymal elements, analysis of the liver plates and sinusoids areas, the cell population of liver plates and sinusoids, the caryometric description of different types of cells. The essential difference of similar quantitative indexes in the biopsy specimens of patients with HCV and НВV was revealed and discussed. In total, the quantitative analysis of cell population structure in liver biopsies during the course of chronic hepatitis, especially in the case of defective biopsies, could be used for diagnostics and prognoses by expert evaluation.

The liver biopsy has long been the gold standard for the evaluation of the state of liver diseases in patients, especially in chronic hepatitis. Hepatic fibrosis plays the most important role in the evolutionary process from chronic hepatitis to cirrhosis. Therefore, accurate assessment of the degree of hepatic fibrosis in chronic viral hepatitis is important to understand not only the clinical condition and prognosis of patients, but also the natural history of hepatitis. Information on a stage of chronic viral hepatitis C is essential to make prognosis and decide antiviral treatment. Semi quantitative scoring systems have been used in most studies that have relied upon liver biopsy to evaluate changes in fibrosis (Knodell et al., 1981, Desmet et al., 1994, Chevallier et al., 1994, French METAVIR, 1994, Ishak et al. 1995). Comparative characteristic of different scoring systems is presented by Brunt (2000) and Goodman (2007). However, these methods cannot completely avoid the observer’s bias. Recent studies have reported that the estimation of fibrosis by semi quantitative scoring system is not always accurate and high rate of interand intraobsrever discrepancies takes place (Scheurer, 2003). The alternative to semi quantitative fibrosis scores is direct measurement of the amount of fibrosis, or necroinflammatory lesions, or portal zones in the biopsy by computer-assisted morphometric image analysis (O’Brien et al., 2000), or stereological morphometric analysis (Filimonova et al., 2010). Cell population analysis gives additional information about structure of parenchyma elements (liver plates and sinusoids).

Quantitative stereological morphometric analysis of liver biopsy was used for more correct evaluation of dynamic of liver damages in patients with chronic viral hepatitis C. The analysis allows to estimate the area (%) of non-parenchymal elements such as portal tracts, bridging and piecemeal necrosis, intralobular focal infiltrates. That is important for estimation of efficiency of antiviral therapy. The investigation showed that the portion of the area of non-parenchymal elements of different patients strongly varied; the interrelations between some morphological parameters and level of serum alanine aminotransferase were established. This study demonstrates that the ratio of the area of non-parenchymal elements to that of entire tissue specimen in the initial biopsy might be a predictive factor for prognosis.

The problems of cell division regulation and tissue growth are very important for the theory and practice of medicine and biology. The main way of the problem solution is uncovering the mechanisms regulating the cell proliferation. The aim of this work is the creation and analysis of the mathematical model of rapidly renewal tissue at perturbation state and its reparation after irradiation. As a sample of cell population, we selected the intestinal epithelium. This tissue is very interesting due to its high radio sensitivity; the intestinal epithelium renewal on the base of stem cells is well studied. In addition, this tissue is very interesting for modeling because of the presence of a special genetic program apoptosis, which leads under distinct conditions to cell death. To create the mathematical model of cell population dynamics we used the kinetic approach based on differential computation. For the model construction, we used the assumption that there exists double mechanism of the feedback between the cells of the population: at the level of stem cells of crypt and at the level of the column cells of villus. The constructed model showed its good correspondence with data received in experiments under sharp and fractionated irradiation in dose range from 0 to 10 Gy.

UNLABELLEDThe aim of the present study was to determine the target site cells in the rat thymus after exposure to the synthetic glucocorticoid, dexamethasone, at therapeutic doses. The findings of histology and histochemistry (Feulgen, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling--TUNEL) with quantification by computerized histomorphometry are described.MATERIAL AND METHODSA quantified investigation of apoptotic and mitotic thymic lymphocytes in 36 young adult Wistar rats was performed at 1-7 days after a 3-day injection of dexamethasone (a total dose of 1.2 mg/rat intraperitoneally).RESULTSAt the first day after dexamethasone administration the moderate involution and atrophy of thymus histology were observed with simultaneous fall in cortical cellularity and mitotic activity of thymocytes. More rapid fall appeared in the inner cortex. The number of apoptotic (TUNEL-positive) cells was significantly increased. On the days 5 and 7 the expression of apoptosis and the cell proliferation were at almost normal level.CONCLUSIONSThe findings suggest that dexamethasone-induced apoptosis of cortical thymic lymphocytes, mainly correlated with synchronous inhibition of mitosis and cell number fall in thymus. The main target sites of dexamethasone injury were cells in the inner cortex of lobuli thymi.

The suggested mathematical model of cell population kinetics in small intestine allows to obtain general picture of the population behaviour and to determine the time of the population repair to normal level after different doses of radiation. The model well corresponds to experimental data.

The mathematical model of the bilirubin transport through liver is suggested. The model is sufficiently reflects the peculiarities of the bilirubin behavior under the different physiological conditions. The model is described by the system of difference-differential equations. The coefficients of the intensity of bilirubin transfer in normal liver are calculated.

Abstract The fine structure of the intestinal epithelium of Cucumaria frondosa has been examined. The digestive cells possessed some specific ultrastructural peculiarities which testify to their participation in the transport of substances and in the processes of intracellular and cavitary digestion. The main units of the intestinal epithelium appear to be multifunctional digestive cells in which the phenomenon of compartmentalization among the organelles' groups, intended to carry out various functions, occurs. The heterogeneity of the structure of populations of the digestive cells is explained. Both primitive and progressive features of the structure of the digestive tract of C. frondosa are indicated.

The fine structure of esophagus, stomach and intestinal epithelial cells of the sea urchin Strongylocentrotus droebachiensis have been examined. Three types of mucosal cells secreting neutral, acid and sulfated mucopolysaccharides have been found in the esophagus. The modes of origin of different types of mucous granules are described. Storage and zymogen cells have been identified in the stomach epithelium. Storage cells are adapted for intracellular digestion and synthesis of nutritive substance. Zymogen cells are adapted for enzyme synthesis. The columnar cells occur in the intestinal epithelium; they are adapted for nutrient absorption and lipid storage. Digestion in different parts of the digestive tract is considered in context with the ultrastructural and histochemical data obtained. Some problems concerning the relation between intracellular and extracellular digestion are discussed.