Role of Ebola Virus vp24 Protein in Inhibition of Interferonogenesis.

The effects of recombinant analog of natural Ebola virus protein vp24 in configurations virulent (vp24-ad) and avirulent (vp24-w) for guinea pigs on interferonogenesis were studied in vivo and in vitro. Amino acid differences were determined by His186 substitution in avirulent (nonlethal) configuration for Tyr in the virulent (lethal) one. Recombinant analogs vp24-w and vp24-ad inhibited interferonogenesis in vivo and in vitro. Inhibition by the two protein configurations was virtually the same.

Development of Endoplasmic Reticulum Stress during Experimental Oxalate Nephrolithiasis.

Morphological and ultrastructural study of the kidney was performed in rats with oxalate nephrolithiasis. Specific features of endoplasmic reticulum stress were evaluated during nephrolithiasis and treatment with α-tocopherol. We observed the signs of endoplasmic reticulum stress with activation of proapoptotic pathways and injury to the cell lining in nephron tubules and collecting ducts. Ultrastructural changes were found in the organelles, nuclei, and cell membranes of epitheliocytes. A relationship was revealed between endoplasmic reticulum stress and oxidative damage, which developed at the early state of lithogenesis.

Expression of B23/Nucleophosphamine Nonribosomal Nucleolar Protein in Smooth Muscle Tumors of the Corpus Uteri.

Immunohistochemical study of B23/nucleophosmine (NPM), the key argyrophilic nonribosomal protein associated with the nucleolus-producing regions, in smooth muscle formations of the body of the uterus and myometrium was carried out. The uterus index, number of positively stained nucleoli in the nucleus, integral optical density, and area of positively stained nucleoli were studied by computer-aided image analysis. The results indicated significant differences in the integral optical density and less so in the areas of NPM+ nucleoli in leiomyosarcomas, but not in intact myometrium and leiomyomas. Coefficients of variations in the integral optical density and NPM area proved to be significant parameters for differentiating leiomyosarcomas by size (≤7 cm or larger), evaluating the malignancy grade and stage of the process. NPM parameters in leiomyomas and leiomyosarcomas differed significantly, depending on the levels of Ki-67 and p53 expression and the receptor status of leiomyosarcoma. The index of area and the integral optical density of NPM could serve a accessory objective differential diagnostic criteria in cellular and mitotically active liomyomas and well-differentiated leiomyosarcoma of the corpus uteri.

Stimulation of Cardiomyocyte Regeneratory Reactions under Conditions of Cytopathic Hypercholesterolemia.

The regeneratory reactions of cardiomyocytes in the heart under conditions of cytopathic exposure to hypercholesterolemia and during verapamil treatment were studied by immunohistochemical detection of proliferation marker Ki-67 and evaluation of cardiomyocyte count. A 30-day exposure of rats to atherogenic diet led to an increase of Ki-67+ cardiomyocytes by 14.5-16.7 times (p<0.05). The Ki-67 label index in cardiomyocytes remained higher than normally (8-9-fold; p<0.05) after 64 days. It remained elevated (8-11-fold; p<0.05) after verapamil treatment. Evaluation of cardiomyocyte count and of their nuclear status detected various regeneratory strategies: increase of the total cardiomyocyte count, with increase of the percentage of mononuclear cardiomyocytes (particularly in response to verapamil in group 1 rats); decrease of total cardiomyocyte count with increase of the percentage of multinuclear cardiomyocytes (particularly in group 2 rats in response to verapamil).

Immunohistochemical Analysis of MMP-2 Expression in the Myocardium During the Postinfarction Period.

Immunohistochemical analysis revealed 2.5-fold increased of expression MMP-2 in myocardium samples during the early period (up to 3 days) of postinfarction reparative regeneration. During this period, MMP-2 was detected mainly in monocytes/macrophages circulating in the blood and migrating to the necrotic zone, while expression in the intermuscular and perivascular connective tissue was lower. At later terms, with development of large focal and diffuse cardiosclerosis, MMP-2 expression significantly decreased (to the initial level) and was detected mainly in the foci of intermuscular and perivascular fibrosis, its area in the sections increased by 1.8 times. Evaluation of MMP-2 expression in the blood vessels showed that the immunohistochemical reaction was the most pronounced in the walls of new sinusoidal vessels and the minimum in the intramural arteries of medium diameter. These results attest to an important role of MMP in connective tissue remodeling (proteolytic degradation) during the early period of postinfarction reparative regeneration. The decrease in MMP-2 expression observed at later terms correlated with myocardial fibrosis progression.

Ultrastructural Analysis of the Prostate Gland under the Effect of Factors of Chemical Industry.

We analyzed ultrastruciture of the cell populations in the prostate gland in chronic nonbacterial prostatitis in a chemical industry worker. It was shown that ultrastructural reorganization of the epithelium consisted in reduction of the secretory compartment of the cytoplasm and dystrophic-degenerative changes in cell organelles. Endothelial cells of the capillaries showed signs of significant degeneration and low intensity of micropinocytosis. Most of the smooth muscle cells underwent dystrophic-degenerative modifications of ultrastructural elements. The dominance of degenerative cell changes in the epithelial and stromal cell populations along with intensification of collagen formation in the absence of inflammatory elements allows us to interpret this pathological condition of the prostate gland as prostate pathology of occupational or mixed genesis.

Content of circulating extracellular DNA, plasma activities of matrix metalloproteinases, and ultrastructure of the myocardium in hypothyroid rats with hypercholesterolemia.

Free circulating extracellular DNA, plasma activities of matrix metalloproteinases in hypothyroid rats, and ultrastructural changes in the myocardium were studied under conditions of experimental hypercholesterolemia. For suppression of thyroid function, the animals received antithyroid drug mercazolyl under conditions of cholesterol diet. Hypercholesterolemia in hypothyroid rats (thyroxine concentration 2-fold below the normal) was paralleled by a pronounced increase of the concentration of free circulating extracellular DNA and total matrix metalloproteinases 2 and 7 activity. These changes were associated with lytic and destructive changes in cardiomyocytes and blood capillary endotheliocytes. Changes in the cardiomyocyte and endotheliocyte ultrastructure were more pronounced in hypothyroid rats.

Expression of ryanodine receptor mRNA and sarcoplasmic Ca²âº-ATPase mRNA in the myocardium and intracellular reorganization of cardiomyocytes in dyslipidemia and during verapamil treatment.

In experiments on rats, atherogenic diet led to hypercholesterolemia, while addition of verapamil to the diet led to the development of hypertriglyceridemia in these animals. Dyslipidemia induced significant changes in the cardiomyocyte ultrastructure (lytic changes in myofibrils and sarcoplasmic matrix and aggravation of autophagocytosis) that were most pronounced after addition of mercazolyl alone to the diet. After 30-day atherogenic diet, we observed a decrease in the relative content of RyR2 mRNA (by 67-73%, p<0.01) and SERCA2a mRNA (by 75-91%, p<0.01) in the myocardium. In 64 days these parameters remained reduced: by 64-72% (p<0.05) and 54-62% (p<0.05), respectively. Verapamil treatment reduced the severity and number of lytic lesions in cardiomyocytes, induced considerable glycogen accumulation in the sarcoplasm and its sequestration, promoted normalization, and prevented pronounced decrease of the relative RyR2 mRNA and SERCA2a mRNA in rat myocardium.

Immunohistochemical reactions of primary neuroendocrine breast cancer.

We performed immunohistochemical analysis of neuroendocrine breast cancer. The significance of this approach to differential diagnosis of this cancer form by detection of specific markers is demonstrated. The incidence of diagnosis of neuroendocrine breast cancer has increased 8-fold over 5 years. Neuroendocrine breast cancer is responsible for 3.4% of all breast cancer cases. The disease is significantly more incident in patients over 60. Neuroendocrine breast cancer is characterized by high expression of neuron-specific enolase (100%) and chromagranin A (57%). The data indicate that irrespective of the location, macroscopic picture, and immunohistochemical profile of cytokeratins and steroid hormones, breast cancer is characterized by clear-cut expression of neuroendocrine markers in different combinations. Immunohistochemical detection of these markers makes it possible to detect the hormone-secreting elements and evaluate their role in the tumor pool, which is essential for the differential diagnosis of neuroendocrine breast cancer, including studies of puncture biopsy specimens.

Relationship of argyrophilic proteins of nucleolal organizer regions in Ki-67+ cells with clinical and morphological parameters in lung adenocarcinoma.

Ninety-four lung adenocarcinoma samples obtained during surgeries were examined using a combination of immunohistochemical staining for Ki-67 antigen and silver nitrate staining for argyrophilic nucleolar organizer (Ag-NOR) proteins. In lung adenocarcinoma, we studied the correlation between the area of Ag-NOR proteins in Ki-67(+) cells and clinical and morphological TNM parameters: maximum tumor diameter (T) and stage of the disease and tumor differentiation degree (N). Survival of patients with small area of Ag-NOR proteins in Ki-67(+) cells was higher that in patients with great area of these proteins. The area of Ag-NOR proteins in Ki-67(+) cells is an independent prognostic factor in lung adenocarcinoma. The area of Ag-NOR proteins in Ki-67(+) cells correlates with clinical and morphological TNM parameters and survival of patients with lung adenocarcinoma.

Immunohistochemical study of apoptotic marker p53 as a prognostic factor in breast cancer.

Histological study of breast cancer specimens obtained from 294 female residents in the Republic of Sakha (Yakutia) of different age and ethnic groups and immunohistochemical analysis of protein expression of mutant suppressor gene p53 in tumor cells were performed. A positive correlation was found between expression of mutant gene p53 protein and the degree of malignancy of breast tumors. The indicator should be considered when assessing malignancy potential and risk of breast cancer, because it is important for optimization and individualization of the treatment.

Correlation of HER-2/Neu protein expression and HER2 gene amplification with clinical morphological parameters of nonsmall cell lung cancer.

A total of 218 operation specimens from patients with nonsmall cell lung cancer were examined. The expression of HER-2 protein was studied by the immunohistochemical method, amplification of HER2 gene and centromere part of chromosome 17 (CEP17) - by the in situ chromogenic hybridization method. Moderate correlation (r=0.38, p<0.001) between the immunohistochemical status and HER2 gene amplification was detected. A relationship between TNM clinical morphological parameters and HER-2 protein content and chromosome 17 polysomia (for N parameter), HER-2 protein content, and HER2 gene amplification (for tumor histogenesis) was detected. The content of HER-2 protein and amplification of HER2 and CEP17 correlated with the clinical morphological parameters of patients with nonsmall cell lung cancer.

Ultrastructural and metabolic features of stress-adaptive rearrangements of myocardium in infants after surgery-induced up-regulation of lipoperoxidation processes.

Surgical correction of complicated congenital heart disease in infants is terminated by postischemic reperfusion of myocardium accompanied with closure of more than 20% microvessels in the coronary bed by edematous endothelial cells, their bleb-like fragments, and aggregates of the blood formed elements. Degradation of the transmission capacity of the coronary microvessels developed in parallel with moderation of activity of the antiradical protection enzymes and positively correlated with the level of LPO secondary products, whose elevation during the surgery stages was not reduced by catalase activation.

Expression of molecular biological markers in breast cancer under conditions of the Sakha Republic (Yakutia).

The results of histological studies of operation material from 294 women with breast cancer and immunohistochemical detection of estrogen and progesterone receptors and evaluation of proliferative activity (Ki-67) and expression of proteins p53 and bcl-2 were analyzed. Breast cancer was more incident (60%) in nonindigenous residents of the Sakha Republic (Yakutia) than in representatives of the indigenous population (40% cases). The disease was more incident in women aged 50-59 years (32.6%) and 60-69 years (21.8%) irrespective of the ethnic appurtenance. The findings indicated the most informative criteria of mammary tumor malignancy: low expression of estrogen and progesterone receptors, high expression of Ki-67 and p53, and low expression of bcl-2.

Morphoimmunohistochemical characteristics of synchronous bilateral breast cancer.

We performed a complex morphoimmunohistochemical study of synchronous bilateral breast cancer (diagnosed in 0.69% cases in women of the postproductive period after the age of 50). The molecular genetic characteristics of this cancer were detected and the algorithm of diagnostic studies was suggested. We have demonstrated that bilateral breast cancer was in 75% cases estrogen- and progesterone-positive, but androgen-negative. The HER-2/neu status of the tumor was negative in 75% cases, but expression of HER-2/neu was detected in the tumor of one mammary gland in 20% cases. Membrane expression of E-cadherin in infiltrative ductal cancer cells significantly decreased up to its complete disappearance with the decrease in tumor cell differentiation degree. The data provide new insight into prognostic value of the studied biomarkers and help to develop treatment protocol with consideration for the molecular biology of this group of tumors.

Argyrophilic proteins of nucleolar organizer regions and proliferative activity of cells in squamous cell carcinoma of the lung.

Argyrophilic proteins associated with nucleolar organizer regions (Ag-NOR proteins) and Ki-67 antigen were analyzed in 118 samples of squamous cell carcinoma of the lungs. Tumors with low and high content of Ag-NOR proteins and Ki-67 index were selected. It was found that the content of Ag-NOR proteins correlated with some clinical and morphological parameters (indexes T and N, tumor size less and more that 3 cm, stage of the disease, and tumor differentiation degree) and survival rate. High survival was associated with low content of Ag-NOR proteins and Ki-67 index and low survival correlated with high content of Ag-NOR proteins and Ki-67 index, while intermediate survival was associated with opposite values of Ag-NOR protein content and Ki-67 index. The tumor size, parameter N, and the content of Ag-NOR proteins had independent effects on patient's survival. In patients with squamous cell carcinoma of the lung without metastases in lymph nodes, survival correlated with the content of Ag-NOR proteins; in patients without metastases, survival correlated with tumor size.

Mathematical simulation of p53-Mdm2 protein biological system regulation.

Digital simulation of disorders in the p53 and Mdm2 protein system, involved in repair of DNA defects and in many vital processes, including tumor formation, neurodegeneration, and aging, was carried out on the base of functional nonlinear octoparametric mathematical model. Stress situations were studied, associated with emergence of imbalance in p53 and Mdm2 generation and degradation rates and with disorders in the mechanism of protein interactions, regulated within the framework of the suggested model via dissociation constants. Variants of the analyzed disorders compensation by repeated stress for the system parameters were studied numerically.

Morphofunctional analysis of the prostate exposed to chemical manufacture factors.

The structural characteristics of the prostate and the blood flow in the organ were studied in chemical plant workers suffering from chronic prostatitis. The prostate echostructure was characterized by vast zones of fibrosis and calcinosis, the hemodynamic ultrasonic parameters were low. Degenerative changes in the acinar structures and stromal fibrosis predominated in the biopsy specimens, these shifts were the most pronounced in the peripheral and transitory zones. Foci of common and small-acinar degeneration of the glands, abundant concrements, and significant collagenization with periglandular and perivascular sclerosis without or with slight inflammatory infiltration were detected. We hypothesize that long exposure to adverse factors of sulfuric acid manufacture contributed to the development of pathological changes in the prostate.

Mathematical modeling of functioning of the p53-Mdm2 protein system.

Mathematical simulation of the time course of p53 and Mdm2 protein concentrations during their interactions was carried out. The system states corresponding to threatened uncontrolled apoptotic cell death accelerating aging processes and to situation with hypersuppression of p53-dependent apoptosis fraught with high risk of cancer were modeled. New data indicate the possibility of description (within the framework of mathematical model used in the study) of the p53 and Mdm2 proteins feedback mechanism, which could guarantee adequate reaction to serious DNA damage.

Microcirculation and structural reorganization of the bladder mucosa in chronic cystitis under conditions of ozone therapy.

Structural reorganization of the bladder mucosa in chronic cystitis and its correction by ozone therapy were studied. A relationship between the epithelial layer restructuring of different kinds (dystrophy, metaplasia, and degeneration), level of cell proliferation, and ultrastructural organization of urotheliocytes was detected. This complex of structural reactions was combined with dysregulation of tissue bloodflow in the bladder mucosa, shown by laser Doppler flowmetry. Positive structural changes were most marked in intravesical and less so in parenteral ozone therapy added to the therapeutic complex and manifested in reduction of inflammation and alteration in parallel with more intense reparative reactions. A special feature of parenteral ozone therapy was a significant improvement of microcirculation in the bladder mucosa.