MAST CELLS IN DAMAGED LYMPH-HEART STRIATED MUSCLE OF ADULT FROG: AN ULTRASTRUCTURAL, IMMUNOCYTOCHEMICAL AND AUTORADIOGRAPHIC STUDY.

Ultrastructural features of mast cell activation were studied in degenerative/reparative experimental model of mechanically damaged lymph-heart striated muscle during the first postoperative week. 24 h after damage, the intracytoplasmic empty degranulation channels were revealed in a certain part of resident and circulating mast cells (MCs) located near the site of injury. These findings are evidence that previously ther was activation and secretory response of MCs by a process known as compound exocytosis which involves not only granule- to-plasma membrane fusions and formation of degranulation channels. We have found rare MCs showing release of single. altered, membrane-free granules into the space restricted by the plasma membrane of mature resident MCs and the basal lamina surrounding them. During the fist postoperative week we have more commonly observed activated lymph-heart MCs with prominent intragranular changes characterized by diminishing of electron-dense component of granules and loss of granule matrix. We have also revealed close association of some unaltered and altered secretory granules with completely or partially empty degranulation channels that permeate the mast cell cytoplasm. Some of these channels remained open directly to the exterior of the cells. It is not improbable that these findings provide the ultrastructural basis for potential discharge of small amounts of granule mediators into the open channels (incomplete exocytosis) duding partial degranulation of activated in vivo frog MCs. Immunocytochemical staining for histamine of activated frog MCs showed the presence of the immunoreactive material both in unaltered and altered cytoplasmic granules. As a rule, gold particles were located over residual electron-dense component of altered granule, whereas little or no gold particles were revealed over the swollen matrix. Ultrastructural features of secretory granule formation in the cytoplasm of some MCs were observed toward the end of the first postoperative week. Electron-microscopic autoradiography has shown that replicative and transcriptional activity of MCs is not observed in the studied postoperative period. No mitoses in MCs have been revealed in this period. A few mast cells were found in necrosis. Taken together, our results indicate that MCs are one the first cell types that respond to injury of lymph-heart striated muscle and release a variety of preformed mediators, including histamine. These findings suggest that activated MCs play an important role in the initiation of an inflammatory process during repair of damaged lymph-heart striated muscle.

Comparison of clinical and laboratory parameters in patients with end-stage renal failure in the outcome of chronic glomerulonephritis and patients with end-stage renal failure in the outcome of other diseases.

BACKGROUND: frequent complications of hemodialysis treatments are coagulation disorders. This is due to activation of the coagulation of blood flow in the interaction with a dialysis membrane material vascular prostheses and extracorporeal circuit trunks. In addition, in hemodialysis patients receiving heparin for years, there is depletion of stocks in endothelial cells in tissue factor inhibitor, inhibits the activity of an external blood clotting mechanism. AIM: the aim of our study was to evaluate the hemostatic system parameters in patients with end-stage renal failure, depending on the cause of renal failure. MATERIAL AND METHODS: to evaluate the hemostatic system parameters in patients with end-stage renal failure, depending on the cause of renal failure and hemodialysis treatment duration conducted a study that included 100 patients observed in the department of chronic hemodialysis and nephrology hospital №1 Republican National Medical Center in the period of 2013-2016. RESULTS: in patients with end-stage renal failure in the outcome of chronic glomerulonephritis, a great expression of activation of blood coagulation confirm increased the mean concentration of fibrinogen, whereas in the group, which included patients with end-stage renal failure in the outcome of other diseases, such is not different from the norm, and a higher rate of hyperfibrinogenemia, identified in 2/3 patients in this group. CONCLUSIONS: it was revealed that the state of homeostasis in patients with end-stage renal failure in increasingly characterizes the level of fibrinogen and the activation of the hemostatic markers: soluble fibrin monomer complexes, D-dimers.

[The Mechanisms of Orientation Sensitivity of Human Vision System. Part II: Neural Patterns of Early Processing of Information about Line Orientation].

The high density EEG was registered in 41 healthy subjects (20 males, 21 females) in the cardinal (horizontal and vertical) and oblique (45 and 135 deg) line orientation identification task. The analysis of the adaptive amplitude maximum (4 ms averaging) of P1 and N1 evoked potentials in the symmetrical occipital, parietal and inferior temporal areas and dipole source modelling showed the anisotropy of cortical responses in the 80-150 ms interval. The amplitude is higher on the oblique orientations as comparison with cardinal ones. The temporal and regional features of cortical answers were discovered. The earlier selective response (~90 ms latency) is registered.in the parietal areas, while the later (~145 ms latency) is found in the occipital ones. We discovered a number of sex-related differences in the early stages of line orientation detection. In males, the amplitude of components is higher; they have broader area of localisation of their dipole sources: in addition to the occipital and parietal regions, cortex of the temporal regions is involved. Theobtained data are discussed in the context of the idea of effective neural coding (Barlow, 1959) and the features of spatial information processing in the visual system of males and females.

[The Modeling of Human Visual ERPs Sources in the Task of Line Orientation Identification].

The high-resolution EEG was taken from 35 healthy subjects (16 males, 19 females) in the task of identification of cardinal (horizontal, vertical) and oblique (45°, 135°) orientations. The integrated approach was developed and used for the ERPs analysis. It includes the modeling of the distributed intracranial dipole sources of the ERPs' components by the weighted Minimum Norm Estimates (wMNE) method, the combination of distributed intracranial dipole sources with the M RI data and the statistical analysis of the current density. It was shown that a multiple network of brain structures was active during the early stage of analysis of line orientation. It includes not only primary striate visual area, but also the parietal, temporal and prefrontal corticies. The new data was obtained about the participation of structures, responsible for navigation (parahippocampal gyrus and entorhinal area) in the line orientation detection. For the first time the gender-related differences in the organization ofthe line detection network were described. The obtained data provides a significant supplement to the knowledge of the mechanisms of the orientation sensitivity and assumes that orientation sensitivity plays an important role in the navigation.

[Mast cells of lymph hearts during ontogenesis of frogs Rana temporaria].

Electron microscopic observations of the lymph hearts of tadpoles and yearling frogs of Rana temporaria showed that mast cells (MCs) were present not only between muscle fibers (population of resident MCs), but in the cavities of lymph heart (population of circulating MCs), too. There were some differences in the ultrastructure of the resident MCs at each studied stage of larval development. The first recognizable MCs were revealed in the lymph hearts at premetamorphosis (stages 39-41). MCs presented as mononuclear relatively small and slightly elongated cells with a few immature secretory granules and numerous free ribosomes, polysomes and short cisternae of rough endoplasmic reticulum (RER) in the cytoplasm. Chromatin of their nuclei was poorly condensed; the Golgi apparatus was moderately developed. At pro-metamorphosis (stages 44-45), we revealed MCs at different levels of their differentiation. Some MCs demonstrated an active process of granulogenesis in their cytoplasm. Among densely packed cytoplasmic organelles, immature secretory granules were closely associated with cisternae of RER and free ribosomes. Other MCs appeared as more differentiated cells. They were characterized by a predominantly heterochromatic nuclei and cytoplasm filled with polymorphic and heterogeneous granules. MCs also showed a reduction in the number of free ribosomes and cisternae of RER in the cytoplasm. On the contrary, the Golgi apparatus was well developed. Stacks of Golgi cisternae, detaching vacuoles, and progranules occupied the perinuclear region. The majority of the outlines above ultrastructural features of differentiated MCs were typical for MCs of yearling frogs. At metamorphic climax (stages 52-53), MCs often tightly contacted with macrophages. We did not reveal apoptotic MCs. However, some MCs exhibited morphological features typical for programmed necrosis-like death, which was characterized by mitochondria swelling, dilatation of cisternae of RER and nuclear envelope, plasma membrane rupture and subsequent loss of intracellular contents. Electron microscopical immunocytochemistry revealed the localization of atrial natriuretic peptide (ANP), substance S (SP) and heat shock protein (Hsp70) in the secretory granules of the resident and circulating MCs at different stages of tadpole development and in yearling frogs.

[Resident and circulating mast cells in propulsative organs of the frog Rana temporaria].

Mast cells (MCs) of the "blood" and lymph hearts of the adult frog Rana temporaria were investigated at histochemical and ultrastructural levels. Two populations of MCs were revealed in these propulsative organs: population of resident MCs and population of circulating MCs. It has been shown that the resident cardiac MCs have an oval or elongated form and are located between atrial or ventricular myocytes and under endocardial endothelium. The resident cardiac MCs are situated in connective tissue of epicardium, too. Avascular myocardium of the frog ventricle consists of a spongy network of muscle trabeculae. We revealed circulating MCs in intertrabecular spaces and clefts of the spongy myocardium and in the blood of the main central cavity. Circulating MCs are round in shape and contain a large central nucleus enriched with condensed chromatin. They resemble the lymphocytes, but show cytoplasm filled with granules. These granules ultrastructure is much like that of the granules of the cardiac resident MCs. In the lymph heart, oval and somewhat elongated resident MCs are located in the interstitial space among cross-striated muscle fibers and among smooth muscle cells of tubular (afferent and efferent) valves. Sometimes lymphocyte-like circulating MCs are revealed in the cavity of lymph heart. Circulating MCs are also present in the lymphatics located adjacent to the lymph hearts. In certain parts of the lymphatic walls MCs are in close adhesion to the mesothelial cells lining the lymphatic cavity. Our histochemical investigation revealed that both the resident and circulating MCs of the propulsative organs give a strongly positive reaction with alcian blue, but weakly red with safranin and weakly metachromatic with toluidine blue. The presence of population of circulating MCs in the frog suggests that there are differences in biology of MCs between lower and higher vertebrates.

[Chromogranin A: immunocytochemical localization in secretory granules of frog atrial cardiomyocytes].

Chromogranin A (CgA) is a member of the granin family of acidic proteins that present in the secretory granules (SGs) of many endocrine, neuroendocrine and neuronal cells. Atrial natriuretic peptide (ANP)-storing SGs in atrial cardiomyocytes of rat heart also contain CgA. Cardiosuppressive effect of CgA-derived peptides (vasostatins) on in vitro isolated and perfused working frog and rat hearts has been shown under both basal conditions and beta-adrenergic stimulation. More recently it has been revealed that rat heart produces and processes CgA-derived vasostatin-containing peptides. Until now nothing has been known about the presence of CgA in an amphibian heart. We have investigated the subcellular localization of CgA in atrial myocytes of adult frog Rana temporaria heart using ultraimmunocytochemical method. Immunocytochemical staining of the frog atrial tissue for CgA and ANP has shown that out of three morphologically different types (A, B and D) of specific cytoplasmic granules (SCGs) present in myocytes only two (A and B)--large (120-200 nm in diameter) granules with more and with less electron dense core--exhibit immunoreactivity (IR) to these two antigens. The third type (D) of granules (80-100 nm in diameter) are small membrane bound granules characterized by highly electron dense core surrounded with a thin halo. These granules revealed negative reaction on immunostaining for both CgA and ANP. The presence of CgA- and ANP-IR in the same SCGs in frog atrial myocytes is consistent with the endocrine nature of these granules. Taking into account our and literature data we propose that CgA present in frog atrial cardiomyocite SCGs might be a precursor of vasostatin-containing peptides, as it takes place in rat heart. It is possible that these CgA-derived peptides together with ANP exert their regulatory function through the autocrine and/or paracrine mechanisms and play important cardioprotective role in frog heart under stress condition.

Secreted aspartate proteinases, a virulence factor of Candida spp.: occurrence among clinical isolates.

Production of secreted aspartate proteinases was determined in a set of 646 isolates of Candida and non-Candida yeast species collected from 465 patients of the University Hospital in Olomouc (Czechia) in the period 1995-2002, and Candida samples obtained from 64 healthy volunteers using solid media developed for this purpose. Using random amplified polymorphic DNA analysis (RAPD) 79 Candida isolates from blood were analyzed to show potential relationships between clustering of the fingerprints and extracellular proteolytic activity of these strains. C. albicans, C. tropicalis and C. parapsilosis possess always proteolytic activity while non-Candida species did not display any proteolysis. A tight relationship between fingerprints and extracellular proteolysis in the Candida isolates was not shown. A remarkable consistency between fingerprint clusters and proteolysis occurred in a subset of C. parapsilosis samples. Suboptimal pH of the growth medium was shown to facilitate the investigation of potential co-incidence of genotypic and phenotypic traits.

[Immunocytochemical localization of atrial natriuretic peptide in endothelial and granular cells of the heart of lophotrochozoa]. / Immunotsitokhimicheskaia lokalizatsiia predserdnogo natriiureticheskogo peptida v éndotelial'nykh i granuliarnykh kletkakh serdtsa lofotrokhoforovykh.

In parallel with contraction, vertebrate cardiomyocytes perform endocrine function and produce natriuretic peptides (NP)--ANP and BNP--involved in cardiovascular homeostasis maintenance. ANP-like peptides have been reported also in hearts of some invertebrates, however, their cellular localization was not determined. By electron microscopical immunocytochemistry with polyclonal monospecific antibodies raised against ANP and protein A-gold technique, we have localized ANP-like immunoreactivity in granules within endothelial cells in the heart of the brachiopod Rhynchonella psittacea, the polychaete Arenicola marina, and the gastropod mollusc Achatina fulica--all being representatives of the major phylogenetic group Lophotrochozoa. ANP-like immunoreactivity was also revealed in one of 3 morphologically distinguishable types of granules in the snail heart granular cells. By electron microscopical autoradiography with the use of [3H]-thymidine, the ability for DNA synthesis was demonstrated in heart endothelial cells of the investigated animals. Forms of NP-system organization in hearts of Lophotrochozoa and Vertebrates, and close histogenetic relationships of endothelial and granular cells in the snail heart are discussed.

[Endothelium and platelets in pregnant women with chronic glomerulonephritis and therapeutic efficacy of acetylsalicylic acid and dipiridamol]. / Sostoianie endoteliia i trombotsitov u beremennykh s khronicheskim glomerulonefritom i lechebnymi vozvozhnosti atsetilsalitsilovoi kisloty i dipiridamola.

AIM: To estimate platelet and endothelial condition in pregnant women with chronic glomerulonephritis (CGN), prognostic value of these changes and efficacy of acetylsalicylic acid (ASA) and dipiridamol in prevention of unfavourable outcomes of pregnancy in CGN. MATERIAL AND METHODS: The examination covered 74 CGN pregnant patients, 14 non-pregnant CGN patients, 11 pregnant women with preeclampsia, 19 healthy pregnant women. The levels of fibronectin, endothelin-1,2, 6-keto-PGF1a, thromboxane B2 (TxB2) secretory beta-thromboglobulin in the blood, activity of intrathrombocytic lactate dehydrogenase (LHG), platelet aggregation. ASA (125 mg/day) was given to 33 CGN pregnant women in combination with dipiridamol (150-225 mg/day). Control group consisted of 32 CGN pregnant women. RESULTS: Content of fibronectin, endothelin, TxB2 and beta-thromboglobulin in blood plasm, aggregation with ADP in CGN pregnant women were higher than in healthy pregnant women and nonpregnant CGN patients. Plasmic 6-keto-PGF1a was low. Preeclampsia was accompanied with elevated fibronectin, TxB2 and beta-thromboglobulin, hyperactive LDH. Platelet aggregation was suppressed. Blood beta-thromboglobulin directly correlated with systolic and diastolic arterial pressure, 24-h proteinuria and blood creatinine. Reverse Correlation was seen in blood beta-thromboglobulin with albuminemia, glomerular filtration rate, body mass of the newborn and term of labor. A direct correlation was found between the activity of intrathrombocytic LDH and systolic and diastolic arterial pressure, a weak reverse correlation--between platelet count in capillary blood and systolic pressure, platelet aggregation with ADP and terms of labor. Of the highest prognostic value were the level of beta-thromboglobulin and fibronectin, the activity of intrathrombocytic LDH and platelet aggregation in response to ADP. ASA and dipiridamol reduced the risk of fetal retardation and fetal loss. CONCLUSION: Pregnant women with CGN have endothelial-thrombocytic dysfunction because of unidirectional influence of both CGN and pregnancy. We think that endothelial-platelet dysfunction connects renal impairment and placental failure in pregnant women with CGN deteriorating a gestational CGN and pregnancy complications. Correction of endothelial-platelet state with ASA and dipiridamol is effective in prevention of fetal retardation and fetal loss in pregnant women with CGN.

[Heart ventricular cells of neonatal rats in a culture: DNA synthesis, ultrastructure and localization of atrial natriuretic peptide]. / Kletki zheludochkov serdtsa neonatal'nykh krys v kul'ture: sintez DNK, ul'trastruktura i lokalizatsiia predserdnogo natriiureticheskogo peptida.

We determined the optimal conditions suitable for expanding cardiac cells in vitro for their future use in experimental transplantation into injured myocardium of adult animals. Ventricular cardiac cells were isolated enzymatically from 2-3 day-old rats and cultured at different cell densities within 5-7 days to 4 weeks. Mixed cultures of muscle and non-muscle cells were examined by light autoradiography, electron microscopy, and immunogold method. The best results were obtained at a density of 3 x 10(5) cells/ml in the medium, consisting of 90% DMEM and 10% fetal calf serum, during 5-7 days of cultivation. In such cultures myocytes made 62.5 +/- 7.9%. After a 24 h incubation with 3H-thymidine, 22.0 +/- 2.2% of myocytes were labeled. Muscle cells contact with each other and with non-muscle cells, contain myofibrils, contract and display atrial natriuretic peptide (ANP)-like immunoreactivity.

[Platelet dysfunction and pregnancy in gestational exacerbation of chronic glomerulonephritis]. / Disfunktsiia trombotsitov i iskhody beremennosti pri gestatsionnom obostrenii khronicheskogo glomerulonefrita.

AIM: To evaluate platelet dysfunctions and pregnancy outcomes in females with gestational exacerbation of chronic glomerulonephritis (CGN) and the disease remission. MATERIALS AND METHODS: Platelet metabolism was studied by activity of intraplatelet LDG, platelet secretory activity by plasm beta-TG and ADP-aggregation in 75 gravidae. Of them 16 had exacerbation of CGN, 40 females were in remission of CGN and 19 healthy pregnant women served control. RESULTS: Enhanced LDG activity and intensity of maximal ADP aggregation, high beta-TG levels compared to control were recorded in gravidae with CGN both in exacerbation and remission. The frequency of preterm deliveries, intrauterine growth retardation, neonatal hypotrophy was greater in women with gestational exacerbation of nephritis compared to pregnant women with stable nephritis. CONCLUSION: Metabolic and functional platelet hyperactivity with platelet intravascular activation in pregnancy in aggravated CGN suggest contribution of platelets to onset of the disease gestational exacerbation. Pregnancy-induced overactivation of platelets in CGN exacerbation stimulates intravascular coagulation in placental circulation with resultant microthrombi in placental vessels responsible for high rate of unfavorable pregnancy outcomes in relevant patients.

[Method of diagnosing liver tumors]. / Metod diagnostiki opukholevykh porazhenii pecheni.

The investigation is concerned with developing a model for using blood serum from patients with hepatic tumors to induce the activity of a gamma-glutamyl transferase enzyme in homogenized bovine hepatic tissue culture. An immunoglobulin protein was found to be responsible for the effect. The technique under discussion (Patent No. 2027997) provides a means for differential diagnosis of hepatoma and liver metastasis, on the one hand, and cholecystitis, pancreatitis, hepatitis, echinococcosis and hemangioma of the liver, on the other. The effectiveness of the method was 94.7; sensitivity--90.8 and specificity--97.9%.

[Reaction of lymphatic heart muscle in the adult grass frog to autotransplantation. Electron microscopic and electron microscopic autoradiographic studies]. / Reaktsiia myshechnoi tkani limfaticheskogo serdtsa vzrosloi travianoi liagushki na autotransplantatsiiu. Issledovanie metodami élektronnoi mikroskopii i élektronno-mikroskopicheskoi avtoradiografii.

Cellular and subcellular responses, related to destruction, survival and regeneration of striated muscle fibers in the autografted intact lymph heart of the adult frog, were examined by electron microscopy and electron microscopical autoradiography. It has been shown that a small number of myofibers in the grafted lymph heart wall undergo an irreversible degeneration. Numerous myofibers retain their almost normal ultrastructure up to the end of the 1st posttransplantation week and later. However, a considerable part of surviving muscle fibers exhibited various degrees of reversible degenerative changes. Prominent morphological features of activation, related to the nuclear and cytoplasmic ultrastructural reorganization, were revealed in these reversibly degenerated myofibers since the middle of the second week after autotransplantation. It is shown that some myonuclei of these reactive fibers are able to synthesize DNA. No mitoses were observed in myonuclei. The number of satellite cells (normally approximately 11%) doubled by the end of the 3rd week of posttransplantation. Evidence was provided for the formation of some myotubes in the interstitium outside the basal membrane of the original muscle fibers. Cellular sources of regeneration are discussed in addition to peculiar reaction of lymph heart muscle fibers to free grafting.

[Thrombocytic disorders in pregnant women with chronic glomerulonephritis and hypertension]. / Trombotsitarnye narusheniia u beremennykh s khronicheskim glomerulonefritom i gipertonicheskoi bolezn'iu.

To assess platelet changes in pregnant women with chronic glomerulonephritis (CGN) and essential hypertension (EH) we estimated platelet lactic dehydrogenase activity (LDH), beta-thromboglobulin and thromboxane B2 (TxB2) plasma levels and ADP-stimulated platelet aggregability. Five groups of gravidae (26-40 weeks of gestation) were studied: with EH (n = 20), with CGN and hypertension (n = 31), with CGN without hypertension (n = 29), with late toxemia (n = 11), nonpregnant CGN women (n = 10) and healthy pregnant women (n = 20). Activation of platelet function was found in gravidae with CGN and EH. Platelet disorders were especially pronounced in pregnant women with CGN and with EH, but they were less pronounced than in control group with late toxemia. We believe that hypertension is more important stimulating factor for platelet activation than renal disease. We suggest that platelet disorders in outpatients are brought about by endothelium damage caused by elevated blood pressure.

[Papillary cancer of the breast (clinico-morphological aspects)]. / Papilliarnyi rak molochnoi zhelezy (kliniko-morfologicheskaia kharakteristika).

Examinations of 23 specimens of papillary carcinoma of the mammary gland helped single out the basic criteria for the morphologic diagnosis of papillary carcinoma: numbers of cystic dilated ducts with ramified true and epitheliocyte-formed papillae. The cells are monomorphic, medium-sized, with large polymorphous and hyperchromic nuclei. Metastases to the regional lymph nodes were detected in 8.7 percent of cases. Total 3-year, 5-year, and 10-year survival in this form are 95.45 +/- 4.44, 89.09 +/- 7.41, and 89.09 +/- 7.41 percent, respectively. Since the prediction is favorable, radical resection of the mammary gland is recommended for patients with Stages I and II A papillary carcinomas of the mammary gland.

Reinfusion of UV-irradiated blood in the treatment of infection. I. Experimental infection with staphylococci.

During the last 10 years, reinfusion of UV-irradiated blood has been rediscovered again as a therapeutic method suitable in the treatment of a variety of diseases. The described series of model experiments on rabbits confirm its beneficial effects in the treatment of staphylococcal infection: the control animals reinfused blood not exposed to UV radiation died all within 48 hours after injected with a suspension of live Staphylococcus aureus culture; all rabbits reinfused UV-irradiated blood (2 ml per kg body weight) survived the whole period of observation (30 days); reinfusion of UV-irradiated blood in a volume reduced to 1 ml.kg-1 body weight prolonged the animals' life-span to 96 or 120 hours.