[Ultrastructural study of podocyte alterations in non proliferative glomerulopathy]. / Ul'trastrukturnoe issledovanie al'teratsii podotsitov pri neproliferativnykh glomerulopatiiakh.

Ultrastructural changes in podocytes are an important diagnostic and prognostic marker for nephropathies. However, the biomedical understanding of detected submicroscopic changes in podocytes remains controversial. OBJECTIVE: To investigate the relationship between the ultrastructural changes of podocytes (fusion of cytopodia and denudation of the basement membrane as a result of their desquamation) with a number of clinical and laboratory indicators of kidney dysfunction in case of non-proliferative glomerulopathies (NPGP). Thirty-seven patients (23 men, 14 women) with NPGP, including 8 with focal segmental glomerulosclerosis (FSGS), 17 with membranous nephropathy (MN), and 12 with minimal change disease (MCD), were examined. SUBJECT AND METHODS: All the patients underwent standard laboratory and instrumental studies: determinations of the levels of total serum cholesterol (mmol/l), total serum protein (g/l); serum albumin (g/l); CKD-EPI glomerular filtration rate (GFR) (ml/min/1.73 m2), and daily protein loss (g/day). Light optical changes were measured; completely sclerotic and/or focally segmentally sclerotic glomeruli were taken into account. Quantitative ultrastructural stereological analysis was carried out estimating the cytopodium width (CPW) and the degree of glomerular basement membrane denudation (GBMD) (%). RESULTS: NPGP cases showed the largest number of sclerotic glomeruli in FSGS, which was accompanied by the lowest level of daily proteinuria and GFR. Quantitative values of CPW were associated with the level of daily protein loss (r=0.47; p < 0.05) and serum albumin (r=-0.57; p <0.05) in patients with nephrotic syndrome. In MN, the absolute value of CPW was larger than that in the other two patient groups. A correlation analysis of CPW and GBMD values among patients with NPGP revealed a statistically insignificant negative relation between these morphometric parameters. However, when a subgroup of patients with podocytopathies (only MCD and FSGS) was identified in the study group, this relationship was found to be significant (r=-0.54; p=0.012). CONCLUSION: The patients with NPGP exhibited a relationship between the severity of nephrotic syndrome and proteinuria/hypoalbuminemia, on the one hand, and CPW, on the other. The established negative relationship between CPW and the percentage of GBMD in the subgroup of patients with podocytopathies may be due to the early stages of podocyte injury, which are accompanied by transient GBMD.

[CALCIFYING NANOPARTICLES IN PATHOMORPHOGENESIS OF STRUCTURAL LESIONS OF HEART VALVES].

Over the last 10 years, calcifying nanoparticles (CNP) have attracted attention as structures detected. together with many other nanostructures in biopsies from patients operated for the correction of aortic valve malformations. The results of the present work performed with the use of high-resolution transmission and scanning electron microscopes agree on the whole with the data of other authors. Some new findings include CNP adhesion to collagen fibers and specifically-shaped, shallow invaginations or craters at their surface. The possible pathophysiological mechanisms that promote involvement of CNP in the development of the disease are considered.

[Plasma membrane focal defects in structurally normal cells].

The technique of perfusion fixation through the rat kidney vasculature was modified to ensure the highest possible level of cell preservation close to that under in vivo conditions. Electron microscopic analysis of the tissue specimens treated in such a way revealed local defects of the plasma membrane in a number of cells than that otherwise looked normal. These findings together with the evidence for reparability of such defects and some data on the purely artificial nature of certain alterations should be taken into consideration in order to avoid misinterpretations while diagnosing the biopsy specimens.

[Type 3 membranoproliferative glomerulonephritis: an unusual variety of well-known pathology].

Light microscopy, immunohistochemistry, and submiscroscopy were used to study a renal biopsy specimen obtained from a 51-year-old old male suffering from type 3 membranoproliferative glomerulonephritis (MPGN-3) concurrent with HCV infection. Along with the signs characteristic of MPGN-3 (glomerulosclerosis; crescents; subendothelial and subepithelial deposits; proliferation of mesangial and endothelial cells), abundant pro- and myelocytes were found in the glomerular capillary lumens with active granular formation in the Golgi apparatus and with granular exocytosis into the glomerular basement membrane. Apoptotic elements were recorded among both glomerular and tubular epithelial cells.

[Choice of tools for ambulatory uvulopalatoplasty on the basis of experimental and clinical studies].

We studied interaction of a radiofrequency scalpel and a semiconductor laser with the tissue phantom. The heated part of the phantom can be easily removed and measured for size of the coagulated area. We investigated to what extent the width of the coagulation necrosis along the incision is influenced by the power and speed of each instrument used for incision. As for the laser, the width of the coagulation zone increased with rise of power and slowing of cutting velocity. For the scalpel, this relationship between incision velocity and coagulation effect was much weaker but had the same trend. We compared hemostasis efficiency during uvulopalatoplasty with both instruments. Much more potent hemostatic effect was registered in laser uvulopalatoplasty.

[Endoplasmic reticulum (a review of the literature)]. / Endoplazmaticheskii retikulum (obzor literatury).

Recent data on the structural and molecular organisation of the endoplasmic reticulum (ER) are reviewed. A special attention is paid to the mechanism of soluble and integral membrane protein translocation across the ER bilayer. A model of phospholipid-coupled polypeptide translocation is introduced served to overcome hydrophobic and (or) conformational constraints during the passage of polar amino acid residues within a polar environment of the ER membrane. In addition the mechanisms of diverse covalent and noncovalent post-translocational polypeptide modifications are considered, together with the process of the sorting events among ER-resident polypeptides and those destined to leave the ER domain for the Golgi apparatus.

[Coated vesicles and their functional role in animal cells]. / Okaimlennye puzyrki (coated vesicles) i ikh funktsional'noe znachenie v kletkakh zhivotnykh.

A review is made of the structural organization of coated vesicles, their contents, mode of formation and of destruction and intracellular routes in different cells of various species. Special attention is given to the participation of coated vesicles in the segregation and transferring of diverse substances in the cytoplasm, as well as to the final biological effects of these substances on cell metabolizm. The participation of coated vesicles in intracellular digestion, secretion and neurosecretion, in vitellogenesis and some other cellular function is considered in details.

[Electron microscopic study of glycogen granules in the epithelial cells of human endometrial glands]. / Elektronnomikroskopicheskoe issledovanie granul glikogena v kletkakh épiteliia zhelez éndometriia cheloveka

At the early secretory stage of the menstrual cycle part of glycogen granules in the human endometrial glandular cells are limited with threelayered membranes 6--9 nm thick. Transitional forms between the above mentioned granules and vesicles derived from the Golgi cisterns were found. On moving away from the Golgi apparatus the density of these granules with the membranes disappearing, thus suggesting that the Golgi apparatus of the human endometrial glandular cells may give rise to beta-particles of glycogen, whose polysaccharid nature is confirmed by amylase digestion.

[Modernization of the methods for detecting NADH diaphorase and glucose-6-phosphatase at the light-optical level]. / Modernizatsiia metodik vyiavleniia NADH-diaforazy i gliukozo-6-fosfatazy na svetoopticheskom urovne.

A technique for NADH-diaphorase and glucose-6-phosphatase activity visualization at the light microscope level has been essentially modified by the use of a short pre-fixation of cryostat sections in a gluteraldehyde-containing fixative followed by a prolonged (18-20 h) incubation in the chilled (4-6 degrees C) standard media. Besides, for revealing NADH-diaphorase Triton X-100 is recommended to add to the incubation medium. The offered technical modifications secure a high staining intensity and specificity of both histochemical reactions tested without any substantial sophistication of the procedure.

[Intensive drying of grids after lead citrate staining lowers precipitate contamination on the surface of ultrathin sections]. / Intensivnoe vysushivanie posle kontrastirovaniia protivodeistvuet vypadeniiu osadkov na poverkhnosti ul'tratonkikh srezov.

Two techniques are proposed to minimize precipitate contamination of ultrathin sections after lead citrate staining. According to the first one, stained and washed grids are placed, sections downward, on a stack of filter papers. The second one involves a consecutive washing of grids with distilled water, 96% ethanol, and n-hexane. Both techniques are equally efficient; the former being simpler in processing, while the latter is superior in the reproducibility of results.

[Electron microscopic study of horseradish peroxidase pinocytosis by white rat peritoneal macrophages in vivo]. / Elektronno-mikroskopicheskoe issledovanie pinotsitoza peroksidazy khrena peritoneal'nymi makrofagami belykh krys in vivo.

Pinocytic activity of rat peritoneal macrophages has been studied morphometrically at the ultrastructural level, 1.5-50 minutes following the intraperitoneal administration of horseradish peroxidase at varying concentrations. Internalization of the tracer was carried out by relatively large vacuoles and coated microvesicles. A probable mechanism underlying the two structurally distinct kinds of pinocytosis by macrophages is discussed.

[Submicroscopic study of adenohypophyseal cells with the hypophysis incubated in a hypertonic solution. I. The distribution of acid phosphatase, thiamine pyrophosphatase and arylsulfatase activities]. / Submikroskopicheskoe issledovanie adenogipofizarnykh kletok pri inkubatsii gipobiza v gipertonicheskom rastvore. I. Raspredelenie aktivnostei kisloi fosfataze, tiaminpirofosfatazy i arilsul'fatazy.

In the rat adenohypophyseal mammotrophs and somatotrophs, activities of acid phosphatase, thiamine pyrophosphatase and aryl sulphatase were defined after the incubation of the pituitary gland in the medium containing 1.65 M sucrose. Following 60--120 minutes of the incubation the above enzymes were found, in addition to their usual sites, in all cisternae of the Golgi apparatus and the rough endoplasmic reticulum as well as within the perinuclear cisternae. The mechanism of the above alterations is discussed, and a suggestion is put forward that the emergence of enzymatic activities in the above structures may reflect their biogenetic pathways in the cell.

[Submicroscopic study of adenohypophyseal cells with the hypophysis incubated in a hypertonic solution. II. Differential analysis of the intracellular distribution of arylsulfatase and thiamine pyrophosphatase within a single cell]. / Submikroskopicheskoe issledovanie adenogipofizarnykh kletok pri inkubatsii gipofiza v gipertonicheskom rastvore. II. Differentirovannyi analiz vnutrikletochnogo raspredeleniia arilsul'fatazy i tiaminpirofosfatazy v predelakh odnoi kletki.

In the rat pituitary gland exposed to an hour incubation in the medium containing 1.65 M sucrose activities of aryl sulphatase (AS) and thiamine pyrophosphatase (TPP) were successively defined within a single specimen. After the detection and registration of both enzyme activities with the electron microscope ultrathin sections were processed with 50% H2SO4 resulting in the complete removal of the reaction products for TPP. In the mammotrophs and somatotrophs, cisternae of the Golgi apparatus and rough endoplasmic reticulum showed either AS, or TTP or both the enzymatic activities. In the latter case the two enzymes could be restricted to a single cistern.

[Spatial distribution of thiamine pyrophosphatase and arylsulfatase activities in the area of the Golgi apparatus of the adenohypophyseal secretory cells in the white rat]. / Prostranstvennoe raspredelenie aktivnostei tiaminpirofosfatazy i arilsul'fatazy v oblasti apparate Gol'dzhi sekretornykh kletok adenogipofiza beloi krysy.

Specimens of albino rat pituitary glands were processed consecutively for demonstration of thiamine pyrophosphatase (TPPase) and aryl sulfatase (ArSase) activities. To differentiate between the structures associated with each particular enzyme studied within a single cell, either somatotroph or mammotroph, ultrathin sections were exposed for 2 minutes to 2--50% H2SO4 which removed the reaction products for TPPase rather than those for ArSase. The comparative study of pairs of micrographs of the same area taken before and after the etching with H2SO4 has shown TPPase and ArSase to reside in the Golgi apparatus and GERL system, respectively. Transitional elements have also been discovered, thus supporting the idea that GERL may be a derivative of the Golgi apparatus.

[Defining the optimal conditions for immunoperoxidase cytochemistry at the submicroscopic level]. / Opredelenie optimal'nykh uslovii immunoperoksidaznoi tsitokhimii na submikroskopicheskom urovne.

The authors tested a number of experimental protocols and chemicals known to facilitate permeabilization of tissues to immunoperoxidase markers without ultrastructural alterations of the cells to be examined. Monoclonal antibodies producing hybridoma lymphomas served as a primary test object. None of the procedures employed (i.e., quenching of the fixative aldehydes by some reducing agents; cryopermeabilization; treatment by detergents) were shown either to intensify stainability or to increase the penetration of immunoreagents into the tissue depth. The diffusion efficiency depended only on the marker molecular mass and the thickness of the vibratome section incubated. The Elder and coworkers (1983) two-step technique has been found superior in the preservation of both immunoreactivity and fine structure of the cell.

[Ultrastructural and metabolic characteristics of the thyrocytes exposed to cyclophosphane]. / Ul'trastrukturnye i metabolicheskie kharakteristiki tirotsitov pri deistvii tsiklofosfana.

Ultrastructural, morphometric and metabolic characteristics were studied in thyrocytes of 96 mice treated with immunosuppressing and cytostatic drug cyclophosphamide (CY), injected intraperitoneally every other day either for a short time (up to 6 days) in high doses (400 mg/kg) or for a long time (up to 70 days) in moderate doses (40 mg/kg). High doses of CY caused the reduction in thyrocyte height, NADH-diaphorase activity in their cytoplasm and protein content in the follicular colloid as compared to these parameters in a control group. The cisterns of rough endoplasmic reticulum (RER) underwent swelling and deformation with the loss of electron density of their contents. This was accompanied by mitochondrial swelling with matrix clarification and disorganization of cristae and an increase in the amount of cytoplasmic lipid droplets. Following long-term administration of CY in moderate doses the thyrocyte height exceeded that one in control group, while the NADH-diaphorase activity changed insignificantly and protein concentration in colloid increased. In the majority of thyrocytes the nucleus and major organelles retained their normal structure, while in some cells the cisterns of RER appeared dilated. Acid phosphatase activity was unchanged in both experimental groups. Thus, despite the approximately equivalent total CY dose in experimental groups, its damaging effect on thyrocytes was expressed much less in animals receiving the moderate doses of CY, that, probably, could be explained by the lack of CY cumulative effect and high rate of its metabolic degradation and clearance.

[Electron histochemical characteristics of laser necrosis]. / Elektronno-gistokhimicheskaia kharakteristika lazernogo nekroza.

Linear or dot-shaped lesions were inflicted on rat liver with Nd:YAG laser, and fine structural alterations of hepatocytes were studied in the specimens processed for an endoplasmic reticulum (ER) marker glucose-6-phosphatase (GP). 5-7 s after irradiation a severe cell damage and GP inhibition occurred near the lesions, with less injured cells located laterally. 24 hr later the zone of the necrosis increased markedly. An autolytic decomposition in the newly formed necrotic area was much more pronounced as compared to the area of the initial necrosis. Phagocytic resorption of the intensively irradiated tissue was retarded this explaining some clinical observations on the long-term healing after Nd:YAG laser surgery. Based on our observations the so-called contact regimen of the irradiation is recommended due to the small size of the initial necrosis produced with this method. The various patterns of cell injuries including some changes in ER and enzyme GP as its marker are described in detail.

[Submicroscopic changes in the mammary gland epithelium of white rats during chronic administration of high doses of estrogen]. / Submikroskopicheskoe izmeneniia v épitelii molochnykh zhelez belykh krys pri dlitel'nom vvedenii vysokikh doz éstrogena.

Ultrastructural alterations of female albino rat mammary glands in normal estrous cycle as well as in lactation, postlactation involution caused by weaning of sucklings from the nursing dams, and after administration of estrogen (synestrol) in a daily dose of 0.1 mg per 100 g body weight were studied. Particular attention was given to histochemical and morphometrical analys is of the secretory and vacuolar apparatus of the epithelial cells. A significant increase in the number of secretory granules in estrus as compared to that in diestrus accompanied by the reverse relationships in primary lysosome (small coated vesicles) count was observed. Similar relationships between the above parameters were found when the mammary glands of the lactating animals and those from the dams with their sucklings weaned were compared. The abundant primary and secondary lysosomes, autophagosomes, and heterophagosomes as well as some Golgi lamellae and smooth cisternae of endoplasmic reticulum filled with acid phosphatase were often observed in the epithelial cells of the involutary mammary glands, indicating the activation of the vacuolar apparatus. In synestrol-treated animals a gradual decrease in both secretory and catabolic activities occurred after transient successive increase of these two activities. The possible role of the relationship between the secretory and catabolic activities of the epithelial cells during mammary gland tumorogenesis is considered.

[In the dying cell secretory protein diffuses through the membranes into the cytosol]. / V gibnushchei kletke sekretornyi protein diffundiruet cherez membrany v tsitozol'.

Anti-horseradish peroxidase IgG (a-HRP) secreting hybridoma lymphoblasts grown subcutaneously in recipient mice have been studied light and electron microscopically 30-120 min following capitation of the animals. Conventional HRP-DAB immunocytochemical staining was performed for demonstration of a-HRP which in the living cells was restricted to the rough endoplasmic reticulum, the perinuclear cisterns, the Golgi apparatus and some microvesicles. 30 min after death in a number of the cells a-HRP began to invade the cytosol leaving, however, the nucleus and mitochondrial matrix free of the secretory marker. 30 to 90 min later staining intensity became similar in all cellular structures thereby making an impression of overall a-HRP spreading throughout the cell. In the light of these findings and the data obtained by other investigators a conclusion is made on the diffusion of macromolecules across intracellular membranes as a result of considerable post-mortem disturbances in membrane permeability.

[Epithelial permeability study of the bronchi in chronic bronchitis using an electron-dense tag]. / Issledovanie pronitsaemosti épiteliia brokhov pri khronicheskom bronkhite s primeneniem élektronno-plotnogo markera.

An electron dense tracer, horseradish peroxidase (HRP), treated with DAB-H2O2 for electron histochemical demonstration of peroxidase was used in an effort to find out whether or not the human bronchial epithelium was permeable to some macromolecules in chronic bronchitis. The biopsy specimens obtained from 5 patients at bronchoscopy and cultured in a medium with 2% HRP for 1-6 hours showed the reaction product to be distributed within intracellular spaces of the epithelium as well as in some vesicles and vacuoles of the apical portion of the ciliated cells. When 2% HRP solution was topically applied on the bronchial epithelium surface of a narcotized patient for 10 min followed by biopsy of this part of the epithelium fixation and treatment for HRP demonstration, electron dense precipitates were identified only within some intercellular spaces. It is suggested that some intercellular contacts of the bronchial epithelium in chronic bronchitis are permeable to macromolecules, and that the ciliated cells of the bronchial epithelium are capable of endocytosing some macromolecules from the surface of the mucosa.