INFLUENCE OF THE ACUTE INFLAMMATION AND ENDOTHELIAL DYSFUNCTION ON THE RETINAL VEIN OCCLUSION FORMATION AFTER CARDIO-SURGICAL INTERFERENCES WITH THE USE OF CARDIOPULMONARY BYPASS

Acute inflammation and endothelial dysfunction (EDF) are typical pathological processes, which determine the development of retinal vein occlusion (RVO) during cardio-surgery with the use of cardiopulmonary bypass (CB), but the connection of seromarkers according to the terms of occlusion appearance remains undefined. The aim – to determine the influence of the acute inflammation and EDF for RVO formation after cardio-surgical interferences with the use of CB according to the terms of occlusion appearance. Material and methods. There were selected for the research the data of 137 eyes (126 patients, the main group) with RVO after the surgery with CB. The comparison group contains the data about examination of 86 eyes (43 patients), who had not any occlusion during all term of examination. The control group consisted of 10 eyes (5 patients) without occlusion, which were examined before surgery. An ophthalmologist 2, 7, 30, 60, 90 and 180 days after cardio-surgical interference, examined patients. The content of IL-6, IL-8 and VE-cadherin in blood serum was determined by immunoenzyme technique (Bender Medsystems, Austria). Statistical data processing was performed with the use of Statistica 10 program (StatSoft, Inc., USA), regression analysis – with the use of the program package GLZ. Results. The conduction of cardio-surgeries with the use of CB caused an increase of the interleukins content in the early period (IL-6 on the 2nd and 7th days, and IL-8 up to 30 days), while the content of VE-cadherin (VE-C) was slightly increased during almost all period of monitoring. With the availability of RVO, the content of IL-6 during all terms of occlusion appearance was significantly higher, the content of IL-6 was up to 30 days, and the content of VE-C in a greater degree was after the 7th day. The regression analysis showed that after 1-2 days RVO appearance was directly related with the content of IL-6 and IL-8 in the blood, on the 3rd and 7th days – only with the content of IL-8, on the 8th and 30th days – with the content of all markers, and then with the content of IL-6 and VE-C. The accuracy of the prediction of the presence or absence of RVO at the appropriate period according to the calculated regression model is at least 78 % (p <0.001), what proves the influence of markers on the development of RVO. Conclusions. The undertaken study shows the meaning of the acute inflammation and EDF by appearance of RVO with the use of CB, what justifies the application of the preventive measures - at the early stages the restriction of activity of the inflammatory process, at the later stages – prevention of EDF development.

Immunocytochemical localization of cytosol 5'-nucleotidase in chicken liver.

We investigated the localization of cytosol 5'-nucleotidase in chicken liver by use of a pre-embedding immunoenzyme technique. Cytosol 5'-nucleotidase was purified from chicken liver and a monospecific antibody to this enzyme was raised in a rabbit. Fab fragments of the antibody were conjugated with horseradish peroxidase. Tissue sections of the fixed chicken liver were incubated with the peroxidase-Fab fragments, followed by DAB reaction for peroxidase. By light microscopy, dark-brown staining was present in the cytoplasm of parenchymal cells, Kupffer cells, and endothelial cells. The latter two types of cells were stained more strongly than the former. By electron microscopy, reaction deposits were present in the cytoplasmic matrix but not in cell organelles, such as mitochondria, endoplasmic reticulum, and peroxisomes, or in nuclei. In control sections incubated with peroxidase-conjugated Fab fragments from non-immunized rabbit, no specific reaction was noted. The results indicate that cytosol 5'-nucleotidase is contained more in the sinus-lining cells and less in the parenchymal cells, and that the enzyme is present in the cytoplasmic matrix of these cells.

Immunocytochemical localization of cathepsin B in rat kidney. I. Light microscopic study using the indirect immunoenzyme technique.

Localization of cathepsin B in rat kidney was studied using immunocytochemical techniques. Cathepsin B was purified from rat liver and antibody to it was raised in rabbits. The antibody reacted with a lysosomal extract of rat kidney to form a single precipitin line in a double-diffusion test. Immunoblot analysis of lysosomal cathepsin B of rat kidney showed two species of 29K and 25K MW. After removal of Epon, semi-thin sections of glutaraldehyde-fixed tissue were stained by the indirect immunoenzyme technique. Dark-brown reaction product, indicating the antigenic sites for cathepsin B, was found in cytoplasmic granules throughout the nephron. Staining intensity and size of the positive granules varied widely in each segment of the nephron. In the glomeruli and distal tubules, a few small cytoplasmic granules were stained. In the proximal tubules, the S1 segment exhibited many large granules which were most heavily stained, whereas the S2 and S3 segments contained few positive granules. All segments of the distal tubules showed the smallest amount of positive granules. A few positive granules were also noted in the cortical and medullary collecting tubules. Control experiments confirmed the specificity of the staining. The results indicate that the major site for cathepsin B in rat kidney is the S1 segment of the proximal tubule which is known to actively take up proteins leaked through the glomerulus.

Immunohistochemical localization of thyroxine (T4) and triiodothyronine (T3) in the rat thyroid gland under various experimental conditions

Detection of T4 and T3 in paraffin sections of the rat thyroid gland in various functional states was attempted by an immunoenzyme technique. Both hormones showed a similar localization, being detected mainly in the colloid in the follicular lumen, and sometimes in certain regions of the cytoplasm of the follicular epithelium. Following administration of TSH, their stainability was increased, and the localization within the cytoplasm became more remarkable. Decrease or disappearance of stainability was observed after hypophysectomy or administration of PTU. These findings support the view that the immunostainability of T4 and T3 is closely correlated with the functional state of the thyroid gland.