A Study of the Cytotoxic Effect of Microcapsules and Their Constituent Polymers on Macrophages and Tumor Cells.

We studied the effect of different concentrations of polyelectrolytes poly(allylamine hydrochloride) (PAH) and polystyrene sulfonate (PSS) as well as the effects of microcapsules coated with these polymers on survival of Ehrlich ascites carcinoma cells and mouse peritoneal macrophages and on ROS production by phagocytes. PAH reduced viability of Ehrlich ascites carcinoma in a concentration-dependent manner (LD50=12-15 µg/ml). This effect was presumably determined by its ability to bind phosphates, thereby depleting the culture medium. At the same time, PAH did not affect the viability of macrophages. PSS produced no cytotoxic effect on the examined cells. Polyelectrolyte capsules with the shell architectonics (PAH/PSS)3 and (PAH/PSS)3PAH in the examined concentration range had no effect on the viability of macrophages and tumor cells. PAH microcapsules with positively charged surface much more rapidly and more intensively activated macrophages. The chemiluminescence response directly depended on the amount of capsules in the solution.

Effects of Polyelectrolyte Microcapsules with Different Surface Charge on Erythrocyte Sedimentation Rate.

Relationship between changes in the erythrocyte sedimentation rate in rats and concentration and charge of polyelectrolyte microcapsules was studied by the Panchenkov method. Positively charged microcapsules reduced erythrocyte sedimentation rate in a concentrationdependent manner. This effect was related to a decrease in the content of high-molecularweight proteins in the plasma due to their adsorption in positively charged microcapsules with polyacrylamide surface layer.

[Distribution of proteins inside polyelectrolyte microcapsules depend on pH of medium].

Distribution of bovine serum albumin and ferritin inside polyelectrolyte microcapsules was studied by transmission electron and confocal microscopy at the pH range 2-5. It was estimate that protein's distribution depends on isoelectric point (pI) and first polyelectrolyte used for preparation of capsule shell. So peptide is placed in the bulk of capsule if pH values of medium are lower isoelectric point of protein and polycation was used as a first polyelectrolyte layer. If the first polyelectrolyte was polyanion, the protein is located near internal surface of the shell. The protein is situated near internal surface of the shell for both polyelectrolytes when pH is equal to pI.

[Investigation of the influence of temperature on polyelectrolyte microcapsules containing and not containing proteins].

Using the methods of light scattering and optical microscopy, data on the thermosensitivity of hollow microcapsules generated by alternative layers of poly(allylamine) and poly(sterenesulfonate) polyelectrolyte and microcapsules with included polyelectrolyte complexes and proteins have been obtained. It has been shown that all three types of capsules shrink with increasing temperature and the time interval of thermal influence, and their diameter decreases. The thermosensitivity has been estimated by means of the temperature factor of shell shrinkage (Ec). For all three types of the microcapsules containing from 6 to 10 layers in the shell, the phenomenon of the thermosensitivity alternation depending on the number of shell layers was revealed. With an odd number of the shell layers, the shrinkage is stronger than with an even number. Using the transport proteins of blood hemoglobin and bovine serum albumin as an example, the dependence of the thermosensitivity of microcapsules on the quantity, the degree of ionization, and the conformational state of the incapsulating protein was investigated.

[An electron microscopy study of the structure of polyelectrolyte microcapsules containing protein and containing no protein].

Electron micrographs of ultrathin sections of polyelectrolyte microparticles containing protein and free from protein for the formation of which CaCO3 spherulites served as a core basis have been obtained and analyzed. Polyelectrolyte microparticles with the number of alternately layered polyelectrolyte layers of polystyrene sulfonate and polyallylamine from 6 to 11 have been studied. It follows from the data obtained that protein-free polyelectrolyte particles having the dimensions 4.5-5 mm are formations of an intricate internal organization, which consist of a set of threadlike and closed nanoelements of polyelectrolyte nature with a thickness of 20-30 nm. The particles containing six to eight polyelectrolyte layers lack the external envelope; therefore, they were called polyelectrolyte microspherulites. With the number of layers nine and more, when a polyelectrolyte envelope appears on the surface, they transfer into polyelectrolyte microcapsules. It was found that, in a protein-containing polyelectrolyte microcapsule, as distinct from protein-free polyelectrolyte microspherulite and microcapsule, polyelectrolytes are located only in the nearsurface layer, and the external spatially organized envelope restricts the internal volume filled with protein solution. As the number of polyelectrolyte layers increases, the thickness of the envelope increases. The reasons for such substantial differences in the structures of polyelectrolyte microcapsules formed on protein-containing and protein-free CaCO3 spherulite are discussed.

[Incapsulation of enzymes into polyelectrolyte nano- and microcapsules and the problem of the development of enzymatic microdiagnostics].

The incapsulation of proteins into polyelectrolyte microcapsules (PE-microcapsules) has been studied with the aim to develop microdiagnostica for the presence of low-molecular-weight compounds in native biological fluids. The problem was solved using two enzymes: lactate dehydrogenase and urease. Polyelectrolyte microcapsules were prepared using two polyanions: polystyrene sulfonate (PSS) and dextran sulfate (DS), and two polycations: polyallylamine (PAA) and polydiallylmethylammonium (PDADMA). CaCO3 microspherulites with the incapsulated enzyme served as a "core" in the formation of polyelectrolyte microcapsules. It was shown that the main problem in the preparation of a polyelectrolyte microdiagnosticum is the selection of an oppositely charged pair of polyelectrolytes optimal for the active functioning of the enzyme. It follows from the results obtained that the best polyelectrolyte pairs for the formation of the envelope of a PE-microcapsule are PAA/DS and PAA/PSS for lactate dehydrogenase and PSS/PDADMA for urease. Taking into account these data, we designed enzyme-containing microcapsules with different polyelectrolyte compositions and different numbers of layers and studied their properties.

[Some aspects of venous surgery based on advanced knowledge of venous anatomy of the lower extremities]. / Nekotorye voprosy flebokhirurgii v svete uglublennykh znanii anatomii venoznogo rusla nizhnikh konechnostei.

Analysis of the errors and faults in the treatment of the main diseases of veins of the vena cava inferior system showed that they can be explained to a great extent by the uncoordinated ideas of the anatomy of the lower limb venous channel. Our experience suggested the expediency of evaluating the condition of venous circulation from the standpoint of comprehensive consideration of the vascular channel structure. The suggested scheme of the structure of the venous channel with all the collaterals and the intravenous connections is systematized and logically completed. We employed it for analysis of some up-to-date problems of the diagnosis and treatment of the most frequently encountered diseases of the lower limb venous system. Knowledge of the general and individual characteristics of the structure of the venous channel opens up new opportunities for their use in surgery and at the same time protect against unreasoned and erroneous decisions.

[Anatomo-topographic basis for operations of transposition of the superficial femoral vein in post-thrombophlebitic disease of the lower extremities]. / Anatomo-topograficheskoe obosnovanie operatsii transpozitsii poverkhnostnoi bedrennoi veny pri posttromboflebiticheskoi bolezni nizhnikh konechnostei.

Post-thrombophlebitic disease of the lower limbs may be occlusive, (2%), recanalized (54%), and mixed (44%) in character. Operations for restoration of the normal flow of blood cannot remove the retrograde flow in veins with destroyed valves. Many variants of valve restoration have been suggested. The simplest among them are transplantation of the superficial femoral vein, the lateral circumflex femoral vein, and the great saphenous vein. The anatomical structure of the lower limb venous system was studied on the corpses of humans without venous pathology. In most cases the deep femoral vein was found to have one valve (46.8%), two (30.6%), and three (12.8%) valves. Valves were found at a distance of 1.0 to 4.5 cm from the site of drainage of the deep femoral vein into the superficial vein in 48.35% of cases, 43.5% had a sufficient diameter (0.6 cm). Thus, implantation of the superficial femoral vein into the deep vein can be conducted in less than half of the patients. Study of the anatomy of the lateral circumflex femoral vein showed that the operation can be performed in 9.6% of cases. The great saphenous vein is the main route of blood drainage in post-thrombophlebitic disease. Its use as a valvular protection is very risky. It should also be taken into account that in post-thrombophlebitic disease the great saphenous vein is dilated and marked by relative venous incompetence. Though they are relatively simple, these operations, can be therefore, performed in less than half of the patients. Examination of patients should include ultrasonic angioscanning and retrograde transfemoral phlebography, which allow precise determination of the anatomical features of the lower limb venous system in each patient.

[Comparative analysis of immune response indices in lung cancer]. / Sravnitel'nyi analiz pokazatelei immunologicheskoi reaktivnosti pri rake legkogo.

In 43 patients with cancer of the lung, a number of immunologic indexes were evaluated including lymphocyte response to PHA, lymphokine production, serum immunoglobulin levels and the regulatory effect of sera and T-lymphocytes on lymphokine production. No significant deficiency in T- and B-lymphocytes was registered in patients with stage I-III cancer, nor any rise in TG cell level was in evidence. The response to PHA was low; however, the capacity to produce lymphokines in the presence of allogeneic or autologous tumor antigens was unimpaired. Purified T- and TG-cells isolated from blood or tumor tissue suppressed immune response in leukocyte migration inhibition test. However, this effect was not related to the increase in levels of these cells in the circulation. The autologous sera of practically all tumor-sensitized patients exerted a blocking effect in vitro. This effect was related neither to levels of serum immunoglobulins, nor to those of blood TG cells.