The aim: to find out the effect of a two-hour tourniquets ischemia of the limb and acute blood loss on systemic disorders of the body in the postperfusion period.
Materials and methods: The experiments were performed on 96 nonlinear male rats weighing 200-220 g. All experiments were performed under sodium thiopental anesthesia.
In the first experimental group, two-hour tourniquets limb ischemia was simulated. In the second experimental group, acute blood loss was modeled. In the third experimental group, these lesions were combined. In 1 and 2 hours, as well as in 1, 7 and 14 days, the biliary function of the liver was determined in the experimental animals.
Results: Two-hour tourniquets ischemia of the limb in the reperfusion period compared with the control was accompanied by a significant decrease in the rate of bile excretion, which reached a minimum value in 3 h – 1 day of the experiment and returned to the control level in 14 days. After the simulation of acute blood loss, the rate disturbance of bile excretion became significantly greater in all observation periods. The complication of acute ischemia-limb reperfusion caused a greater decrease in the rate of bile excretion with a maximum in 1 day of the experiment. Under these conditions, at 1, 7, and 14 days, the indicator was statistically significantly lower than in the other study groups.
Conclusions: In the case of acute blood loss, complicated by two-hour tourniquets ischemia of the limb, the reperfusion period is accompanied by a summation of the negative impact of blood loss, ischemia and reperfusion of the limb on the functional state of the liver, which is a significant decrease in the rate of bile excretion with a maximum in 1 day, which further increases, but up to 14 days does not reach the level of control.
 KEY WORDS: blood loss, tourniquets, ischemia, reperfusion, bile excretion

   We studied the influence of maternal hypothyroidism on progeny skin morphogenesis by means of histological, histochemical and lectinhistochemical methods. Hypothyroid conditions in rats were achieved by daily food supplementation with antithyroid drug Mercazolil. The experiment was conducted on 10 control and 10 hypothyroid rats, which delivered 70 and 46 offsprings, respectively. We discovered that maternal hypothyroidism induces the accumulation of mast cells (MCs) in the skin of progeny on the 1st, 10th and 20th postnatal days, with decrease of these cell’s count returning to control level on 40th postnatal day. These results indicate that offsprings developing under conditions of maternal hypothyroidism are a risk group for changes in immune status and the occurrence of allergic reactions. The stratum corneum of epidermis, its lipid barrier as well as pilosebaceous units, in both control and experimental group animals, at the early stages of postnatal ontogenesis are enriched with carbohydrate determinants of αDMan, βDGal, βDGal(1–3)DGalNAc, αLFuc, αDGalNAc, αDGlcNAc, Neu5Ac. Galanthus nivalis agglutinin (GNA) is a selective histochemical marker of MCs, while Lactarius torminosus fungus agglutinin (LTFA) is a selective label of Langerhans cells. Maternal hypothyroidism resulted in reduction of lectin binding with the structural components of progeny skin and its derivatives. We speculate that alterations in glycoconjugate processing and degradation sequences have an impact on the cell signaling, formation of adhesive contacts, cellular proliferation and differentiation. The lectin set we used clearly demonstrated specific labeling of cellular subpopulations, monitoring glycoconjugates processing and degradation under physiological and pathological conditions in all skin components.
Keywords: skin, Mercazolil, hypothyroid rats’ progeny, lectin histochemistry.

UDC 577.615.324-027.2.615.076

   Creation of novel remedies efficient in supporting wound healing remains an actual task in pharmacology. Hydrogels showed high efficiency in wound healing and tissue regeneration due to viscosity, elasticity and fluidity that provide them with functional characteristics similar to that in extracellular matrix. The aim of the study was to create chitosan-based hydrogels functionalized with different components (chondroitin-6-sulfate, hyaluronic acid, N-stearoylethanolamine) and to estimate their biocompatibility and biodegradability in vitro. For the first time, a lipid substance N-stearoylethanolamine (NSE) known as suppressor of pro-inflammatory cytokines expression was used as hydrogel component (1.95 mg/g). FTIR analysis confirmed the complexation of chitosan molecule with hyaluronate, chondroitin-6-sulfate, NSE. MTT-test and Trypan blue exclusion test were used to study hydrogels cytotoxicity towards human cells of different tissue origin. Biodegradability of hydrogels was evaluated using direct hydrogel contact with cells and cellindependent degradation. It was shown that chondroitin-6-sulfate (<2 mg/ml), hyaluronic acid (<2 mg/ml) and NSE (26 μg/ml) did not demonstrate significant toxic effects towards pseudonormal human cells of the MCF10A, HaCat, HEK293 lines and mouse cells of the Balb/3T3 line. The studied hydrogels were stable in saline solution, while in a complete culture medium containing 10% fetal bovine blood serum they underwent degradation in >24 h. The identified biodegradability of the chitosan-based hydrogels is important for the release of noncovalently immobilized NSE into biological medium. Further studies on laboratory animals with experimental wounds are expected to explore the potential of created hydrogels as anti-inflammatory
and wound-healing agents.
K e y w o r d s: chitosan hydrogels, chondroitin-6-sulfate, hyaluronic acid, N-stearoylethanolamine, FTIR analysis, human pseudonormal cells, toxicity, biodegradability