[The kinetic parameters of retainment and release of deposited in human tissue structures calcium].

The analysis of retainment and release kinetics of deposited in tissue structures calcium was made in the hypercalcemic conditions in 28 healthy volunteers (22 males and 6 females) of the age of 33 ± 6.5 years via drip infusion (Groups 1, 2) and in 9 individuals (3 males and 6 females) in 12 trials via stream infusion (Group 3). By the end of each hour after the termination of calcium infusion the amount of calcium retained in tissues was calculated (Mtis./kg); the parameters of its binding (specific buffer volume--ß3 sp, association constant--Ka, number of binding centers--n) were established using the Langmuir and Scetchard coordinates. The Group 1 volunteers (n = = 12) showed a section of positive cooperativity (a direct regression on Sketchard coordinates, Hill coefficient 3.36 ± 1.63) and 2 sections of the consecutive calcium separation from one set of noninteracting centers. 5 volunteers of Group 2 and 8 volunteers of Group 3 demonstrated a slight calcium delivery to tissues after 1 hour of observation which then followed for 2 volunteers of Group 2 and for 2 volunteers of Group 3. Other volunteers of Groups 2 and 3 showed a release of tissue-deposited calcium via the mechanism of the consecutive separation from one set of noninteracting centers with ßsp 3 times less and Ka 7 times higher than with the calcium infusion. The excretion of calcium in urine was the highest in Group 1 and the lowest in Group 3. The [Ca2+] and Mtis./kg values were normalized in Groups 1 and 2 the next morning and in Group 3 after 2-3 hours of observation. An assumption was made about the relationship between the tissue and kidney [Ca2+] normalizing mechanisms with hypercalcemia.

[Correction of cronic liver failure by transplantation of liver cells suspension and cell-engineering designs (experimental investigation)].

On an experimental model of chronic fibrotic liver damage (male rats Wistar (n-60), damage of CCl4, the duration of the experiment 90 days) it was studied the effectiveness of cell therapy for the correction of chronic liver failure. These rats were divided into 3 experimental groups: in the Ist-group (control, n=10) isotonic saline (650 mkl.) was injected; in the IInd-group (n=20) suspension of liver cells was applicated in a dose 8 - l0 x 10(6) cells; in the IIIrd-group (n=30) suspension of liver cells and bone marrow cells (mesenchymal stromal cells) in ratio 5:1 were used as cell associates on microparticles intjectable heterogeneous biopolymer hydrogel "SpheroGEL" (cell-engineering design) in common dose 8 - l0 x 10(6) It was ascertained that in the 2nd and in the 3rd groups the accelerated normalization of disturbed liver functional indices (ALT, AST, ALP) took place - to 30 days, but in the control group only to 90 days. The reliable differences in rats ofnormalization offunctional indices were absent between the IInd and the IIIrd groups. But in 90 days by using special histological dyeing it was found out that defibrotic processes in liver tissue were more expressed in the IIIrd group in comparison with the IIIrd group. Received results were consequence of prolonged vital activity of cells (liver cells and mesenchymal stromal bone marrow cells) into cell-engineering designs, which were transplanted in the IIIrd group. The obtained effect can be explained by that the developed cell-engineering designs provide adequate conditions for prolonged vital activity of the transplanted cells.

[Kinetic parameters of calcium binding to tissue structures in human acute hypercalcaemia].

Calcium binding kinetic by tissue structures was analysed in 19 health volunteers (13 men and 6 women) of the age group of 33 +/- 6.5 under conditions of acute hypercalcaemia followed by a drip i.v. infusion of calcium gluconate over 2.5 hours. At the end of each 30-minute period the calcium amount retained by tissue structures was recorded and the kinetic parameters of calcium binding were determined according to Langmuir and Scatchard. In all volunteers there was a segment of binding isotherm with positive cooperativity (direct regression in Scatchard) with analogous buffer capacity (beta) for calcium in Langmuir (0.58 +/- 0.24 L kg). One half of volunteers demonstrated cooperativity at [Ca++] 1.3-1.5, while another--at [Ca++] 1.0-1.3 mmol/L which corresponded to the differences in the association constant (Ka) and the number of interactive sites (n) with [Ca++] 1 mmol/L. Additionally, two segments of binding isotherm were detected with the successive binding of calcium to one set of noninteractive sites with similar kinetic parameters of calcium binding (beta, K(a), n). Four different curves of calcium binding in healthy volunteers were established. This study may serve as the basis for a functional diagnostic test of disorders of the tissue calcium-binding properties in different pathological conditions.

[Clinico-experimental basis of application of biodegradable implants in surgical treatment of peripheral nerve lesions].

Aim of this study was to provide clinico-experimental basis of application of biodegradable implants in surgical treatment of peripheral nerve lesions. Experimental investigations were performed on non-pedigree female rats with body weight of 200-250 grams. 10 animals were used, 20 sciatic nerves were operated. Morphological examinations were done on 21st and 101st days after surgery. Clinical part of the study was performed after finishing of experimental block and included 20 patients with peripheral nerve lesions. Information about patients included: sex, age, type and localization of lesion, preoperative neurological status, data of electrophysiological and ultrasonographic studies and MRI. Follow-up period varied between 3 and 12 months. The series included 13 male and 7 female patients, age was 20-63 years. Injection of "Sphero(r)Gel" in the area of nerve suture enables more effective regeneration of axons and their growth through the injury site. Application of "ElastoPOB(r)" membrane favors demarcation of peripheral nerve from surrounding scar tissues in the wound. Acquired data demonstrate decreased influence of scarring process on the nerve trunk in the postoperative period according to neurological examination, electroneuromyography, ultrasonography and MRI.

[Experimental study of the pharmacokinetics of acetylsalicylic acid upon transdermal administration].

A comparative analysis of the concentrations of acetylsalicylic acid (ASA) and its metabolite, salicylic acid (SA), in the blood was performed and the other pharmacokinetic parameters were studied after conventional intragastric and transdermal administration in rabbits. It is established that the ASA concentration in the blood upon transdermal administration in a dose of 100 mg/kg is above the minimum therapeutic level during 24 h. Prolonged action of a transdermal therapeutic system (TTS) in comparison to intragastric administration was proved. After the transdermal administration via TTS, the ASA half-elimination time and the mean retention time increased in comparison to the values upon intragastric administration. The ASA bioavailability upon the transdermal administration increased by a factor of 4.5 in comparison to the case of peroral administration.

[The influence of multipotent mesenchymal stromal cells of bone marrow on process of local radiation injury in rats after local beta-irradiation].

The influence of multipotent mesenchymal stromal cells (MMSC), got from allogenic bone marrow, on local radiation injury in rats after beta-irradiation by a source 90Sr/90Y in a dose 140 Gy under various conditions and ways transplantation was studied. It was established, that transplantation MMSC, allocated on biodegradative membranes, which have been carried out for 21 day after an irradiation in conditions minor surgical of ulcers, resulted in reduction of the area of local beam defeats and acceleration of healing of skin, in comparison with the control of an irradiation. The introduction of suspension MMSC subcutaneous around of the center of a defeat at 8 day after an irradiation caused earlier healing of ulcers. It was concluded, that application of MMSC is perspective for treatment of local radiation injury and necessity of development of optimum conditions of their use at cell therapy of radiation injury of skin.

[Matrix transdermal systems for caffeine delivery based on polymer and emulsion compounds].

The goal of this work was to develop and test transdermal therapeutic systems for caffeine delivery. In vitro experiments showed that the rate of caffeine diffusion through untreated rabbit skin from a transdermal therapeutic systems based on polymer compound containing 50 mg medicine was 67.2 (9.1 microg/cm2h; for a system based on emulsion compound it was 173 (19 microg/cm2h. Methods for studying the caffeine release rate and quantitative measurement of caffeine content in the emulsion-based transdermal therapeutic system were developed. These methods are required to obtain data for standard drug documentation. The results of in vivo experiments in rabbits showed the absence of irritating effect of the emulsion-based transdermal therapeutic system. The obtained data on the specific efficiency of the transdermal therapeutic systems for caffeine delivery (50 mg) in healthy volunteers showed that this medicine could be used as a nonnarcotic psychoactivator for improving mental and physical activities and attention concentration.

[A comparative study of the dynamics of deep burns healing in using medullary allogenic fibroblast-like mesenchymal stem cells from bone marrow immobilized on biodegrading membranes or taken from cultural plastic].

Transplantation of suspention of allogenic fibroblast-like mesenchymal stem cells (AFMSC) of the bone marrow and AFMSC immobilized on the biodegradable membrane (BM) stimulates healing of deep burn wounds compared to control (without cell transplantation) because these cells produce biologically active substances into the wound. AFMSC immobilized on BM activate repair processes in the wounds earlier. This may be due to their higher functional activity created in the monolayer by adequate intercellular interactions. To avoid complications caused by low diffuse properties of BM (protein precipitation), BM should be removed from the surface of the burn wound maximum 3 days after its application.

[Transdermal delivery of insulin]. / Transdermal'nye sistemy vvedeniia insulina.

A possibility is studied of the transdermal delivery of insulin by using a mixture of synthetic analogues of phosphoglycerides (SAP), as a potential activator of hormone diffusion, through the skin. Experimentally in vitro, it was proven that the diffusion of insulin through the skin of two types of transdermal therapeutic form (TTF)--matrix-type and matrix-hydrogel-type--is possible only in presence of activator SAP-M-99. The detected optimal composition of insulin matrix TTF with the area of 40 sq cm enabled a trandermal hormone diffusion speed of 0.26 UNITS/h, which is compatible with the secretion of insulin by the pancreas of an adult (0.25-1.5 UNITS/h). A change-over for the matrix-hydrogel system of insulin delivery based on a 40 sq cm collagenous sponge enabled to increase the insulin diffusion up to 0.54 UNITS/h.

[A comparative analysis of the two approaches to the evaluation of thrombogenic qualities of the biomaterials in vitro]. / Sravnitel'nyi analiz dvukh podkhodov k otsenke trombogennosti biomaterialov v usloviiakh in vitro.

The paper contains an analysis of thrombogeneticity of biomaterials in vitro with regard for the changing time of recalcification of the platelet-free and platelet-related human blood plasma after incubation with a studied sample. Two sizeless thrombogeneticity parameters are suggested for describing the thrombogeneticity of studied biomaterials irrespective of a incubation-medium composition. It was demonstrated by the example of glass granules, which were modified by various immobilized functional groups, that the measuring of re-calcification time performed after samples' incubation secures a more objective assessment of platelet-resistant properties of materials. The relative index of thrombogeneticity of biomaterials (with the above glass granules being the controls), unlike the standardized parameter, opens up the possibility to study comparatively the activating action exerted on the coagulating and anti-coagulating blood systems of the studied samples, which differ by the ratios of their surface squares to the incubation-medium volume.

[Interaction of heparin with amino group-containing materials]. / Vzaimodeistvie geparina s aminosoderzhashchimi materialami.

The influence of the glass surface immobilized aminogroups nature and the experiments conditions on the interactions between heparin and aminogroups containing surface was studied using total internal reflections fluorescence. It was shown, that the nature of the terminating aminogroup, but not its mobility or the amount of the aminogroups in the main chain, is the major factor influencing the heparin adsorbtion from individual solution. Both the nature of terminating aminogroup and its mobility determine heparin adsorption on the aminogroups containing surfaces from the blood plasma proteins containing solution. Heparin irreversibly adsorbed from individual solution is not replaced by blood plasma proteins. The tertiary aminogroups containing surface adsorbs maximal quantity of the heparin.

[Competitive adsorption of human fibrinogen on an amorphic quartz surface]. / Konkurentnaia adsorbtsiia fibrinogena cheloveka na poverkhnosti amorfnogo kvartsa.

The kinetics of adsorption of fibrinogen on the surface of amorphous quartz from binary solutions containing human serum albumin and gamma-globulin was studied by the method of fluorescence of total internal reflection. A model of energetically nonhomogeneous interactions in the surface/protein system was used to explain the mechanism of competitive adsorption of proteins.

[Mathematical processing of human platelet distribution according to size for determination of cell heterogeneity]. / Matematicheskaia obrabotka raspredeleniia trombotsitov cheloveka po razmeram dlia vyiavleniia geterogennosti kletok.

The paper proposes a method for mathematical treatment of the distribution of human platelets by sizes to detect the heterogeneity of cell populations. Its use allowed the authors to identify three platelet populations that have different parameters of size distribution. The proposed method opens additional vistas for analyzing the heterogeneity of platelet populations without sophisticating experimental techniques.

[Comparative effectiveness of the use of hematological methods to analyze the procoagulant activity of biological materials]. / Sravnitel'naia éffektivnost' primeneniia gematologicheskikh metodov dlia analiza prokoaguliantnoi aktivnosti meditsinskikh materialov.

Whether clinical hematological methods by employing two parameters: (plasma recalcification time (PRT) and activated partial thromboplastin time (ACTT) can be used to examine the procoagulant activity of the surface of biological materials was comparatively studied. The method PRT was shown to be sensitive and reproducible while ACTT failed to significantly record differences in the procoagulant activities of various materials. The authors chose the optimal protocol for evaluating the blood compatibility of biological materials: the surface area/plasma volume ratio is more than 150 cm2/ml, the incubation time of samples is no less than 180 sec at 37 degrees C and continuous mixing.

[Use of the fluorescence method in the study of protein adsorption onto the surface of intraocular lenses]. / Primenenie metoda fliuorestsentsii dlia issledovaniia adsorbtsii belkov na poverkhnosti intraokuliarnykh linz.

The paper shows it possible to use the method of total internal reflection-fluorescence for study of the kinetics of protein adsorption onto the surface of intraocular lenses (IOL) made from optically transparent hydrophobic polymers and hydrogels. The nature of the materials (polymethylmethacrylate, silicone, p-HEMA, p-HEMA-collagen copolymer) used for IOL on the adsorption and desorption of human serum albumin was studied.

[The use of laser diffraction spectroscopy for measuring the size distribution of human thrombocytes]. / Primenenie lazernoi difraktsionnoi spektroskopii dlia izmereniia raspredeleniia po razmeru trombotsitov cheloveka.

The proposed procedure for obtaining the distribution of human platelets by their sizes by using laser diffraction, followed by computer processing of the results permits studies by defining quantitative parameters, such as the pattern of distribution, the count of platelet populations and the variations in platelet counts in the populations. This procedure may be used to examine the functional status of cells in health and in disease.

[Methodology of obtaining and digital processing of adhesive cells]. / Metodika polucheniia i tsifrovoi obrabotki izobrazhenii adgezirovannykh kletok.

The paper deals with the methodological features of investigations of the interaction of cells and foreign material surfaces by scanning electron microscopy. The proposed technique for obtaining electron microscopic images results in their easy binarization and further treatment using computer software. The specificity of obtaining electron microphotographs permits detailed quantitative image analysis. This approach enables one to study both quantity of objects and their individual morphologies, as well as to define distributions of objects by their diameter, area, etc. Human platelet adhesion to glass and metal is considered as an example.

[Adsorption of human serum albumin on the original and albuminized surface of quartz]. / Adsorbtsiia chelovecheskogo syvorotochnogo al'bumina na iskhodnuiu i al'buminizirovannuiu poverkhnosti kvartsa.

The method of TIRF is used for obtaining kinetic curves of HSA adsorption on coated and uncoated silica surfaces. Using the model of energetic heterogeneous surface an analysis of positive effect of passivation as a result of transformation of energetic heterogeneity of quartz surfaces from rectangular to exponential is performed.