[Reaction of population of pulmonary mast cells in rat bronchial asthma under the effect of ß-adrenoreceptor antagonists].

Multifunctional granular mast cells (MCs) are among targets in bronchial asthma (BA) therapy. We studied pulmonary MC population in a rat model of BA under the effect of ß-adrenoreceptor antagonists and of the latter combined with the standard therapy (glucocorticoid budesonide + ß2-adrenergic agonist salbutamol). MCs of different degrees of maturity were identified on paraffin section of lung stained with Alcian blue and Safranin. MC density in the lung of rats with BA increased 1.9 times. Alcian blue-positive immature cells predominated in the lungs of both intact rats and rats with BA. In response to pharmacological agents, the mean MC densities were reduced in 2-2.7 times in all the variants of experiments and were close to the norm. It allows us to suppose that MCs migration from the outside was suppressed and, in consequence of the decline of MC densities, the release of the mediators involved in the progression of BA may be diminished.

[A sharp increase in the density of pulmonary and pericardial mast cells under monocrotaline-induced pulmonary hypertension in rats].

Multifunctional granular mast cells (MCs) are involved in various pathological processes. The response of MC populations of myocardium, pericardium and lung to pulmonary hypertension (PH) has been studies 8 weeks after injection of monocrotaline. Five intact and five experimental rats were used. The density of MCs of different maturity was estimated on paraffin sections stained with Alcian blue and Safranin. Expressiveness of PH was estimated by functional parameters with the help of echocardiograms and by morphological markers. The MC density in myocardium of the intact and experimental rats was relatively low: 2 to 4 cells/mm2. MC density in the pericardium of intact rats was 14 times higher than in myocardium and increased 3 times for PH. The mature Safranin-positive cells predominated (70-80%) in myocardium and pericardium of intact and experimental rats. The MC density in the lungs of intact rats was about 30 cells/mm2; 98% of these cells were immature Alcian-positive cells. The mean density of MCs in the lungs of rats with PH increased 5.6 times. The mature Safranin-positive cells appeared in the lungs of rats with severe pathology. The greatest number of MCs in lungs was in the rats with the most pronounced disorders of myocardium function and marked histological damages (injuries) of myocardium and lungs. The finding show active response of MC population to monocrotaline-induced PH that stimulates migration of immature MCs into pericardium and lungs from the outside. Our data indicate the important role of MCs in the pathogenesis of PH.

[Effect of renin-angiotensin system inhibitors on the density of myocardial, pericardial and pulmonary rat mast cells under experimental heart failure].

Activation of the renin-angiotensin system (RAS) plays a critical role in the pathogenesis of heart failure (HF). We studied the effect of lisinopril (LP) and fosinopril (FP), the inhibitors of angiotensin-converting enzyme, and of losartan (LT), the antagonist of Angiotensin II receptors, on the behavior of multifunctional mast cells (MCs) under experimental HF. The inhibitors of RAS were daily injected during 4 weeks in 4 weeks after two (at 24-h interval) isoproterenol injections. MCs of different degrees of maturity were identified on paraffin sections stained with Alcian blue and Safranin. Expressiveness of HF was estimated by functional parameters with the help of echocardiogram and by morphological markers. The MC density in the myocardium of the intact rats as well as of the rats with HF, both treated and untreated with the preparations, was relatively low: from 3 to 4 cells/mm2. The MC density in the pericardium of the intact rats was several times higher than in the myocardium: 35 +/- 7 cells/mm2. The density ofpericardial MC under HF was 1.7 higher than that in the intact rats at the expense of the increase in the density of Alcian-positive immature cells (P < 0.05). The injections of LP increased the MC density still in 1.4 times at the expense of the density of Safranin-positive mature cells (P < 0.01). The injections of FP and LT had no influence on the MC density and the balance of cells of different degrees of maturity in the pericardium. 96-99% of MCs in lung were Alcian-positive cells. The density of such cells in the intact rats, in the rats with HF, and in the rats with HF treated with FP was 30 cells/mm2. The injections of LP and LT decreased the density of pulmonary MCs up to 7 cells/mm2 (P < 0.01) and 19 cells/mm2 (P < 0.05), respectively. Functional parameters of the hearts were consistent with the data of morphological analyses. Myocardium function improvement was noted only in the rats with HF treated with FP and LT. The reaction of MCs (as cell elements of "tissue" RAS) to injections of inhibitors of RAS was various in the myocardium, pericardium and lung of the rats with HF. The injections of LP stimulated maturation of the resident MCs in the pericardium and the replenishment of the population through immature cells migrating from the outside. It allows us to suppose an intensification of secretory activity of the cells. In contrast, the injections of LP and LT reduced the pulmonary MC population.

[The density of myocardial and pericardial rat mast cells in isoproterenol-induced heart failure].

Myocardial mast cells (MC) respond to cardiovascular pathology. The behavior of MC population in myocardium and pericardium of rats has been studied 24 h, 14, 28 and 60 days after two isoproterenol injections (at 24 h intervals). The extent of heart failure has been estimated by supersonic inspection 28 and 60 days after isoproterenol injections. The density of MCs of different degrees of maturity was estimated on paraffin sections stained with Alcian blue--Safranin. The MC density in myocardium of intact and experimental rats was relatively low: from 4 to 6 cells/mm2. The MC density in pericardium of intact rats was several times higher than in myocardium: 48.6 +/- 13.0 cells/mm2. In 24 h and 14 days after isoproterenol injections the pericardial MC density was 1.5 times higher than in control rats (P < 0.05) at the expense of increase in the number of mature MCs with Safranine-positive granules without the increase in the number of immature cells with Alcian blue-positive granules. In 28 days the pericardial MC density was 2 times higher than in intact rats (P < 0.05) at the expense of increase in number of immature and mature cells. In 60 days after isoproterenol injections the pericardial MC density and the ratio of immature and mature cells compared with control did not reach statistical significance. The changes in pericardial MC population corresponded to severity of heart failure according to functional indices. The findings show active reaction of pericardial MCs on myocardium dysfunction that stimulates the maturation of resident immature MCs in pericardium and migration of immature cells to pericardium of damage heart.

[Comparative characteristics of some pharmacological models of chronic heart failure].

Effectiveness and informativity of various experimental pharmacological models of the chronic heart failure (CCF) in rats have been evaluated by means of ultrasonic cardiography. It has been established that isoproterenol (INN isoprenaline) (80 mg/kg, s.c., twice with an interval of 24 h) produces compensated CCF. Monocrotaline (single dose 60 mg/kg, i.p.) caused the development of pulmonary hypertension, right ventricle hypertrophy, and pronounced right ventricle failure in a fraction of experimental rats. Doxorubicin administration (cumulative doze 15 mg/kg, i.p., divided into 6 injections within 14 days) resulted in decreased left ventricle contractility, eccentric heart remodeling, and CCF development in most cases. Thus, the doxorubicin-induced disorder in rats can be considered the most effective pharmacological model of CCF, leading to the development of pronounced and progressive CCF in the majority of experimental animals.

[Activation of mast cells after experimental myocardial infarction in 3 week-old rats].

It is known that mast cells (MC) take an active part in regeneration processes in postinfarction heart in adult rats and humans. Behaviour of population of cardial MCs has been studied 20, 60, 75 and 90 days after experimental myocardial infraction induced in 3 week-old and adult rats by ligation of left coronary artery. The density of MC of different degrees of maturity was estimated in atrium and ventricle on paraffin sections stained with Alcian blue - Safranin. Findings were compared with MC density obtained in hearts of intact rats. The MC density in intact 1.5-2.5 month-old rats in atrium and ventricle was about 0.6 cells/mm2, in intact 3.5-4.0 month-old rats in atrium--1.2 cells/mm2, in ventricle--0.6 cells/mm2. The MC density in 3 week-old rats with infarction was significantly higher than in intact rats: 5-fold increase in 20 and 60 days in atrium, and 2-fold increase in 60 and 75 days in ventricle. In 60 days after infarction the MC density in adult rats was 3 times lower in atrium and 2 times lower in ventricle than in the same heart compartments of 3 week-old rats with infarction. After infarction in 3 week-old rats, a relative share of young cells with alcian-positive granules sharply increased in 20 days and then decreased by 60-75 days. This indicates a migration of immature MCs to infracted myocardium and their subsequent differentiation. The MC activation after infraction in young rats may result from a more active immune reaction in younger rats and/or functional peculiarities of their MC.

[Human fetal ventricular cardiomyocytes in vitro: proliferation and differentiation].

In this study, in the primary cell culture of human fetal cardiomyocytes proliferation of myocytes combines with their differentiation. The cells were isolated enzymatically from 19-22 week-old human fetuses and cultured for 14 days. DNA synthesis, ultrastructure and presence of atrial natriuretic peptide (ANP) were examined. In 7 day-old culture, the myocytes make about 60%, in 14 day-old culture--about 50%. Myocytes synthesize DNA and divide mitotically. After a 24 h incubation with 3H-thymidine in 7 day-old culture 1.8 +/- 0.5% of muscle and 25.2 +/- 11.7% of non-muscle cells are labeled, in 14 day-old culture--2.5 +/- 0.5 and 8.1 +/- 1.7% of cells are labeled, respectively. In 7 and 14 day-old cultures the degree of redifferentiation of contractile apparatus in myocytes varies from scattered actin and myosin filaments surrounded by ribosomes to differentiating myofibrils with distinct sarcomeres and Z-discs. Single electron-dense granules, morphologically similar to secretory atrial granules, display ANP-immunoreactivity. Thus, human fetal ventricular cardiomyocytes in cell culture proliferate, differentiate and synthesize ANP for 14 days; this is indicative of vitality of these cells.

[Clinical value of morphological analysis of radial and internal mammary arteries in patients with ischemic heart disease]. / Klinicheskoe znachenie morfologicheskogo analiza luchevoi i vnutrennei grudnoi arterii u bol'nykh ishemicheskoi bolezn'iu serdtsa.

Morphological study of intraoperative biopsies of radial and internal mammary arteries taken from patients with ischemic heart disease during coronary bypass surgery revealed structural alterations of intima and media mostly in radial artery. These alterations depended on severity of arterial hypertension, presence of diabetes and extent of atherosclerosis. Hyperplasia of intima could be a basis for radial artery spasm and cause of graft stenoses and occlusions. Therefore protocols of pre and post operative care should take into consideration individual risk factors and morphological and functional features of an artery used for grafting.

[Heart ventricular cells of neonatal rats in a culture: DNA synthesis, ultrastructure and localization of atrial natriuretic peptide]. / Kletki zheludochkov serdtsa neonatal'nykh krys v kul'ture: sintez DNK, ul'trastruktura i lokalizatsiia predserdnogo natriiureticheskogo peptida.

We determined the optimal conditions suitable for expanding cardiac cells in vitro for their future use in experimental transplantation into injured myocardium of adult animals. Ventricular cardiac cells were isolated enzymatically from 2-3 day-old rats and cultured at different cell densities within 5-7 days to 4 weeks. Mixed cultures of muscle and non-muscle cells were examined by light autoradiography, electron microscopy, and immunogold method. The best results were obtained at a density of 3 x 10(5) cells/ml in the medium, consisting of 90% DMEM and 10% fetal calf serum, during 5-7 days of cultivation. In such cultures myocytes made 62.5 +/- 7.9%. After a 24 h incubation with 3H-thymidine, 22.0 +/- 2.2% of myocytes were labeled. Muscle cells contact with each other and with non-muscle cells, contain myofibrils, contract and display atrial natriuretic peptide (ANP)-like immunoreactivity.

[Mitotic activity, ploidy and ultrastructure of cardiomyocytes from human embryo and fetuses]. / Mitoticheskaia aktivnost', ploidnost' i ul'trastruktura kardiomiotsitovémbrionov i plodov cheloveka.

There is an evidence that mitotic activity of human cardiomyocytes in late fetal and early postnatal ontogenesis is very low. But little is known of the division of human cardiomyocytes at earlier stages of development. In this study mitotic activity of ventricular and atrial human cardiomyocytes of 4-8-week-old embryos and 17-32-week-old fetuses has been studied. On these stages the mitotic index is relatively low to reduce moderately within the 1st to the 3rd trimester of pregnancy from 1.4 to 0.7%. These findings are consistent with the data on cell ploidy demonstrating the presence of relatively small share of myocytes with 3c and 4c DNA in ventricles of 6-8-week-old embryos and 12-22-week-old fetuses. The share of such cells in the 1st and 2nd trimesters of pregnancy varies from 19 to 24% and from 8 to 18%, respectively. Cells with 3c and 4c DNA are most likely to be in mitotic cycle. This assumption is supported by electron microscope pictures showing all phases of typical mitosis. Cyclic changes of myofibrillar ultrastructure during mitosis of prenatal human cardiomyocytes are the same as during mitosis of low differentiated myocytes in mouse and rat hearts. These results suggest that in prenatal human cardiomyogenesis the level of myocyte differentiation and the cell number increase at slow rate.

[Correlation between the level of polyploidy and hypertrophy and degree of human atrial cardiomyocyte damage in certain congenital and acquired heart pathologies]. / Korreliatsiia mezhdy urovnem poliploidii i giptertrofii i stepen'iu povrezhdeniia kardiomiotsitov predserdiia cheloveka pri nekotorykh brozhdennykh i priobretennykh patologiiakh serdtsa.

Cardiomyocytes of right auricle of human heart were obtained during open heart surgery from 5 patients (aged from 3.6 to 6 years) with congenital heart diseases (atrial septal defect and tetralogy of Fallot, group I), from 6 patients (aged from 20 to 49 years) with mitral valve disease (group II) and from 5 patients (aged from 40 to 54 years) with ischemic heart disease (group III). Ploidy and area of cells were measured in isolated myocytes after the Feulgen staining. The degree of ultrastructural changes in myocytes was evaluated by means of scoring system (0 to 3). The hearts of group I patients contain 55 to 82% of cells with 2c DNA, 18 to 41% of cells with 4c DNA and 0 to 4% of cells with 8c DNA. In the hearts of group II patients the level of cell ploidy is high: the share of 2c-cells varies from 1 to 24%, that of 4c-cells--from 1 to 55%, that of 8c-cells--from 13 to 57%, that of 16c-cells--from 1 to 39%, the share of cells with 32c and more DNA--from 3 to 47%. The ploidy of myocytes of group III patients is not high: 2c-cells make 35 to 55%, 4c-cells--39 to 55%, 8c-cells--5 to 15%. The average cell areas in group II are 4 times larger than in group I, and twice as large as in group III. In most cases, cells of all ploidy levels are larger in patients with higher average cell sizes. In all patients with increased ploidy levels, cells grow in size, but the rate of this growth is lower than that of cell DNA content. In all patients the myocytes with different degrees of organelle damage are observed. In patients of groups I and III there is a direct correlation between the extent of myocyte ploidy and hypertrophy, on the one hand, and the degree of cell degeneration, on the other hand. In contrast, in group II patients an inverse relation between ploidy and extent of degenerative changes of cells is evident.

[Antibodies to skeletal muscle myofibrillar proteins in the diagnosis of rheumatoid arthritis]. / Antitela k miofibrilliarnym belkam skeletnoi myshtsy v diagnostike revmatoidnogo artrita.

One of the mechanisms of muscular involvement in RA patients was elucidated by identification of antibodies to skeletal muscle components (actin and tropomyosin). Clinical symptoms of skeletal muscles involvement in RA proved to be associated with elevated count of tropomyosin antibodies against an insignificant informative value of actin. Enzyme immunoassay with tropomyosin has established a direct correlation between the number of the antibodies and RA activity.

[The DNA and protein content of atrial cardiomyocytes and the size and ultrastructure of the cardiomyocytes in children with congenital heart defects]. / Soderzhanie DNK i belka v kardiomiotsitakh predserdiia, razmery i ul'trastruktura kardiomiotsitov u detei pri nekotorykh vrozhdennykh porokakh serdtsa.

Right atrium myocytes were obtained during open heart surgery from 4 patients with atrial septal defect, and from 1 patient with Fallot's tetrad, aged from 3.5 to 6 years. DNA and total protein contents were measured consecutively in isolated cells using two-wave-length scanning cytophotometry following double staining after Feulgen and with naphthol yellow S. All the biopsies contained polyploid myocytes: the number of 4c cells varied from 18 to 41%, that of 8c cells from 0 to 4%. The average values of cells area and protein content for tetraploid cells were 1.5 times higher that for diploid ones. In all the cases normal and little changed myocytes were dominating (69-91%). The highest number of cells with moderate and severe degenerative changes was registered in the cases of maximum degree of cell hypertrophy and polyploidization.

[Cytophotometric, morphometric and electron microscopic studies of the cardiomyocytes of the human atrium in ischemic heart disease]. / Tsitofotometricheskoe, morfometricheskoe i élektronno-mikroskopicheskoe issledovanie kardiomiotsitov predserdiia cheloveka pri ishemicheskoi bolezni serdtsa.

During an open-heart surgery, right atrium myocytes were obtained from 5 patients (aged from 40 to 54 years) with ischemic hearts. The DNA and total protein contents were measured in isolated cells using two-wavelength scanning cytophotometry after the double staining: the Feulgen nuclear reaction followed by naphthol yellow S staining. The degree of ploidy in the investigated myocytes appeared not high. The number of 2c, 4c and 8c cells varied, respectively, from 35 to 55%, from 39 to 55%, and from 5 to 15%. The share of 2-nuclear cells varied from 0 to 6%. The average square of tetraploid cells is more than that of diploid cells. In cells of a given size the protein content is higher in cells with higher ploidy, but the difference is not of 2-fold value. Larger 2c and 4c cells had more protein. In all the patients examined, along with myocytes with normal ultrastructure, some myocytes with different degree of organelle damage were observed, whose share varied in different patients. Myocytes with moderately to heavily damaged organelles are of more frequent occurrence in patients who displayed as well the highest myocyte hypertrophy and the largest share of polyploid cells.

[DNA synthesis in myocytes of the heart conduction system in myocardial infarction induced in young rats]. / Sintez DNK v miotsitakh provodiashchei sistemy serdtsa pri infarkte miokarda, indutsirovannom u molodykh krys.

The myocardial infarction was induced in 13- and 21-day-old rats. The extent of the DNA synthesis activation in myocytes of conduction system compartments compared with atrial and ventricular myocytes was evaluated by cumulative indices of labeled nuclei after 30-times-repeated 2H-thymidine injections at 12h intervals. Labeling indices in atrial myocytes are two to six times higher (0.01 < P < 0.001) in rats with infarction than in control. On the contrary, in conduction system compartments there are no statistically significant differences between control and experimental groups (P > 0.05). An activation of DNA synthesis can occur in myocytes of His bundle in some animals.

[The properties and areas of use of an immobilized granulated streptolysin O]. / Svoistva i oblasti primeneniia immobilizovannogo granulirovannogo streptolizina-O.

The heat-resistant preparations of immobilized granulated streptolysin O were obtained with the use of emulsion polymerization techniques. In experiments the presence of gangliosides and cholesterol affected the hemolytic activity of immobilized streptolysine O. The preparations thus obtained were used in the enzyme-linked immunosorbent assay as antigens for the determination of specific antibodies in rheumatic fever patients and healthy persons.

[Changes in the ultrastructure and DNA and protein content in human atrial myocytes in cardiac hyperfunction due to mitral valve defects]. / Izmeneniia ul'trastruktury, soderzhaniia DNK i belka v miotsitakh predserdiia cheloveka pri giperfunktsii serdtsa, vyzvannoi porokami mitral'nogo klapana.

Biopsies from human right auricles were obtained during open heart surgery, prior to valve replacement, from six patients (aged from 20 to 49 years) with rheumatic heart disease. DNA and the total protein contents were measured in isolated myocytes by means of the two wave-length scanning cytophotometry after the double Feulgen and Naphthol yellow S staining procedure. In all the biopsies polyploid hypertrophied myocytes predominate. The hypertrophic, nondegenerated cells and the cells with degenerative changes of varying severity (in the first place, changes of contractile apparatus and membranes) are present. The highest degree of cell ploidy occurs in patients of functional class IV according to the New York Heart Association classification, 72 to 98% of cells displaying octaploid and higher DNA values. With the increase in ploidy of myocytes in series 2c----4c----8c----16c----32c----64c the protein content increases only as 2.0----3.0----5.8----7.8----13.0----16.8. Neither direct correlation between the ploidy level and the degree of cell degeneration, no inverse correlation between the degree of degeneration and the value of ejection fraction was observed.

[Immobilization of myocardial tropomyosin]. / Immobilizatsiia miokardial'nogo tropomiozina.

Immobilized granulated myocardial tropomyosin with magnetic properties of prolonged period of storage was obtained. Tropomyosin inclusion into granules emulsion polymerisation is 89.5%. The largest amount of protein is extracted from granules after 2-3 days, and later this process is slowed down. Studying temperature effect, the maximum protein output was also shown to be reached after 2-3 days and depends on the temperature. Processing of granules, containing tropomyosin, with a solution with high concentration of hydrogen ions, the same results were obtained as under the influence of temperature. The use of 3 mol potassium rodanide did not show the peak of protein washing out of granules.

[The use of immobilized granulated G-actin with magnetic properties in patients and experimental animals]. / Ispol'zovanie immobilizovannogo granulirovannogo G-aktina s magnitnymi svoistvami v klinike i éksperimente.

G-actin was isolated from human myocardium, purified by means of repeated polymerization-depolymerization and immobilized in polyacrylamide gel by the procedure of "emulsion granulation". The granules acquired magnetic properties after addition of iron protoxide hydrophilic particles into the polymerizating mixture. Immobilized G-actin in granules with magnetic properties was used for production of specific antisera, for isolation of pure antibodies by means of an immunosorption procedure, for estimation of specific antibodies in blood sera of patients with rheumatism, of healthy persons and of experimental animals using immunofluorescent, immunoenzymatic and radioimmune procedures.

[Sensitivity to chlorine-containing disinfectants of Salmonella typhi strains with transmissible drug resistance]. / Chuvstvitel'nost' k khlorsoderzhashchim dezsredstvam shtammov Salmonella typhi s transmissivnoi lekarstvennoi ustoichivost'iu.

Sensitivity of 201 S. typhi strains to antibiotics and chlorine containing disinfectants was studied. The strains were isolated in the regions of the Left Bank Ukraine. Among the typhoid cultures the proportion of R+ strains was on the whole equal to 29.3 per cent. Chloramine resistant strains amounted to 21.8 per cent. Among chloramine resistant strains the number of the cultures carrying R plasmids was 3 times higher than that of the strains sensitive to disinfectant. Simultaneous transfer of the R plasmid and chloramine resistance to the recipient strain suggested that resistance of the S. typhi strains to chloramine was of plasmid nature.