[Influence of activator and inhibitors of Ca2+ channels on proliferative activity in Tetrahymena pyriformis infusoria].

It was determined that change in DNA content in macronuclei occurs in the T. pyriformis infusoria under the influence of an activator (caffeine) and inhibitors of Ca2+ channels (verapamil), NiCl2, and CdCl2. Caffeine (10 mM) stimulates DNA synthesis. Verapamil (5 microM), CdCl2 (125 microM), and NiCl2 (100 microM) decrease DNA content in macronuclei by 30 min after proliferative stimulation. By 4 h of incubation, there is, on average, 10% less DNA in macronuclei of Tetrahymena preprocessed with verapamil than in the control cells. The cells preprocessed with CdCl2 and NiCl2 differ from the control cells by lower DNA content almost at all studied periods, but they restore the level of nuclear DNA by 4 h. It is assumed that transmission of proliferative signals in the T. pyriformis has a Ca2+ -dependent character.

Up-to-date concentrations of long-lived artificial radionuclides in the Tom and Ob rivers in the area influenced by discharges from Siberian chemical combine.

The Siberian Chemical Combine (SCC) is located in Seversk (formerly known as Tomsk-7) in the Tomsk Region of the Russian Federation. The main contribution of radionuclides in the SCC process water discharged into the Tom River was from the single-pass reactors, now removed from service (the last SCC reactor was shutdown on June 5, 2008). The data on the concentrations of (90)Sr, (137)Cs, (239,240)Pu and other artificial radionuclides in water, bottom sediments and flood-plain soils of the Tom and Ob rivers from Tomsk to the confluence of the rivers, are presented and discussed. The results of measurements carried out after shutdown of the last SCC single-pass reactor indicated no radiologically significant consequences of SCC activities for the studied water environment compartments. Contemporary activity concentrations of long-lived artificial radionuclides (3)H, (90)Sr, (137)Cs and (239,240)Pu in river water were below the intervention levels established by current regulations of the Russian Federation for these radionuclides. The results of (3)H analysis in water from the Tom and Samuska rivers demonstrated no inflow of contaminated formation water to surface water from the sites where liquid radioactive wastes of the SCC were injected below the surface. However, the density of flood-plain soil contamination by long-lived (137)Cs in the area influenced by SCC liquid discharges was higher than regional technogenic background. There were local flood-plain areas contaminated not only by (137)Cs, but also other gamma-emitters, such as (60)Co and (152)Eu.

[Changes in the DNA content in EGF-stimulated ciliate Tetrahymena pyriformis under effect of caffeine, KCl, and inhibitors of PLC and PKC].

It has been established that DNA content in ciliata Tetrahymena pyriformis changes under effect of the caffeine, and PLC and PKC inhibitors (U 73122 and chelerythrine accordingly). 10 mM caffeine stimulated DNA synthesis in these cells, 1 microM U 73122 and 25 microM chelerythrine, on the contrary, blockades the synthesis. 50 and 500 mM KCl, the agent which causes depolarization of plasma membrane stimulating Ca2+ entry into the cells, has no effect. The transduction of the proliferative signals in T. pyriformis cells has been proposed to be of Ca(2+)-dependent nature.

[Cellular aspects of hypertrophic cardiomyopathy pathogenesis: the role of cardiomyocyte polyploidy and activation of the proliferating cell nuclear antigen in the myocardium].

Mechanisms of hypertrophy development in hypertrophic obstructive cardiomyopathy (HOCM) have not been enough investigated. In our study, there have been examined patients with severe HOCM at different ages, including children, and patients with essential arterial hypertension (EAH). There was found, that HOCM in children compared to adults was characterized by considerable interventricular septum (IVS) hypertrophy and it was accompanied by the acceleration of cardiomyocyte polyploidy. The average ploidy level of cardiomyocytes in children with HOCM was higher than analogous indices in adults. The average ploidy level of nuclei, the part of PCNA-positive nuclei and polyploidic nuclei of cardiomyocytes in aduls with HOCM were authentically higher than in patients with EAH. Activation of the nuclear antigen in stromal cells was detected only in patients with HOCM. Our findings provide evidence of an important role of cardiomyocyte polyploidy and activation of the proliferating cell nuclear antigen in development of the myocardial hypertrophy in patients with HOCM.

[The influence of the EGF on proliferative signal transduction in ciliate Tetrahymena pyriformis].

It has been shown that the transduction of the proliferative signal induced by EGF in ciliata Tetrahymena pyriformis cells is not connected with autophosphorylation of the receptor tyrosine kinases. The results obtained indicate that EGF in ciliata cells initiates the mitogenic pathway including the membrane proteins of the tyrosine kinases-like type (without tyrosine phosphorylation sites), adenylate cyclase, tyrosine- and Ca2+ -dependent ERK-like kinases.

[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.

[A cytophotometric study of the influence exerted by epidermal growth factor on RNA and protein synthesis in the ciliate Tetrahymena pyriformis]. / Tsitofotometricheskoe issledovanie vliianiia épidermal'nogo faktora rosta na sintez RNA i belka v infuzoriiakh Tetrahymana pyriformis.

The influence of EGF (10(-8) M) on RNA and protein synthesis in the ciliate Tetrahymena pyriformis was studied. It was found that EGF stimulated RNA and proteins synthesis in T. pyriformis within 3 h after addition of EGF.

[Epidermal growth factor stimulates proliferation and DNA synthesis in ciliate cells]. / Stimuliatsiia épidermal'nym faktorom rosta proliferatsii i sinteza DNK v kletkakh tsiliat.

We studied the effect of murine epidermal growth factor on cell proliferation and DNA synthesis in macronuclei of ciliate Tetrahymena pyriformis G1. Mitogenic effect of epidermal growth factor on proliferation-induced tetrahymena cells has been revealed. This effect is due to the induced progression of cells at G1 and, consequently, their earlier entering DNA synthesis phase of the first cell cycle. Epidermal growth factor had no mitogenic effect on the resting cells from stationary culture (G0 phase) whose development is independent of the growth factors in the medium.

Ploidy levels and the number of nuclei in cardiomyocytes of the lamprey and fish.

It is well known that polyploidization of cardiomyocytes (CMC) is an essential component of heart growth in the warm-blooded vertebrates. Using the Feulgen cytophotometry of alkali-dissociated cells, we determined the ploidy in CMC of the lower vertebrates: lamprey Lampetra fluviatilis (Cyclostomata), skate Bathyraja maculata (Chondrostei), sterlet Acipenser ruthenus, and Russian sturgeon Acipenser güldenstädti (Ganoids), as well as paradise fish Macropodus opercularis, Amur sleeper Perccottus glehni, and Atlantic salmon Salmo solar (Teleostei). The data obtained have demonstrated a wide variety in CMC ploidy of both cyclostomata and fishes. About 85% of the lamprey CMC contain 2 or more (up to 17) nuclei per cell; with 90 and 10% of the nuclei being, respectively, diploid and tetraploid. Hearts of the skate and sturgeons contain mononucleated diploid CMC. In the perch-like fishes, mononucleated diploid and mononucleated tetraploid CMC make, respectively, 95 and 5%. The salmon heart contains near 50% of mononucleated diploid CMC, 13% of mononucleated tetra- and octaploid CMC, the rest CMC being multinucleated (up to 6 nuclei per cell). In all the examined species, the increased nuclear ploidy is accompanied with a significant increase in the nuclear volume. The number of nucleoli per nucleus does not correlate with the nuclear ploidy level. Evolutionary aspects of CMC polyploidy in chordates are discussed.

[The influence of epidermal growth factor and insulin on proliferation and DNA synthesis in ciliates Tetrahymena pyriformis]. / Vliianie épidermal'nogo faktora rosta i insulina na proliferatsiiu i sintez DNK v kletkakh tsiliat Tetrahymena pyriformis.

The influence of the epidermal growth factor (EGE) (10(-8) M), insulin (10(-6) M) and EGF (10(-8) M) in combination with insulin (10(-6) M) on proliferation and DNA synthesis in the nuclei of ciliates Tetrahymena pyriformis GL was studied. Insulin and EGF, known to stimulate growth of many types of mammalian cells revealed a mitogen influence on the unicellular eukaryotes. This effect involves stimulation of DNA synthesis, rising synchronization of cell division (upon the influence of EGF), and increase in cell number during the exponential growth. The mitogen effect may be evoked by cell progression in G1-phase under the action of growth factors and, consequently by earlier entry of cells into S-phase of the first cell cycle. Insulin repressed division of cells that entered into the generative cycle. These cells were delayed in late S-phase and G2-phase of the cycle. Part of these cells perished, while other cells could successively overcome the cell block to start their division by the 4th hours of cultivation. A collateral cytotoxic effect of insulin was found, being most prominent in early periods of Tetrahymena cultivation.

[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.

[Modelling of myocardial hypertrophy in vitro for solving problems of medicinal correction]. / Modelirovanie gipertrofii miokarda in vitro dlia resheniia voprosov medikamentoznoi korrektsii.

The work has been done on primary heart culture from neonatal rat ventricle. Cardiomyocyte hypertrophy was modelled using noradrenaline (NA), angiotensin II (AII) and fetal serum, respectively. Cell hypertrophy of primary heart cultures was assessed by measuring the surface area, the scope of protein synthesis estimated by 3H-leucine autoradiography and the contents of nucleic acids in gallocyanin-chromalum stained cardiomyocytes. The structure of myofibrillar apparatus was studied by rhodamine-conjugated phalloidin and indirect immunofluorescence of muscle alpha-actinin. Treatment with 10(-6) M NA increased 3H-leucine incorporation in 9-day old heart culture by 42% without changing cell size. AII in a dose 1 microM stimulated protein synthesis activity by 1.3 fold and the surface area by 1.7 fold, both in 2- and 9-day old primary heart cultures. The maximum stimulation of cell hypertrophy was provided by the medium supplemented with fetal serum. RNA contents in the cytoplasm of cardiomyocytes increased by 7.8 fold and the myocardial cell size by 2.9 fold in serum-supplemented culture by 9 days of cultivation. In the medium with fetal serum, amounts of cardiomyocytes with tetraploid nuclei reached 33%, against 14% in control. Coculturing of myocardiocytes and fibroblasts rendered effects of fetal serum on the growth of myocardiocytes. Cultivation in the presence of 1 microM enalapril, an ACE inhibitor, suppressed the development of cardiac muscle cells hypertrophy. The effect of enalapril depended on the degree of cellular hypertrophy. Addition of 10 microM amiloride to the medium lowered the protein synthesis by 29% independently on the initial cellular hypertrophy.

[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.

[Comparative study of the level of cardiomyocyte hypertrophy in patients suffering from ischemic heart disease with normal arterial pressure and in hypertension]. / Sravnitel'noe issledovanie urovenei gipertrofii kardiomiotesitov patsientov, stradaiushchikh ishemicheskoi bolezn'iu serdtsa pri normal'nom arterial'nom davlenii i v sostoianii gipertenzii.

A study was performed on the human atrial myocytes, isolated by means of alkaline dissociation of bioptates, obtained in clinics during open heart surgery, from two groups of patients. The first group consisted of 8 men aged from 40 to 62 years, with ischemic heart disease and normal arterial blood pressure. The second group involved 10 men, aged from 43 to 67 years, with ischemic heart disease and arterial hypertension. The nuclear DNA and the total protein in the cytoplasm of isolated cardiomyocytes were revealed by means of the two consecutive tests: the Feulgen and Naphthol yellow S staining. DNA and protein contents were determined by two wave-length scanning cytophotometry. It is ascertained that the temperate arterial hypertension exerted no appreciable influence on the polyploidy level of atrial myocytes, which correlated with the age of patient; in both groups (coefficients 0.7 and 0.4 for the first and second groups, resp.). The myocyte areas correlate with their ploidy (coefficients--0.7 and 0.7, accordingly). The protein contents in myocytes also correlate with their ploidy (0.5 and 0.7). The average protein content in myocytes of patients in the second group is higher than in the first group, the difference is most distinct when calculating the protein quantity per 2c-genome (717 +/- 34 a. u. and 517 +/- 51 a. u., resp.). A comparison of cytochemical and morphometric indices with some clinical findings obtained by means of echocardiography allowed to reveal the presence of correlative connections among them. The protein contents in myocytes correlate with the left ventricle mass (coefficients 0.6 and 0.5 for the first and second groups, resp.). The correlation between the rate of transmitral blood flow and myocyte ploidy (0.8 and 0.9), and the correlation between the myocyte area and the rate of transmitral blood flow (0.8 and 0.8) look convincing.

[A cytophotometric study of the amount of DNA in the macro- and micronuclei of the infusorian Nyctotherus cordiformis from the gut of tadpoles and juveniles of the common frog]. / Tsitofotometricheskoe issledovanie kolichestva DNK v makro- i mikronukleusakh infuzorii Nyctotherus cordiformis iz kishechnika golovastikov i segoletok travianoi liagushki.

The investigations of DNA amounts in the nuclei of the ciliate Nyctotherus cordiformis was continued. The measured ciliates were collected from guts of tadpoles and the most small frogs just from the water. The DNA content was measured (in arbitrary units--a.u.) in the nuclei of cysts, precysts, vegetative ciliates and small frogs. This paper is the second part of the investigation. The first part dealt with DNA amounts of the nuclei of the vegetative ciliates tested in spring, autumn and winter. The amount of the Feulgen-DNA complex was measured with a two-wave microcytophotometer MCFU-1. The average content of the DNA ranged from 1.7 +/- 2 a.u. in the Mi of cysts to 2.6 +/- 0.1 a.u. in the Mi of the youngest frogs. The DNA content of the Ma ranged from 275 +/- 21 a.u. in cysts to 479 +/- 25 a.u. in vegetative ciliates. The DNA content in presynthetic Mi (G1) is supposed to be approximately 1.3 and approximately 2.6 a.u. in postsynthetic (G2) Mi. Using nuclei of erythrocytes of Rana temporaria as internal standard, the DNA content of a 2 c Mi from tadpoles of N. cordiformis is supposed to amount to 0.57 pg or approximately 350 gDa. The DNA amount in Ma of N. cordiformis is at average 140-220 times as that of Mi.

[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.

[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.

[Changes in the cytochemical and morphometric characteristics of the myocytes in the right heart of the rat in adrenal-regeneration hypertension]. / Izmeneniia tsitokhimicheskikh i morfometricheskikh kharakteristik miotsitov pravykh otdelov serdtsa krysy pri adrenalregeneratornoi gipertenzii.

A study was made on myocytes of the right atrium and the right ventricle of rats with experimental arterial hypertension due to adrenal regeneration. The nuclear DNA and the total protein in the cytoplasm were revealed using two consecutive tests: the Feulgen reaction and Napththol yellow S staining. A two-wavelength scanning cytophotometry was used for measuring DNA and protein contents. It has been ascertained that the polyploidy levels in the myocytes of the right atrium and the right ventricle, measured 6, 12 and 26 weeks after nephradrenalectomy by the Skelton technique, differ from the control level negligibly. A small increase in the share of tetraploid myocytes occurred in 12 weeks in the right atrium, and in 26 weeks in the right ventricle. The mean nuclear volume in atrial myocytes decreased gradually within 12 weeks, to increase afterwards but did not reach to control level in 26 weeks. The cytoplasm volume of atrial myocytes decreased in 6 weeks, and then was seen to increase gradually approaching the norm within 26 weeks. At the same time the mean cytoplasm volume of ventricular myocytes, which remained practically unchanged within 6 weeks, decreased within 12 weeks more than twice as compared to the control level, but in 26 weeks started to rise, although less quickly than the volume of atrial myocytes. The common protein content was seen to decrease 6 weeks after the operation both in atrial and ventricular myocytes, then gradually approached the control within 26 weeks, these changes in atrial myocytes being sharper than in ventricular myocytes.

[The effect of adrenal-regeneration hypertension on protein-nucleic acid metabolism in rat atrial cardiomyocytes]. / Vliianie adrenalregeneratornoi gipertenzii na belkovo-nukleinovyi obmen v predserdnykh kardiomiotsitakh krys.

A study was made on cardiomyocytes of the left atrium of rats with experimental arterial hypertension due to adrenal regeneration. The nuclear DNA and the total protein in the cytoplasm were revealed using two consecutive tests: the Feulgen reaction, and Naphthol yellow S staining. The two-wavelength scanning cytophotometry was used for measuring DNA and protein contents. It has been ascertained that the polyploidy level in the nuclei of the left atrium myocytes, measured 6 and 9 weeks after nephradrenalectomy, remained the same as in the intact animals. The increase in the share of tetra- and octaploid nuclei occurred by 12 weeks. The mean nuclear volume decreased gradually throughout 9 weeks and subsequently was seen to increase, but by 12 weeks it did not yet reach the control level. The cytoplasm volume decreased by 6 weeks, then started to increase by 9 weeks to reach almost the standard level. The common protein content in the atrial myocytes decreases also by 6 weeks, but restores more rapidly than the volume, and reaches the standard level by 9 weeks, exceeding the same by 12 weeks. Thus, the prolonged hyperfunction of the atrial cardiomyocytes, associated with arterial hypertension, results in their hypertrophy which becomes apparent both in the increased polyploidy and in the augmented protein mass of the cells.

Ultrastructure and biosynthetic activity of polyploid atrial myocytes in patients with mitral valve disease.

Biopsies of right auricle of human heart have been obtained during open heart surgery from 6 patients aged 23 to 49. The DNA and total protein content have been determined in isolated myocytes by two-wavelength scanning cytophotometry after double staining: Feulgen and naphthol yellow S. In all the biopsies predominant are polyploid hypertrophied myocytes. Both hypertrophied non-degenerating cells and cells with different extent of degenerative changes, primarily of myofibrils and membranes, are present. The highest extent of cell ploidy is in patients belonging to functional class IV according to the classification of New York Heart Association (NYHA); in these cases 72 to 98% of cells have nuclei with 8 c and more DNA content. With an increase in ploidy level, cells grow in size and in protein content, however the rate of this growth is much lower than that of DNA content in cells. There is no direct relation between ploidy and cell degeneration extent and no inverse relation between degeneration extent and ejection fraction.