[Interaction of C60 fullerene-polyvinylpyrrolidone complex and brain Aß(1-42)-peptide in vitro].

Using a spectrophotometric method changes occurring in solution containing brain Aß(1-42)-peptide, fullerene C60, and polyvinylpyrrolidone were analyzed. Using the Bent-French method relative binding constants of fullerene C60 with Aß(1-42)-peptide and polyvinylpyrrolidone with Aß(1-42)- peptide were determined. These data suggest that Aß(1-42)-peptide interacting with the C60 fullerene-polyvinylpyrrolidone complex partially displaces polyvinylpyrrolidone and generates a new three molecular compound.

[Comparative studies of amyloid properties of muscles proteins and brain Abeta-peptides and identification of approaches to destruction of their amyloids in vitro].

In this review our data on the comparative study of amyloid properties of titin family proteins and brain Abeta-peptides are represented. Approaches to the destruction of amyloid fibrils of muscle X-protein and brain Abeta(1-42)-peptides by various chemical compounds are also described.

[Changes in gene expression and content of titin (connectin) in striated muscles of chronically etihanol-fed rats].

Changes in gene expression and isoform composition of giant sarcomeric protein titin (connectin) in cardiac muscle, as well as changes of its isoform composition in skeletal muscle (m. soleus) of chronically ethanol-fed rats have been studied using real-time RT-PCR and low percentage SDS-gel electrophoresis. The decrease of titin content in examined muscles and the decrease in titin gene expression in myocardium of chronically ethanol-fed rats have been shown. These changes indicate the development of pathologic process.

[Changes in titin and myosin heavy chain isoform composition in skeletal muscles of Mongolian gerbil (Meriones unguiculatus) after 12-day spaceflight].

Changes of titin and myosin heavy chain isoform composition in skeletal muscles (m. soleus, m. gastrocnemius, m. tibialis anterior, m. psoas major) in Mongolian Gerbil (Meriones unguiculatus ) were investigated after 12-day spaceflight on board of Russian space vehicle "Foton-M3". In m. psoas and m. soleus in the gerbils from "Flight" group the expected increase in the content of fast myosin heavy chain isoforms (IIxd and IIa, respectively) were observed. No significant differences were found in the content of IIxd and IIa isoforms of myosin heavy chain in m. tibialis anterior in the gerbils from control group as compared to that in "Flight" group. An unexpected increase in the content of slow myosin heavy chain I isoform and a decrease in the content of fast IIx/d isoform in m. gastrocnemius of the gerbils from "Flight" group were observed. In skeletal muscles of the gerbils from "Flight" group the relative content of titin N2A-isoform was reduced (by 1,2-1,7 times), although the content of its NT-isoform, which was revealed in striated muscles of mammals in our experiments earlier, remained the same. When the content of titin N2A-isoform was decreased, no predictable abnormalities in sarcomeric structure and contractile ability of skeletal muscles in the gerbils from "Flight" group were found. An assumption on the leading role of titin NT-isoform in maintenance of structural and functional properties of striated muscles of mammals was made.

[Dopamine as a possible substance for oncotherapy and for quantitative evaluation of cytosolic G-actin].

Viability, histology and ultrastructure of normal cells and cells of different degrees of malignancy after interaction with dopamine as well as the ability of these cells and isolated G-actin in model experiments to stain by Falck technique were studied. It is shown that dopamine, virtually having no effect on the viability of the "normal" non-tumorigenic transformed cells, noticeably reduces cell viability of slightly tumorigenic cells, causes a significant reduction in viability of attachable cancerous cells and a very significant decrease in cell viability of cancerous cells growing in suspension. The intensity of fluorescence of the cytosole in cells treated with dopamine, has been very high and varied in different cultures, and that of isolated actin directly depended on its concentration. Common to all cell morphological feature of damage from the action of dopamine and the putative substrate of fluorescence was actodopamine filaments network strands (identified on the structure and size), which appears in the cytosole loci, where they were absent in control. The data show that dopamine can be used as an oncotherapeutic remedy and diagnostic tool interacting with G-actin as a cellular target.

[Seasonal changes in the isoform composition of the myosin heavy chains in skeletal muscles of hibernating ground squirrels Spermophilus undulatus].

Seasonal changes of the isoform composition of myosin heavy chains in skeletal muscles (m. triceps, m. longissimus dorsi, m. soleus, m. gastrocnemius, m. vastus lateralis) of hibernating ground squirrels Spermophilus undulatus were studied. Functional properties of myosin (the actin-activated ATPase activity and its Ca(2+)-sensitivity in vitro) were also examined. It was observed that the content of slow myosin heavy chain I isoform increased and the content of fast IIx/d isoform decreased in muscles of torpid ground squirrels and animals which are active in autumn and winter. In muscles of these animals the content of N2A-titin isorfom decreased although the relative content of NT-titin isoform, observed in striated muscles of mammals in our previous experimental works, increased. Actin-activated ATPase activity and Ca(2+)-sensitivity of myosin isolated from skeletal muscles of torpid and interbout ground squirrels were found to reduce. The changes observed are discussed in the context of adaptation of skeletal muscles of ground squirrels to hibernation conditions.

[Role of light chains of myosin in the regulation of contraction of vertebrate striated muscles].

The data of the study on Ca2+ sensitivity of ATPase activity of myosin from vertebrate striated muscles in the presence of actin and the conditions of its manifestation and disappearance are presented. The role of Ca2+ sensitivity of actin-activated myosin ATPase in the regulation of contraction of vertebrate striated muscles is discussed.

[Expression of titin in the myocardium of spontaneously hypertensive rats during in the course of hypertrophy].

Changes in the expression of N2B- and N2BA-isoforms of titin in the left ventricle of the myocardium of spontaneously hypertensive rats during the development of hypertrophy have been analyzed by the methods of real-time scale polymerase chain reaction and SDS gel electrophoresis. It was shown that, in early development of hypertrophy (15-week-old rats), an increase in the expression of mRNA of the titin gene and a decrease in the content of the protein itself occur. At a later stage of development (26-week-old rats), a decrease in the expression of titin at the level of both mRNA and the protein per se was observed. The results obtained can be used in the development of methods for diagnosing the development of myocardium hypertrophy.

[Effect of vibrostimulation on support zones of rat's feet, and support loading on titin N2A-isoform and T2-fragment in m. soleus under the conditions of simulated microgravity].

Effects of daily 3-hr vibrostimulation of foot support zone and 3 hrs. of usual locomotion on titin N2A-isoform and proteolytic T2-fragment were studied in m. soleus of rats during 7-d suspension. Besides, effects of single one-hour support loading following 7-d weight-deprivation on titin N2A- and T2-forms were also evaluated in the muscle. Relative N2A content in m. soleus of the suspended group was noted to reduce approximately 25% as compared with the group of control. In the meantime, relative T2 content grew 2-3 times. Along with other changes in the muscle apparatus, N2A reduction contributes to the decline of m. soleus contractility in suspended rats, and development of the "hypogravity muscular syndrome" The daily 3-hr vibrostimulation of the foot support zones substantially (almost twice) decreased N2A disintegration with parallel reduction in approximately 1.5 times relative T2 content comparing with the Suspension group. Three hours of locomotion a day during suspension did not appear to decrease the relative N2A content in m. soleus. However, in this group of animals the relative T2 content in m. soleus was twice as high as in the control. Single-time presentation of support load for one hour soon after 7-d suspension resulted in more significant (approximately 30%) reduction in relative N2A content and twofold fall of relative T2 content as compared with m. soleus of the rats in group Suspension. These data attest to the leading role of support stimulation in maintaining the structural and functional properties of the musculoskeletal system, specifically the titin N2A-isoform content in m. soleus of rat deprived of gravitational loading. Besides, it was also shown that tracking changes in titin makes possible to follow the course of hypogravitational muscle syndrome and to evaluate effectiveness of preventive methods.

[Contractile properties of fibers and cytoskeletal proteins of gerbil's hindlimb muscles after space flight].

The work had the goal to compare the microgravity effects on gerbil's muscles-antagonists, m. soleus and m. tibialis anterior. The animals were exposed in 12-d space microgravity aboard Earth's artificial satellite "Foton-M3". Findings of the analysis of single skinned fibers contractility are 19.7% diminution of the diameter and 21.8% loss of the total contractive force of m. soleus fibers post flight. However, there was no significant difference in calcium sensitivity which agrees with the absence of changes in the relative content of several major cytoskeletal proteins (titin and nebulin ratios to heavy chains of myosin were identical in the flight and control groups) and a slight shifting of the myosin phenotype toward the "fast type" (9%, p < 0.05). These parameters were mostly unaffected by the space flight in m. tibialis anterior. To sum up, the decline of contractility and diminution of gerbil's myofibers after the space flight were less significant as compared with rats and did not impact the sytoskeletal protein ratios.

[Composition of the titin isoforms in muscles in pathology].

An electrophoretic study of changes in the composition of titin isoforms in skeletal and cardiac muscles of human and rat in pathology has been carried out. A more considerable decrease in the content of intact titin isoforms in comparison with the content of N2BA- and N2B-titins in hypertrophic heart of spontaneously hypertensive rats, N2A-titin in back muscle from patients with the "stiff-man syndrome", and in m. soleus of human and rats during the development of "muscle hypogravity syndrome" has been observed. In the last case, the relation between the reduction of titin content in m. soleus and the increase in the time the rats were in conditions of simulated microgravity has been revealed. On electrophoregrams of the left ventricle myocardium of patients with the terminal stage of dilated cardiomyopathy, intact titin and N2BA-titins were absent, and a considerable decrease in the content of N2B-titins has been observed, which could be a consequence of the terminal stage of pathology. Our results show that the development of the diseases is accompanied by a greater destruction of intact titin than of its other forms, which may be important for diagnostics of the pathological process.

[Seasonal changes in the composition of the titin isoforms in muscles of hibernating ground squirrels].

An electrophoretic study of changes in the content of intact titin isoforms, N2B-, N2BA-, N2A-titins and T2 in skeletal and cardiac muscles of ground squirrel (Spermophillus undulatus) in different periods: summer activity, autumn activity, hibernation, arousal, and winter activity has been carried out. In atriums and ventricles of autumn activity ground squirrels, an increase (approximately 1.5 times) in the N2BA-to-N2B ratio in comparison with that in cardiac muscle of summer activity ground squirrels has been observed. During hibernation, the decrease in the relative content of N2B,- N2BA-titins and T2 in cardiac muscle and of N2A-titin and T2 in skeletal muscles has been tested against the background of preservation of the relative amount of intact titin isoforms. During arousal of ground squirrel and in short-time period of winter activity, a rapid restoration in the content of N2B,- N2BA-, N2A-titins and T2 in muscles has been observed. In this case, in cardiac muscle of hibernating, arousing and winter active ground squirrels, the increased N2BA-to-N2B ratio has been preserved. The changes in the titin content are discussed in the context of adaptation of ground squirrels to hibernation.

[Contractile properties of the isolated rat musculus soleus and single skinned soleus fibers at the early stage of gravitational unloading: facts and hypotheses].

The contractile properties of the postural rat soleus muscle at the early stage of the gravitational unloading (3-day rat hindlimb suspension) have been studied using different modes of muscle contraction (twitch and tetanic contraction of the isolated muscle, Ca-induced contraction of isolated skinned fibers). A significant enhancement of the twitch maximal tension of unloaded muscles without changes in time-dependent characteristics was observed, although the half-relaxation time tended to increase. The fiber diameter did not change (42.37 +/- 0.76 vs 43.43 +/- 1.15 microm in controls). The Ca-induced maximal isometric tension in unloaded soleus was significantly decreased (32.1 +/- 1.05 vs 37.6 +/- 1.52 mg in controls, p < 0.05). The maximal specific tension was respectively decreased (23.14 +/- 0.77 vs 27.6 +/- 2.36 kN/m in controls). The pCa50 in unloaded muscle decreased from 6.05 +/- 0.02 in controls to 5.97 +/- 0.02 (p < 0.05), indicating the loss of myofibrillar calcium sensitivity. The analysis with the calcium probe Fluo-4AM demonstrated that the intracellular [Ca2+] was sufficiently increased after hindlimb suspension. At the same time, the relative content of titin and nebulin did not change.

[Structure and functions of titin, a giant protein of skeletal and cardiac muscles: evidence and suppositions].

The structural and functional properties of skeletal and cardiac titin are described according to the recent experimental evidence and suppositions.

[Titin isoforms].

The titin isoform composition in skeletal and cardiac muscles of humans and animals has been studied using SDS elecrtophoresis in agarose-strengthened 1.3-2.3% polyacrylamide gels modified by us and immunoblot analysis in order to reveal new titin isoforms with molecular weights of more than 3700 kDa. The experimental data obtained have provided a basis for the suggestion that the titin bands of high molecular weights discovered by us are intact titin isoforms, while the titin bands designated on electrophoregrams as N2A, N2B and N2BA isoforms are their fragments.

[Seasonal changes in phosphorylation of myosin regulatory light chains and C-protein in myocardium of hibernating ground squirrel Citellus undulatus].

A comparative study concerning the extent of phosphorylation of myosin regulatory light chains and C-protein from the left ventricle of hibernating ground squirrel Citellus undulatus during the periods of hibernation and activity was carried out. During hibernation, regulatory light chains of ground squirrel were found to be completely dephosphorylated. In active animals, the share of phosphorylated light chains averages 40-45% of their total amount. The extent of phosphorylation of the cardiac C-protein during hibernation is about two times higher than that in the active state. Seasonal differences in phosphorylation of the two proteins of ground squirrel myocardium are discussed in the context of adaptation to hibernation.

[Creatine as a metabolic controller of skeletal muscles structure and function in strength exercises in humans. The cellular mechanisms].

The effects of creatine oral supplementation combined with a 10-week resistive training of morphometric, contractile and molecular characteristics of human vast lateral muscle fibers were studied. 2 groups consisting of 9 young healthy men each were involved in resistive training of knee extensors for 10 weeks. Volunteers of the first group received per os 20 g of creatine for the 1st week of training and 5 g for the rest of the experimental training period. We found a significant increase of slow and fast-twitch fiber size in both trained groups and a significant increase of Ca-sensitivity of skinned single fiber contractility in creatine-supplemented group. The serum creatine phosphokinase activity in blood samples taken 24 hours after exercise session increased in all stages of the experimental training in both groups. At the same time, the adaptive decrease of the after-exercise CK concentration was observed in the placebo but not in the creatine-supplemented group. The altered integrity of the subsarcolemmal dystrophin layer was revealed in both groups after training.

[Myosin light chains of skeletal and cardiac muscles of ground squirrel Citillus undulatus in different periods of hibernation].

The isoform composition of myosin light chains and the extent of their phosphorylation in skeletal and cardiac muscles of ground squirrel Citellus undulatus in different periods of hibernation were studied. Regulatory myosin light chains of skeletal muscles of hibernating ground squirrels were completely dephosphorylated, while 25% of these light chains in active animals were phosphorylated. During hibernation, a shift of isoform composition of essential and regulatory skeletal muscle myosin light chains toward slower isoforms was observed, which is evidenced by the data obtained on m. psoas and on the totality of all skeletal muscles. In the atrial myocardium of hibernating ground squirrels, ventricular myosin light chains 1 (up to 60%) were registered. In contrast, during arousal of ground squirrels, in ventricular myocardium the appearance of atrial myosin light chains 1 (up to 30%) was revealed. A possible role of posttranslation changes in myosin light chains and their isoform shifts in the hibernation scenario is discussed.

[The behavior of titin and the proteins of its family from skeletal muscles of ground squirrel (Citellus undulatus) during hibernation and rats under conditions of simulated microgravity]. / Povedenie taitina i belkov ego semeistva v skeletnykh myshtsakh suslikov (Citellus undulatus) pri zimnei spiachke i krys v usloviiakh modeliruemoi mikrogravitatsii.

By the use of SDS PAGE, the behavior of titin and MyBP-C in fast (m. psoas) as well as titin and MyBP-X in slow (m. soleus) muscles of ground squirrels (Citellus undulatus) during hibernation was compared with the behavior of titin and MyBP-X in rat m. soleus under conditions of simulated microgravity. A decrease in the amount of titin 1 and MyBP-C relative to that of myosin heavy chains by approximately 30% and approximately 40%, correspondingly, in muscles of hibernating and arousing ground squirrels was revealed in comparison with active animals. No differences in the relative amount of MyBP-X in m. soleus of hibernating, arousing and active ground squirrels were found. Under conditions of simulated microgravity, a decrease in the amount of titin 1 by approximately 2 times and MyBP-X by approximately1.5 times relative to that of myosin heavy chains in rat m. soleus was observed. By the method of SDS PAGE modified by us, an almost twofold decrease in the amount of short isovariants of the titin N2A isoform relative to that of myosin heavy chains was shown in muscles of hibernating and arousing ground squirrels, whereas no changes were found in the amount of long titin isovariants. The conditions of simulated microgravity resulted in a twofold decrease in the relative amount of both short and long titin isovariants in rat m. soleus. The results indicate that hibernating ground squirrels have an evolutionarily determined adaptive mechanism of selective degradation of fast muscle fibers and preservation or increase of slow fibers, as the most economic and energetically advantageous, with proteins typical of them. The microgravitation of nonhibernating animals (rats) leads to a non-selective degradation of MyBP-X and titin isovariants, which contributes to considerable atrophy of soleus fibers.

[Human soleus fibers contractile characteristics and sarcomeric cytoskeletal proteins after gravitational unloading. Contribution of support stimulus]. / Sokratitel'nye kharakteristiki i belki sarkomernogo tsitoskeleta volokon m. soleus cheloveka v usloviiakh gravitatsionnoi razgruzki. Rol' opornogo stimula.

The effects of support withdrawal and support stimulation on the contractile characteristics of human soleus fibers and cellular factors which influence them were studied. The experimental model of the "dry" head-out water immersion was used in the study. In this model, the hydrostatic pressure on different sites of the body surface are equal so that the experimental conditions are close to the complete supportlessness. A 7-day exposure to dry immersion resulted in a decrease in the maximal isometric tension of the skinned fibers, a decline in the myofibrillar Ca2+-sensitivity, and the relative loss of the titin and nebulin content. A significant decrease in the percentage of fibers containing slow myosin heavy chains was also observed after dry immersion. The application of the mechanical stimulator influencing the plantar support zones with a pressure of 0.2 +/- 0.15 kg/cm2 6 times a day for 20 minutes of each hour brought about a complete prevention of the above listed effects of dry immersion. The data obtained allow one to conclude that the decline in maximal tension and Ca2+-sensitivity as well as myosin shift and loss of sarcomeric cytoskeletal proteins are associated with the support withdrawal during the exposure to dry immersion.