Role of L1CAM in the Regulation of the Canonical Wnt Pathway and Class I MAGE Genes.

Molecule L1CAM is specific for nerve cells and tumors of various localizations. The expression of L1CAM is significantly higher in melanoma in comparison with benign nevi and correlates with the progress of melanoma and transition from radial to vertical growth. Monoclonal antibodies to L1CAM effectively and specifically attenuate melanoma growth, though stimulates the epithelial-mesenchymal transition. shRNA-mediated knock-down of L1CAM showed the involvement of L1CAM in regulation of activity of the canonical Wnt pathway and expression of genes of class I melanoma-associated antigens (MAGE).

[Exogenic proteins in the human exhaled breath condensate].

The analysis of the protein composition of exhaled breath to diagnose diseases of the respiratory system raises a problem of differentiation proteins of expressed in the tissues of the lungs and respiratory tract (endogenous) and got in the respiratory system from the ambient air in the process of respiration (exogenous). In this work an attempt was made to estimate a set of exhaled exogenic proteins by mass spectrometry coupled with nanoflow HPLC. Six-month isolation of healthy donors indoors with air cleaned of dust leads to removal from the spectrum of exhaled proteins of some keratins that are considered therefore to be exogenic. Non-keratin proteins may also circulate between the ambient air and human respiratory ways, but their concentration appears to be significantly lower the keratin concentrations (especially epidermis keratin). Among non-keratins dermcidin seems to be the most significant exogenic protein of exhaled air. The conclusion of the diagnostic value of exhaled proteins can be done only after careful comparison of the results of quantitative and qualitative analysis of their composition in norm and pathology for a statistically significant sample of donors.

[Human temperature portrait and its relations with aerobic working capacity and the level of blood lactate].

In study with participation of 53 healthy men volunteers and infra-red thermograph application we obtained data confirming thermal portrait (i.e. skin temperature distribution in muscle rest conditions with minimal thermoregulatory activation) interrelations with maximal aerobic capacity (r = +0.6) and lactate level after critical muscle load (r = -0.7). Acute regional cooling (by 1 minute feet placing in ice water) led to temperature rise in certain breast and back skin local areas and increase in oxygen uptake, pulmonary ventilation and respiratory coefficient. Moreover lactate level in peripheral blood reduced. Summarizing obtained results we assume brown adipose tissue (BAT) activity influencing on thermal portrait formation in conditions with environmental temperature below thermoneutral. This hypothesis permits to explain negative correlation between skin temperature and body mass index and 2-fold increase in oxygen uptake during acute cold exposure. Nevertheless further investigations are needed to clarify physiological mechanisms providing significant correlation between skin temperatures in rest thermoneutral conditions on the one hand and maximal aerobic capacity, anaerobic threshold and lactate content after critical muscle load on the other hand.

[Genetic and social factors in developing of aggression].

The association of 5-HTTLPR gene polymorthism and aggression was studied in control group of males and females and in the athletes. The sport activities were found to decrease the aggression: the effect persist for the synchronized swimmers and for the wrestlers. Control group of males were characterized by higher aggression scores (Assault, Negativism, Suspicion and Verbal aggression scales of Buss-Durkee Hostility Inventory) compared to females. For all female-subjects irrespective of sport activities and age, the association between the variants of 5-HTTLPR gene and the Indirect Aggression and Negativism scores were found: carries of SS genotype has higher scores on Indirect Aggression and lower scores on Negativism. For the males the association was different: The averaged Hostility scores were higher for the carriers of LL-genotype. The brain processes, presumably underlying the association between aggression and 5-HTLPR gene, were studied in male control group. The increased MMN component of ERP, which responsible for the automatic change detection, and decreased P3a component, related to involuntary attention and cognitive control were found in LL-carries. It might be considered as a sign that SS-carries process the information with more cognitive resources. Probably they perceive the stimulus as more complicated, which lead to activation the additional resources of frontal cortex. It might be also suggested that the carries of SS-genotype tend to deeper processing of the incoming information. Probably, it is this more "serious" analysis of external information, which underlies the rejection of impulsive aggressive actions.

[Molecular mechanisms of stress adaptation: immediate early genes].

Immediate early gene (IEG) expression is the important part of adaptive reactions caused by short-term high-intensity physical activities. IEG leads to activation of transcription, protein neo-synthesis and form-specific stress response. We were looking for human body physical exercise adaptation leukocytes IEG expression markers. Gene expression profiles were measured using microarray technology and validated by real time PCR. In this study, we describe several cell stress adaptation regulators groups: stress response, splicing and cell signaling.

Short-term highly intense physiological stress causes an increase in the expression of heat shock protein in human leukocytes.

Extracellular heat shock protein with molecular weight of 70 kDa is a signal molecule of the immune system. It is secreted by the peripheral blood, liver and muscle cells in response to physiological, thermal, and mental stresses. The main goal of our study was to compare the levels of expression of heat shock protein (70 kDa) matrix ribonucleic acid in leukocytes and serum concentrations of the protein before and after physiological stress. In order to solve this problem, we developed enzyme immunoassay of serum heat shock (70 kDa) protein concentration and a method for evaluating the expression of matrix ribonucleic acid of this protein in leukocytes by the real time PCR. The concentration of 70 kDa heat shock protein in the serum increased 1.7 times as a result of even a short-term highly intense physiological stress, while the expression of its matrix ribonucleic acid in leukocytes increased 1.5 times. The individual features determine the response to physiological stress. Probable sources of 70 kDa heat shock protein are discussed.

Comparative study of stabilometric parameters in sportsmen of various disciplines.

Stabilometric parameters were compared in sportsmen of various disciplines (biathlon, boat racing, judo, and water polo). A decrease in the role of gravitational factor in sport activity was accompanied by the impairment of balance characteristics, which remained within normal limits of the mean population level.

[Linear B-cell epitope prediction].

Combination of sliding window method with physical properties scale of amino acids is a classical approach for linear B-cell epitope prediction. But it was shown that accuracy of these methods is poor. We reviewed classical and new algorithms of epitope prediction and present own implementation of one of them. The AAPPred software is available online at http://www.bioinf.ru/aappred/.

[Modulation of cardiac rhythms in humans exposed to extremely weak alternating magnetic fields].

The influence of extremely weak alternating magnetic fields (EW AMF) with amplitudes of < or = 2 microT on the heart rate variability in humans has been studied. The volunteers were placed in a large- volume square coils system (2x2x2 m), which provided the exposure of the whole body to extremely weak alternating magnetic fields homogeneous in amplitude. It was shown that the exposure of volunteers to different types of extremely weak alternating magnetic fields can both increase and decrease the magnitude of stress. In particular, the field tuned to the nuclear spins of hydrogen atoms (amplitude 1.6 microT, frequency 76 Hz) induces a decrease in the Baevsky's stress index, while the field tuned to the magnetic moments formed by the orbiting electrons in some atoms (amplitude 0.192 microT, frequency 3000 Hz) increases the stress index. The results obtained provide a possible explanation for the mechanism of adverse effects of some particular types of technogenic and natural extremely weak alternating magnetic fields on the human cardiovascular system.

[Genetic basis of time perception in athletes].

The association between the subjective time perception and polymorphism of some genes, regulating activity of serotonin and dopamine, was studied in 89 synchronized swimmers. COMT gene, responsible for dopamine destruction, influences on reproduction of short time intervals (1-2 s). 5-HT2A and MAOA genes, regulating activity of serotonin, influence on subjective time flow. 5-HTT and COMT genes, regulating activity of serotonin and dopamine respectively, are related with accuracy of orientation in time. Association of time perception with different genes and mediators suggests different perception mechanisms, in different time ranges, in concordance with the previous physiological studies. The current study reveals that these physiological mechanisms have different molecular-neurochemical basis that helps to overcome the gap between the investigation on systemic and molecular levels.

[C-reactive protein level and rate of detection of autoantibodies to beta(1)-adrenoreceptors in patients with supraventricular tachyarrhythmias].

In order to assess concentration of C-reactive protein (CRP) and prevalence of autoantibodies against beta(1)-adrenoreceptors (abeta(1)-AR) in patients with cardiac supraventricular arrhythmias we studied 53 patients with arrhythmias and 20 healthy control subjects. Patients with idiopathic arrhythmias (atrial fibrillation or flutter and atrial tachycardia, n=35) comprised group I. Group II was formed of 15 patients with supraventricular arrhythmias and dilated cardiomyopathy (DCM) or chronic myocarditis. Patients of group III (n=23) had supraventricular arrhythmias and arterial hypertension (AH). CRP concentration was determined by recently developed well standardized high sensitivity method. abeta(1)-AR were detected in blood serum by direct immunoassay. Synthetic fragment containing 26 amino acids of abeta(1)-AR second loop was used as antigen. Patients with supraventricular arrhythmias and DCM or chronic myocarditis had higher median CRP (8.0 mg/1) than patients with idiopathic arrhythmias (0.78 mg/l), with supraventricular arrhythmias and AH (1.57 mg/l), or control group (0.6 mg/l). Groups I, II and III showed similar prevalence of ab1-AR (51.4, 40.0, 52.2%, respectively), that was significantly higher than in control subjects (10%) (p<0.005). These results provide evidence of the possible presence of autoimmune and/or inflammatory processes that may be involved in the genesis of supraventricular arrhythmias.

[Investigation of ribosomes of E. coli and T. maritima by atomic force microscopy].

Subunits 70S, 50S, and 30S of ribosomes of E. coli and T. maritima have been studied by atomic force microscopy. A considerable heterogeneity of structures was visualized when 70S and 30S subunits were sorbed on mica. The linear size and the height of molecules were estimated. It was found that the heights of ribosomes of E. coli and T. maritima substantially differ. The average height of 70S ribosomes of E. coli was 9.4 + 0.01 nm and that of T. maritima was 10.35 +/- 0.02 nm. The differences in the dimensions were probably determined by special organization of the mobile ribosomal element the L7/L12-stalk.

[The role of anti-beta1-adrenergic receptor autoantibodies in development of heart failure].

Epitopic analysis of anti-beta1-adrenergic receptor autoantibodies in the serum of patients with cardiomyopathy was performed. An immunoenzyme test system for detection of autoantibodies was developed using peptide containing 26 amino acids of the second extracellular loop of the beta1-adrenergic receptor as antigen. It was demonstrated that removal of autoantibodies against beta1-adrenergic receptor in patients with cardiomyopathy by immunoadsorption and plasmapheresis metodes led to improvement of the heart pumping capacity.

[Spectral parameters of heart rate variability and frequency of detection of autoantibodies to beta(1)-adrenoreceptors in patients with tachyarrhythmias: idiopathic and at the background of primary myocardial diseases].

In order to assess parameters of heart rate variability (HRV) and prevalence of autoantibodies against the beta(1)-adrenoreceptors in patients with cardiac arrhythmias we studied 42 patients with arrhythmias and 20 healthy control subjects. Thirty one patients with idiopathic arrhythmias were included in group I: with paroxysmal atrial fibrillation or flutter (n=13), paroxysmal atrial tachycardia (n=2) and paroxysmal ventricular tachycardia (n=16). Group II was formed of 11 patients with paroxysmal ventricular tachycardia and dilated cardiomyopathy or chronic myocarditis. ab1-AR were determined in blood serum by direct immunoassay. Synthetic fragment containing 26 amino acids of ab1-AR second loop was used as antigen. Groups I (54.8%) and II (63.6%) showed similar prevalence of ab1-AR, which was significantly higher than in control subjects (10%) (p<0.005). HRV parameters in I group were lower in ab1-AR-positive compared with ab1-AR-negative patients. At the same time HRV parameters in ab1-AR-positive patients were significantly different from those in controls (p<0.05). In group II HRV parameters of ab1-AR-positive and ab1-AR-negative patients were significantly lower than in control subjects (p<0.05). We suppose, that ab1-AR could participate in dysfunction of chronotropic heart regulation and contribute to development of arrhythmias in patients with structurally normal hearts.

Humoral immune response to recombinant viral NS3 protein in patients with hepatitis C.

Full-length recombinant NS3 protein was used in a test system for detection of specific antibodies in the sera from patients with hepatitis C. Possible antigenic determinants in NS structure were predicted. It was demonstrated that serological analysis requires enzyme immunoassay with full-length NS3.

[A quantitative analysis of complex formation between IgM and immobilized ligand using atomic force microscopy]. / Kolichestvennyi analiz obrazovaniia kompleksov IgM s immobilizovannym ligandom s pomoshch'iu atomno-silovoi mikroskopii.

Specific interaction between human IgM and polyclonal antibodies immobilized on support was studied by atomic force microscopy. Human IgMs are responsible for a number of side effects arising during the xenotransplantation of mammalian organs to man. On the basis of atomic force microscopy, a quantitative analysis of complexes with IgM was performed. The data of the analysis agree well with the results of enzyme immunoassay. It was shown that the method of detection of immune complexes based on atomic force microscopy is able to detect specific antibodies/antigens in serum.

[Detection of immune complexes using atomic force microscopy]. / Vyiavlenie immunnykh kompleksov s pomoshchiu atomno-silovoi mikroskopii.

Complex formation between immunoglobulins and ligands immobilized on mica was studied by atomic force microscopy in two different systems. In the first system, 60-kDa ligands possessing only one site for antibody recognition were used. In the other system, a more complex interaction of human immunoglobulin with immobilized polyclonal antibodies was studied. In both systems, specific complexes with proper ligand appeared, and unspecific interaction was not detected. The method of revealing immunocomplexes by image atomic force microscopy can be used in the development of modern diagnostic systems.