[A. N. Bakoulev Scientific Center for Cardiovascular Surgery].

OBJECTIVE: to assess dynamics of diastolic function for detection of development of diastolic dysfunction (DD) and it's causes, to evaluate the effect of DD on prognosis in the postoperative period in patients with acquired heart diseases. MATERIALS AND METHODS: We included in this study 112 patients with aortic and mitral valve diseases (90 men, 22 women, median age 51 [35; 57] years). All patients underwent echocardiography (echo), tissue Doppler, speckle tracking echo prior to surgery, in the early postoperative period (8-14 days) and in 12-36 months after surgery. In 28 patients dynamic contrast-enhanced magnetic resonance imaging was also performed. Patients were divided into groups according to prognosis: group 0 - without complications; group 1 - with postoperative heart failure (HF) and preserved left ventricular ejection fraction (EF); group 2 - with HF and EF <45 %. The following parameters were used for identifying left ventricular (LV) DD: septal velocity es <7 cm / sec, lateral el <10 cm / sec, average E / e ratio >14, left atrial (LA) volume index >34 ml / m2, peak tricuspid regurgitation velocity >2.8 m / sec. RESULTS: Initially diastolicLV function was normal in 34 of 112 patients (30.4 %), in early postoperative period DD emerged in 9 (26.5 %) of these patients. The appearance of LV DD was associated with decrease of septal es immediately after surgery and its subsequent progressive decline in the long-term postoperative period from 8.5±0.71 to 4.6 ±0.53 cm / sec (p=0.005). Worsening of diastolic function and lowering of septal velocity was detected namely in patients with presence of fibrosis. In the group of other patients in whom fibrosis was not studied and the degree of DD increased there was a transient decrease of lateral el (from 10.2±3.1 to 7.5±2.43 cm / sec, p=0.035) and an increase of the E / el (from 10.53±4.07 to 14.5±5.23, p=0.05) in the early period after the operation. There were no correlations between DD and LV EF,LV volumes, and development of arrhythmias. The prognostic model for DD included average longitudinal deformation of LA (global LA longitudinal strain) and E / e ratio on the tricuspid lateral annular velocity. CONCLUSIONS: Appearance of DD in postoperative period after correction of acquired heart defects was due to damage of the septal diastolic function which correlated with fibrosis and was indicative of inadequate myocardial protection. The model of development of heart failure with normal EF after operation was designed.

[Visualization of sinus node arteries by multislice spiral computed tomography coronary angiography].

OBJECTIVE: To assess the capabilities of multislice spiral computed tomography coronary angiography (MSCT-CA) to visualize the anatomy of the sinus node artery (SNA). MATERIAL AND METHODS: The retrospective analysis of coronary artery examinations covered 46 patients with the referral diagnosis of coronary heart disease. MSCT-CA showed no evidence of coronary artery stenosis. This sample included 23 (50%) men and 23 (50%) women; the mean age of the patients was 52.4 +/- 9.1 years; the mean height was 170 +/- 6.67 cm; the mean weight was 80.7 +/- 12.1 kg. X-ray computed tomography was carried out using a SOMATOM Definition AS+ 128-slice computed tomography scanner with retrospective ECG synchronization, reconstructed slice thicknesses of 1 and 3-mm. The Spearman correlation test was used for statistical data analysis. RESULTS: The SNA was visualized in 83% of the patients. It originated from the right coronary artery (RCA) and the circumflex branch of the left coronary artery in 84 and 16% of cases, respectively. No significant association was found between the type of heart blood supply and that of sinus node one (r = 0.06). In 18% of cases, the SNA was visualized only at the level of the ostium, allowing the assessment of the origin of the artery, and at the level of its mid-third in 32%; the distal SNA bed was visualized up to its division; in 10% of them the artery could be visualized all the way, including the division (the dissipation site). Unclear visualization of the proximal SNA was observed among 17% of the patients in whom the SNA could not be visualized with a heart rate (HR) of more than 80 beats/min in 62.5% of the patients, less than 41 beats/min in 12.5%, and 60-61 beats/min in 25%. HR was not found to be associated with the quality of SNA visualization (r = 0.09). CONCLUSION: MSCT can assess the anatomy of SNA up to the distal bed and dissipation site. In the overwhelming majority of the patients, the SNA originated from the RCA (84%) regardless of the type of heart blood supply. The best SNA visualization was noted with a HR of 50 to 80 beats per minute. There was no statistical relationship of the quality of visualization to HR.

[Reorganization of actin cytoskeleton in the initial stage of transendothelial migration of bone marrow multipotent mesenchymal stromal cells].

The analysis of actin cytoskeleton reorganization in rat bone marrow multipotent mesenchymal stromal cells after one hour adhesion to a monolayer of endothelial cell line EA.hy 926 allowed us to identify three types of cells interacting with the endothelial cells. Approximately half of multipotent mesenchymal stromal cells retained a rounded shape, most of them contained large round actin aggregates, had irregular borders and contacted with the surface of the endothelial cells by microvilli or protrusions similar to small lamellae. Almost all other cells were surrounded by narrow lamellae along the entire perimeter. In addition, a small amount.of elongated flattened cells that contacting with endothelial cells by means of focal contacts was observed. Microenvironmental factors such as proinflammatory cytokine tumor necrosis factor α or plasma proteins affected the ratio of stromal cell types, with different types of organization of the actin cytoskeleton in multipotent mesenchymal stromal cells population.

[Arrhythmogenic right ventricular dysplasia: polymorphism of clinical manifestations].

We observed 15 patients with arrhythmogenic right ventricular dysplasia (ARVD): 9 with definite and 5 with probable ARVD (modified European Criteria, 2010). Eight patients had typical ARVD (frequent right ventricular extrasystoles, nonsustained right ventricular tachycardia without heart failure with or without myocarditis). Five patients had ARVD with progressive heart failure (right- or biventricular with or without myocarditis). Two patients had full scale arrhythmic form (sustained right ventricular tachycardia without or with right ventricular dilation, with or without myocarditis). In 3 cases diagnosis was confirmed morphologically or with DNA-diagnostics. This material allowed us to highlight the following specific points related to diagnostics of ARVD. Detection of fat at MRT is not obligatory for diagnosis, fat can be detected by MSCT; ventricular arrhythmias can move backwards in the picture of the disease; leading clinical manifestation can be unexplained right ventricular insufficiency; ARVD can be combined with other genetic cardiomyopathies as well as with infectious immune myocarditis (up to 50% of patients); elevated titer of anticardiac antibodies is not characteristic for isolated ARVD; myocardial biopsy allows to verify both ARVD and concomitant myocarditis. The paper also contains discussion of the role of myocarditis in various forms of ARVD and possibilities of its diagnosis and treatments.

[Role of proteins in Fas-mediated apoptosis in tumor cells and lymphocytes co-cultured in vitro].

The investigation deals with the role of Fas, FasL, RIP, caspase 3, and PARP taking part in Fas-mediated apoptosis, and contributing to in vitro interaction of hepatoma MH-22a and histiocytic sarcoma J-774 in mice with syngenic splenocytes. Protein expression was identified by means of indirect immunofluorescence. There were two patterns of interaction of tumor cells and splenocytes: apoptosis occurred either in 80% or in an insignificant number of tumor cells. In the latter case, high Fas expression was identified before and when it dropped after the experiment. FasL expression in tumor cells often peaked before the experiment and then it decreased after contact with lymphocytes. That mechanism was reversed in splenocytes: contact with tumor cells boosted expression. RIP, caspase 3 and PARP expression was very low and failed to show until the experiments on both patterns of cells were undertaken. After the experiments, it either remained latent or soared up. In the latter case, simultaneous expression of all proteins took place both in tumor cells and lymphocytes. A second battery of experiments demonstrated maximum rates of apoptosis both of tumor cells and splenocytes. However, the situation was different: Fas expression intensified in both patterns of cells after their interaction which was followed by post-experimental drop in RIP, caspase 3, and PARP expression in tumor cells; hence, the importance of perforin/granzyme-mediated apoptosis which occurred at the early stages of tumor growth in the midst of interaction with immune system cells. That pattern of apoptosis was highly cytotoxic. It is suggested that Fas-mediated apoptosis or any other receptor-sensitive pathway might take place during tumor progression, i.e. at a stage when tumor is most susceptible to change.

[The RT-PCR-identified Fas and FasL expression and apoptosis in mouse hepatoma MH-22a and histiocytic sarcoma J-774 clonal lines coupled with in vitro cultivation with syngenic splenocytes].

Malignant growth is associated with various patterns of interaction between tumor cells and those of the body immune system, interaction between Fas-receptor (Fas) and Fas-ligand (FasL) expression being one of them. These mechanisms were simulated in vitro using the main cell populations from murine hepatoma MH-22a, histiocytic sarcoma J-774 and their clonal lines obtained from cocultivation of tumor cells and syngenic splenocytes. Fas and FasL expression was identified by the RT-PCR method while apoptosis--by electrophoresis of low molecular DNA fractions and clonogenic survival.

[RAPD-polymerase chain reaction analysis of the genetic structure of tumor hepatocyte population MH-22A]. / Analiz geneticheskoi struktury populiatsii opukholevykh gepatotsitov MG-22A metodom RAPD-polimeraznoi tsepnoi reaktsii.

The viability and genetic variability of mouse hepatoma MH-22a were studied by RAPD-PCR method using 3 primers, and the genetic structure of the tumor hepatocyte population based on the number of clonal fingerprint rearrangements was identified: 56% of hepatocytes made up clones with 0-3 rearrangements (the stem line of the population); 32%--the variable part with 4-7 rearrangements per clone and 12%--the aberrant part with 8-12 rearrangements per clone. The stem line appeared the most viable featuring zero or least genetic variability while the clones characterized by the greatest genetic variability were least viable.

[Clonal analysis of the heterogeneity of neoplastic hepatocytes]. / Klonal'nyi analiz geterogennosti opukholevykh gepatotsitov.

The heterogeneity of hepatoma MH-22a cell line was studied by clonal analysis on the basis of the following factors: splenocyte-induced apoptosis of tumor hepatocytes, tumor hepatocyte-induced apoptosis of splenocytes and profile of B1-associated DNA fragments. Induction of apoptosis of tumor hepatocytes by splenocytes and of splenocytes--by tumor hepatocytes was carried out in the main population of hepatocytes and in five clonal lines of hepatoma MH-22a. Genetic heterogeneity was studied using the same material and PCR primed for murine B1-elements. It was shown that apoptosis of hepatocytes of the main population and two clonal lines of hepatoma MH-22a was induced by splenocytes. The ability to induce apoptosis of splenocytes was observed in the same clonal lines and population of tumor hepatocytes. The latter involved enhanced genetic heterogeneity which was identified by B1-PCR analysis. Thus, a correlation was established in tumor hepatocytes between apoptosis-related characteristics and frequency of genome rearrangements.

Spectrophotometric quantitative analysis of the main hemoglobin derivatives.

A spectrophotometric multiwavelength method for the simultaneous determination of the main hemoglobin derivatives including oxyhemoglobin, deoxyhemoglobin, carboxyhemoglobin, and methemoglobin has been developed. The choice of analytical wavelengths was made by the methods of information factors and linear programming. The software for the quantitative analysis was developed considering the least square method, linear programming, algebraic background correction, and combined methods including linear programming and algebraic background correction.

[Polymorphism of B1-associated DNA fragments on various stages of ontogenesis of mice in tissues of different histogenesis]. / Polimorfizm B1-assotsiirovannykh fragmentov DNK na raznykh stadiiakh ontogeneza myshi v tkaniakh razlichnogo gistogeneza.

Pattern of B1-associated DNA fragments was studied by means of polymerase chain reaction (PCR) in the mouse tissues of different histogenesis of 15 and 20 day old embryos and of adult mice C57B1/6. As many as 20 DNA fragments were revealed on electrophoregrams differing in their molecular masses (m. m.) and amounts of amplified products. DNA fragments varied within a 100-10,000 bp range. The clusters of B1-associated DNA fragments, containing 100-200, 300-400 and 800-1000 bp, were most intensive in all studied electrophoregrams. The B1-associated DNA fragments from muscles of adult mice differed from those of other tissues by the presence of a DNA fragment with 800 bp. A comparative analysis of the spectra of B1-associated DNA fragments from hepatocytes of two inbred strains, C57B1/6 and C3HA, has shown their general similarities in m. m. values. But a significant distinction, that was found, involved the presence of a DNA fragment with m. m. approximately 6000 bp in the spectra of B1-associated DNA fragments from C3HA strain mice, that is absent in the spectra of respective fragments from hepatocytes of C57B1/6 strain mice. The obtained results allow to use the B1-PCR method for studying genome recombination during ontogenesis, intraspecies divergence and also at malignant cell transformation.

[Polymorphism of B1-associated DNA fragments from normal and transform ed murine hepatocytes]. / Polimorfizm B1-assotsiirovannykh fragmentov DNK v normal'nykh i transformirovannykh gepatotsitakh myshi.

Our study of B1-associated DNA fragment polymorphism in murine hepatocytes by means of polymerase chain reaction (PCR) allowed to reveal as many as 20 DNA fragments differing in their molecular masses (m. m.) and amount of amplified products varying within the range of 100-1000 bp. Within the same inbred strain of mice (C57B1/6), spectra of B1-associated DNA fragments were similar in different periods of ontogenesis (embryos of 15 and 20 days, adult mice), in different mice of the same or different litters. A comparative analysis of the spectra of B1-associated DNA fragments from hepatocytes of two inbred strains, C57B1/6 and C3HA, has shown in general their similarities in m. m. values. But a significant distinction, that was found, involved the presence of DNA fragments with m.m. approximately 600 bp in the spectra of B1-associated DNA fragments from C3HA strain mice, that is absent in the spectra of respective fragments from hepatocytes of C57B1/6 strain mice. The spectra of B1-associated DNA fragments from transformed hepatocytes of murine hepatoma MH-22a, in general, were the same as those from hepatocytes of C3HA strain mice. At the same time, a DNA fragment with m.m. of 450 bp, not detected in normal hepatocytes, was revealed in transformed ones. Nevertheless a DNA fragment with m.m. of 600 bp, characteristic of normal hepatocytes, was not observed in the transformed hepatocytes. The B1-PCR method can be used for studying genomic polymorphism both in different populations of mice, and during malignant growth.

[The use of B1-PCR method for studying the genomic polymorphism involved in malignant growth]. / Ispol'zovanie metoda v1-ptsr dlia izucheniia genomnogo polimorfizma pri zlokachestvennom roste.

The BI-PCT method showed the profile of BI-associated fragments of LNA in the cell line of the mouse hepatoma MH-22a to differ from that of the liver cells of C3HA mice, hepatoma cells incorporating the DNA fragments with 450 bp and those with 600bp disappearing. Application of the same method failed to reveal any differences in the profiles of BI-associated DNA fragments in the differentiated and non-differentiated cells of the embryonal carcinoma F9 induced by retinoic acid and cAMP dibutyryl treatment. It is suggested that the spectra of BI-associated DNA fragments might correlate with genetic stability in tumor cells.