The Study of Efficiency of the Approach to Prevent the Adhesions in the Abdominal Cavity of Rats.

Adhesions in rat abdominal cavity were studied after laparotomy and subsequent single intraperitoneal injection of 2 ml of 5% aqueous solution of oxidized dextran (OD) with a molecular weight of 40 kDa (oxidation degree 10%). On days 7 and 21 after laparotomy, the number of adhesions in OD-treated rats was lower by 7.5 and 4 times than in animals not receiving OD. The number of neutrophils in adhesions on day 21 was manyfold lower in OD-treated rats. In 7 and 21 days after laparotomy, the number of fibroblasts in the adhesions of rats receiving and not receiving OD was similar, but 2-fold higher than in the peritoneum of non-operated rats. The content of collagen in adhesions on day 21 after laparotomy in OD-treated rats was 10-fold lower than in animals no receiving OD.

Destructive Processes and Fibrotic Complications in the Liver of Mice with BSG-Induced Granulomatosis Treated with Anti-Tuberculous Drugs.

Three months after infection with Mycobacterium tuberculosis (MBT) from BCG vaccine, male BALB/с mice were treated with isonicotinic acid hydrazide, dextrazide (oxidized dextran), and liposome-encapsulated dextrazide intraperitoneally or in inhalations in a dose of 14 mg/kg (calculated for isoniazid) twice a week for 6 months. All these drugs exhibit different antimycobacterial efficiency. In the liver parenchyma, an up to 5-fold decrease in the number of destructed hepatocytes was observed depending on the efficiency of treatment. No destructive processes were observed in granulomas. Type I and III collagens were revealed around the granulomas; their content in the liver parenchyma was negligible. TNFα, IL-6, MMP-1, ТIMP1 were expressed only by granuloma macrophages. As the number of damaged hepatocytes and size of inflammatory infiltrates in the liver parenchyma decreased, the content of both types of collagen decreased. No evidence of hepatotoxicity of MBT degradation products in macrophages in vivo was obtained; the assumption that fibrotic complications are only the post-destruction process was not confirmed. Fibrotic complications are supposed to be an "excessive" systemic nonspecific adaptive process aimed at the maintenance the so-called structural homeostasis initiated by activated М2-macrophages in granulomas.

Analysis of the Efficiency of Different Antituberculous Drugs and Approaches to Treat BCG-Induced Granulomatosis in Mice and Abundance and Localization of Mycobacterium tuberculosis in the Liver.

In 3 months after infection with Mycobacterium tuberculosis (MBT) from BCG vaccine, male BALB/с mice received intraperitoneal injections of isonicotinic acid hydrazide, dextrazide, or liposome-encapsulated dextrazide, or inhalation of liposome-encapsulated dextrazide 2 times a week for 6 months. In 6 months, no MBT were detected in macrophages outside granulomas in treated mice. Macrophages containing MBT can incorporate into granulomas and leave them after suppression of MBT persistence. Liposome-encapsulated dextrazide showed the maximum therapeutic efficiency: the total MBT level in granuloma macrophages and volume density of destruction foci in the liver parenchyma decreased by 5.1 and 5.3 times, respectively, in comparison with the corresponding parameters in mice treated with isonicotinic acid hydrazide. Inhalations of liposome-encapsulated dextrazide prevented the destructive processes in liver granulomas due to macrophage migration from granulomas, which reduced granuloma sizes and destructive potential of granuloma lysosomes and therefore improved their diffusion-dependent trophics.

Structural Changes in the Lungs and Liver of Mice with Experimental Tuberculosis Treated with Liposome-Encapsulated Dextrazide.

Male BALB/с mice were intravenously infected with Mycobacterium tuberculosis H37Rv (0.5 ml of 2-week culture). One month later, treatment with liposome-encapsulated dextrazide (LEDZ, a conjugate of isonicotinic acid hydrazide (INH) and 40 kDa oxidized dextran encapsulated in phosphatidylcholine liposomes), INH, or a combination of LEDZ with INH was started. The doses of LEDZ (liposome suspension) and INH were 0.025 ml/10 g body weight and 5 mg/kg body weight, respectively. All the substances were administered 2 times a week via inhalation or intraperitoneally (a total of 40 doses). We studied the number and the size of tuberculous granulomas, the size of destruction foci and inflammatory infiltrates in the lungs and liver, the amount of fibrous connective tissue, and the dynamic of these parameters. LEDZ+INH inhalations were most effective by the therapeutic ratios in comparison with inhalation and intraperitoneal injections of INH.

Surgical treatment of atrial fibrillation: technique of thoracoscopic radiofrequency fragmentation of the left atrium. / Khirurgicheskoe lechenie fibrilliatsii predserdii: tekhnologiia vypolneniia torakoskopicheskoi radiochastotnoi fragmentatsii levogo predserdiia.

Presented in the article is a detailed description of a modified technique of minimally invasive surgical treatment of patients with atrial fibrillation - thoracoscopic radiofrequency fragmentation of the left atrium. This modification differs from the prototype GALAXY procedure by a significant increase of the 'quantitative' rather than 'qualitative' parameter of surgical aggression in relation to the left atrium. This technique results in creation of multiple transmural continuous closed lines of lesion to the left atrium and, consequently, a reduced risk of inadequate surgical treatment for atrial fibrillation. Besides the radiofrequency action on the wall of the left atrium, the protocol of the operation included destruction of the ligament of Marshall and resection of the left atrial appendage. An indication for performing this operation is the presence of various forms of atrial fibrillation.

[Biological aspects and clinical application of bone-marrow stem cells in critical lower limb ischaemia]. / Biologicheskie aspekty i klinicheskoe primenenie stvolovykh kletok pri kriticheskoi ishemii nizhnikh konechnostei.

Critical lower-limb ischaemia (CLLI) is associated with high risk of limb loss and a lethal outcome, as well as with decreased quality of life. The underlying cause of the disease is imbalance between blood supply of ischaemized tissues and their metabolic demands. Restoration of adequate perfusion with the help of standard medicamentous therapy or operative treatment is often inefficient or impossible. Cell therapy (CT) is a novel strategy making it possible to stimulate the growth of the microvascular bed and the mechanisms of molecular-cellular repair. One of the most promising trends is the use of bone marrow stem cells (BMSCs). Sufficient evidence concerning safety and relative efficacy of CT has by now been accumulated. The existing differences in the results of studies are related to numerous peculiarities of both CT itself and methods of its application. The present review is dedicated to contemporary notions of the biology of BMSCs, assessment of safety of CT and the results of its application in treatment of CLLI.

[Results of hybrid loop endarterectomy from the superficial femoral artery with the MultiTASC]. / Rezul'taty gibridnoi petlevoi éndarteréktomii iz poverkhnostnoi bedrennoi arterii petlei MultiTASC.

The purpose of this study was to analyse the results of hybrid loop endarterectomy from the superficial femoral artery (SFA) in its occlusion, preformed in a total of forty-two patients. Of these, 27 patients had prior to the intervention been diagnosed with stage II B ischaemia and 15 patients had been diagnosed as having critical ischaemia. Technical success of the operation amounted to 88%, with the frequency of early thromboses equalling 2.7%. In the early postoperative period, one patient died of acute myocardial infarction. The 1-, 2- and 3-year remote cumulative primary patency rate amounted to 81, 74 and 74%, respectively. There were no amputations performed within the timeframe of the follow-up period. The technique of hybrid loop endarterectomy with the MultiTASC loop followed by stenting of the proximal portion of the popliteal artery in occlusion of the SFA and stenoses of the common femoral artery has proved to be a highly efficient intervention yielding good immediate and remote results.

Study of Antiviral Efficiency of Oxidized Dextrans In Vitro and In Vivo.

Antiviral efficiency of oxidized dextrans (OD) with different molecular weights and oxidation degree (OD40min, OD70min, OD40max, and OD70 max) was studied in vitro and in vivo. Dextrans OD40max and OD70max prevented the development of the cytopathic effect of influenza A(H1N1)pdm09 virus in more than 50% MDCK cells vs. control (no OD). Four intranasal doses of OD40min, OD40max, and OD70min and one intranasal dose of OD70max before infection of BALB/c mice with A(H1N1)pdm09 influenza virus significantly reduced mortality and prolonged life span in comparison with controls receiving saline. These and our previous data attest to clear-cut preventive effect of OD in influenza infection.

Cell Death and Development of Fibrotic Alterations in Lung Granuloma of BALB/c Mice during Chronic BCG-Induced Granulomatosis.

Light microscopy, immunohistochemistry, and morphometric examinations established that cell death in lung granulomas of BCG-infected mice resulted mainly from activation of receptor-mediated apoptosis, which did not prevent the persistence of the causative agent in macrophages of the granulomas and promoted the formation of pronounced fibrosis in granulomas and pulmonary interstitium.

Preventive Effects of Oxidized Dextran on Functional Activity of Pulmonary Macrophages in Mice Infected with Influenza A Virus.

We analyzed cytokine profile of pulmonary macrophages in mice infected with highly pathogenic influenza A/H5N1 virus after preventive injections of oxidized dextran. Light microscopy, immunohistochemistry, and morphometric examinations showed that preventive injections of oxidized dextran led to more effective virus elimination, modulation of the proinflammatory cytokine response, and host antiviral response and reduce animal mortality. Our findings allow recommending oxidized dextran for further studies in order to create a vaccine with antiviral and adjuvant potencies.

Effect of Oxidized Dextran on Cytokine Production and Activation of IRF3 Transcription Factor in Macrophages from Mice of Opposite Strains with Different Sensitivity to Tuberculosis Infection.

We studied differences in the production of pro- and anti-inflammatory cytokines and IRF3 transcription factor by peritoneal macrophages from mice of opposite strains CBA/J and C57Bl/6 and the effect of 60-kDa oxidized dextran on these parameters. Macrophages from C57Bl/6 mice were mainly characterized by the production of proinflammatory cytokines TNFα, IL-12, and MCP-1 (markers of M1 polarization). By contrast, CBA/J mice exhibited a relatively high level of anti-inflammatory cytokine IL-10 and lower expression of proinflammatory cytokines (M2 phenotype). IRF3 content in peritoneal macrophages of CBA/J mice was higher than in C57Bl/6 mice. Oxidized dextran decreased the expression of IRF3 upon stimulation of cells from CBA/J mice with LPS, but increased this process in C57Bl/6 mice. Despite a diversity of oxidized dextran-induced changes in cytokine production, the data confirm our hypothesis that this agent can stimulate the alternative activation of macrophages.

Oxidized Dextran Enhances Alternative Activation of Macrophages in Mice of Opposite Lines.

Differences in peritoneal macrophage polarization in mice of opposite lines CBA and C57Bl/6 and the effects of 60 kDa oxidized dextran were studied. Macrophages of C57Bl/6 mice demonstrated a phenotype close to M1, with increasing expression of CD86 costimulatory molecule and unchanged CD206 expression in response to activation. Macrophages of CBA mice demonstrated higher plasticity in response to activating agents; expression of the markers increased irrespectively on stimulated receptor (TLR-4 or mannose receptor) and both CD86 (classical activation) and CD206 (alternative activation) increased. Macrophage response to addition of oxidized dextran (60 kDa) to the culture medium could be characterized as potentiation of their alternative activation: expression of CD86 in CBA mice in response to LPS and LPS+IL-4 and in C57Bl/6 mice in response to IFN-γ and LPS+IFN-γ decreased, while expression of CD206 by intact macrophages of CBA mice and by macrophages stimulated by IFN-γ and IL-4 increased under the effect of 60 kDa oxidized dextran.

[STUDYING THE ACUTE TOXICITY OF DEXTRAN POLYALDEHYDE CONJUGATE WITH ISONICOTINIC ACID HYDRAZIDE.]

The acute toxicity of a new antubercular drug, dextran polyaldehyde conjugate with isonicotinic acid hydrazide (dextrazide), has been studied on animals, HepaRG cell line, and FRSN human skin fibroblasts. Average tolerable doses (LD(10), LD(16)), half-lethal doses (LD(50)), and lethal doses (LD(84)) of dextrazide have been determined for intraperitoneal introduction in mice and rats and intravenous injection in rabbits.

Effects of Liposomal Compositions with Oxidized Dextrans on Functional Activity of U937 Macrophage-Like Cells In Vitro.

We studied the effects of liposomal pharmaceutical compositions with oxidized dextrans on functional activity of U937 monocyte/macrophage-like cells. Liposomes in the emulsion contained oxidized dextran with a molecular weights of 40 kDa or 70 kDa or isonicotinic acid hydrazide (INAH) conjugated with oxidized dextran (40 kDa). Cell viability was evaluated by MTT test; mitochondrial transmembrane potential and production of superoxide anion and H2O2 were studied by fluorescent methods. The studied compositions exhibited no cytotoxic effect and even improved cell viability and mitochondrial respiration. Liposomes with oxidized 40 kDa dextran, including those with INAH-conjugated dextran, inhibited production of superoxide anion, but increased H2O2 generation.

Modifying Effects of Nanosized Diamonds on Hydrolytic Potential of Macrophages In Vitro.

The effects of nanosized particles (4-6 nm) of synthetic diamonds on mouse macrophage expression and secretion of lysosomal cathepsins (B and D) and MMP-1 and MMP-9 were studied in vitro. Culturing of peritoneal macrophages in medium with diamond nanoparticles led to an increase in the counts of macrophages expressing the above enzymes and to stimulation of their secretion. However, the manifestations of these effects varied significantly for various enzymes. The data indicate modulation of macrophage functions by nanodiamonds. These results help better understand the possible role of the "corpuscular" xenobiotic factors in the pathogenesis of diseases associated with macrophage capturing of these factors irrespective of their chemical "activity".

Effectiveness of composition based on oxidized dextran in the treatment of grade IIIB skin burns.

Grade IIIB skin burns were treated with a composition based on oxidized dextran with a molecular weight of 40 kDa (oxidation of 7% glucose residues). On day 32 after burn infliction and from the start of the treatment, the area of skin defect in rats was 30% less than in the group without treatment and by 2.3 times less than in rats treated with panthenol. In rats treated with dextran-based composition or panthenol, the eschar was absent on day 21 after the start of the treatment; by day 32, we found cells of surface epithelium, hair follicles, and sebaceous glands above the scar tissue that were absent in untreated animals; in rats treated with the composition, their number was higher by 2.5 times than in animals treated with panthenol. Treatment with the composition increased volume density (by 2.5 times) and numerical density (by more than 3 times) of blood vessels in the wound and reduced signs of inflammation and fibroplastic activity of fibroblasts in comparison with the corresponding parameters in untreated animals or animals treated with panthenol.

In vitro effects of nanosized diamond particles on macrophages.

The effects of synthetic diamond nanoparticles (4-6 nm) on mouse macrophage biotropism and biocompatibility and the modulation of the macrophage functions (expression of IL-1α, TNF-α, GM-CSF, bFGF, and TGF-ß) by nanoparticles in different concentrations were studied in vitro during exposure of different duration. Macrophage endocytosis of nanodiamonds increased with increasing the concentration of nanoparticles in culture and incubation time. Nanodiamonds exhibited high biotropism and biocompatibility towards macrophages; in doses of 10-20 µg/ml, they induced expression of GM-CSF and TGF-ß, inhibited expression of bFGF, and did not stimulate IL-1α and TNF-α. These data indicate that nanodiamond capture by macrophages in the studied experimental model led to modulation of the functional status of macrophages that determine their capacity to stimulate reparative processes without increasing proinflammatory and profibrogenic status.

Effects of preventive administration of oxidized dextran on liver injury and reparative regeneration in mice infected with influenza A/H5N1 virus.

Intranasal infection of outbred male mice with influenza A/H5N1 A/goose/Krasnoozerskoye/627/05 virus led to high (85%) mortality of animals. Morphological studies of liver specimens showed destructive changes in the parenchyma (93.5% hepatocytes), caused by long persistence of the virus in the liver. The virus persistence was conjugated with activation of cellular immunity, manifesting by an increase in the counts of cells with high expression of proinflammatory cytokines (TNF-α) and lysosomal enzymes (lysozyme, cathepsin D). Injections of oxidized dextran 3 and 1 days before infection reduced mortality and 2-fold attenuated destructive changes in the liver, presumably due to prevention of virus penetration into the target cells, modulation of immune reactions, and stimulation of reparative plastic processes.

Experimental study of the efficiency of oxidized dextran for prevention of influenza A/H5N1.

Oxidized dextran is suggested for prevention of infection induced by influenza A/H5N1 viruses, methods of its use and doses are determined. Two intravenous injections of dextran 3 and 1 days before experimental infection of outbred mice by influenza A/H5N1 A/goose/Krasnoozerskoye/627/05 virus resulted in a high preventive dose-dependent effect: the mean lifespan was 25% prolonged, the mortality decreased 3-fold.

[Comparison of morphological features of carotid arteries atherosclerotic plaques with clinical-instrumental data in symptomatic and asymptomatic patients with severe carotid atherosclerosis].

A complex histomorphometric and clinical-instrumental analysis of atherosclerotic lesions of the carotid arteries obtained during carotid endarterectomies (CEE) of patients with hemodynamically significant stenoses was conducted. Two groups of patients were compared: symptomatic, which earlier underwent cerebral vascular accident (CVA) or transitory ischemic attacks (TIA), and asymptomatic ones with no complications of the disease. Statistical analysis of clinical and laboratory data showed no significant differences between two groups except for the level of lipoprotein(a) [Lp(a)] in the blood plasma, which was higher (p<0.05) in asymptomatic patients compared with symptomatic ones. Statistical analysis of carotid arteries ultrasound duplex scanning (USDS) in the preoperative period did not reveal significant differences in the degree of maximum vessels stenosis between the compared groups of patients. Surface defects of atherosclerotic plaques (ASP) were shown to be significantly more common (p<0.05) in the group of symptomatic patients compared with asymptomatic ones. According to histological analysis 88% of extracted ASP was unstable in symptomatic patients and 77% of ASP - in asymptomatic patients. This may indicate high risk of CVA/TIA in both groups of patients. Statistical evaluation of magnetic resonance tomography (MRT) and USDS techniques in comparison with abilities of the most reliable histological analysis showed that both non-invasive diagnostic methods are highly sensitive in detecting unstable ASP, though MRT showed higher level of specificity compared with USDS.