[The increase in receptor-mediated endocytosis of drugs in the composition of nanoparticles with the address fragment]. / Uvelichenie éndotsitoza protivoopukholevykh lekarstv v sostave nanochastits s adresnym fragmentom.

It is known that disorders in the cell functioning of the organs/tissues is accompanied by increased expression of certain receptors. A modern approach to improve the specificity of the drug accumulation in the affected area is to construct the delivery nanosystems with the address fragments. Active tagged transport may help to reduce the dose of the drug, minimizing the impact on healthy cells and organs (reduced adverse events). This approach is particularly important in oncology because of the high toxicity of the drugs used. In this work we have obtained and characterized the pharmaceutical composition of doxorubicin and chlorine e6 into colloidal nanoparticles with synthesized previously targeted conjugates based on folic acid and biotin. On the cell culture Hep G2 it was shown an increase in the internalization of drugs when they were introduced in the incubation medium in the form of drug compositions with transport nanosystems and targeted fragments.

[The in vivo study of the medicinal composition property of doxorubicin as a part of colloidal nanoparticles with the address fragment].

The use of targeted transport systems for drug delivery is a promising approach to improve pharmacokinetics of drug substances, accumulation in the lesion. In this study we have obtained and characterized the pharmaceutical composition of doxorubicin in colloidal nanoparticles equipped with targeted conjugates based on folic acid and biotin with dodecylamine. The inclusion of the address fragments into colloidal nanopartical was carried out without surface and drug substance modification The accumulation of anthracycline antibiotic doxorubicin in tumor tissue was compared in Lewis lung carcinoma mouse models after intravenous administration of the composition of colloidal nanoparticles with targeted conjugates biotin-dodecylamine and folic acid-dodecylamine or free doxorubicin. It was shown that the doxorubicin accumulation in tumor tissue when administered in drug compositions with targeted fragments are 2 times higher for the folic acid-dodecylamine conjugate and 1.4 times higher for the biotin-dodecylamine conjugate.

[Pharmacological agents and transport nanosystems based on plant phospholipids].

A new generation of plant phosphatidylcholine (PC)-based pharmacological agents has been developed under academician A.I. Archakov leadership at the Institute of Biomedical Chemistry (IBMC). For their production a unique technology allowing to obtain dry lyophilized phospholipid nanoparticles of 30 nm was elaborated. The successful practical application of PC nanoparticles as a drug agent may be illustrated by Phosphogliv (oral and injection formulations). Being developed at IBMC for the treatment of liver diseases, including viral hepatitis, Phosphogliv (currently marketed by the "Pharmstandard" company) is approved for clinical application in 2000, and is widely used in medical practice. Based on the developed and scaled in IBMC technology of prerparation of ultra small size phospholipid nanoparticles without the use of detergents/surfactants and stabilizers another drug preparation, Phospholipovit, exhibiting pronounced hypolipidemic properties has been obtained. Recently completed preclinical studies have shown that PC nanoparticles of 20-30 nm activate reverse cholesterol transport (RCT) and in this context it is more active than well known foreign preparation Essentiale. Phospholipovit is now at the stage of clinical trials (phase 1 completed). PC was also used as a basis for the development of a transport nanosystem with a particles size of 20-25 nm in diameter and incorporation of various drug substances from various therapeutic groups. Using several drugs substances as an example, increased bioavailability and specific activity were demonstrated for the formulations equipped with such transport nanosystem. Formulations equipped with the transport nanosystems have been developed for such pharmacological agents as doxorubicin, rifampin, budesonide, chlorin E6, prednisone, and others.

Role of Small Noncoding RNAs in Bacterial Metabolism.

The study of prokaryotic small RNAs is one of the most important directions in modern molecular biology. In the last decade, multiple short regulatory transcripts have been found in prokaryotes, and for some of them functional roles have been elucidated. Bacterial small RNAs are implicated in the regulation of transcription and translation, and they affect mRNA stability and gene expression via different mechanisms, including changes in mRNA conformation and interaction with proteins. Most small RNAs are expressed in response to external factors, and they help bacteria to adapt to changing environmental conditions. Bacterial infections of various origins remain a serious medical problem, despite significant progress in fighting them. Discovery of mechanisms that bacteria employ to survive in infected organisms and ways to block these mechanisms is promising for finding new treatments for bacterial infections. Regulation of pathogenesis with small RNAs is an attractive example of such mechanisms. This review considers the role of bacterial small RNAs in adaptation to stress conditions. We pay special attention to the role of small RNAs in Mycobacterium tuberculosis infection, in particular during establishment and maintenance of latent infection.

[Clinical profile and treatment of left main coronary artery disease in a real-world practice].

Aim of this study was to assess clinical profile, treatment and long-term results in patients with left main coronary artery disease in a real-world practice. 225 cases were analyzed. Long-term results were evaluated from 213 (97.7%) patients. Median follow-up period was 49 months. Fifty two (23.9%) patients received nonsurgical treatment, coronary artery bypass grafting (CABG) was performed in 106 (48.6%) patients, percutaneous coronary interventions (PCI) in 60 (27.5%) patients. Patients of nonsurgical group had more severe clinical profile compared with PCI group. There was no differences between nonsurgical and CABG groups as well as between CABG and PCI groups in clinical profile. Major adverse cardiac and cerebrovascular event rate was higher in nonsurgical group. There was no difference between CABG and PCI groups. Survival in CABG and PCI groups was higher compared with nonsurgical group. Survival effect of revascularization was observed immediately (before 6 months) and continued long-term (more than 60 months). Revascularization was independent predictor of improved long-term survival. After diagnosis of left main coronary artery stenosis PCI was performed more quickly than CABG.

[Development of targeted drug delivery system: synthesis of conjugates of address fragment (RA-COOH) with ligand (R-NH2)].

One of the main ways to increase the effectiveness of well-known medical formulations well-established in clinical medicine - development of delivery systems using new technological approaches and nanomaterials. Currently, much attention is given to targeted delivery systems. At the same time drug carrier has in addition to medication the so-called vector/address with a high affinity for binding to specific receptors on cells/tissue target. In this paper it is described the method for producing of address conjugates to over-expressed receptors on the tumor cells. As address fragment it was folic acid and as a linker was dodecylamine, causing inclusion the conjugate into lipid nanoparticles.

[Expression of Mycobacterium tuberculosis small RNAs in mice models of tuberculosis].

Identification of mechanisms that contribute to bacterial pathogens survival in the infected organism is a powerful approach to influence the pathogenic bacteria. Recently it was established that bacteria use small RNAs to regulate their metabolism. We studied the expression level of the three most highly expressed M. tuberculosis small RNAs MTS0997, MTS 1338 and MTS2823 at different stages of infection in. mice with different genetic resistance to tuberculosis. The maximum expression level of these small RNAs was observed at the early stages of infection.

Mycobacterium tuberculosis Transcriptome Profiling in Mice with Genetically Different Susceptibility to Tuberculosis.

Whole transcriptome profiling is now almost routinely used in various fields of biology, including microbiology. In vivo transcriptome studies usually provide relevant information about the biological processes in the organism and thus are indispensable for the formulation of hypotheses, testing, and correcting. In this study, we describe the results of genome-wide transcriptional profiling of the major human bacterial pathogen M. tuberculosis during its persistence in lungs. Two mouse strains differing in their susceptibility to tuberculosis were used for experimental infection with M. tuberculosis. Mycobacterial transcriptomes obtained from the infected tissues of the mice at two different time points were analyzed by deep sequencing and compared. It was hypothesized that the changes in the M. tuberculosis transcriptome may attest to the activation of the metabolism of lipids and amino acids, transition to anaerobic respiration, and increased expression of the factors modulating the immune response. A total of 209 genes were determined whose expression increased with disease progression in both host strains (commonly upregulated genes, CUG). Among them, the genes related to the functional categories of lipid metabolism, cell wall, and cell processes are of great interest. It was assumed that the products of these genes are involved in M. tuberculosis adaptation to the host immune system defense, thus being potential targets for drug development.

[Identified small RNAs of Mycobacterium avium].

Posttranscriptional regulation of gene expression by small RNAs was shown for multiple pathogenic microorganisms and plays an important role in virulence. 4 putative sRNA genes located in intergenic loci were identified: MAV_0380-0381 (4.5S RNA), MAV_1034-1035 (trans-encoded sRNA), MAV_1415-1416 (antisense or trans-encoded sRNA) and MAV_1531-1532 (processed 5' UTR of 16S rRNA gene). The revealed sRNAs represent the first small noncoding RNAs identified in M. avium.

Adaptive Changes in Mycobacterium avium Gene Expression Profile Following Infection of Genetically Susceptible and Resistant Mice.

We performed a comparative analysis ofMycobacterium aviumtranscriptomes (strain 724R) in infected mice of two different strains- resistant and susceptible to infection. Sets of mycobacterial genes transcribed in lung tissue were defined, and differentially transcribed genes were revealed. Our results indicate thatM. aviumgenes coding for enzymes of the Krebs cycle, oxidative phosphorylation, NO reduction, fatty acid biosynthesis, replication, translation, and genome modification are expressed at high levels in the lungs of genetically susceptible mice. The expression of genes responsible for cell wall properties, anaerobic nitrate respiration, fatty acid degradation, synthesis of polycyclic fatty acid derivatives, and biosynthesis of mycobactin and other polyketides is increased in the resistant mice. In the resistant host environment,Mycobacterium aviumapparently transitions to a latent state caused by the deficiency in divalent cations and characterised by anaerobic respiration, degradation of fatty acids, and modification of cell wall properties.

Blood coagulation initiates respiratory burst in neutrophils.

Using the in vitro reaction of nitroblue tetrazolium reduction we showed that blood coagulation stimulates production of reactive oxygen species by human neutrophils. Heparin and chondroitin sulfate produced by thrombin-activated basophils are good candidates for inductors of these processes. Similar activation probably occurs in vivo under the influence of inductors secreted by mast cells.

[Nitroblue tetrazolium reduction by human blood neutrophils. I. The influence of pH].

Activity of blood neutrophils of 14 healthy volunteers (8 women, 6 men, aged from 21 to 37 years) was investigated in the Nitroblue Tetrazolium (NBT) reduction reaction at pH 7.0, 7.3, and 7.6. The dependence of the percentage of NBT-positive neutrophils on the reaction time was described by a kinetic equation with constants k1 and k2, characterizing, accordingly, speeds of NBT absorption and reduction by neutrophils. A strong positive correlation (r = 0.99; P < 0.0001), was found between k1 and k2, which pointed to dependence of the speed of NBT restoring by neutrophils on the speed of its entry to the cell. Both the constants and speeds of successive process stages have maximum values at pH 7.3 (corresponding to physiological value of blood pH), and decrease at medium acidification or alkalization. The k1/k2 ratio does not depend on the time (0-6 h) or pH (7.0-7.6) of blood incubation before the reaction. The most active neutrophils (potential pH agocytes) have a lesser membrane permeability than do neutrophils of another class, and their activity more increases at medium acidification in vitro and, probably, in vivo. The activation of neutrophils leads to an increase in medium pH that alongside with an increase in the number of active potential phagocytes, evidently has a physiological meaning.

[Nitroblue tetrazolium reduction by human blood neutrophils. II. The influence of sodium and potassium ions].

Activity of blood neutrophils of 18 healthy volunteers (4 men and 14 women aged from 20 to 32 years) was investigated in the Nitroblue Tetrazolium (NBT) reduction reaction in Na- and K-phosphate buffers (0.1 M, pH 7.3) after a preliminary incubation of heparinized blood (37 degrees C, 0-7 h). It was found that the percentage of NBT-positive neutrophils in the presence of Na+ is higher than in the presence of K+. The dependence of percentage of NBT-positive neutrophils on the reaction time is described by a kinetic equation with constants k1 and k2, characterizing accordingly, speeds of NBT absorption and reduction by neutrophils. It has been shown that k1, k2 and k1/k2 values differ in potential phagocytes and in neutrophils, which are not capable of adherence. The kinetics constants of the latter cells do not depend on both cations and incubation time, while the percentage of NBT-positive potential phagocytes, and their k1 and k2 in the presence of Na+ are higher than in the presence of K+, and the values of described parameters are changed during incubation. During incubation the percentage of NBT-positive neutrophils increases to result in a partial aggregation of thrombocytes, as a consequence of excretion in the blood of products involved in this process.

[Neutrophil activation in vitro]. / Osobennosti aktivatsii neitrofilov in vitro.

Kinetics of spontaneous and induced reactions of Nitroblue tetrazolium (NBT) restoration by neutrophils was investigated in vitro. The index of activated neutrophils (IAN), commonly characterizing cell activation degree depending on the reaction time (t), was described by the equation: IAN = [NBT0](1-exp[-k1t]) + k0t; where (NBT0) is intracellular concentration of NBT, which reflects membrane permeability, k1 and k0--speed constants of the first and zero order reactions, characterizing, respectively, stimulation and spontaneous activities of enzymatic systems involved in NBT restoration. We detected the ability of heparin (2.0-7.5 mkg/ml) or chondrotin sulphate (0.25-2.5 mkg/ml) to activate neutrophils and presumably to maintain in vivo their spontaneous activity. The values of kinetics equation parameters enable us to speculate about relative hyper-, hypo- and normal function of neutrophils, related to the functional state of the whole organism. The value of [NBT0] changes from 8 to 103 conventional units, and a high correlation exists between this index and the reaction speed constants, differing in stimulated and spontaneously activated cells. The latter are represented by neutrophils only, which, as shown before, produce active forms of oxygen, used, probably, not for phagocytosis needs. The share of such cells makes approximately 30-40% of the neutrophil recirculating pool, and positively correlates with [NBT0], (r = 0.58; P < 0.05). This is determined by a very high membrane permeability of this neutrophil subpopulation. The detected different spontaneous and stimulated ability, various membrane permeability and enzyme system activity confirm the neutrophil functional unequality, which defines their activation peculiarity.

[Palmitates of isomeric 15-oxygenated delta8(14)-sterols]. / Pal'mitaty izomernykh 15-oksigenirovannykh delat 8(14)-sterinov.

3 beta-Hexadecanoyloxy-5 alpha-cholest-8(14)-en-15-one, 3 alpha-hexadecanoyloxy-5 alpha-cholest-8(14)-en-15-one, 15 beta- hexadecanoyloxy-5 alpha-cholest-8(14)-en-3 beta-ol, 15 alpha-hexadecanoyloxy-5 alpha-cholest-8(14)-en-3 beta-ol, 15 beta-hexadecanoyloxy-5 alpha-cholest-8(14)-en-3-one, and 15 alpha-hexadecanoyloxy-5 alpha-cholest-8(14)-en-3-one were synthesized and their chromatographic and 1H NMR characteristics were determined. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 2; see also http://www.maik.ru.

[A simple method for preparation of 18alpha-glycyrretinic acid ant its derivatives]. / Prostoi metod polucheniia 18alpha-glitsirretinovoi kisloty i ee proizvodnykh.

Treatment of 18 beta-glycyrrhizic acid with a methanolic solution of HCl resulted in a 1:1 mixture of methyl esters of 18 alpha- and 18 beta-glycyrrhetinic acids. Benzoylation of the mixture led to methyl esters of 3-benzoyl-18 alpha-glycyrrhetinic acid and 3-benzoyl-18 beta-glycyrrhetinic acid, which were separated by chromatography on silica gel. 18 alpha-Glycyrrhetinic acid was prepared by alkaline hydrolysis of methyl 3-benzoyl-18 alpha-glycyrrhetinate and was further used for the syntheses of 3-keto-18 alpha-glycyrrhetinic acid and methyl esters of 18 alpha-glycyrrhetinic acid and 3-keto-18 alpha-glycyrrhetinic acid.

[Transformation of 3beta-(2-hydroxyethoxy)-5alpha-cholest-8(14)-en-15-one in a polar metabolite in HepG2 hepatoma cells]. / Prevrashchnie 3beta-(2-gidroksiétoksi)5alpha-kholest-8(14)-en-15-ona v poliarnyi metabolit v kletkakh gepatomy HepG2.

Incubation of 3 beta-(2-hydroxy-2[3H]-ethoxy)-5 alpha-cholest-8(14)-en-15-one with Hep G2 cells led to the accumulation of a radioactive polar product in the culture medium, which was identified as 3 beta-(2-hydroxyethoxy)-15-keto-5 alpha-cholest-8(14)-ene-24-oic acid. Its structure was confirmed by a chemical counter synthesis. The labeled ketosterol was rapidly (tau 1/2 = 6 min) and reversibly bound by Hep G2 cells. The intracellular concentration of 15-ketosterol decreased during incubation mainly due to the formation of a polar metabolite, secreted to the medium. The level of cholesterol biosynthesis was 22 +/- 5% of the control value in Hep G2 cells at a 15-ketocholesterol concentration in the medium of 30 microM. However, further incubation for 3 h in the medium without the ketosterol led to restoration of the level of biosynthesis to 85 +/- 11% of the control value. These results suggest that inhibition of the cholesterol biosynthesis by 15-ketocholesterol in Hep G2 cells depends on the intracellular concentration of the inhibitor, which, in turn, is determined by the rate of its conversion into the polar metabolite. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2003, vol. 29, no. 6; see also http://www.maik.ru.

[Stereospecific reduction of keto group in 3beta-triphenylmethoxy-5alpha-cholest-8(14)-en-15-one]. / Stereospetsifichnoe vosstanovlenie ketogruppy v 3beta-trifenilmetoksi-5alpha-kholest-8(14)-en-15-one.

The reduction of 3 beta-triphenylmethoxy-5 alpha-cholest-8(14)-en-15-one with lithium aluminum hydride resulted in a quantitative yield of 3 beta-triphenylmethoxy-5 alpha-cholest-8(14)-en-15 beta-ol.