[Adherence to guidelines on management of acute coronary syndrome in Russian hospitals and outcomes of hospitalization (data from the RECORD-2 Registry)].

Background: Complete following existing guidelines for management of acute coronary syndrome (ACS) is known to be associated with better outcomes. Partly this is explained by lesser adherence to recommendations in high risk patients. Aim of our study was to assess relationship between degree of following current guidelines and in hospital outcomes independently from initial assessment of risk.

[Preparation of prokaryotic cDNA for high-throughput transcriptome analysis].

High contents of non-coding RNA in total bacteria RNA complicates considerably transcriptome analysis using standard approaches like high-throughput sequencing, gene expression profiles, subtractive hybridization. We suggest a procedure of preparation of bacterial cDNA for transcriptomics that includes rRNA and tRNA depletion with preservation of relative abundance of coding sequences. The method is based on the second order hybridization kinetics and unique properties of Kanchatka crab duplex-specific nuclease.

[Application of the duplex-specific nuclease for fast analysis of single nucleotide polymorphisms and detection of target DNA in complex PCR products].

We have developed a simple method for fast analysis of single nucleotide polymorphisms and identification of target clones from cloned complex PCR products. The method utilizes Kamchatka crab duplex-specific nuclease and universal fluorescent probe and is alternative to laborious screening procedures using radioactive probes, restriction analysis followed by gel electrophoresis or expensive sequencing. The method efficacy was demonstrated in several model experiments.

[Signaling function of phagocytic NADPH oxidase: activation of MAP kinase cascades in phagocytosis].

Until recently, the production of reactive oxygen species by NADPH oxidase has been considered only in the context of the oxidative damage to pathogens inside the phagosome. However, homologues of phagocytic NADPH oxidase have been found in almost all cell types, where they produce hydrogen peroxide and thereby regulate the initial intracellular stages of MAP kinase cascades. In the present work, the activation of two MAP kinase cascades, p38 and Erk1/2, during phagocytosis has been studied.

[A method for the preparation of normalized cDNA libraries enriched with full-length sequences].

We developed a new method for the preparation of normalized cDNA libraries enriched with full-length sequences. It is based on the properties of the recently characterized duplex-specific nuclease from the hepatopancreas of the Kamchatka crab. The duplex-specific nuclease is thermostable, it effectively cleaves double-stranded DNA and is inactive toward single-stranded DNA (Shagin et al.

[A family of genes of multidomain free lectins from a planarian: structure, expression, and use as markers for regeneration monitoring ].

A family of genes of the agamic race of planarian Girardia tigrina were described that encode proteins that belong to the superfamily of C-type lectins and were demonstrated to have a unique domain organization. The genes are differentially expressed in the planarian body. The protein products of at least two genes (scarf2 and gtlec1) are expressed in specifically differentiated gland cells of the planarian and secreted into the environment through long cell necks.

[Cloning and analysis of a new neurogene in the mouse].

The previously described gene sip1 belongs to transcription factors of the zinc finger family. It has been ascertained recently that this gene is involved in TGF signaling cascade. Mutations in human gene sip1 cause Hirschprung syndrome.

[A natural fluorescent protein that changes its fluorescence during maturation].

The gene of a new red fluorescent protein zoan2RFP from a coral polyp Zoanthus sp., a homologue of the known green fluorescent protein from the Aequorea victoria jellyfish, was cloned. At early maturation stages, zoan2RFP exhibits a green fluorescence, which then turns into the red one.

[A new planarian extrachromosomal virus-like element revealed by subtraction hybridization].

A combination of suppression subtraction hybridization (SSH) and a new technique of mirror orientation selection (MOS) was used to compare the total DNA for two, sexual (SR) and asexual (AR), races of freshwater planarian Giradia tigrina. Several race-specific DNA fragments were found. A new element termed planarian extrachromosomal virus-like element (PEVE) was revealed in AR.

[Cloning and structure of gene encoded alpha-latrocrustoxin from the Black widow spider venom].

The primary structure of the crusta gene encoding alpha-latrocrustoxin (alpha-LCT), a high molecular mass neurotoxin specific to crustaceans, was determined in the black widow spider Latrodectus mactans tredicimguttatus genome. The total length of the sequenced DNA was 4693 bp. The structural part of the black widow spider chromosome gene encoding alpha-LCT does not contain introns.

[Selective suppression of polymerase chain reaction].

The selective suppression of the polymerase chain reaction and methods based upon it (construction of cDNA libraries from low amounts of biological material, subtractive hybridization and differential display of mRNA, fast cloning of full-size cDNA, chromosome walking, cloning in vitro, and others) are reviewed. These methods display a high effectiveness and, taken together, enable intricate DNA analyses to be performed--from the search for nontrivial sequences to the total sequencing of the corresponding genes.

[Cloning of region-specific genetic markers of planaria using a new method--ordered differential display].

A new method for finding differentially expressed genes, termed ordered differential display of mRNAs (ODD), was used in the search for region-specific molecular markers of freshwater planarian Dugesia tigrina. In this method, the effect of selective suppression of a polymerase chain reaction (PCR) is used for the differential amplification of a pool of 3'-terminal cDNA fragments generated by digestion of cDNAs with a restriction endonuclease. In the resulting amplified cDNAs, every mRNA is represented by a cDNA fragment whose length is determined by the position of the restriction site nearest to the 3'-terminus.

[In vitro cloning of DNA fragments by one polymerase chain reaction].

An improved version of the in vitro DNA cloning method described earlier is proposed. The method allows amplification by polymerase chain reaction (PCR) of individual DNA molecules of an unknown primary structure followed by sequencing. The modifications described here provide for in vitro cloning by means of a 40-45-cycle PCR (the original protocol required two consecutive amplifications).

[An analysis of the expression of the genes containing the LeR-1 and VeR-1 sequences in the embryogenesis, regeneration and in the intact tissues of newts].

A method of construction of amplified cDNA libraries was developed on the basis of selective inhibition of cDNA amplification and modified for the studied models for analysis of expression of the genes containing LeR-1 and VeR-1 sequences. Time-related changes in expression of these genes were studied during regeneration of the adult lens and during embryogenesis of newts. The pattern of expression of the LeR-1 and VeR-1 genes proved to be not tissue-specific.

[Differential gene expression during the reparative regeneration of differing polarities in planarians].

We identified two new genes (scarf and collar) of planarians using subtracting hybridization. mR-NAs of these genes are distributed in different ways in regeneration blastemas of different polarity. Zone-specific expression of these genes in non-regenerating planarians has been demonstrated.

[A novel method of subtraction cDNA hybridization allowing cloning of genes showing differential expression in cell micropopulations].

A novel, efficient and simple technique that combines subtractive hybridization with kinetic enrichment is proposed for obtaining enriched cDNA. The method is based on the use of a set of special primers that allow for the selective amplification by PCR only of differentially distributed sequences. Using the proposed technique, cDNA of a new gene XEp-1, specifically expressed in the presumptive epidermis of Xenopus laevis, was cloned, starting with the stage of midgastrula.

[A method for obtaining the normalized cDNA libraries based on the effect of suppression of polymerase chain reaction].

A new efficient method for obtaining cDNA libraries with equal representation of all cDNA types (equalized libraries) in a single round of equalization was developed. The method is based on differences in the renaturation kinetics of double-stranded cDNAs of different genes and allow the selection of the equalized single-stranded fraction resulted from the incomplete reassociation of the total cDNA without laborious and inefficient physical separation. The equalized single-stranded fractions are selectively amplified by polymerase chain reaction (PCR).

[Cloning cDNA for the ha-SDGF gene from a Syrian hamster cell line with increased metastatic potential using subtractive hybridization].

Using subtractive hybridization, a cDNA library containing over 50% of clones specific for a highly metastatic cell line was obtained from two hamster embryo fibroblast lines with different metastatic potentials. Most of the clones (83%) contained new sequences. One clone contained the ha-SDGF gene cDNA homologous to SDGF cDNA from rodents.

[Selective amplification of evolutionally conserved expressed sequences].

An efficacious method of cloning the sequences common for two cDNAs was proposed. The method was used for constructing a library of expressed sequences evolutionarily conserved for human and hamster.

[Molecular-biological approaches to studying gene expression during regeneration].

This paper constitutes a review of the methodical approaches allowing analysis of the mechanisms underlying development and differentiation. Progress in investigation of the mechanisms underlying embryogenesis is related to the discovery of genic families in the Drosophila genome, which are responsible for different periods of embryogenesis. The true revolution in studies of developmental mechanisms began with the application of molecular-genetic methods for analysis of Drosophila mutant lines.

[Identification of new genes and analysis of their expression during lens and retinal regeneration in adult newts].

During lens regeneration in Pleurodeles waltl, the dorsal iris zone is the cell source of the lens regeneration, while the ventral iris zone can serve as the cells' source of lens regeneration only under certain experimental conditions. The method of subtractive hybridization was used for the identification of genes responsible for the different proliferative potential of these zones. Differential screening of the enriched cDNA libraries, which were obtained as a result of subtractive hybridization of the cDNA samples of the ventral and dorsal iris zones 14 days after lens removal, revealed four clones specific to the dorsal iris and six clones specific to the ventral iris.

[The cloning of fragments of homeobox genes expressed during planarian regeneration].

The polymerase chain reaction with degenerate primers corresponding to the most conservative amino acids 16-21 (ELEKEF) and 49-54 (WFQNRR) of the Antennapedia class homeodomain was used for the amplification of cDNA from regenerating planarians (asexual race of Dugesia tigrina). A total of six new Antennapedia-like homeobox sequences, designated Dutarh-1-Dutarh-6 (Dugesia tigrina asexual race homeobox gene), were obtained. Their comparison with other homeobox genes using a "Genebee" software (the EMBL Data Library) showed that all sequences except Dutarh-6 belong to the Antennapedia class.

[Highly-effective subtractive hybridization of cDNA].

A scheme for subtractive hybridization is described allowing for a 500-1000 fold enrichment of low abundant cDNA. The scheme is based on the previously described principle of normalization of an initial mixture of differently represented cDNAs in the single-stranded portion of a tracer after the first round of subtraction and the principle of a trapper excluding the fraction of the double-stranded cDNAs formed during the first round from the subsequent PCR-amplification. The technique is simple and makes unnecessary the separation of the tracer, driver and hybrids formed after the subtraction.

[Excretion of pyridinoline with urine in children with Ehlers-Danlos syndrome and Marfan's syndrome and the effect of beta blocker and complex vitamin therapy on it].

Excretion of pyridinoline and polypeptide-bound hydroxyproline with urine was studied in 27 children with hereditary impairment of connective tissue. At the same time, effects of beta-adrenoblocking agents and vitamin complex, prescribed during preoperation period before thoracoplasty in hereditary chest deformation, were investigated. Clinical efficiency of the treatment depended distinctly on the initial value of ratios pyridinoline/creatinine and polypeptide-bound hydroxyproline/creatinine.

[Anesthesiologic and postoperative complications in children with isolated forms of funnel chest].

In thoracoplasty for isolated funnel-shaped deformity of the chest in children, the main anesthesiologic complications which develop are the following: delayed restoration of muscular tonicity and adequate respiration and disorders in cardiac rhythm. At the postoperative period, the complications, are mainly caused by injury to parietal pleura at intervention. The pulmonary, cardiac, gastroenterologic and hemorrhagic complications were observed less often.