OPTIMUM LEVEL OF POSITIVE END-EXPIRATORY PRESSURE IN ACUTE RESPIRATORY DISTRESS SYNDROME CAUSED BY INFLUENZA A(H1NI)PDM09: BALANCE BETWEEN MAXIMAL END-EXPIRATORY VOLUME AND MINIMAL ALVEOLAR OVERDISTENSION.

THE AIM: to determine optimum level ofpositive end-expiratory pressure (PEEP) according to balance between maxi- mal end-expiratory lung volume (EEL V)(more than predicted) and minimal decrease in exhaled carbon dioxide volume (VCO) and then to develop the algorithm of gas exchange correction based on prognostic values of EEL K; alveolar recruitability, PA/FiO2, static compliance (C,,,) and VCO2. MATERIALS AND METHODS: 27 mechanically ventilatedpatients with acute respiratory distress syndrome (ARDS) caused by influenza A (HINJ)pdm09 in Moscow Municipal Clinics ICU's from January to March 2016 were included in the trial. At the beginning of the study patients had the following characteristic: duration offlu symptoms 5 (3-10) days, p.0/FiO2 120 (70-50) mmHg. SOFA 7 (5-9), body mass index 30.1 (26.4-33.8) kg/m², static compliance of respiratory system 35 (30-40) ml/mbar: Under sedation and paralysis we measured EELV, C VCO and end-tidal carbon dioxide concentration (EtCO) (for CO2 measurements we fixed short-term values after 2 min after PEEP level change) at PEEP 8, 11,13,15,18, 20 mbar consequently, and incase of good recruitability, at 22 and 24 mbar. After analyses of obtained data we determined PEEP value in which increase in EELV was maximal (more than predicted) and depression of VCO2 was less than 20%, change in mean blood pressure and heart rate were both less than 20% (measured at PEEP 8 mbar). After that we set thus determined level of PEEP and didn't change it for 5 days. RESULTS: Comparision of predicted and measured EELV revealed two typical points of alveloar recruiment: the first at PEEP 11-15 mbar, the second at PEEP 20-22 mbar. EELV measured at PEEP 18 mbar appeared to be higher than predicted at PEEP 8 mbar by 400 ml (approx.), which was the sign of alveolar recruitment-1536 (1020-1845) ml vs 1955 (1360-2320) ml, p=0,001, Friedman test). we didn't found significant changes of VCO2 when increased PEEP in the range from 8 to 15 mbar (p>0.05, Friedman test). PEEP increase from 15 to 18 mbar and more lead to decrease in VCO2 (from 212 (171-256) ml/min to 200 (153-227) ml/min, p<0,0001, Friedman test, which was the sign of overdistension. Next decrease of VCO2 was observed at PEEP increase from 22 to 24 mbar (from 203 (174-251 ml/min) to 185 (182-257) ml/min, p=0.0025, Friedman test). Adjusted PEEP value according to balance between recruitment and overdistension was higher than the one initially set (16(15-18) mbar vs 12(7-15) mbar, p <0.0001). We observed increase of SpO2 from 93 (87-96) to 97(95-100)% (p<0.0001 followed by decrease in inspiratory oxygen fraction from 60(40-80) to 50(40-60)%(p<0.0001). Low EELV VCO2 and VCO2/EtCO2 at PEEP 8 mbar has low predictive value for death (AUROC 0,547, 0706 and 0.596, respectively).Absolute EELV value at PEEP 18 and 20 mbar were poor predictors of mortality (AUROC 0.61 and 0.65 respectively) Alveolar recruit ability was measured by subtraction of EELV at PEEP 20 and at PEEP II mbar - value below 575 ml was a good predictor of death (sensitivity 75%, specificity 88%, AUROC 0.81). Lowering of VCO2 at PEEP 20 mbar to less than 207 ml/min was a marker of alveolar overdistension and associated with poor prognosis (sensitivity 83%, specificity 88%, AUROC 0,89). C has poor predictive value at PEEP 8 and 20 mbar (AUROC 0,58 and 0,74 respectively. Conclusion: PEEP adjustment in ARDS due to influenza A (H1N1) pdm09 in accordance with balance between recruitment and overdistension (based on EELV and VCO measurements) can improve gas exchange, probably, not leading to right ventricular failure. This value of "balanced" PEEP is in the range between 15 and 18 mbar: Low lung recruitabiilty is associated with poor prognosis. Measurements of EELV and VCO2 at PEEP 8 and 20 mbar can be used to make a decision on whether to keep "high" PEEP level or switch to extracorporeal membrane oxygenation in patient with ARDS due to influenza A (N1H1).

[The role of recipient cells in the mechanism of pathological calcification of heart valve and vascular transplants].

It was found using the model of subcutaneous implantation in rats that the calcification of the aorta wall occurs by two mechanisms of which one is dependent on, and the other independent of the migration of recipient cells to the transplant.

[Prooxidant and cytotoxic action of N-acetylcysteine and glutathione combined with vitamin Bl2b].

We studied the prooxidant and cytotoxic action of thiols N-acetylcystein (NAC) and glutathione (GSH) combined with vitamin Bl2b. The synergism of action of the thiols and Bl2b resulted in human carcinoma cell damage was found. It was shown that GSH and NAC in physiological doses combined with Bl2b caused the initiation of apoptosis. It was established that prooxidant action of the thiols combined with vitamin Bl2b, i. e. generation and accumulation of hydrogen peroxide in culture medium, led to intracellular oxidative stress and injury of cell redox system. These effects were completely abolished by nonthiol antioxidants catalase and pyruvate. The chelators of iron phenanthroline and deferoxamine did not suppress the H2O2 accumulation in culture medium but significantly inhibited the cell death induced by the thiols combined with Bl2b. Therefore, the thiols GSH and NAC widely used as antioxidants, in combination with vitamin Bl2b show prooxidant characteristics and induce, with the participation of intracellular iron, apoptotic HEp-2 cell death.

Functional activity of hepatocytes in tissue fragments in a new bioreactor biological artificial liver.

Functional activity of hepatocytes in a new bioreactor designed for culturing of liver tissue fragments under perfusion conditions was tested. Specific hepatic functions such as ammonium detoxification, urea and protein synthesis, and P-450-dependent metabolism of p-nitroanisole were maintained for 1.5 days. The bioreactor can be used as a bioartificial liver support apparatus.

[WWWMGS: an integrated server for molecular-genetic studies]. / WWWMGS: integrirovannyi server dlia molekuliarno-geneticheskikh issledovanii.

We report an integrative technology for molecular biology studies in the field of transcription regulation by using Internet. A set of databases, programs, and systems are included into WWWMGS Web server. For example, the use of TRRD database information for site prediction is described. Using this method, the computer system SeqAnn was developed. The system performs the "real time" searching for prediction of initiation transcription site position according to database information. WWWMGS is available at URL: http://wwwmgs.bionet.nsc.ru/.

[Experience in the undergraduate training of ophthalmologists]. / Opyt podgotovki spetsialistov-oftal'mologov v subordinature.

The authors present their experience gained in the training of ophthalmologists at the Chair of Ocular Diseases of the Krasnoyarsk Medical Institute. In 1991 the interns, instead of traditional state examinations, maintained their diploma research, which was assessed as their skills and knowledge in the field of ocular diseases, social hygiene, and public health organization. The authors prove that such diploma research is an integral part of undergraduate training and one of the types of individual work at higher educational institutions, and enumerate the topics of diploma research.

[Modular evolution of Bsp-repeats: formation of subrepeats and monomers preceded the divergence of four species of Canidae]. / Blochnaia évoliutsiia Bsp-povtorov: formirovanie subpovtorov i monomerov predshestvovalo raskhozhdeniiu chetyrekh vidov Canidae.

Determination of two nucleotide sequences of satellite-like DNA (Bsp-repeats) segments from the raccoon-like dog genome and analysis of these and three other homologous sequences from distantly related genomes of Canidae (silver fox, domestic dog, and gray fox) show that all these DNA fragments consist of five types of subrepeats (A, B, C, D, and E) and two types of monomers-ACB and ACED. Raccoon-like dog repeated segment of 1620 bp is composed of both type of monomers: ACBACED. Different monomers of the same type could be distinguished by subtypes of subrepeats. Building of a cladogram of 18 subrepeats and analysis of the structures of monomers and higher-order repeated units allowed us to reveal probable evolution events which have determined the hierarchical order in Bsp-repeat organization. Approximate periods of time for these evolution events were calculated from the number of substitutions per branch. Five main types of subrepeats and two types of monomers have been generated and propagated through several steps before divergence of the main phylogenetic trees of Canidae. Combinatorial variability appears to play a fundamental role in forming the hierarchical layers in Bsp-repeats structure.

[A new method of global search for functional DNA segments using a fractal representation of nucleotide texts]. / Novyi metod global'nogo poiska funktsional'nykh uchastkov DNK s ispol'zovaniem fraktal'nogo predstavleniia nukleotidnykh tekstov.

New global method for computer prediction of functional sites in nucleotide sequences, based on the fractal representation, is presented. Fractal representation of set of sequences (FRS) provides simple way for generating recognitions matrix of functionally similar sequences and simple estimations of its efficiency for searching homologous regions in new sequences. Other advantages of the method are absence of the necessity of sequences alignment during generating based set and searching new homologous regions and small CPU time. Usage of the method illustrated for searching globin and histone genes, for ALU repeats in human genome and long terminal repeats in virus genome.

[Search for functional domain boundaries and localization of functional sites based on oligopeptide vocabularies]. / Poisk granits funktsional'nykh domenov i lokalizatsiia funktsional'nykh saitov na osnove oligopeptidnykh slovarei.

A new method based on the analysis of oligopeptide composition of the amino acid sequences from different protein families is presented. We assume, that any protein family can be characterized by the set of oligopeptides (oligopeptides vocabulary). We demonstrate, that oligopeptides vocabulary comparison can distinguish different families from each other and from random sequences. It should be noted, that this comparison can be successfully performed on the set of only 25 dipeptides and without preliminary alignment. We demonstrate, that characteristic peptides are localized in the regions of functional significance, as shown on the example of GTP-binding domain of translation elongation factors. We suggest how to use this method to localize the boundaries of functional domains in amino sequences. On the example of few functional domains we demonstrate, that the average error of prediction does not exceed 3-4 amino acid residue.

["DIROM"--an interactive system for planning experiments on directed mutagenesis and design of artificial genes]. / "DIROM"--interaktivnaia sistema dlia planirovaniia éksperimentov po napravlennomu mutagenezu i konstruirovaniiu iskusstvennykh genov.

We present a computer system "DIROM" for oligonucleotide-directed mutagenesis and artificial gene design experiments planning and support. "DIROM" allows to search for optimal oligonucleotides according to such parameters as sufficient energy of oligonucleotide-target hybridization, secondary structure of oligonucleotide and target DNA, presence of alternative attachment sites in target DNA, terminal G/C pairs presence. Both single-stranded and double-stranded vector mutagenesis methods are implemented. It can be also used for optimal primer selection for polymerase chain reaction, sequencing etc. "DIROM" can search for both existent and potential carry out vector+target sequence construction. Both amino acid and nucleotide sequences can be operated.

[Study of binocular vision by the binary metric method]. / Issledovanie binokuliarnogo zreniia metodom binarimetrii.

The authors describe a method for binocular vision studies with the use of spatial visual effects emerging when double images are fused in physiologic diplopia. Various forms of binocular cooperation were detected and a clinical classification of these forms developed, based on the ability to obtain a binocular visual image. One of the distinctions of the suggested method is the detection of not only binocular fusion, but of the ability to in-depth perception. The authors analyze the status of binocular function in subjects with various forms of binocular sensor cooperation, basing on binary metric data and the findings of the color test and synaptophore studies. Binocular cooperation impairments, detected by binary-metry, appear less severe than if detected by the color test or synaptophore studies in the same patients.

[A method of calculating the discrete secondary structures of globular proteins]. / Metod rascheta diskretnykh vtorichnykh struktur globuliarnykh belkov.

The model of formation of alpha-helices and beta-structures determined by joint action of the three elements: N-terminal, internal and C-terminal fragments are presented. Algorithm for calculation of their localization in a given amino acid sequence was constructed on the base of this model. The preference of the fragments of the amino acid sequence to a definite type of the secondary structure was estimated on the base of corresponding average values of linear discriminant functions dsk (s = alpha, beta, k = N, in, C). The latter were constructed in the previous paper on the base of the revealed significant characteristics. These integral characteristics are used for calculating the localisation of discrete secondary structures. The total prediction for 3 states (alpha, beta, c) given 71% correctly predicted residues (for 4 states alpha, beta, c, t) 62% for the training set, consisting of 72 proteins. For the control set (15 proteins) the accuracy of prediction is about 65%. The essential advantages of this method are: 1) the possibility to localize the discrete secondary structures; 2) the high accuracy of prediction of long secondary structures (for alpha-helices approximately 90%, for beta-structures approximately 80%), which is important for the determination of the protein folding. The influence of mutation on the secondary structure of proteins was investigated. The anormally high stability of the secondary structures of immunoglobulins to mutations was revealed. This probably results from the selection during evolution of such variants of amino acid sequences, which are able to provide the functional variability of antigenic determinants, but keep invariant the tertially structure of protein.

[Features of the structure and evolution of complex tandemly organized Bsp-repeats in the fox genome. II. Tissue-specific and recombinant BamHI-dimer sites]. / Osobennosti struktury i évoliutsii slozhnykh tandemno organizovannykh Bsp-povtorob genoma lisitsy. II. Tkanespetsificheskie i rekombinatsionnye saity BamHI-dimera.

The complex structure of the clustered Bsp-repeats in fox genome seems to have evolved throughout a long period of time as a result of multiplication, recombination and divergence events. The sequence of the subrepeat (SR) approximately 245 b.p long is the basic substructure for the hierarchically arranged Bam HI-repeat 1468 b.p. long. The monomer consists of 3 SRs with a 43-59% homology. A dimer is composed of 2 monomers with a 93% homology. Amplification of the Bsp-repeats during evolution seems to have occurred at least twice: first--on the SR ancestral form level, second--on the monomer level. Despite profound divergence, there are still conservative regions in SRs with sequences homologous to known functional sites in eukaryotes. However qualitative and quantitative composition of most functional motifs is stringently individual in every SR. The performed analysis revealed that throughout evolution SRs acquired significant amount of motifs homologous to promoter and enhancer regions in tissue-specific genes and virus regulatory regions. Functional motifs in separate SRs are being differently grouped. Most inducible motifs are located in the III and II subrepeats, putative promoters--in the II one; elements participating both in transcriptional and replicational processes--mainly in the I subrepeat. A few ensembles of functional motifs remotely resemble extended regulatory regions of some tissue-specific genes. The monomers are potentially capable of ensuring diverse aspects of transcriptional regulation. As a whole, motifs of the 3 SRs are potentially capable of regulating the RNA synthesis periodicity with respect to the cellular cycle, activation and repression of genetical material in response to signals from the environment (AP-1, AP-2, AP-4, T-antigen, etc) and temporal ("octamers") etc. Apart from the BamHI-dimer, a few homologues fragments were isolated from fox genome and sequenced. Some of them were rearranged with respect to the BamHI-dimer. Inversion locally alters the composition of motifs and the sequence acquires new functional potential. Thus, the analysis of the emergence and development of Bsp-repeat structural variations allows us to consider repetitive DNA sequences as an ideal material in constructing multiprofile regulatory sequences.

[Features of the structure and evolution of complex, tandemly organized Bsp-repeats in the fox genome. I. Structure and internal organization of the BamHI-dimer]. / Osobennosti struktury i évoliutsii slozhnykh, tandemno organizovannykh, Bsp-povtorov genoma lisitsy. I. STruktura i vnutrenniaia organizatsiia BamHI-dimera.

A 1468 b.p. DNA BamHI-fragment homologous to the Bsp-repeat was isolated from the fox genome and sequenced. This fragment is an hierarchically arranged dimer. Its 734 b.p.-monomers consist of three subrepeats (SR), each 245 b.p. long, abundant with overlapping imperfect tandem repeats which in turn are rich in short direct related repeats (each 4-7 b.p. in size). The latters are mainly composed of AG, TG dinucleotides and their complements CT, CA. All subrepeats in the BamHI-dimer are flanked by motifs homologous to Jeffreys' sites. At certain points the sites are doubled. The above data allow to assume that the Bsp-repeat complex structure is likely to have developed throughout long multi-step evolution of relatively simple DNA sequences which had emerged de novo. Single substitutions, small inserts and deletions, multiple duplication and recombination events seem to have most contributed to the evolution of the Bsp-repeats. Single substitutions in SRs with respect to the consensus are not equally distributed along their length. A wave-like pattern of this distribution is the evidence for non-random character of mutations accumulation. A correspondence was noted between conservative regions in SR and the presence therein of functional motifs homologous to the binding sites of already known regulatory proteins.

[Platelet aggregation and plasma lipid spectrum in patients with myocardial infarct developing on the background of first-time appearing angina pectoris]. / Agregatsiia trombotsitov i lipidnyi spektr plazmy krovi u bol'nykh infarktom miokarda, razvivshimsia na fone vpervye voznikshei stenokardii.

Patients with myocardial infarction in the presence of first angina pectoris have been found to display less platelet aggregability in the acute period of the disease than those with recurrent myocardial infarction, which is associated with a lower incidence of postinfarction complications and smaller plasma triglyceride levels. The changes observed in plasma lipid concentrations and platelet aggregation in different periods of the disease have been shown to be similar to those seen in patients with myocardial infarction concurrent with sustained angina.