Specific Antibodies to the Fragments of Meningococcal IgA1 Protease during the Formation of Immunity to Bacterial Infections.

The features of individual fragments of IgA1 protease of Neisseria meningitidis serogroup B during the formation of immunity to bacterial infections in animals and humans were studied. The antibodies to the immunogenic regions of the studied proteins are also detected in mice infected with some bacterial pathogens and in humans with bacterial meningitis. A region of IgA1 protease was identified that is not capable of producing antibodies during immunization of animals, but that detects homologous antibodies in the blood of humans and animals recovered from bacterial infections. It has been suggested that this fragment plays a regulatory role in the process of immunogenesis.

[Telemonitoring in patients with chronic heart failure].

Actuality. High risk of hospitalisation and death in patients with heart failure highlight the importance of developing methods to moni­ tor weight, diuresis, heart rate and other parameters and provide the physicians with an ability to change the therapy immediately if needed. The aim of this work is an analysis of clinical trials which investigate telemonitoring in patients with heart failure. Discussion. The Cochrane meta-analysis is also discussed in this work. Main conclusions. Our analysis showed that there is no consistency among trials. Not all trials have demonstrated that telemonitoring can reduce the risk of death and heart failure hospitalisations. Potentials explanations are lack of compliance with systems which didn't include the direct contact between the patient and the caregivers, using parameters with low sensitivity in some of the methods and including of stable patients in some of the studies. It is also seeming that effect of telemonitoring is low in regions with existing programs to treat heart failure.

Peculiarities of the Formation of Antimeningococcus Immunity in Mice Immunized with Fragments of N. meningitidis IgA1 Protease.

We studied immunogenicity of two recombinant proteins FR.9 and FR.11-3 created on the basis of fragments of the primary structure of N. meningitidis IgA1 protease with different molecular weights containing different sets of T and B epitopes. The proteins actively protect animals infected with live virulent culture of meningococci, serogroups A, B, and C. Analysis of CD4+, CD8+, and CD19+ lymphocyte populations in mouse blood showed predominant contribution of different cell populations to the formation of immune response to different proteins. Injection of FR.11-3 protein to animals did no affect the immunoregulatory index, hence, this protein can be used for creation of immunologically safe vaccine preparation.

[Palliative surgical correction of respiratory insufficiency in diffusive pulmonary emphysema]. / Palliativnaia khirurgicheskaia korrektsiia dykhatel'noi nedostatochnosti pri diffuznoi émfizeme legkikh.

AIM: To analyze early postoperative period in patients with diffuse pulmonary emphysema after palliative surgical correction of respiratory failure. MATERIAL AND METHODS: The study included 196 patients who underwent bullectomy (n=111) and surgical reduction of pulmonary volume (n=85). RESULTS AND DISCUSSION: Overall morbidity and mortality were 40.8% and 12.2% respectively. Among patients older than 60 years these values were significantly higher (58.0% and 22.6% respectively). It was shown that age over 60 years is associated with high risk of complications and mortality after excision of large and giant bulls. In patients <60 years morbidity is comparable after bullectomy and surgical reduction of pulmonary volume. CONCLUSION: Selection of patients for palliative surgical correction of respiratory failure is generally corresponded to that for lung transplantation. However, these methods should be considered complementary rather competing.

Closed Vitrification System as A Platform for Cryopreservation of Tissue Engineered Constructs.

BACKGROUND: Cryopreservation of mesenchymal stromal cells (MSCs) and MSCs-based tissue engineered constructs (TECs) is a promising strategy for regenerative medicine. OBJECTIVE: To examine vitrification system consisting of multicomponent vitreous solution, closed type container, human adult MSCs and two-step exposure procedure as a platform for cryopreservation of MSCs-based TECs. MATERIALS AND METHODS: Vitrification properties of solutions were studied by visual analysis and calorimetry. Viability (trypan blue, MTT-test), metabolic activity (Alamar Blue assay) and adhesion of cells were assessed both after exposure with vitreous solutions and following rapid cooling-thawing in standard cryovials. RESULTS: The feasibility of the vitrification system was tested on MSCs suspensions (S-MSCs) and alginate encapsulated MSCs (AE-MSCs). The minimal concentrations of cryoprotectants, which allowed avoiding ice formation during rapid cooling and rewarming comprised 10 % for dimethylsulfoxide, 20 % for ethylene glycol, 20 % for 1.2-propanediol and 0.5 M sucrose. To achieve viability and metabolic activity rates of AE-MSCs comparable to S-MSCs after vitrification the extension of the exposure time within the same vitreous solution was sufficient. After vitrification both S-MSCs and AE-MSCs retained the capacity to osteogenic and adipogenic differentiation. CONCLUSION: Data demonstrate that this vitrification system can be used as a platform for development of effective protocols for cryopreservation of MSCs-based TECs.

Serological Analysis of Immunogenic Properties of Recombinant Meningococcus IgA1 Protease-Based Proteins.

Using the genome sequence of IgA1 protease of N. meningitidis of serogroup B, four recombinant proteins of different structure and molecular weight were constructed. These proteins were equal in inducing the formation of specific antibodies to IgA1 protease and had protective properties against meningococci. In the sera of immunized mice, anti-IgA1 protease antibodies were detected by whole-cell ELISA, which indicated the presence of IgA1 protease on the surface of these bacteria. We hypothesized that the protective properties of IgA1 protease-based antigens and IgA1 protease analogs could be realized not only via impairment of bacterium adhesion to the mucosa, but also via suppression of this pathogen in the organism. The presented findings seem promising for using these proteins as the basis for anti-meningococcus vaccine.

Truncated Variants of Serratia proteamaculans Oligopeptidase B Having Different Activities.

Treatment of native psychrophilic oligopeptidase B from Serratia proteamaculans (PSP, 78 kDa) with chymotrypsin (soluble or immobilized on modified porous glass MPG-PA) in the presence of 50% glycerol leads to production of a truncated enzyme form (PSP-Chtr, ~66 kDa), which retains activity toward the low molecular weight substrate of PSP, BAPNA, but in contrast to PSP, is active toward the protein substrate azocasein. It has been shown by MALDI-TOF mass-spectrometry that PSP-Chtr lacks the N-terminal region of the molecule that envelops the catalytic domain of PSP and supposedly prevents hydrolysis of high molecular weight substrates. It has also been established that the lacking fragment corresponds to the N-terminal highest rank element of the informational structure of PSP. This finding confirms the usefulness of the method of informational structure analysis for protein engineering of enzymes. A similar treatment of PSP with immobilized trypsin also led to production of a stable truncated enzyme form (PSP-Tr, ~75 kDa) which lacked 22 C-terminal amino acid residues and completely lost enzymatic activity, presumably because of changes in the nearest environment of His652 of the catalytic triad.

A study of the influence of the T2DL.2DS-2SS translocation and the 5S(5D) substitution from Aegilops speltoides on breeding-valuable traits of common wheat.

The use of the gene pool of wild relatives for expanding the genetic diversity of common wheat is an important task of breeding programs. However, the practical application of common wheat lines with alien genetic material is constrained by the lack of information on chromosomal rearrangements and the negative impact of the transferred material on agronomically important traits. This research is aimed at studying 14 introgression lines with the T2DL.2DS-2SS translocation and the 5S(5D) substitution from Aegilops speltoides obtained from crossing common wheat varieties (Aurora, Krasnodarskaya 99, Nika Kubani) with the genome-substituted form Avrodes (BBAASS). Hybrid lines with different combinations of T2DL.2DS-2SS and T1BL.1RS translocations and 5S(5D) substitution were characterized by resistance to leaf and yellow rusts, productivity components and technological qualities of grain. The assessment of the varieties' resistance to rust diseases showed that Krasnodarskaya 99, Nika Kubani and the Aurora variety, which is a carrier of the T1BL.1RS translocation, are highly susceptible to diseases, while the presence of the T2DL.2DS-2SS translocation and the 5S(5D) substitution, both together and separately, provides resistance to fungal pathogens. The analysis of the lines using markers designed for known resistance genes of Ae. speltoides did not reveal the presence of the Lr28, Lr35 and Lr51 genes in the lines. The results suggest that the genetic material of Ae. speltoides transferred to chromosomes 2D and 5D contains new resistance genes. To determine the effect of the T2DL.2DS-2SS translocation and the 5S(5D) substitution on the productivity and technological qualities of grain, the lines were assessed by weight of 1000 grains, grain weight and number of ears per 1 m2, by protein and gluten content, gluten quality and general baking evaluation. A positive effect was determined upon the weight of 1000 grains, protein and gluten content. There were no significant differences in other characteristics. The T2DL.2DS-2SS translocation and the 5S(5D) substitution did not have a negative effect on the productivity and technological quality of grain, and are of interest for breeding practice.

Analysis of the Effects Produced by Trade Liberalization in the Republic of Uzbekistan and the EAEU.

This study quantitatively assesses the consequences arising from the liberalization of the regional market in the EAEU member states for the exports of the Republic of Uzbekistan using a number of trade indicators, including Balassa index of revealed comparative advantage, the regional orientation index and others. It is shown that for a significant part of the exports of the Republic of Uzbekistan, the reduction in trade barriers (in particular, nontariff measures of customs regulation) will have a positive effect, i.e., it will contribute to the growth of exports to the Member States of the Union. It is also demonstrated that the main part of the export from the Republic of Uzbekistan does not compete with the export of the EAEU member states in the markets of third countries.

Comparative Analysis of Two Enoxaparin Sodium Drugs by Heptest.

The optimal conditions for analysis of low-molecular-weight heparin (LMWH) drugs by the Heptest and the metrological characteristics of the Heptest method were determined. The effects of two LMWH preparations of enoxaparin sodium, the domestic drug Enoxaparin sodium (Technologiya Lekarstv, Russia) and the original drug Clexane (Sanofi, France), on the clotting time of human plasma in the Heptest were comparatively analyzed. The mean blood clotting times were calculated for each dilution of each series of Enoxaparin sodium and Clexane based on the results of the Heptest. Graphs of the dependence of the plasma clotting time on the degree of dilution of these drugs were plotted. The shapes of the curves of the dependence of the blood plasma clotting time on the activity of enoxaparin sodium were shown to be similar for both Enoxaparin sodium and Clexane. The Heptest method showed that Enoxaparin sodium (Technologiya Lekarstv, Russia) was comparable to the original drug Clexane (Sanofi, France).

Using the synthetic form RS5 to obtain new introgressive lines of common wheat.

The use of the gene pool of wild relatives, which have a signif icant reserve of genetic diversity, is of immediate interest for breeding common wheat. The creation and use of synthetic forms as "bridges" is an effective method of transferring valuable genetic material from wild relatives to cultivated wheat. For this purpose, genome addition, genome substitution and recombinant "secondary" synthetic forms have been created in the P.P. Lukyanenko National Center of Grain. The synthetic recombination form RS5 (BBAASDt), in which the third genome consists of chromosomes of Aegilops speltoides (S) and Aegilops tauschii (Dt), was obtained from crossing the synthetic forms Avrodes (BBAASS) and M.it./ Ae. tauschii (BBAADt Dt), in which the D genome from Ae. tauschii was added to the BBAA genomes of the durum wheat cultivar Mutico italicum. Introgression lines resistant to leaf rust, yellow rust and powdery mildew have been obtained from backcrosses with the susceptible common wheat cultivars Krasnodarskaya 99, Rostislav and Zhirovka. Twelve resistant lines that additionally have high technological characteristics of grain and f lour have been selected. The cytological study (С-banding) has revealed chromosomal modif ications in 6 of 8 lines under study. The rearrangements mainly affected the chromosomes of the D genome, 1D, 3D, 4D, 6D and 7D. It was found that in most cases the genetic material from the synthetic form RS5 in the studied lines was represented by substituted chromosomes from Ae. tauschii. In line 5791p17, the substitution of chromosomes 6D from Ae. tauschii and 7D from Ae. speltoides was revealed. Substitutions 4D(4Dt), 6D(6Dt) from Ae. tauschii and 7D(7S) from Ae. speltoides were obtained for the f irst time. Molecular analysis of 12 lines did not reveal effective leaf rust resistance genes, presumably present in synthetic forms of M.it./Ae. tauschii and Avrodes. It is assumed that the lines may carry previously unidentif ied genes for fungal disease resistance, in particular for resistance to leaf rust, from Ae. tauschii and Ae. speltoides.

[Features of binding of proflavine to DNA at different DNA-ligand concentration ratios].

The binding of proflavine to calf thymus DNA has been studied using the methods of differential scanning calorimetry and spectrophotometry. It was shown that proflavine can interact with DNA by at least 3 binding modes. At high DNA-ligand concentration ratios (P/D), proflavine intercalates into both GC- and AT-sites, with a preference to GC-rich sequences. At low P/D ratios proflavine interacts with DNA by the external binding mode. From spectrophotometric concentration dependences, the parameters of complexing of proflavine with DNA were calculated. Thermodynamic parameters of DNA melting were calculated from differential scanning calorimetry data.

[Effect of freezing on cord blood serum proteins].

The effect of freezing regimes and storage temperatures on protein conformation and the spectrum of cord blood serum has been investigated. Changes in the parameters of ESR spectra of spin probes in cord blood serum after slow freezing and subsequent thawing were established, indicating protein conformational changes characterized by loosening. This fact is confirmed by an earlier process, the first stage of albumin heat denaturation, as indicated by calorimetric data. It was shown that slow cooling results in the aggregation of serum protein in which serum albumin and immunoglobulins play an important role. It was concluded that, for retaining the properties, of cord blood serum proteins, it is preferable to perform cooling at a rate not lower than 100 degrees C/min and a storage temperature of -80 degrees C and lower.

Development and validation of reversed phase high performance liquid chromatography method for determination of dexpanthenol in pharmaceutical formulations.

This paper describes the validation of an isocratic HPLC method for the assay of dexpanthenol in aerosol and gel. The method employs the Vydac Proteins C4 column with a mobile phase of aqueous solution of trifluoroacetic acid and UV detection at 206 nm. A linear response (r>0.9999) was observed in the range of 13.0-130 microg mL(-1). The method shows good recoveries and intra and inter-day relative standard deviations were less than 1.0%. Validation parameters as specificity, accuracy and robustness were also determined. The method can be used for dexpanthenol assay of panthenol aerosol and gel with dexpanthenol as the method separates dexpanthenol from aerosol or gel excipients.

[Development of a rapid method for the detection of prostate-specific antigen by immunochromatography].

A single-step qualitative rapid test for the determination of prostate-specific antigen (PSA) in samples of human blood serum by immunochromatography using a complex of colloidal gold with monoclonal antibodies to PSA as the detection agent was developed. The determination limit for PSA in serum blood samples is 10 ng/ml; the analysis time, 15-25 min; the sensitivity of the method, 100%; and its specificity, 92.5%.

[Interaction of actinocin derivative with different poly(rC) structures].

The interaction of actinocin derivative Act III with single- and double-stranded poly(rC) has been investigated by the methods of differential scanning microcalorimetry and UV-vis absorption spectroscopy. It was shown that, after the addition of the ligand, the temperature, enthalpy and entropy of poly (rC) melting decrease. The analysis of poly(rC)-ActIII absorption spectra indicated that the conformation of polynucleotide differs from that of free poly (rC) in the presence of ActHI at pH 4.46 and pH 6.0. Using the DALSMOD optimization program, the parameters of interaction of Act III with poly (rC) were calculated. It was found that the binding constant of ActHI with double-stranded poly (rC) is essentially higher than that with the single-stranded one upon monomeric binding. On the basis of these data, we conclude that the conformation changes of the matrix are the main cause of the decrease in melting temperature and enthalpy observed by calorimetry. Possible mechanisms of interaction of actinocin derivative with poly (rC) are discussed.

[Comparison of the antigenic spectrum A1, A2, and Ax erythrocytes: inhibition of mab with glucoconjugates of lipid and protein nature with different isoelectric properties].

The inhibition of monoclonal antibodies (MAbs) (anti-A1 2-24, anti-A 2-22, anti- 2-19, and anti- A 2-18; Workshop IV) by glycotopes (glycoconjugates) obtained by the enzymatic treatment of A1, A2, and Ax erythrocytes and by chloroform-methanol method from the membranes of these erythrocytes, followed by ion-exchange gel chromatography was evaluated. According to specificity and isoelectric properties, glycoconjugates (in the sequence of desorption from an anion exchanger) of glycoprotein (A(pr-00), A(pr-0), A(pr-1), A(pr-3)) and glycosphingolipid (A1p,00, A(1p-0), A(1p-1), A(1p-3), A(1p-3)) origin were identified. A1 erythrocytes showed the broad-spectrum inhibiting activity of glycoconjugates, which being at maximum with acid A(1p-00) and A(pr-3) (the ionic points of pH 6.55 and 6.45), as well as with A(lp-00) and A(1p-00) 1 The inhibition of A(pr-3) from A2 was significantly weaker that that from A1. An A(pr-3) fraction from Ax erythrocytes inhibited as strongly as in A1 and A2. The inhibition of A(pr-3p) from Ax either failed to develop at all (MAb 2-19) or turned out be weakened (MAbs 2-22 and 2-18). Thus, the cascade decrease in agglutinogenicity in the order of A1, A2, and Ax correlated with the reduction in the inhibiting capacity of A(pr-3) in all the study groups, including Ax.

[Modified method for determining circulating immune complexes in the complement fixation test with polyethylene glycol precipitate].

The authors have modified a method for determining circulating immune complexes in the complement fixation test. It is shown that with the 7% concentration of polyethylene glycol (PEG)-6000, there is a more complete precipitation of low- and medium-molecular weight immune complexes. The time and temperature of serum incubation were optimized when PEG-6000 was obtained. The use of the soluble circulating immune complexes (sCIC) prepared from human serum as a standard for the construction of a standard plot could substantially enhance the sensitivity of the method (0.325 microg/ ml). The content of circulating immune complexes was studied in donors and in patients with connective tissue dysplasia (CTD) by the improved procedure. In the group of donors, the mean level of sCIC was 0.62 +/- 0.24 mg/ml. In the CTD group, that was 2.32 +/- 0.93 mg/ml; the serum concentrations of sCIC ranging from 1.1 to 5.0 mg/ml. In the donors and patients, the detection rate of serum sCIC was 100%. The proposed method may be clinically used to measure the human serum levels of sCIC.

Content specificity of attentional bias to threat in post-traumatic stress disorder.

BACKGROUND: Attentional bias to affective information and reduced cognitive control may maintain the symptoms of post-traumatic stress disorder (PTSD) and impair cognitive functioning. However, the role of content specificity of affective stimuli (e.g., trauma-related, emotional trauma-unrelated) in the observed attentional bias and cognitive control is less clear, as this has not been tested simultaneously before. Therefore, we examined the content specificity of attentional bias to threat in PTSD. METHODS: PTSD participants (survivors of a multistory factory collapse, n=30) and matched controls (n=30) performed an Eriksen Flanker task. They identified the direction of a centrally presented target arrow, which was flanked by several task-irrelevant distractor arrows pointed to the same (congruent) or opposite direction (incongruent). Additionally, participants were presented with a picture of a face (neutral, emotional) or building (neutral=normal, emotional=collapsed multistory factory) as a task-irrelevant background image. RESULTS: We found that PTSD participants produced overall larger conflict effects and longer reaction times (RT) to emotional than to neutral stimuli relative to their healthy counterparts. Moreover, PTSD, but not healthy participants showed a stimulus specific dissociation in processing emotional stimuli. Emotional faces elicited longer RTs compared to neutral faces, while emotional buildings elicited faster responses, compared to neutral buildings. CONCLUSIONS: PTSD patients show a content-sensitive attentional bias to emotional information and impaired cognitive control.