Preparation a Recombinant Form of Pneumolysin Protein from Streptococcus pneumoniae.

A recombinant form of pneumolysin from Streptococcus pneumoniae was obtained. By using Vector NTI Advance 11.0 bioinformatic analysis software, specific primers were designed in order to amplify the genome fragment of strain No. 3358 S. pneumoniae serotype 19F containing the nucleotide sequence encoding the full-length pneumolysin protein. A PCR product with a molecular weight corresponding to the nucleotide sequence of the S. pneumoniae genome fragment encoding the full-length pneumolysin was obtained. An expression system for recombinant pneumolysin in E. coli was constructed. Sequencing confirmed the identity of the inserted nucleotide sequence encoding the full-length recombinant pneumolysin synthesized in E. coli M15 strain. Purification of the recombinant protein was performed by affinity chromatography using Ni-Sepharose in 8 M urea buffer solution. Confirmation of the recombinant protein was performed by immunoblotting with monoclonal antibodies to pneumolysin.

High intensity focused ultrasound (HIFU) for the treatment of symptomatic breast fibroadenoma.

PURPOSE: The aim of the study was to evaluate the efficacy of high intensity focused ultrasound (HIFU) in the treatment of symptomatic breast fibroadenomas (FA) after 6 and 12 months. MATERIALS AND METHODS: Between December 2013 and November 2014, 27 patients with histologically confirmed FA received one application of HIFU under local anesthesia (NCT02011919). Follow-up visits occurred after 6 and 12 months measuring the FA volume and clinical symptoms. A volume reduction of more than 65% was defined as success. Core needle biopsy (CNB) was offered after 12 months if indistinct residuals were visible on ultrasound (US). RESULTS: A successful reduction in FA volume after 12 months was achieved in 24/27 patients (89%). At baseline 16 patients (59%) had pain, which was resolved in 63% (10/16). All patients were satisfied with the cosmetic related outcome. Twenty-four patients (89%) would repeat the procedure. After 12 months 21 patients with sonographically indistinct residuals underwent a CNB. There were no vital cells in 86%. Three cases showed vital cells of FA. Retrospectively possible reasons in these three cases were an insufficient treatment due to bad visibility and insufficient fixation of the FA during HIFU and/or a too short follow-up time. CONCLUSION: US-guided HIFU is an effective procedure and a minimally invasive alternative for the treatment of breast FA.

Disseminated tumor cells from the bone marrow of patients with nonmetastatic primary breast cancer are predictive of locoregional relapse.

BACKGROUND: Disseminated tumor cells (DTCs) are detectable in the bone marrow (BM) of patients with primary breast cancer (PBC) and predictive of an impaired prognosis. This large trial aimed to analyze the impact of DTC detection on locoregional relapse (LR). PATIENTS AND METHODS: Patients with nonmetastatic PBC were eligible for this analysis. BM aspiration (BMA1) was carried out during primary surgery and DTCs were detected by using immunocytochemistry (A45-B/B3 antibody against pancytokeratin) and morphological criteria. At the time of LR, a subgroup of patients with nonmetastatic and operable LR received a secondary BM aspiration (BMA2). RESULTS: A total of 3072 patients were included into the analysis. Of these, 732 (24%) presented with DTCs at BMA1. One hundred thirty-nine patients experienced LR and 48 of these (35%) were initially DTC positive. DTC detection was significantly associated with an increased risk of LR in univariate (P = 0.002) and multivariate analysis (P = 0.009) with a hazard ratio of 1.65 (95% confidence interval 1.13-2.40). Of the patients with LR, 55 patients were available for BMA2 and 17 of these (32%) were DTC positive. DTC detection at the time of LR was indicative of impaired overall survival (univariate analysis, P = 0.037). CONCLUSIONS: DTC detection in patients with PBC is associated with an increased risk of LR, indicating that tumor cells may have the ability to recirculate from the BM to the site of the primary tumor. The impaired prognosis associated with DTC detection at the time of LR may help to identify patients that are in need for additional or more aggressive treatment.

Treatment with Ca2+ ionophore improves embryo development and outcome in cases with previous developmental problems: a prospective multicenter study.

STUDY QUESTION: Does calcium ionophore treatment (A23187, calcimycin) improve embryo development and outcome in patients with a history of developmental problems/arrest? SUMMARY ANSWER: Application of A23187 leads to increased rates of cleavage to 2-cell stage, blastocyst formation and clinical pregnancy/live birth. WHAT IS KNOWN ALREADY: Studies on lower animals indicate that changes in intracellular free calcium trigger and regulate the events of cell division. In humans, calcium fluctuations were detected with a peak shortly before cell division. Interestingly, these calcium oscillations disappeared in arrested embryos. Mitotic division blocked with a Ca(2+) chelator could be restored by means of ionophores in an animal model. STUDY DESIGN, SIZE, DURATION: This prospective, multicenter (five Austrian centers), uncontrolled intervention study (duration 1 year) includes 57 patients who provided informed consent. PARTICIPANTS/MATERIALS, SETTING, METHODS: Inclusion criteria were complete embryo developmental arrest in a previous cycle (no transfer), complete developmental delay (no morula/blastocyst on Day 5), or reduced blastocyst formation on Day 5 (≤15%). Severe male factor patients and patients with <30% fertilization rate after ICSI were excluded because these would be routine indications for ionophore usage. The total of the 57 immediately preceding cycles in the same patients constituted the control cycles/control group. In the treatment cycles, all metaphase II-oocytes were exposed to a commercially available ready-to-use ionophore for 15 min immediately after ICSI. After a three-step washing procedure, in vitro culture was performed as in the control cycles, up to blastocyst stage when achievable. MAIN RESULTS AND THE ROLE OF CHANCE: Fertilization rate did not differ (75.4 versus 73.2%); however, further cleavage to 2-cell stage was significantly higher (P < 0.001) in the ionophore group (98.5%) when compared with the control cycles (91.9%). In addition, significantly more (P < 0.05) blastocysts formed on Day 5 in the study compared with the control group (47.6 versus 5.5%, respectively) and this was associated with a significant increase (P < 0.01) in the rates of implantation (44.4 versus 12.5%), clinical pregnancy (45.1 versus 12.8%) and live birth (45.1 versus 12.8%). All babies born at the time of writing (22/28) were healthy. LIMITATIONS, REASONS FOR CAUTION: The frequency of patients showing embryo developmental problems was expected to be low; therefore, a multicenter approach was chosen in order to increase sample size. In one-third of the cycles, the clinician or patient requested a change of stimulation protocol; however, this did not influence the developmental rate of embryos. WIDER IMPLICATIONS OF THE FINDINGS: This is the first evidence that developmental incompetence of embryos is an additional indication for ionophore treatment. The present approach is exclusively for overcoming cleavage arrest. STUDY FUNDING/COMPETING INTERESTS: No funding received. T.E. reports fees from Gynemed, outside the submitted work. All co-authors have no interest to declare.

[Immunogenic properties of experimental production series of Staphylovac-2 vaccine].

AIM: Study experimental production series of Staphylovac-2 by accumulation of specific IgG and safety. MATERIALS AND METHODS: Experimental production samples of staphylococci vaccines were studied by the accumulation of specific IgG in sera of immunized BALB/c line mice in EIA. Safety was evaluated in tests of acute and chronic toxicity including pathomorphologic and histologic, hematologic and biochemical studies, studies of the effect on central nervous system. RESULTS: A statistically significant (2.6 - 3.0 times) increase of IgG levels in sera of immunized mice compared with control was noted. In the experiments studying acute and chronic toxicity the increase in body mass and mass of internal organs differed from data obtained from control animals at no observation periods. None of the studied methods of safety evaluation showed differences of the studied vaccine series from the control. CONCLUSION: The recommended dose for subcutaneous administration into human of 200 µg is experimentally justified and could be the basis for carrying out clinical studies of staphylococci vaccines in humans.

[Study of protective activivty of "Staphylovac-2" vaccine].

AIM: Study the protective properties of "Staphylovac-2" vaccinie. MATERIALS AND METHODS: Samples of the vaccine manufactured by SPA "Microgen" based on the developed technology were studied in balb/c mice during 3- and 6-fold immunization schemes. Protective activity of the preparation was determined in experiments with active and passive protection during intraperitoneal infection, seeding of the causative agent from spleen and kidneys during intravenous infection, of animals. RESULTS: In experiments with active protection of mice for both 3- and 6-fold immunization schemes, a significant protective activity of the studied series was determined, compared with the control group of mice. Sera obtained after animal immunization (rabbits, mice) by staphylococcus vaccine had protective properties. A reduction of spleen and kidneys seeding by Staphylococcus aureus in immunized mice compared with the control group was detected in the model of generalized staphylococci infection. CONCLUSION: The preclinical studies carried out with the "Staphylovac-2" vaccine, developed baed on the complex of protective staplylococci antigens, have confirmed the high protective activity of the preparation.

[Genetic characteristics of Staphylococcus aureus No 6. strain--producer of secreted protein-containing compounds possessing protective properties].

AIM: Genotype characteristics of Staphylococcus aureus No 6: strain that is a producer of a protective protein complex. MATERIALS AND METHODS: Features of structure of 9 genes, that code synthesis of pathogenicity factors, of S. aureus--spa, coa, sea, seB, sec, pvl, tst-h, mecA and scc-mecA, that are responsible for synthesis of protein A, coagulase, enterotoxins A, B and C, Panton-Valentine toxin (PVL), heat shock syndrome protein, resistance to methicillin and staphylococci chromosomal cassette, respectively, were studied by amplification in PCR of the respective gene fragments with subsequent conduction of direct sequencing. RESULTS: The S. aureus No 6 strain under study possesses pvl gene fragments, as well as Spa and coagenes, detected in all the studied strains, that belong to t12507 and EMRSA-16 types, respectively. Sea, seb, sec genes responsible for.the synthesis of enterotoxins A, B and C were not detected in it, tst-h, mecA and scc-mecA gene fragments were not present. CONCLUSION: The detection of pvl gene fragment in the strain under study, on the one hand, and protective properties of the secreted protein-containing compound, on the other hand, give evidence in favor of the necessity of further analysis of extracellular proteome of S. aureus No 6.

[Analysis of extracellular proteome of Staphylococcus aureus 6 at the end of exponential growth phase].

AIM: Determine protein specter that Staphylococcus aureus synthesizes and secretes at early growth phase--the exponential phase. MATERIALS AND METHODS: Proteins secreted by S. aureus strain 6 into cultivation medium at the end of exponential growth phase (4.5 hours) were studied. 11 proteins were identified by liquid chromatography--mass-spectrometry method. RESULTS: Only in 3 of these proteins the presence of signal peptides was predicted, which indicates their extracellular localization; the rest of the proteins were localized predominantly in bacterial cytoplasm. 5 of 11 proteins function as enzymes of carbohydrate metabolism. Other extracellular proteins that could indicate its contamination with proteins from disrupted bacterial cells were not detected in S. aureus cultural liquid filtrate. It has been suggested that enzymes of carbohydrate metabolism can provide bacterial cells with energy necessary for passage from lag-phase into exponential growth phase. Superoxide dismutase enzyme probably provides the course of oxidation-reduction processes. Synthesis of other proteolytic enzymes and toxins is carried out at later stages of development of bacterial population. Immunization of mice with a mixture of 11 identified proteins showed their protective properties after infection by S. aureus 6 strain. CONCLUSION: Based on the above-mentioned, the complex of isolated proteins may be perspective in development of a new strategy of prophylaxis and therapy of staphylococcus infections.

Ultrasound guided cryoablation of fibroadenomas.

PURPOSE: Fibroadenoma (FA) of the breast is the most common disorders in young women. The aim of the study was to evaluate cryoablation (CA) under ultrasound guidance in the office setting for patients with FA. MATERIALS AND METHODS: For this prospective multicenter trial an office-based cryosurgical system was used to treat histological confirmed benign FA with a maximum dimension of 3 cm. 23 CA procedures were performed under ultrasound guidance. The cryoprobe was inserted into the center of the FA guided by real-time ultrasound. A freeze-thaw-freeze treatment cycle was performed according to the size of the FA. During the CA procedure continuous ultrasound monitoring of the ice ball was performed, verifying engulfment of the FA. Patients attended 4 follow-up visits at 1 week, 3 months, 6 months and 1 year and underwent ultrasound and physical examinations. RESULTS: Data was collected from 23 procedures. The ice ball engulfed the treated FA in 91.3 %. A sharp reduction in volume was observed at 6-month follow-up and continued more moderately up to 1 year. No serious but four minor adverse events occurred. At 1 year follow-up, lumps that were assessed pre-treatment as hard were assessed as soft or not palpable. 7 patients complained of pain caused by the lump prior to cryotherapy, while 5 of these patients felt transient pain during the follow up period. It is reasonable to assume that the pain was not related to the CA procedure as it was not consistent. In 96 % of the cases patients and physicians rated the cosmetic results of the procedure as excellent or good. CONCLUSION: The cryodestruction proved functional and safe, while showing reduction in palpability and pain caused by the FA in the treated patients.

Interdisciplinary consensus recommendations for the use of vacuum-assisted breast biopsy under sonographic guidance: first update 2012.

PURPOSE: The vacuum biopsy of the breast under sonographic guidance (VB) was introduced in Germany in the year 2000 and the first consensus recommendations were published by Krainick-Strobel et al. in 2005. Since then, many clinical studies on this technique have been published. The purpose of this publication is to update the consensus recommendations from 2005 regarding the latest literature. MATERIALS AND METHODS: The consensus statements were the result of two preliminary meetings after the review of the latest literature by members of the Minimally Invasive Breast Intervention Study Group from the German Society of Senology. The final consensus text was review by all members of the working group. The statements listed under results obtained complete acceptance (consensus 100 %). RESULTS: The consensus recommendations describe the indications, investigator qualifications, technical requirements, documentation, quality assurance and follow-up intervals regarding the latest literature. CONCLUSION: The VB is a safe method for extracting breast tissue for histological workup. The technique allows the resection of breast tissue up to 8 cm3. Besides the diagnostic indications, the method qualifies for a therapeutic resection of symptomatic benign lesions (e. g. fibroadenomas). The technique should be used in specialized breast centers working in a multidisciplinary setup. This paper is an expert's recommendation for the use of VB under sonographic guidance. It is not formulated as a nationwide guideline.

Reliability of constant-load treadmill testing in patients with intermittent claudication.

AIM: Constant-load treadmill testing is frequently used to estimate walking ability of patients with peripheral arterial disease and to assess changes in functional capacity following therapeutic interventions or through disease progression. The value of this test has frequently been disputed based on doubts concerning its reproducibility. The aim of this study was to investigate the reliability of treadmill testing at a speed of 3 km/h and a constant inclination of 12%. METHODS: Sixty-eight patients with stable peripheral arterial disease who were familiar with treadmill testing underwent two sessions of walking exercise at 3 km/h and 12% inclination within three weeks. Initial claudication distance (ICD) and maximum walking distance (MWD) on the treadmill as well as reported walking distances on level ground at individual speed were recorded. RESULTS: Correlation coefficients of 0.8 and within-subject variation coefficients of 15% for ICD and 13% for MWD demonstrated low variability of walking distances between visits. No learning effects were observed. Intra-class correlation coefficients of 0.78 for ICD and 0.83 for MWD proved good reproducibility of treadmill testing. Correlation of measured and reported walking distances was poor. CONCLUSION: Constant-load treadmill testing at 3km/h and 12% inclination is a well reproducible method to investigate walking capacity of patients with stable intermittent claudication who are familiar with this test method. From treadmill walking distances, no conclusions should be drawn concerning the actual walking range of the patient.

Relevance and methods of interventional breast sonography in preoperative axillary lymph node staging.

Reduction of therapy-induced morbidity is an important goal for the improvement of the quality of breast cancer treatment. The introduction of sentinel lymph node biopsy (SLNB) significantly contributed to the reduction of surgery-induced morbidity in the shoulder-arm region. However, a clinically positive nodal status is still considered a contra-indication for SLNB. The current data constellation clearly shows that the clinical and also the sonographic malignancy assessment is insufficient for the accurate identification of the nodal status. A merely clinical and/or image-based appraisal of the axillary lymph node status would lead to overtreatment due to unnecessary axillary dissection in approximately 40 % of patients. In order to reduce the rate of unnecessary axillary dissection (AD), pretreatment interventional clarification is necessary to provide more detailed information about the histological condition of the lymph node. Comparing the currently available methods, fine needle aspiration (FNA) is the best in terms of cost and time requirement, practicability and complication rate. However, considering the sensitivity, it is inferior to ultrasound-guided core needle biopsy (CNB). Thus, a negative FNA outcome requires further clarification, which possibly can be performed with CNB. With a specificity of nearly 100 % and therefore a low rate of false positive cases for FNA, complete AD can be indicated by a positive FNA result. In the context of the interventional procedure, it must be stressed that FNA requires a high level of expertise on the part of both the examiner and the cytologist. The prerequisite for optimal interventional diagnostics of lymph nodes is an adequate sonographic assessment on the basis of standardized sonomorphological criteria.

[Sonographic criteria for the differentiation of benign and malignant breast lesions using real-time spatial compound imaging in combination with XRES adaptive image processing]. / Dignitätskriterien der Mammasonografie unter Anwendung des Real-Time Compound Image in Kombination mit dem XRES Adaptiv Image Processing.

PURPOSE: The purpose of the study was to analyze sonographic criteria for the differentiation of benign and malignant breast lesions using real-time spatial compound imaging (CT) in combination with adaptive image processing (XRES). MATERIALS AND METHODS: In a retrospective analysis of 460 patients, the sonographic criteria: shape, orientation, margin, echo pattern and posterior acoustic features were determined using CT and XRES. All investigations were performed using a 12 MHz linear transducer. The findings were classified according to the DEGUM criteria analogous to BIRADS and were histologically confirmed by core needle or vacuum biopsy. Statistical analysis was performed using a Chi-square test, logistic uni- and multivariate regression analysis and an ROC-curve analysis to detect the false-positive rate. RESULTS: All investigated diagnostic criteria were significant in the descriptive analysis (Chi-squared). The multivariate analysis showed that the criteria of irregular shape versus round, and not circumscribed margin versus circumscribed margin as well as the posterior acoustic features of enhanced versus reduced have a significant influence on the prediction of a malignant finding. The univariate analysis also showed a statistical significance using the indifferent and not parallel orientation versus parallel. To attain a detection rate of > 95 %, a false-positive rate of 60 % must be expected. CONCLUSION: This study shows that when using CT in combination with XRES, the analyzed sonographic criteria for the differentiation of benign and malignant breast lesions are still of diagnostic value. In particular, the shape, margin and posterior sonographic features are important.

[Novel type of vaccine with a combination of Toll like receptor agonists--TLR 1/2, 4, 5/6, 9].

AIM: Proof of therapeutic efficacy of a novel type of vaccine with a combination of natural Toll like receptor agonists (TLR) 1/2, 4, 5/6, 9 in infectious and noninfectious human pathology. MATERIALS AND METHODS: Immunovac-VP-4 vaccine, containing antigens of opportunistic microorganisms that are TLR 1/2, 4, 5/6, 9 ligands, was used for experiments and clinical trials. RESULTS: Immunovac-VP-4 activates innate immunity by inducing maturation of dendritic cells with expression of costimulating molecules on their membrane (CD40+, CD80+, CD86+), terminal differentiation molecules--CD83+ and antigen-presenting molecules (MHC class I and II); activates proinflammatory (TNFalpha, IL-6) and regulatory (IL-12, INFgamma) cytokine synthesis that programs T-lymphocyte differentiation by Th1 pathway; increases cytotoxic activity of natural killer cells, inhibits melanoma B16 growth and Lewis lung carcinoma metastasis. Therapeutic effect of Immunovac-VP-4 was evident regardless of pathology by a significant decrease of quantity and severity of recidives, improvement of all clinical parameters, reduction of quantity of administered pharmaceutical preparations including antibiotics and glucocorticosteroids. The rate of intercurrent acute respiratory viral illnesses and their bacterial complications decreased. Immunovac-VP-4 therapy modified course of illness from severe into milder forms. Positive therapeutic effect was 69.2 - 100%. CONCLUSION: High therapeutic effect of vaccine therapy is based on innate immunity activation by combining TLR agonists. Immunovac-VP-4 contains an optimal combination of natural TLR agonists that ensure high therapeutic effect in various nosological forms of infectious and noninfectious human pathology.

Growth-dependent release of carbohydrate metabolism-related and antioxidant enzymes from Staphylococcus aureus strain 6 as determined by proteomic analysis.

Proteins released into the culture medium by Staphylococcus aureus (S. aureus) strain 6 were determined at the end of the exponential growth phase (4.5 h). Eleven proteins were identified by liquid chromatography coupled with mass spectrometry. Three proteins were predicted to have signal peptides indicating their extracellular localization. The other proteins were presumably located in the cytoplasm of the bacteria. Five out of the 11 proteins were involved in carbohydrate metabolism. Other intracellular proteins of S. aureus were not detected in the culture medium. This indicates that the release of these 11 proteins was specific and that unspecific protein release due to damaged or dying bacteria did not play a role. It is suggested that enzymes associated with carbohydrate metabolism may provide the energy necessary for the transition of bacteria from a resting to a proliferative state. Another enzyme released by S. aureus, superoxide dismutase, may catalyze redox reactions in this context. The production of other proteolytic enzymes and toxins may take place at later stages of bacterial growth. A cocktail of these 11 proteins was used for the immunization of mice. Indeed, vaccination with these proteins prolonged the survival times of mice upon infection with S. aureus strain 6. Therefore, these proteins may have implications for the development of novel strategies for the prevention and therapy of S. aureus infections.

[Prevention and immunotherapy of staphylococcal infections with bacterial vaccines].

Pathogenesis of staphylococcal infection both local and systemic is associated with many pathogenicity factors, which in foreign literature are called virulence factors of Staphylococcus aureus, which were studied as potential candidates for vaccine development. Much difficulties are related to use of known experimental models, which virtually do not allow to determine direct appropriate effect by survival of animals, as well as to data about absence of correlation between increase of antibody titers in animals and protective effect of studied preparations. Despite the importance of the problem of prevalence and severity of staphylococcal infection and intensive research in order to determine protective components able to protect from infection caused by S. aureus, there are no licensed prophylactic preparations with proven efficacy.

[Characteristics of antigenic complexes of Staphylococcus aureus vaccine strains obtained in different cultivation conditions].

AIM: Assessment of characteristics of antigenic complexes of Staphylococcus aureus vaccine strains in different cultivation conditions. MATERIALS AND METHODS: S. aureus vaccine strains (No. 5, 9, 1986, 1991) were grown in liquid nutrient media--full value and semi-synthetic--as well as on solid medium. Reactor cultivation was performed in the fermenter ANKUM-2M. Complex of antigens were obtained by water extraction method applied to staphylococcal biomass inactivated with acetone and assessed by common methods on protein and carbohydrate content; specific activity was assessed by minimal inhibitory dose in passive hemagglutination inhibition assay. Study of acute toxicity was performed on outbred mice. RESULTS: Using strain no. 1991, model of reactor cultivation in full value medium with separation of biomass by microfiltration was validated on the basis of biomass and semiproduct of antigenic complex (acetone powder) yield as well as productivity of biomass cumulation. Study of antigenic complexes obtained from biomass of 4 strains during reactor cultivation compared with complexes extracted from cultures grown on solid medium revealed increased protein and decreased carbohydrate content but similar specific activity. It was demonstrated that complex of antigens obtained from cultures grown either by reactor cultivation or on solid medium were non-toxic. CONCLUSION: New technology for manufacturing staphylococcal complex of antigens with reactor cultivation of vaccine strains in full value medium with subsequent purification of antigenic complex from the biomass by microfiltration was developed. Results of the study demonstrated the usefulness of the developed technology for both further studies on a cellular staphylococcal vaccine and manufacture of staphylococcal component of "Immunovac" vaccine.

[Immunovac-VP-4 for prevention of acute respiratory diseases in children's communities].

AIM: Assessment of prophylactic effect of Immunovac-VP-4 against acute respiratory diseases (ARDs) in collectives of children. MATERIALS AND METHODS: Immunovac-VP-4 was used for prophylaxis of ARDs in communities of children (daycare centers, boarding school). During 3 consecutive controlled studies performed during peak incidence of epidemic influenza and ARD (1st study) and during pre-epidemic period (2nd and 3rd studies) the incidence of ARDs was studied in groups of vaccinated and non-vaccinated children. Criteria for assessment of ARDs incidence and their severity were as follows: number of ARD episodes per child, number of children with recurrent episodes of ARD during period of follow-up, duration of ARDs, and number of children with bacterial complications (bronchitis, otitis, etc.). Effect of intervention on immunologic parameters was assessed on levels of sIgA, IgG, IgA, and antibody titers in saliva. Schedule of vaccine administration was 3 daily intranasal and 6 - 9 oral administration of the vaccine with 3 - 4 days interval. RESULTS: Results of all three studies were virtually the same: decrease of number of ARD episodes and their duration; 3.1-fold reduction of number of children with recurrent infections during 14 months of follow-up, and 10.9--12-fold reduction during first 7 months of follow-up; decrease of bacterial complications rate (1.9% in children in intervention group, 11.9%--in control group). Increase of total immunoglobulin level and antibody titers to antigenic components of the vaccine from low baseline level was observed in saliva of vaccinated children. CONCLUSION: Immunovac-VP-4 results in enhanced and prolonged prophylactic effect on ARDs incidence and recommended for building of optimal system of prevention of polyetiologic complex of ARDs.

[Effect of acellular staphylococcal vaccine "Staphylovac" on several indicators of immune system in mice].

AIM: To study molecularand cellular mechanisms of action of "Staphylovac" vaccine on effectors of innate immunity in mice. MATERIALS AND METHODS: Dynamics of changes of mice spleen mononuclear leukocytes immunophenotype under the influence of the vaccine at determined time intervals was assessed in the study. RESULTS: In response to antigenic stimulation mice immune system reacted by increasing number of cells expressing molecules of T-cell subpopulations--CD3, CD4, CD8; B-cell subpopulation--CD19 and natural killers--NK. Activation of immune system of mice was noted which expressed as increased levels of molecules for antigenic presentation (MHCI, MHCII), regulatory molecules (CD4/CD25) as well as enhanced cytotoxicity potential and phagocytic activity. CONCLUSION: Staphylococcal vaccine as well as studied antigens of Staphylococcus aureus promote activation of immune system. Cytotoxic potential obtained by NK cells under their influence as well as enhancement of phagocytic activity of macrophages point to activation of innate immune mechanisms.