Analysis of Toxicity of Recombinant Pneumolysin from Streptococcus pneumoniae.

We studied toxicity of recombinant Streptococcus pneumoniae pneumolysin protein in experiments on mice and its cytopathogenic effect on cultures of Vero green monkey kidney cells and human lung carcinoma A549 cells in vitro. In vivo and in vitro experiments proved the absence of compromised toxicity and direct cytopathogenic action of the recombinant protein.

Experimental rat models for Hashimoto's thyroiditis.

PURPOSE: Hashimoto's thyroiditis (HT) is an autoimmune thyroid disease characterized by T lymphocyte-mediated destruction of thyroid follicles. To study the pathogenesis of HT and the efficacy of new substances for its treatment, an easily obtained and adequate to the human disease experimental model is needed. The aim of our study was to find out whether it is possible to induce experimental autoimmune thyroiditis (EAT) similar to Hashimoto's thyroiditis by injecting with thyroglobulin (Tg) without using agents that enhance its thyroiditogenicity and without taking into account the genetic sensitivity of animals. METHODS: Wistar rats were immunized with freshly isolated rat Tg or porcine Tg. In 8 weeks, histological studies of the thyroid and parathyroid glands were performed. Thyroid function and total serum calcium level were also evaluated. RESULTS: Immunization with both rat and porcine freshly isolated Tg caused T lymphocytic infiltration of the thyroid gland, thyroid follicle atrophy and degradation in Wistar rats. EAT caused by porcine Tg was characterized by greater severity than EAT induced with rat Tg. In 55% of rats with porcine Tg-induced EAT, oxyphilic metaplasia was detected in the parathyroid glands. In addition, low total serum calcium was observed in these rats. CONCLUSION: Two rat models of autoimmune thyroiditis were obtained. EAT caused in Wistar rats by immunization with rat Tg is similar to Hashimoto's thyroiditis. EAT induced with porcine Tg was accompanied by oxyphil cell metaplasia in the parathyroids and hypocalcemia.

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.

Immunization with Recombinant Pneumolysin Induces the Production of Antibodies and Protects Mice in a Model of Systemic Infection Caused by Streptococcus pneumoniae.

Immunogenic and protective activity of recombinant pneumolysin was studied in experiments on male BALB/c mice. The mice were immunized intraperitoneally with recombinant pneumolysin sorbed on Al(OH)3 (200 µg per mouse). In 2 weeks after immunization, the isotypes of antibodies to recombinant pneumolysin in the serum of immunized mice were determined by ELISA. The animals were infected with Streptococcus pneumoniae serotype 3. Immunization with recombinant pneumolysin induced the production of anti-pneumolysin antibodies, mainly of IgG1 subisotype. On day 21 after intraperitoneal infection with S. pneumoniae serotype 3 in a dose of 106 microbial cells, the survival rate of animals immunized with recombinant pneumolysin in a dose of 25 µg/mouse was 67% vs. 0% in the control (p<0.001). Recombinant pneumolysin could be considered as a promising protective antigen for inclusion in the serotype-independent vaccine against S. pneumoniae.

[Quincke's edema or angioedema. A contemporary view of the problem]. / Otek Kvinke ili angionevroticheskii otek. Sovremennyi vzglyad na problemu.

The article describes statistical data on patients with angioedema (AE) in the structure of ambulance services and four large multidisciplinary hospitals with otorhinolaryngological departments of the city of Moscow for 2017. The ratio of the number of calls to the NSR in Moscow for AE to the total number of calls was analyzed; the ratio of the number of calls to the ambulatory system for AE to the total number of calls to patients with allergic reactions; the ratio of patients with AE among the total number of treated patients in 3 hospitals in Moscow; the ratio of patients with AE of the upper respiratory tract (URT) among the total number of patients with AE; the ratio of patients with AE URT among the total number of patients with ENT pathology. As a result of the study, it was found that the share of AE in the structure of emergency calls of ambulance teams (n=3 070 491) was 0.7% (n=20 040) and 38.4% of the total number of calls for allergic reactions (n=giant urticaria 52 153). Patients with AE make up 0.34% (n=919) of the total number of treated patients in hospitals (n=269 980). The proportion of patients in whom AE was manifested with a lesion of URT among patients with AE is 1 1.5%. The frequency of occurrence of AE URT among patients hospitalized with pathology of ENT organs (n=14 395) is 0.7% (n=106).

T lymphocyte dependence of the immune response to immunosuppressive neoantigen-exposing Fc fragments of IgG.

We recently identified a rheumatoid factor associated with autoimmune disease resistance and remission, and have named it regulatory rheumatoid factor (regRF). Epitopes recognized by regRF can be induced in papain Fc fragments of IgG. Immunization of arthritic rats with homologous Fc fragments that expose neoepitopes recognized by regRF reduces the symptoms of collagen-induced arthritis. Therefore, regRF-producing lymphocytes are a promising therapeutic target in arthritis, and Fc fragments are a means of stimulating this target.

[Oncolytic Properties of a Mumps Virus Vaccine Strain in Human Melanoma Cell Lines].

The oncolytic potential of the attenuated mumps virus (MV) vaccine strain Leningrad-3 (L-3) was evaluated in a panel of four human metastatic melanoma cell lines. The lines were shown to be susceptible and permissive to MV infection. Efficient MV replication led to death of melanoma cells, but the effect differed among the cell lines. Possible mechanisms mediating the selectivity of MV L-3 towards the cell lines were explored. Replicative and oncolytic activity of MV was found to depend on the expression pattern of type I interferon genes. None of the melanoma cell lines showed induction of expression of the total spectrum of genes required to inhibit virus replication. Based on the results, MV L-3 was assumed to be a promising oncolytic agent for human melanoma cells.

Migration of silver on the nanoporous glasses surface under the action of an electric field.

Experimentally, the formation of micro- and nano-labyrinths in grained silver film on the surface of nanoporous glass was shown. The movement of silver ions from the bulk of the nanoporous glass towards the glass surface gradually occurs under direct current application. A similar effect was observed under direct electron irradiation of grained silver film. The proposed mechanism assumes that observed effects are caused by the formation of "hot spots" with a strong electric field, by ionic field emission of positive silver ions from the small charged nanoparticles and by trapping these ions by large nanoparticles.

[The concentration of cerebral natriuretic peptide in tear and blood serum.]

The natriuretic peptides regulate pressure and water metabolism in human organism. The concentration of N-thermal cerebral natriuretic propeptide (NT-proBNP) in tear and blood serum was analyzed. The sampling included 49 patients divided on three groups. The group I (main) included 14 patients with primary open-angle sub-compensated glaucoma degree I-III. The group II (comparative group) included 15 patients with age-related immature cataract. The group III (control) included volunteers without any visual organ diseases. The concentration of NT-proBNP in tear and blood serum was detected by solid-phase enzyme-linked immunosorbent assay technique using test system NT-proBNP - IFA - BEST¼ («Vektor-Best¼, Russia). The median of content of NT-proBNP in tear turned out to be less than in blood serum in all groups (р1 = 0,00763; р2 = 0,00452; р3 = 0,00029) and made up to in patients with glaucoma 41,3 pg/ml, with cataract - 37.5 pg/ml, in healthy people 0 25.0 pg/ml (inter-quartile range made up to 20-60 pg/ml, 20-65 pg/ml, 10-49,5 pg/ml correspondingly). No differences in concentration of NT-proBNP were established in all groups both for blood serum (p = 0,494) and tear (p = 0,388). The concentration of NT-proBNP in tear correlated with its concentration in blood serum (r = 0,694). However, no dependencies were established from value of intra-ocular pressure (r = 0,168). Therefore, the concentration of NTproBNP in tear is not a perspective laboratory arker for diagnostic of increased intra-ocular pressure and stratification of development of glaucoma.

[Endoprosthetic repair with stent grafts in coronary artery restenosis].

Timely performed reconstructive operations on carotid arteries considerably decrease the incidence rate of ischaemic-type acute impairments of cerebral circulation. Nevertheless, restenoses developing in the zone of reconstruction of carotid arteries level the positive result of the primary operation in remote terms of follow up. In such cases it is more preferable to perform stenting of the damaged portions. The article deals with cases of successful use of self-expandable stent grafts in restenoses of carotid arteries after carotid endarterectomy and operations of prosthetic repair of carotid arteries. The analysis was carried out by international publications concerning this problem.

[Roentgen endovascular treatment of common carotid artery dissection after eversion carotid endarterectomy]. / Rentgenendovaskulyarnoe lechenie dissektsii obshchei sonnoi arterii posle eversionnoi karotidnoi endarterektomii.

The article deals with the use of self-expanding stents for endovascular treatment in patients presenting with dissection of the common carotid artery, exemplified by two clinical case reports. In both cases, dissection of the common carotid artery developed after eversion carotid endarterectomy. Intimal detachment was eliminated in both cases by implanting self-expanding stents. These cases demonstrated possibilities of roentgen endovascular methods of treatment making it possible to successfully replace a repeat, technically difficult surgical intervention for iatrogenic dissection of the common carotid artery.

[Hygienic assessment of conditions of the exploitation of facilities of the urban drainage system].

The aim of the study was the hygienic characteristics of the location of sewage pumping stations (SPSs) in the residential area of the city and the assessment of working conditions for the staff. The features of the technological process at the SPSs resulted in the formation of specific working conditions characterized by the presence of a cooling microclimate, noise production, technological vibration, air pollution by microorganisms. The assessment of working conditions has allowed to refer them to the 3 class (harmful working conditions) of the 2 and 3 degree. Preventive measures for SPSs should include the use of equipment for cleaning the air of working zone, having a combined odourremoving and microbicidal action; the automation of the labor process; the installation of additional equipment for filtering air circulating in workplaces and emitting into the atmosphere; provision of sealing equipment with the use of noise insulating materials.

[POSSIBILITY OF USING NESTED POLYMERASE CHAIN REACTION FOR DIAGNOS- TICS OF DISEASES CAUSED BY VARICELLA ZOSTER VIRUS].

AIM: Demonstrate the possibility of using nested PCR method for determination of Varicella Zoster virus (VZV) in clinical samples of peripheral blood of patients. MATERIALS AND METHODS: Material from 35 patients with clinical manifestations of herpes zoster and control group of 20 healthy donors was used in the study. Monocyte fraction of venous blood cells, pretreated with heparin, was isolated by centrifugation in ficoll-verografin density gradient, total DNA was then isolated from cells by phenol-chloroform extraction with subsequent precipitation with alcohol. Polymerase chain reaction was carried out in thermocyclers Tercyc and TProfessional Gradient (Biometra), amplified DNA was analyzed by electrophoresis on 1.6% agarose gel in the presence of ethidium bromide. RESULTS: Data on detection of viral DNA in blood monocytes in 17 (49%) of ill patients, as well as in 1 (out of 20 in control group) practically healthy donor were obtained. A possibility of a subclinical reactivation of the virus is discussed in the latter case. CONCLUSION: A possibility of viral DNA determination in monocytes of patient blood without using expensive equipment is shown, that could find application in clinical practice, especially for diagnostics of patients with non-characteristic clinical manifestations, as well as patients with subclinical forms of the disease.

[Use of self-expandable stent grafts in aneurysms of internal carotid arteries].

Open surgical intervention is an operation of choice in lesions of coronary arteries. In some cases, however, performing reconstructive surgery is either impossible or associated with high risk of both intra- and postoperative complications. In such cases it is possible to perform an endovascular intervention, including endoprosthetic repair. The Russian and foreign literature contains either sporadic or small-in-number case reports concerning the use of stent grafts in carotid arteries aneurysms. The article describes cases of successful use of self-expandable stent grafts in three patients with a true aneurysm and pseudoaneurysms of internal carotid arteries.

[Features of arterial blood pressure in elderly persons of different ethnic groups in Yakutsk].

The differences in arterial blood pressure in the sample of population in the age of 60 and older of different ethnic groups in Yakutsk, as well as its connection with the other cardiovascular diseases risk factors have been analyzed. It was shown that the average values of systolic and diastolic blood pressure in subsample of the Yakuts appeared to be lower than in Caucasoid gerontic persons. The average values of systolic arterial blood pressure both in the Yakuts and in the Caucasoids were detected higher than normal values in all age-dependent subgroups. The average values of diastolic blood pressure in both ethnic groups were within the limits of high normal level. From 60 to 90 years and older the decrease in systolic and diastolic arterial blood pressure was detected; it was more marked in Caucasoid gerontic persons. The average values of pulse pressure in the Yakuts and in the Caucasoids appeared to be higher than the existing standard and didn't have any differences in ethnic groups. In both ethnical subsamples, pulse pressure values increase was observed in persons of 60-89 years old and its decrease after 90. Persons with overweight, obesity, central (abdominal) obesity, dyslypoproteidemias irrespective of belonging to ethnical group were characterized as having higher levels of arterial blood pressure. Statistically significant differences in the levels of arterial blood pressure in the Yakuts and in the Caucasoids depending on hyperglycemia, smoking, the presence of burdened anamnesis, educational level, marital status was not detected.

Effect of long-term treatment with ß-adrenoceptor antagonists on blood lipid composition and survival of rats with heart failure of different severity.

Plasma LDL content increased in rats with mild chronic heart failure, but decreased in animals with severe pathology. The lipid profile of ß-adrenoceptor antagonists depended on their selectivity. The most selective antagonists of ß-adrenoceptors had little effect on the lipid composition. Nonselective ß-adrenoceptor antagonists increased the index of plasma atherogenicity, which did not depend on the severity of a pathological process. In mild chronic heart failure, these changes were associated with a decrease in HDL concentration, while in severe chronic heart failure; they were related to a tendency towards a decrease in HDL content and an increase in LDL concentration. The increase in survival rate was associated with true antagonism of the agent to ß-adrenoceptors (the absence of intrinsic sympathomimetic activity) and, to a lesser extent, with nonselective blockade of ß-adrenoceptors.

[Effect of angiotensin-converting enzyme inhibitors, beta-adrenoblockers, and their combinations on survival and plasma catecholamine levels in rats with chronic heart failure with induced exacerbations].

By using model of chronic heart failure (CHF) occurring with induced decompensations, it was shown that inhibitors of angiotensin-converting enzyme (ACE) and beta-adrenoblockers promoted to different degree the correction of hypercatecholaminemia and an increase of survival of rats. The most pronounced effect was characteristic of beta-adrenoblockers atenolol, propranolol, and carvedilol. The lower activity was demonstrated by other adrenoblockers that had no intrinsic sympathomimetic activity, as well as captopril. However, positive effect of these drugs on rat survival occurred regardless of their action on the sympathoadrenal system. The ACE inhibitors, particularly hydrophilic lysinopril, to the lesser degree than beta-adrenoblockers, prevented death of animals at the period of the CHF decompensation. The combined use of ACE inhibitors with beta-adrenoblockers, especially with carvedilol, promoted the maximally expressed decrease of the sympathoadrenal activity and an increase of the rat survival, including that at the CHF decompensation.

[Changes in lipid spectrum in rats with chronic heart failure of different severity on the background of angiotensin-converting enzyme inhibitors and beta-adrenoblockers].

The character of lipid spectrum changes in rats with experimental heart failure of non-ischemic genesis depended on its severity. In mild heart failure there was found an increase in the low density lipoprotein levels, whereas in the severe one--a decrease of total cholesterol levels at the expense of the low density lipoproteins. The ACE inhibitors produced no significant changes in the lipid spectrum. The effect of beta-adrenoblockers was determined by their selectivity for beta1-adrenoreceptors. The most selective drugs were distinguished by the metabolically neutral activity. On the contrary, nonselective beta-adrenoblockers increased the degree of plasma atherogeneity.

[Plasma lipid spectrum and survival in rats with heart failure varied by severity under the influence of neurohormonal modulators].

In rats with relatively mild chronic heart failure (CHF) there was an increase in plasma LDL levels. In turn, in severe CHF it was revealed a significant decrease in total cholesterol concentration due to LDL levels decline. ACE inhibitors (captopril, enalapril and lisinopril) and the most selective beta1-adrenoblockers (metoprolol, nebivolol) had not any significant influence on plasma lipids. Less selective and especially nonselective beta-adrenoblockers pindolol and propranolol brought up the atherogenic potential of plasma because of HDL levels reduction in mild CHF whereas in case of severe model--due to the trends toward decrease in HDL concentration and toward increase in LDL levels. Lipophilic ACE-inhibitors and beta-adrenoblockers without intrinsic sympathomimetic activity improved survival in rats with mild CHF to the equal extent. On the contrary in severe CHF beta-adrenoblockers, especially non-selective, had an advantage over ACE inhibitors on the survival effect.

[Medical service and health state of vehicle drivers of city water supply and sewerage enterprise].

The article covers data on medical service system and health state of vehicle drivers on major water supply and sewerage enterprise.