[Experience of the military sanatorium "Arkhangelsk" on medical rehabilitation of patients after surgery].

Experience of the military sanatorium <> on medical rehabilitation of patients after surgery in the treatment of coronary heart disease. Showing features of the organization of medical rehabilitation of patients with coronary artery disease after surgical treatment at a sanatorium stage. The technology of application of restorative treatment, physical and psychological recovery, natural and premature medicinal factors are given. Based on the dynamics of the functional status of patients given control of complex clinical and instrumental studies in comparison with analogous-tech at discharge, to translate them into functional class, characterized with greater functionality is installed, the high efficiency of rehabilitation.

Performance Assessment of the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism Method for Rapid Detection of Susceptibility to Ethambutol and Molecular Prediction of Extensively Drug-resistant Tuberculosis in Clinical Isolates of Mycobacterium tuberculosis.

INTRODUCTION: The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was employed for rapid detection of ethambutol (EMB) resistant clinical isolates of Mycobacterium tuberculosis. MATERIALS AND METHODS: From 182 clinical isolates of M tuberculosis collected from different regions, 103 strains were entered in the investigation. DNA was extracted by Chelex 100 method and PCR was performed using specific primers for embB gene. Polymerase chain reaction products were digested with HaeIII and NlaII restriction endonucleases and the patterns of restriction fragments were analysed. Some randomly selected samples were sequenced. RESULTS: Out of 103 studied strains, 52 were resistant to EMB. The cases of secondary tuberculosis were 53 (51.50 ± 1.77%), and primary cases 50 (48.50 ± 1.77%; p > 0.05). From 63 extensively drug-resistant (XDR), pre-XDR and multidrug-resistant (MDR) isolates, 27 (87%), 18 (81.8%) and 7 (70%) strains were resistant to EMB, respectively. Results of PCR-RFLP method showed that from 27R EMB XDR isolates, 13 (sensitivity 48% with CI: 0.307, 0.66 and specificity 100%), from 18R EMB pre-XDR strains, 4 (sensitivity 22% with CI: 0.09, 0.45 and specificity 100%) and of 7R EMB MDR, 2 (sensitivity 28% with CI: 0.082, 0.64 and specificity 100%) had mutation in ATG-Met codon 306. Results of sequencing were concordant with RFLP method. Overall, sensitivity of the molecular method was 36.5% (CI: 0.09, 0.45) and specificity 100%. None of the 40 pansusceptible strains was embB306 mutants. Extensively drug-resistant strains had a higher proportion of embB306 mutants (43%) than pre-XDR and MDR isolates (odds ratio 6.78; p < 0.001). CONCLUSION: Fast detection of susceptibility to EMB drug is possible by PCR-RFLP. The embB306 locus is a candidate marker for rapid prediction of high resistance consisting of MDR and XDR forms to anti-tuberculosis drugs using this method.

[Polymorphism of the genes IL-1RA and TNF-alpha in patients with gastritis and duodenal ulcer associated with Helicobacter pylori].

The goal of this work was to determine the rate of the polymorphism of the genes-antagonists of receptor IL-1 (IL-1RA) and TNF-alpha in patients with gastritis and duodenal ulcer associated with Helicobacter pylori. The receptors were tested for the clinical manifestation of the disease. A total of 126 patients with different gastroduodenal pathology and H. pylori in autopsy were tested. The results of this work demonstrated a correlation between the risk of duodenal ulcer and allele A of gene TNF-alpha in position 308 in the patients. The analysis of the gene-IL-1RA polymorphism demonstrated statistically significant difference between the patients in the frequency of the genotype 2/l. The results of this work showed that parallel typing of the genes of H. pylori in virulence was required for characterization of the bacteria-patient association. The correlation between the results of the typing with polymorphism of genes of cytokines in patient autopsy was also required.

[A new approach to postexposure treatment of rabies by complex of immuno- and chemotherapy in belarus].

A method for preventive treatment of rabies with a complex of immuno- and chemotherapeutics was developed. Rifampicin was used a an etiotropic drug. In the experiments on laboratory animals infected with fixed and street strains of rabies virus it was shown to prolong the incubation period and to increase the survival rate. The protective mechanisms of rifampicin against rabies should be associated with inhibition of RNA transcription, as well as immunomodulating function of macrophages, dendritic cells, B- and T-cells. Since 1992, after the approval of the Ministry of Health of Belarus rifampicin is used in complex with antirabic vaccine for postexposure treatment of rabies in people after severe bites by infected animals (wolves, foxes, dogs). For an 18-year period (1992-2009) of integrated application of chemo- and immunotherapy in Belarus there was not registered any case of hydrophobia in people even after the heaviest wolf bites, incompatible with life (penetrating injuries of the skull, scalping, multiple bites).

Study of promoter and structural gene sequence of whiB7 in MDR and XDR forms of Mycobacterium tuberculosis.

Resistance phenomenon in M tuberculosis is mainly based on decreased permeability of the bacterial envelope and function of effluent pumps. The regulatory gene of the whiB7 transcription determines drug resistance in these bacteria. Increases in WhiB7 protein activity induce transcription of resistance genes leading to intrinsic multidrug resistance. The aim of this work was to evaluate the whiB7 gene sequence in susceptible, MDR and XDR clinical isolates of M tuberculosis in order to further design an inhibitor. Thirty-three clinical isolates of MTB identified as susceptible, MDR and XDR-TB were investigated by PCR for sequencing of the entire promoter (429 bp), structural gene (279 bp) and the end of the upstream gene uvrD (265 bp). No differences were detected in the sequences of the structural gene in susceptible and MDR with XDR isolates and all of them terminated at TGA as stop codon. Examination of sequence profiles of the promoter part of whiB7 by several sets of primers proved that there were no differences between sequence of susceptible, MDR and XDR isolates by type strain (H37Rr). Furthermore, the structure of WhiB7 protein was studied in achieved sequences from clinical isolates. We found that the promoter and structural gene of whiB7 are highly conservative in clinical susceptible and resistant isolates. It is a key finding that would assist in the design of an inhibitor for the WhiB7 protein in all clinical forms in further studies.

[Impact of a magnetic field on blood separation kinetics in patients with joint diseases].

The paper gives the results of experiments on phase separation of blood in the constant magnetic field that allows the structure of blood to be regulated, without changing its cellular and chemical composition. Blood deposition kinetic relationships were obtained for patients with joint diseases of various etiology (osteoarthritis, osteoarthrosis deformans, endoprosthesis instability, contusions, and joint wounds). They correlate with the severity of an inflammatory process in the joint and its adjacent tissues, with a patient's resistance to the development of pathology, and with red blood cell mobility in the biophysical field of a living organism. Analysis of relationships gives information on concentrations in plasma and hence synovial fluid (the basis of which is blood dialysate) in the liquid-crystalline phospholipid and cholesterol phase that determines the lubricity of synovial fluid and a low friction in the joints. The method may be used for the primary evaluation of efficacy of drugs for joint treatment, which is made in vitro on the blood taken from the patients rather than on the latter.

Novel macroarray-based method of Corynebacterium diphtheriae genotyping: evaluation in a field study in Belarus.

In this study, we evaluated a novel macroarray-based spoligotyping method for Corynebacterium diphtheriae strain typing. A total of 20 C. diphtheriae biotype gravis toxigenic isolates collected in Belarus from suspected foci of diphtheria infection (diphtheria cases, carriers, or contacts) were subjected to DNA fingerprinting. All strains had an identical ribotyping profile that was identified as ribotype 'Rossija' by comparison with the international ribotype database at the Institut Pasteur of Paris. A spoligotyping method based on simultaneous reverse-hybridization analysis of two CRISPR (clustered, regularly interspaced short palindromic repeats) loci differentiated these strains into three spoligotypes. Comparison of the spoligotyping results with the epidemiological linkage network helped us to resolve suspected links in the chains of transmission. To conclude, the C. diphtheriae spoligotyping method demonstrated its utility in the field study, in particular, underlining the importance of the use of both CRISPR loci. The generated discrete data can be presented in digital binary format and be easily exchanged between laboratories and stored in local and global databases.

[Synthesis of olean-18(19)-ene derivatives from betulin].

The rare olean-18(19)-ene triterpenoids moradiol and moronic acid were synthesized from betulin, and their antiviral properties were investigated.

[Biological and immunogenic characteristics of Chlamydia trachomatis strain MT-2A (serovar D) and its use for development of experimental inactivated vaccine].

Biological characteristics of C. trachomatis author's strain MT-2A (serovar D) is presented. Stages of development on its basis the experimental formalin-inactivated vaccine against Chlamydia were described. Humoral and cellular immune response to the vaccine administered on 3-dose immunization schedule in conjunction with polyoxidonium as adjuvant was studied. Significant immunological efficacy of the vaccine was shown. T- and B-cell immune responses were characterized. Titer of IgG antibodies against Chlamydia in blood serum after 3rd dose of the vaccine was 10,880+/-1,817.76. Assessment of T-cell response showed that reaction of delayed hypersensitivity with formation of granuloma presented in 60% of animals. Proportion of immunoblasts in reaction of blast-transformation was 29.3+/-2.8%. Perspectives of further studies of the developed corpuscular vaccine against Chlamydia are discussed.

Genomic analyses reveal two distinct lineages of Corynebacterium ulcerans strains.

Corynebacteriumulcerans is an important zoonotic pathogen which is causing diphtheria-like disease in humans globally. In this study, the genomes of three recently isolated C. ulcerans strains, 4940, 2590 and BR-AD 2649, respectively from an asymptomatic carrier, a patient with pharyngitis and a canine host, were sequenced to investigate their virulence potential. A comparative analysis was performed including the published genome sequences of 16 other C. ulcerans isolates. C. ulcerans strains belong to two lineages; 13 strains are grouped together in lineage 1, and six strains comprise lineage 2. Consistent with the zoonotic nature of C. ulcerans infections, isolates from both the human and canine hosts clustered in both the lineages. Most of the strains possessed spaDEF and spaBC gene clusters along with the virulence genes cpp, pld, cwlH, nanH, rpfI, tspA and vsp1. The gene encoding Shiga-like toxin was only present in one strain, and 11 strains carried the tox gene encoding the diphtheria-like toxin. However, none of strains 4940, 2590 and BR-AD 2649 carried any toxin genes. These strains varied in the number of prophages in their genomes, which suggests that they play an important role in introducing diversity in C. ulcerans. The pan-genomic analyses revealed a variation in the number of membrane-associated and secreted proteins that may contribute to the variation in pathogenicity among different strains.

[Point mutations in tox promoter/operator and diphtheria toxin repressor (DTXR) gene associated with the level of toxin production by Corynebacterium diphtheriae strains isolated in Belarus].

DNA fragments 129 bp in length containing promoter region of the tox gene from 81 toxigenic strains Corynebacterium diphtheriae were analyzed using the SSCP (single strand conformational polymorphism). We found that only two strains had mutations; the strains also had highest levels of toxin production (over 5120 Vero CD50/ml). Other strains were characterized either as high-level toxin-producing (640-5120 Vero CD50/ml, 41 strains) or low-level toxin-producing (40-320 Vero CD50/ml, 38 strains). Nucleotide sequence analysis revealed single T to C mutations at positions -54 and -184 within -232 - +85 region of tox operon. The first mutation at the -184 position was mapped outside the tox promoter/operator, whereas the second substitution at the -54 position modified the 9-base-pair interrupted palindromic sequence of the tox promoter/operator from ATAATTAGG in the wild-type bacteriophage (to ACAATTAGG in strains with enhanced level of toxin production. Nucleotide sequence analysis of -76 - +681 region of diphtheria toxin repressor (dtxR) gene from 15 strains of C. diphtheriae revealed two missense mutations resulting in amino acid substitutions A 147 V; and L 214 I in the C-terminal region of the DtxR protein. Seven of these strains were identified as high-level toxin-producing and 4 strains, as low-level toxin-producing. In addition, one low-level toxin-producing strain was shown to contain a missense mutation leading to amino acid substitution I 221 T. Three strains, including two highest-level toxin producing strains contained no nucleotide substitutions, as well as the C7(-) strain. The 10 strains belonging to the Sankt-Peterburg and Rossija epidemic ribotypes as well as NCTC 13129 strain (etiologic agent of the diphtheria epidemic outbreak in the Eastern Europe) was shown to contain two mutations A 147 V and L 214 I in the C-terminal region of the DtxR protein.

[The molecular epidemiology of HIV-1 in Belarus in (1996-2004): a predominance of subtype A variants and circulation of subtype B variants].

To study the molecular epidemiology of HIV-1 in Belarus, the genetic sequences of HIV-1 variants were obtained from 50 infected persons, which represented the main stages, risk groups, and geographic areas of the epidemic. The env and gag sequences were studied for HIV-1 variants from 31 persons, the env sequences were for HIV-1 variants from 18 persons, and the gag sequence was for HIV-1 variant from 1 person. Phylogenetic analysis indicated that the sequences of HIV-1 variants from 46 persons were homogenic and evolutionally closely related to IDU-A strains specific for other epidemics in the former Soviet Union are dominating in the epidemic in Belarus. Circulation of epidemiologically unrelated subtype B viruses was also established.

[Photodynamic inhibition of infection caused by herpes simplex virus type 1 in the cultured cells, by using 5-aminolevulinic acid-induced porphyrins].

On simulating infection caused by different herpes simplex virus type 1 (HSV-1) variants responsive and unresponsive to the drugs acyclovir and phosphonoacetic acid in the cultured Vero and C6 cells has revealed the higher ability of target cells to accumulate 5-aminolevulenic acid (ALA)-induced endogenous porphyrins, which determines the selectivity of their photo damages. Optimal conditions have been defined for all the studied HSV-1 variants to show a virus-inhibiting effect upon photodynamic exposure of infected and ALA-treated cell cultures.

[Changes in vaginal microbiocenosis in patients with chronic pelvic inflammatory diseases following antibiotic therapy].

The species composition of the vaginal microorganisms in healthy women and in patients with chronic pelvic inflammatory diseases before and after treatment in a gynecological hospital was studied. The study revealed that antibiotic therapy did not lead to complete clinical convalescence. During bacteriological investigation of patients changes in vaginal microbiocenosis, manifested by a decreased number of microbial species, an increased proportion of Escherichia coli, the occurrence of Staphylococcus aureus, a decreased number of Lactobacillus ssp., were observed. Antibiotic therapy aggravated the dysbiotic microbial picture of the vagina.

[Specific clinical, epidemiological patterns and laboratory diagnostics of enterovirus infection in the Republic of Belarus].

The clinical and epidemiological patterns as well as the results of the laboratory verification of the outbreak of enterovirus infection (EVI) in Minsk during the period of summer-autumn, 2000, are presented. During this outbreak a variety of clinical forms were observed, the serous meningitis being prevalent (57.5%). Practically simultaneous occurrence of infection on the territory of all administrative districts of the city, the predominant involvement of children aged up to 14 years into the outbreak, a high proportion of simultaneous casualities in the multiple foci. A number of circulating enteroviruses (EV)--ECHO 30, ECHO 6 of three serotypes and Coxsackie B5--were simultaneously isolated from clinical material. EV of the same serotypes were isolated from tap drinking water, and neutralizing antibodies to these serotypes were often detected in the patients blood sera. Infectious EV were also present in samples of bottled water and in water reservoirs used for bathing. The routes of EV transmission and the improvement of EVI control are discussed.

Virologic and serologic investigations of West Nile virus circulation in Belarus.

In 1985-1994 virologic and serologic investigations were performed for the purposes of West Nile (WN) virus circulation establishment on the territory of Belarus. Blood-sucking mosquitoes, midges, wild small mammals, birds as well as blood and cerebrospinal samples from patients with nondifferentiated fevers and from healthy individuals were under studies. Four virus strains were isolated in Belarus for the first time, namely: 1--from birds (48-WN Tremlya); 2--from Aedes mosquitoes (319 and 2438); 1--from a febrile patient (Win). Their antigenic and biological properties were examined in cell cultures and laboratory animals. The isolates turned to be identical with each other and closely related to reference Egypt strain Eg 101, that is a topotype for the African virus group. One more WN virus strain (8891) was isolated from Anopheles mosquitoes in 1999. Specific antibodies to the virus in human blood sera were identified by immunological and serologic assays in 1.7% of Belarusian population. In Gomel and Brest Regions the percentage of seropositive individuals reached 5.8 and 15.4, respectively. WN virus antibodies prevailed in 0.6-5.8% of cattle, in 2.9-6.8% of wild small mammals and in 6.5-16.7% of birds. Thus, the conclusion was made on the existence of favourable conditions for the virus spread throughout the whole country and in the south in particular. Blood-sucking mosquitoes and birds are principle vectors in WN virus circulation in Belarus. 16 serologically confirmed cases of WN encephalitis were revealed in patients with fever of obscure etiology. In the view of the given data, reports on the reemergence of the pathogen in different countries and the tendency in global warming WN virus monitoring should become a subject of concern for Belarusian public medical care services.

Vaccine-associated paralytic poliomyelitis and other diseases with acute flaccid paralysis syndrome in Belarus.

According to the WHO global polio eradication initiative acute flaccid paralysis (AFP) surveillance has been conducted in Belarus since 1996. For the period 1996-2002, 295AFP cases were reported. The main indices ofAFP surveillance in Belarus met the WHO criteria. A11 AFP cases, with the exception of one, were virologically examined. Polioviruses (PV) were isolated from 28 (9.5%) of them. Results of intratypic differentiation (a neutralization test with type-specific monoclonal antibodies and a restriction fragment length polymorphism assay) proved vaccine origin of all isolated PV. According to the final classification, 11 AFP cases were classified as vaccine-associated paralytic poliomyelitis (VAPP). Nine VAPP cases were recipient [six of them developed after the first, two--after the third and one--after the fourth oral poliovirus vaccine (OPV) dose] and two cases in non-vaccinated children were classified as contact VAPP cases. PV of all three serotypes were isolated with an equal frequency from the recipient cases and only PV2--from contact ones. Immunological investigations of children with VAPP showed that the majority of them had disorders in B-cell immunity. A risk of one VAPP case per 96,000 first OPV doses and per 745,000 distributed ones was estimated. The other 284 AFP cases were classified as AFP of non-polio etiology (non-polio AFP). Among them Guillain-Barré syndrome (118 cases, 41.5% of all non-polio AFP cases), traumatic neuritis (63 cases, 22.2%), transient monoparesis of limb (35 cases, 12.3%), myelitis (26 cases, 9.2%) were registered most frequently. Vaccine PV were isolated from 19 (6.7%) children with non-polio AFP, 28 (9.9%) children excreted non-polio enteric viruses. In contrast to VAPP, other AFP with PV isolation had no clinical picture typical of poliomyelitis, and had no any residual paralysis 60 days after the onset of paralysis. PV isolation from them seemed to be not related to the etiology of the disease, but was a mere coincidence of paralysis with the recent vaccination. Results of AFP surveillance supported the previous data on the absence of classical poliomyelitis cases caused by wild PV in Belarus for more than 35 years.

The immunogenicity and reactogenicity of the trivalent vaccine, Trimovax, indicated for prevention of measles, mumps, and rubella, in 12-month-old children in Belarus.

In the Republic of Belarus, immunization of children against measles and mumps had been carried out using monovalent preparations according to the national schedule of measles vaccination at 12 months of age and mumps vaccination at 24 months of age. A rise of rubella incidence in the last few years (i.e., for the official registration period 1980 to 1998, there was an increase from 72.2 to 607.5 cases per 100,000 population) made it necessary to implement immunization against this infection, as well. Therefore, in 1996, combined vaccination against measles, mumps, and rubella of 12-month-old children was carried out for the first time in a clinical trial that used the vaccine Trimovax [Aventis Pasteur (formerly, Pasteur Mérieux Connaught), Lyon, France]. The reactogenicity of the vaccine was investigated in 372 children. Post-vaccination reactions were noted in 5.6% of children; in 1.3% of children the reactions were classified as severe [i.e. associated with body (axillary) temperature > or = 38.6 degrees C]. For the evaluation of immunogenicity, sera from 324 children were obtained 2 to 2.5 months after inoculation, and serum antibody levels were measured by enzyme immunoassays. Among the vaccines, protective antibody titers (expressed in inverse of dilution units) were observed to measles (> or = 1:50) in 97.8%, to mumps (> or = 1:50) in 93.8%, and to rubella (> or = 1:100) in 96.0% of children. Antibodies to all three components of the vaccine were mainly present in intermediate (1:200-1:800) or high (> or = 1:1600) titers: to measles in 96.3%; to mumps in 75.8%; and to rubella in 73.5% of vaccines. The results of these trials are evidence of the good safety and immunogenicity of this MMR vaccine, which provides an alternative to the currently used measles and mumps monovaccines, with the additional benefit of providing immunity against rubella, as well.

Enteroviruses identifications and differentiation on the basis of the selective group--specific inhibitory effect of chemical compounds.

Two new enteroviruses (EV) inhibitors with the selective group-specific effect were detected and studied representing the products of the original chemical synthesis. One of them--nifan (arylfuran derivative) inhibits poliomyelitis virus replication, the other one--belvtazide (synchonic acid derivative) blocks non-poliomyelitis EV (ECHO and Coxsackie B) replication. The study of the reference strains of poliomyelitis virus type 1-3, twenty-three ECHO virus types (from the 1st to the 33rd), Coxsackie B virus type 1-6 and 288 primary EV isolates did not reveal type or strain specific variability in the inhibitors effect. Nifan and belvtazide supress the replication of both EV monostrains and their mixtures. The isolates of mixed nature are inhibited by the mixture nifan + belvtazide. At the same time neither separate chemicals nor their blend affects viruses from other families (Adenoviridae, Orthomyxoviridae, Herpesviridae etc.). The mechanism of nifan and belvtazide action is intracellular EV replication inhibition (they do not affect the process of virus adsorption and penetration into the cell), suppression of de novo virus synthesis by 7.0-2.25 lg (tissue culture infective dose 50 per cent) TCID50/ml and of virus-induced RNA synthesis. The drugs feature is high selectivity (90-91%) regarding RNA polioviruses (nifan) and RNA non-poliomyelitis EV (belvtazide). Nifan and belvtazide antiviral effect selectivity allows unknown cytopathic agents (CPA) belonging to the EV to be established with the high degree (over 98%) of reproducibility at the stage of primary identification with the differentiation of poliomyelitis and non-poliomyelitis viruses.