Analysis of Polymorphism of Angiotensin System Genes (ACE, AGTR1, and AGT) and Gene ITGB3 in Patients with Arterial Hypertension in Combination with Metabolic Syndrome.

Changes in the frequencies of genotypes and mutant alleles of ACE, AGTR1, AGT, and ITGB3 genes were analyzed in patients with arterial hypertension coupled with metabolic syndrome (N=15) and compared with population data and corresponding parameters in patients with isolated hypertension (N=15). Increased frequency of genotype ID of ACE gene (hypertension predictor) was confirmed for both groups. In case of isolated hypertension, M235M genotype (gene AGT) was more frequent, in case of hypertension combined with metabolic syndrome, the frequency of genotypes A1166C and C1166C of the gene AGTR1 was higher in comparison with population data. Comparison of mutant allele frequencies in the two groups showed that at the 90% significance level allele T of the AGT gene was more frequent in hypertension coupled with metabolic syndrome (OR=1.26) and genotype A1166A of the AGTR1 gene was more frequent in the group with isolated hypertension.

[Current approaches to the morphologic diagnosis of different types of congenital epidermolysis bullosa]. / Sovremennye podkhody k morfologicheskoi diagnostike razlichnykh tipov vrozhdennogo bulleznogo epidermoliza.

Congenital epidermolysis bullosa (CEB) is an extensive group of hereditary skin diseases, the differential diagnosis of which is a challenge due to the rarity of this pathology and the diversity of its clinical manifestations. The determination of the type of CEB makes it possible to estimate its prognosis and to facilitate a prenatal diagnosis. AIM: to optimize the morphological diagnosis of different types of CEB. MATERIAL AND METHODS: 28 skin biopsies from 14 patients with different types of CEB were investigated. The investigators performed routine histological examination of skin fragments taken from a bullous area and immunofluorescence antigen mapping using the indirect immunofluorescence test (IIFT) with antibodies against structural proteins of the dermal-epidermal junction (laminin α3, ß3, and γ2 chains, keratins 5 and 14, types VII and XVII collagen, α6 and ß4 integrin subunits, desmoplakin, plectin, kindlin-1, and plakophillin) of the apparently unaffected skin. The intact skin of healthy individuals, which had been obtained during cosmetic operations, was used as controls in IIFT. RESULTS: Immunofluorescence antigen mapping could determine the type of CEB in all cases and in 86% of cases identify the protein, the impaired production of which was responsible for the development of the disease. CONCLUSION: Immunofluorescence antigen mapping is an integral part of the comprehensive morphological diagnosis of CEB, acting as an intermediate between the morphological verification of CEB diagnosis and the targeted search for mutations by a molecular genetic method.

[Role of the skin expression of neuropeptides, neurotrophins and their receptors in the pathogenesis of dermatoses].

OBJECTIVE: to define the role of neurotransmitters and their receptors in the development of itch and in the maintenance of a skin inflammatory response in patients with psoriasis and atopic dermatitis. MATERIAL AND METHODS: Skin biopsy specimens from 30 patients with psoriasis and 30 patients with atopic dermatitis were investigated by histological, immunoperoxidase, and indirect immunofluorescence assays. The investigators determined the expression of protein gene product 9.5 (PGP9.5), amphiregulin, semaphorin 3A, calcitonin gene-related peptide (CGRP) and its receptor (CGRP-R), nerve growth factor (NGF) and its receptor TrkA, and substance P (SP) and its receptor SP-R. The indirect immunofluorescence assay was used for quantitative analysis. The findings were statistically analyzed using a Statistica 10 program. RESULTS: Immunoperoxidase examination of the skin biopsy specimens from patients with atopic dermatitis and psoriasis revealed enhanced expression of amphiregulin, NGF, and PGP9.5, appearance of positively stained epidermal nerve fibers, and decreased expression of the nerve reduction factor semaphorin 3A in all cases. Some patients with atopic dermatitis and psoriasis showed increased expression of CGRP and CGRP-R, SP, SP-R, and TrkA. A pronounced inflammatory response was generally observed in these cases. CONCLUSION: The investigation performed suggests that atopic dermatitis and psoriasis are characterized by a larger number of epidermal nerve fibers and by a direct correlation between this indicator, disease severity, and itch intensity. The production of neuropeptides and neurotrophins is closely related to the development of a skin inflammatory response irrespective of its cause and dysregulation of these processes is likely to favor the body's sensitization and the chronic pattern the course of diseases.

[Future Therapeutic Targets in Management of Autoimmune Skin Diseases].

Pemphigus is a severe, potentially fatal bullous skin disease, caused by desmoglein autoantibody production and immune-mediated regulation of T-cells subsets. Conventional therapy including systemic corticosteroids with or without other immunosupressants causes numerous adverse effects and becomes inefficient in refractory patients. In this work, the authors showed a modern view on the pathogenesis ofpemphigus. This article describes the detailed action mechanism of rituximab, a chimeric monoclonal antibody directed against CD20 antigen of B-cells. The authors conduct the results of meta-analyses of rituximab's efficiency in pemphigus patients. Moreover, in this article, the authors consider new promising treatment tions and potential targets for biological therapy of pemphigus diseases.

[The role of some individual amino acid substitutions in penicillin-binding protein (PBP2) of Neisseria gonorrhoeae in the emergence of resistance to ceftriaxone].

The goal of the study was to identify amino acid replacements in the structure of penicillin-binding protein PBP2, which may influence on the development of resistance N. gonorhoeae to the III cephalosporins generation. The gene penA of 50 strains of N. gonorrhoeae was sequenced: 20 strains with high sensitivity to ceftriaxone (MIC, Minimum Inhibitory Concentration, = 0.002 mg/L) and 30 strains with decreased sensitivity to ceftriaxone (MIC = 0.03-0.25 mg/L). The difference of MIC sensitivity between these strains was 30-250 times. Then nucleotide sequence was transformed into the amino acid sequence of PBP2 protein. Mutations in the gene penA and amino acid replacements in the protein PBP2 were found in 16 of 20 strains (80%) with high sensitivity to ceftriaxone and in all strains with decreased sensitivity to ceftriaxone. Amino acid replacements in the PBP2 protein were compared with amino acid replacements in groups, which characterize the PBP2 structure in accordance with the international classification Ito M. The amino acid replacement of PBP2 at positions 346, 505, 511, 517, 543, 567, 575, 576 are associated with V group by Ito M and have features of resistance of N. gonorrhoeae to ceftriaxone authentically (OR = 3.9 ± 2.5; χ2 = 4.9; p < 0.05). It was shown that the replacement of glycine to serine at position 543 of PBP2 in the analyzed strains induced the multiple increase of resistance to ceftriaxone. These data may be significant as showing strong influence of amino acid replacements at positions 346, 505, 511, 517, 567, 575 and, in particular, 543 for development of resistance N. gonorrhoeae strains to ceftriaxone.

[Rhinosinusogenic orbital complications: the prevalence and principles of treatment].

Results of retrospective analysis of 8397 medical records accumulated in the otorhinolaryngological department of a regional hospital for the last 15 years are presented. The patients were admitted for the treatment of rhinosinusitis. A total of 269 (3.5%) orbital complications were documented. All age groups of the patients were dominated by males. Children most frequently developed orbital complications in case of acute sinusitis and adults in case of exacerbation of the chronic disease. The largest number of orbital complications were associated with the inflammatory process in the frontal sinus. The authors emphasize the necessity of accurate differential diagnosis between complications and propose a strategy for their treatment.

The Russian gonococcal antimicrobial susceptibility programme (RU-GASP)--national resistance prevalence in 2007 and 2008, and trends during 2005-2008.

Antimicrobial resistance (AMR) in Neisseria gonorrhoeae is a major problem worldwide. In the former Soviet countries including Russia, the knowledge regarding AMR has been highly limited. However, in 2004 the Russian gonococcal antimicrobial susceptibility programme (RU-GASP) was initiated. The aims of this study were to examine and describe the prevalence of N. gonorrhoeae AMR in 2007 and 2008 in Russia, and reveal trends in the period from 2005 to 2008. Gonococcal isolates (660 in 2007 and 900 in 2008) from 36 surveillance sites were examined using agar dilution method. From 2005 to 2008, the proportion of isolates resistant to spectinomycin increased from 0% to 7.2%, and remained high for those resistant to ciprofloxacin (approximately 49%). The resistance to azithromycin was 2.3% and 0.4% in 2007 and 2008, respectively. All isolates between 2005 and 2008 were susceptible to ceftriaxone. In conclusion, the AMR of N. gonorrhoeae in Russia is high, as in most countries in the European Union, and ceftriaxone should be the first line for treatment. If there is no access to ceftriaxone or in the presence of severe beta-lactam antimicrobial allergy, spectinomycin should be used; however, the resistance to spectinomycin has increased. Regular, quality-assured national and international surveillance of AMR in N. gonorrhoeae is crucial globally for public health.

National surveillance of antimicrobial susceptibility in Neisseria gonorrhoeae in 2005-2006 and recommendations of first-line antimicrobial drugs for gonorrhoea treatment in Russia.

OBJECTIVES: To investigate comprehensively the antimicrobial susceptibility and resistance of Neisseria gonorrhoeae during 2005-2006 in a national survey and to recommend effective antimicrobial drugs for the treatment of gonorrhoea in Russia. METHODS: The susceptibility of N gonorrhoeae isolates, cultured mainly from consecutive gonorrhoea patients (n = 1030) during the period January 2005 to December 2006 in Russia, to penicillin G, ceftriaxone, ciprofloxacin, tetracycline and spectinomycin was analysed using the agar dilution method. Nitrocefin discs were used for beta-lactamase detection. RESULTS: All isolates were susceptible to ceftriaxone. During 2005 and 2006, however, 5%, 50%, 70% and 77% displayed intermediate susceptibility or resistance to spectinomycin, ciprofloxacin, tetracycline and penicillin G, respectively. Furthermore, 4% of the isolates were beta-lactamase producing during these years. The different federal districts of Russia displayed substantial heterogeneities with regard to the prevalence of gonorrhoea and antimicrobial resistance among N gonorrhoeae isolates. CONCLUSIONS: In Russia, penicillins, ciprofloxacin, or tetracycline should definitively not be used in the empirical treatment of gonorrhoea. The recommended first-line antimicrobial drug should be ceftriaxone. If ceftriaxone is not available, spectinomycin ought to be used. Increasing levels of intermediate susceptibility and resistance to spectinomycin have, however, been observed during recent years and, accordingly, great care and monitoring should be undertaken when using this agent. Continuous local, national and international surveillance of N gonorrhoeae antimicrobial susceptibility, in order to reveal the emergence of new resistance, to monitor changing patterns of susceptibility and to be able to update treatment recommendations on a regular basis, is crucial.

Analysis of the contribution of molecular mechanisms into formation of gonoccocal resistance to tetracycline.

We applied complex genetic analysis for evaluation of tetracycline-resistance markers in 129 clinical strains of Neisseria gonorrhoeae from Central, Privolzhskii, and Siberian regions. For detection of mutations in rpsJ gene and MtrRCDE locus we first used minisequence reaction followed by identification of products by MALDI-TOF mass spectrometry. The incidence of detection of resistance markers among the analyzed strains were: tetM--3.1%, mutations in genes rpsJ--82.2%, penB--62.8%, and mtrR--54.3%. The analyzed genetic markers were not detected in 17.5% strains. tetM gene was detected in only 12.5% strains from the Central Region. No differences were revealed in regional distribution of other genotypes. Genotypes tetM(pres), rpsJ(mut), mtrR(mut), and rpsJ(mut), penB(mut), mtrR(mut) reliably predict tetracycline resistance. Microbiological and genetic testing of tetracycline resistance yielded similar results.

Direct evaluation of drug resistance parameters in gonococcus.

We carried out complex genetic analysis of clinical samples containing N. gonorrhoeae DNA, the genotype and profile of drug resistance of this agent were evaluated. Changes in genes responsible for the formation of N. gonorrhoeae resistance to penicillins, fluoroquinolones, and spectinomycin were detected during minisequencing with subsequent MALDI-TOF mass spectrometry. The sensitivity of gonococcus was evaluated directly in the clinical sample without culturing.

Analysis of genetic markers of N. gonorrhoeae resistance to beta-lactam antibiotics.

A complex method for detection of genetic markers of N. gonorrhoeae resistance to penicillin was developed. Mutations in penA and ponA genes were detected by minisequencing reaction with subsequent detection of reaction products by MALDI-TOF mass spectrometry. This approach was tested on 31 clinical strains of N. gonorrhoeae with minimum inhibitory concentration of penicillin from 0.03 to 8 microg/ml and higher. Mutations in penA and ponA genes in moderately resistant strains were shown (minimum inhibitory concentration up to 0.5 microg/ml) and mutations in penA, ponA, and penB genes in resistant strains (minimum inhibitory concentration more than 1.0 microg/ml). beta-Lactamase genes were detected in 4 strains with high resistance (minimum inhibitory concentration 4-8 and more microg/ml). Correlation between microbiological resistance and presence of respective mutations in the studied locuses was detected.

A multi-centre evaluation of nine rapid, point-of-care syphilis tests using archived sera.

OBJECTIVES: To evaluate nine rapid syphilis tests at eight geographically diverse laboratory sites for their performance and operational characteristics. METHODS: Tests were compared "head to head" using locally assembled panels of 100 archived (50 positive and 50 negative) sera at each site using as reference standards the Treponema pallidum haemagglutination or the T pallidum particle agglutination test. In addition inter-site variation, result stability, test reproducibility and test operational characteristics were assessed. RESULTS: All nine tests gave good performance relative to the reference standard with sensitivities ranging from 84.5-97.7% and specificities from 84.5-98%. Result stability was variable if result reading was delayed past the recommended period. All the tests were found to be easy to use, especially the lateral flow tests. CONCLUSIONS: All the tests evaluated have acceptable performance characteristics and could make an impact on the control of syphilis. Tests that can use whole blood and do not require refrigeration were selected for further evaluation in field settings.

[Detection of fluoroquinolone resistance SNPs in gyrA and parC genes of Neisseria gonorrhoeae using MALDI-TOF mass-spectrometry].

For many known mechanisms of the drug resistance in microorganisms are described genetic markers (specific changes in the genome of microorganism, in the majority of the cases representing single nucleotide polymorphism). The search for the new methods, which make possible to identify single nucleotide changes with the greater effectiveness and at smaller prime is actual for the solution of the problem of the identification of the resistant strains. In this work a new approach of the determination of single nucleotide polymorphisms is proposed. It is based on the reactions of mini-sequencing and/or sequencing with the subsequent Matrix-Assisted Laser Desorption/Ionisation Time Of Flight Mass-Spectrometry (MALDI-TOF MS) of the reaction products. The approach was tested on a clinical group of Neisseria gonorrhoeae strains to investigate specific single nucleotide polymorphisms in genes gyrA and parC (the genetic markers of the bacterium fluoroquinolone resistance). The results of the nucleotide polymorphism deter- mination was completely agreed with the data, obtained earlier with the use of a "gold standard" (sequencing with the classical gel-electrophoresis separation of the reaction products). There is specific interest in the method of sequencing of the short DNA sequences using MALDI-TOF MS. The new high-throughput approach of the single nucleotide polymorphisms determination in bacterial genes considerably increases the effectiveness of the methods of microorganism's identification, genotyping and determining the genetic markers of the drug resistance.

[Resistance of Russian isolates of Neisseria gonorrhoeae to fluoroquinolones].

Fluoroquinolones still belong to the drugs of choice in the treatment of uncomplicated gonorrhea. At the same time, there have been more data on the spreading N. gonorrhoeae strains resistant to fluoroquinolones. A variety of mechanisms, like modification of the target of antibiotic's action (point mutations in genes gyrA and parC), a decreasing permeability of the bacterial cell membrane (amino-acid changes Por protein) and a growing efflux of antibiotic (mutations in the promoter or in the coding region of mtrR) mediate in the shaping resistance of the drugs. The MIC values for four fluoroquinolone-series antibiotics were determined and the gyrA, parC, por and mtrR genes were examined for resistance-responsible mutations in 32 studied clinical strains of N. gonorrhoeae. Strains with high resistance to fluoroquinolones were detected; 3 of them had no common changes in GyrA or ParC, however, amino acid changes and mutations were detected in Por protein and promoter or gene mtrR encoding region, respectively. The paper contains priority data on the detection (in Russia) of N. gonorrhoeae strains with high resistance to fluoroquinolones. Involvement of different mechanisms in the process of resistance shaping is discussed. The results are of practical importance for planning the antibacterial therapy of gonorrhoeae; they point out the need in regional testing of resistance in the N. gonorrhoeae population encountered in Russia.

Fluoroquinolone-resistant Neisseria gonorrhoeae isolates from Russia: molecular mechanisms implicated.

OBJECTIVES: During a longitudinal study of the prevalence of antimicrobial resistance in Neisseria gonorrhoeae, a number of high-level fluoroquinolone-resistant isolates were obtained from the sexually transmitted diseases clinic in the Moscow region in 2002. The aim of the present study was to determine the molecular mechanisms of resistance and to assess the clonal relationship of these strains METHODS: For the 32 clinical strains of N. gonorrhoeae studied, the MIC values were determined for four fluoroquinolones. The gyrA, parC, por and mtrR genes were studied for the presence of mutations associated with fluoroquinolone resistance. RESULTS: We detected strains of N. gonorrhoeae showing high-level resistance to fluoroquinolones (21 strains, with MICs 1-32 mg/L). Mutations in gyrA and parC known to cause fluoroquinolone resistance were detected in a majority of strains. There were four strains (among 21) without known changes in gyrA and parC. However, amino acid changes in the Por protein and mutations in the promoter or encoding region of the mtrR gene were detected in three of them. One strain had no alteration in gyrA, parC, por or mtrR. CONCLUSIONS: The present study documents the first case of fluoroquinolone-resistant N. gonorrhoeae in Russia.

Molecular typing of N. gonorrhoeae strains prevalent in the Russian Federation.

Genetic polymorphism of Russian population of N. gonorrhoeae was detected and a system for genotyping of its clinical strains was introduced into practice. Comparative analysis of the prevalence of N. gonorrhoeae genotypes in Russia and abroad was carried out. For adaptation of the methods of molecular typing of N. gonorrhoeae strains and its approbation on clinical strains isolated in Russia 41 clinical strains of N. gonorrhoeae were typed. The predominance of PIB serovar (83%) was demonstrated.

[The problem of diagnosis and treatment of urogenital chlamydiosis in Russia]. / Problema diagnostiki i lecheniia urogenital'nogo khlamidioza v Rossii.

The problems associated with the distribution, diagnosis, classification and treatment of urogenital tract chlamydiosis in Russia are discussed. Some arrangements for the improvement of the activities of the laboratory diagnostic services, the use of the International Classification of urogenital tract chlamydioses and the treatment optimization are offered. The drug of choice in the treatment of urogenital tract chlamydiosis is azithromycin (Sumamed, Pliva). Doxycycline, erythromycin and ofloxacin are recommended as the reserve drugs.