[Narrow-band medium-wave ultraviolet therapy in patients with atopic dermatitis: efficacy and safety]. / Uzkopolosnaya srednevolnovaya ul'trafioletovaya terapiya bol'nykh atopicheskim dermatitom: effektivnost' i bezopasnost'.

For the treatment of patients with atopic dermatitis of moderate and heavy severity level, narrow-band medium-wave ultraviolet therapy (narrow-band phototherapy) can be used. An analysis of the results of studies of the efficacy and safety of narrow-band medium-wavelength ultraviolet therapy in patients with atopic dermatitis is presented, and a characteristic of the regimens of the phototherapy carried out is given. It has been shown that narrow-band phototherapy is an effective and safe method of treating patients with atopic dermatitis, but its effectiveness varies widely. Data were obtained on the absence of an increase in the effect during therapy with higher doses of radiation, about the higher efficiency of narrow-band phototherapy with concurrent medication, with an increase in the number of irradiation procedures, as well as in patients with a higher minimum erythemal dose, which indicates the possible existence of factors characterizing the individual characteristics of the response of patients to narrow-band phototherapy.

The Effect of Janus Kinase Inhibitors and Phosphodiesterase-4 Inhibitors on Skin and Plasma Cytokine Levels in Patients with Psoriasis.

In patients with moderate-to-severe and severe psoriasis and high efficacy of therapy (PASI≥75) with signaling pathway inhibitors (apremilast, tofacitinib), cytokine spectra in the skin and blood plasma were studied using xMAP technology at baseline and on weeks 14 and 26 of treatment. Comparison of cytokine levels in psoriatic lesional skin and plasma samples of patients treated with apremilast or tofacitinib revealed statistical difference only for IFNγ level (р<0.05) at week 26.

Analysis of the Specificity of Auto-Reactive Antibodies to Individual Fragments of the Extracellular Domain of Desmoglein 3 in Patients with Pemphigus Vulgaris.

A method for the analysis of the epitope specificity of auto-reactive antibodies to desmoglein 3 (Dsg3) using competitive ELISA has been developed. It is based on a two-stage solid-phase ELISA with initial "depletion" of auto-reactive antibodies against the studied epitope and subsequent quantitative assessment of antibodies against full-length extracellular domain Dsg3. The proposed approach for assessing the specificity of the autoimmune response in patients with pemphigus vulgaris can provide in the future the possibility to personalize the therapy using plasmapheresis by preliminary selection of the antigenic composition of the extracorporeal immunosorbent.

Recombinant Fragment of the Extracellular Domain of Human Desmoglein 3 Fused with the Fc-Fragment of Human IgG1 Selectively Adsorbs Autoreactive Antibodies from the Sera of Pemphigus Patients.

Using the recombinant second fragment of the extracellular domain (EC2) of human desmoglein type 3 (Dsg3) as an affinity ligand, an immunosorbent was obtained that selectively binds autoreactive antibodies to this domain from the immune sera of patients with pemphigus. The EC2 protein was obtained in the form of a fusion protein with the Fc-fragment of human IgG1. The production was carried out in CHO cells using the method of transient expression.

Sustained efficacy of secukinumab in patients with moderate-to-severe palmoplantar psoriasis: 2·5-year results from GESTURE, a randomized, double-blind, placebo-controlled trial.

BACKGROUND: Secukinumab has shown sustained efficacy and safety in several manifestations of psoriasis. OBJECTIVES: GESTURE investigated the long-term (2·5-year) safety and efficacy of 150 mg and 300 mg subcutaneous secukinumab in 205 patients with moderate-to-severe palmoplantar psoriasis. METHODS: GESTURE was a randomized, double-blind, placebo-controlled, multicentre, phase IIIb trial conducted across 15 countries. The study was 140 weeks long and consisted of four periods: screening (up to 4 weeks), treatment period 1 (16 weeks), treatment period 2 (116 weeks) and post-treatment follow-up (8 weeks). Eligible patients were aged ≥ 18 years with moderate-to-severe palmoplantar psoriasis and at least one plaque outside of the palms and soles. Efficacy was assessed via a palmoplantar Investigator's Global Assessment (ppIGA) and the palmoplantar Psoriasis Area and Severity Index (PASI). RESULTS: The primary end point, a ppIGA score of 0 or 1, was met at week 16. The effect was sustained over 2·5 years with 59% [95% confidence interval (CI) 43·5-74·1] and 53% (95% CI 35·1-69·6) of patients in the secukinumab 300 mg and 150 mg groups, respectively, achieving clear or almost clear palms and soles (ppIGA 0 or 1). At 2·5 years, the mean palmoplantar PASI percentage was reduced in both the secukinumab 300 mg group (-74·7%) and the secukinumab 150 mg group (-61·6%). A total of 17% (secukinumab 300 mg group) and 18% (secukinumab 150 mg group) of patients experienced no difficulty in hands and feet functionality, as indicated by the palmoplantar quality of life instrument overall scores. The safety profile was favourable. CONCLUSIONS: GESTURE revealed that secukinumab provides a strong and sustained response over 2·5 years in challenging-to-treat palmoplantar psoriasis.

Differential Serodiagnostics of Latent Stages of Syphilis Based on Measuring IgG and IgM Levels towards Extended Panel of Recombinant Antigens of T. pallidum.

Immunochips containing 12 recombinant antigens of T. pallidum (Тр15, Тр17, Тр47, TmpA, Тр0163, Тр0277, Тр0319, Тр0453, Тр0684, Тр0965, Тр0971, and Тр1038) were prepared to assay for IgG and IgM in serum samples (n=68) of healthy individuals and patients with the latent stages of syphilis. The linear discriminant analysis of detected IgG and IgM differentiated three groups of serum samples as 1) early latent syphilis; 2) seroresistant early latent syphilis; and 3) late latent syphilis with overall differentiation potency of 95.6% (88.9-100%). The samples of all syphilis patients were differentiated from the samples of healthy individuals with 100% specificity.

Experimental Modeling of Leprosy in BALB/c, BALB/c Nude, CBA, and C57BL/6ТNF-/- Mice.

Leprosy was modeled in an experiment on BALB/c, BALB/cNude, CBA, and C57BL/6ТNF-/- mice using three Mycobacterium leprae strains obtained from patients with a diagnosis of A30 according to ICD-10 from different regions of the Russian Federation. Proliferation of M. leprae of the used strains showed a temporal-quantitative dependence on the used mouse line. CBA and BALB/cNude mice were optimal for strain R and BALB/c and BALB/cNude lines were optimal for strain I. BALB/cNude mice infected with strain I had low lifespan. M. leprae strain M showed low proliferation activity in BALB/cNude and C57BL/6ТNF-/- mice.

[Biochemical atypia in the modern russian strains of Neisseria gonorrhoeae.] / Ð‘Ð¸Ð¾Ñ Ð¸Ð¼Ð¸Ñ‡ÐµÑÐºÐ¸Ð¹ атипизм Ð¾Ñ‚ÐµÑ‡ÐµÑÑ‚Ð²ÐµÐ½Ð½Ñ‹Ñ ÑˆÑ‚Ð°Ð¼Ð¼Ð¾Ð² Neisseria gonorrhoeae.

A total 267 strains of Neisseria gonorrhoeae obtained in 2016 from 16 regions of the Russian Federation in six federal districts: Southern, Central, Northwestern, Volga, Ural and Siberian were investigated. All microorganisms were identified by biochemical profile on the Vitek 2 Compact analyzer. Matrix-assisted laser desorption ionization-time of flight mass spectrometry(MALDI-ToF MS) was used as an alternative method of identification. Biochemical typing revealed an atypical indistinctive enzymatic profile of N. gonorrhoeae(loss of D-glucose fermentation abilityand reducing of specific enzymes: ProA, TyrA, APPA in 49.1% of studies (131 strains), resulting in 39 strains (14.6%) were assigned to other types of microorganisms. Additional biochemical typing reduced the percentage of error by almost five times (from 14,6 to 3), but 100% confirmation of N. gonorrhoeae was not received.However, verification by mass spectrometer study showed 100% affiliation of the microorganism to N. gonorrhoeae. Biochemical atypia of N. gonorrhoeae represented by the loss of a number of taxonomically significant characters determines the need for an integrated approach to its identification which includes proteomic (massspectrometry) and/or genomic (PCR) studiesalong with biochemical typing.

[Сomparison of immunoarrays for syphilis diagnostics produced by co-polymerization immobilization and non-contact printing techniques.]

The aim of the study was to investigate the characteristics of immunoarrays (microarrays) produced by co-polymerization immobilization and non-contact printing techniques for enhancing the capacities of syphilis diagnostics. In diagnostic context immunoarrays of both protein immobilization techniques have shown high sensitivity and specificity together with potency to differentiate syphilis stages in serologic assays. The article discloses the advantages and limitations of non-contact printing techniques as well as the results and problems revealed in the study. Solution of these problems in future may provide the development of new serodiagnostic tools with higher accuracy of the results.

[Diagnostic imminoarray assay for characterization of immunoglobulin igg and igm level in syphilis patients serum towards 12 recombinant antigens of t. pallidum before and after the therapy.]

The aim of the study was to characterize the dynamics of immunoglobulin IgG and IgM level in syphilis patients serum at different stages of the disease before and after the therapy towards 12 diagnostic antigens of T. pallidum in an microarray assay and to evaluate these data as possible prognostic markers. The dynamics of immunoglobulin IgG and IgM level was measured in the reaction of indirect immunofluorescence using microarray and compared to the results of non-treponemal RPR test and treponemal tests as EIA and reaction of passive hemagglutination. In microarray assay diagnostically high level of IgM in patients with primary, secondary and early latent and late latent syphilis decreased dramatically to zero after the successful therapy. Continuously high level of IgM after the therapy proposes the persistence of infection agents in the organism and points out the need of additional antimicrobial treatment. In most of the cases anti-treponemal IgG level also declined after the successful therapy and this confirms the appropriate treatment. The results of microarray assay coincide with the results of other mentioned laboratory tests for syphilis diagnostics. Microarray assay with the recombinant T. pallidum antigens gives the perspective for creating methods with wider spectrum of diagnostic and therapy control options using the IgM immunoglobulin level as a marker for successful syphilis treatment.

Immunochip for Syphilis Serodiagnostics with the Use of Extended Array of Treponema pallidum Recombinant Antigens.

An immunochip for multiple parallel detection of specific serum IgG in serological screening for syphilis is based on the use of an extended array of Treponema pallidum recombinant proteins and includes traditionally used immunodominant antigens (Tp15, Tp17, Tp47, and TmpA) and new synthetic proteins (Tp0277, Tp0319, Tp0453, Tp0684, Tp0965, and Tp1038). The use of individual antigens has demonstrated high analytical value of Tp0277 (periplasmatic C-terminal protease), Tp0319 (cytoplasmic membrane-associated lipoprotein TmpC), and external membrane-associated protein Tp0453 with transporting function, all of them improving significantly the efficiency of screening for syphilis in comparison with the traditional array of antigens. Multiparametric analysis of the results obtained on the immunochip with the use of linear discriminant analysis confirmed the efficiency of extended array of T. pallidum diagnostic antigens. Due to proposed modification, the "positive" and "negative" sera are clearly differentiated: the serological study showed 94.1% sensitivity and 100% specificity.

[The refinement of leprosy PCR diagnostics by the amplification of specie-specific repeated fragment of the Mycobacterium leprae genome.]

Certain level of new registered cases of leprosy in a number of endemic countries in the world, as well as growing rate of transboundary migratory flows, raise the issue of effective diagnosis of this disease in countries with sporadic incidence of leprosy, including the Russian Federation. The purpose of the study was to develop a highly sensitive PCR test for detecting the genetic material of Mycobacterium leprae and to compare the test robustness and sensitivity with the commercially available Leprosy Genesig Standard Kit (Primerdesign Ltd., UK). The proposed approach uses real time PCR of non-coding repeating element RLEP, unique for the M. leprae genome, using TaqMan probe. The high test specificity was shown using the reference DNA samples of pathogenic and conditionally pathogenic mycobacterium, as well and its comparison with single-copy genes of M. leprae (rrs, fbp, MntH) PCR detection. The use of a commercially available test system based on the single-copy rpoB gene detection provided 59.4% sensitivity to the detection of M. leprae in the clinical material, while the application of the developed approach increased this index to 96.8%. The developed PCR diagnostics test of leprosy is submitted for state clinical approval process, whereupon the practical use of the test diagnostics allows solving a wide range of tasks to identify and confirm new cases of leprosy, and monitoring both the effectiveness of leprosy treatment, and epidemiological (including transboundary) the spread of the disease.

[Drug resistance mutations and susceptibility phenotypes of Neisseria gonorrhoeae isolates in Russia].

Steady growth in the degree of antimicrobial resistance in Neisseria gonorrhoeae calls for the control of the spreading of resistance mutations. Here we present the data describing drug resistance mutations, the results of antimicrobial susceptibility tests, and molecular genotypes of 128 recent N. gonorrhoeae isolates collected across 9 regions of the Russian Federation. The mutations in chromosome genes penA, ponA, rpsJ, gyrA, parC, which determine the susceptibility of N. gonorrhoeae to penicillins, tetracyclines, and fluoroquinolones were detected by multiplex amplification followed by hybridization on a hydrogel microarray. The most frequent mutation was an insertion of an aspartate at position 345 of penA gene (76.6%), whereas mutations Leu421Pro in ponA gene, Val57Met in rpsJ gene, Ser91Phe in gyrA gene, Asp95Gly in gyrA gene, and Ser87Arg in parC gene were detected in 32.8-36.7% of strains. One third of studied N. gonorrhoeae isolates harbored multiple drug resistance mutations in bacterial chromosome, resulting in the bimodal distribution of mutation profiles and related patterns of antimicrobial susceptibility. The spread of multiple resistance could be explained by the vertical transfer of the mutations resulting in the clonality of the N. gonorrhoeae population.

MOLECULAR EPIDEMIOLOGY OF TREPONEMA PALLIDUM IN BORDER REGION OF RUSSIAN FEDERATION (TUVA REPUBLIC).

The 111 strains of T. pallidum subsp. pallidum collected in Tuva Republic in 2013-2016 were typed by the arp, tpr (E, G, J) and tp0548 genes. The 7 subtypes were identified, in which the 14 d/f type was predominant (90.1%). The minor subtypes 14 b/f, 14 c/f, 14 d/g and 14 i/f constituted 0.9-1.8%. Single strains of 4 d/f H 9 d/f types (each 0.9%) previously described in China were detected in 2015. Both 9 and 14 arp gene variants were found in 3 clinical specimens for the first time in 2015-2016. Similarities in the molecular epidemiology of syphilis in Tuva Republic and Russia were demonstrated as well as differences from the T. pallidum subsp. pallidum population in China and Western Europe.

Oligonucleotide Microchip for the Identification of Infectious Agents of Reproductive System with Simultaneous Analysis of Determinants of Resistance to Antimicrobial Substances.

We developed a multiplexed DNA microarray-based assay allowing identification of 12 causative agents of reproductive tract infections with the simultaneous detection of 47 genetic determinants of resistance to antimicrobial substances. The microarray was tested on 93 isolates of Neisseria gonorrhoeae, 32 isolates of Treponema pallidum and 29 samples of Ureaplasma spp./Mycoplasma spp. The N. gonorrhoeae isolates had multiple mutations in the penA, ponA, rpsJ, gyrA, parC, and mtrR genes; their prognostic value significantly increased when combinations of mutations were detected. In the analyzed T. pallidum isolates, single A2058G substitution in the 23S rRNA gene responsible for macrolide resistance was found. DNA sequences of Ureaplasma spp./Mycoplasma spp. were determined as wild type, which was not fully consistent with the results of analysis of their antimicrobial susceptibility.

[Evaluation of the Recombinant Protein Tp0965 of Treponema Pallidum as Perspective Antigen for the Improved Serological Diagnosis of Syphilis].

UNLABELLED: BACKGRAUND. Treponemal tests based on the detection of antibodies against the Treponema pallidum antigens are the most specific methods for serological diagnosis of syphilis. Due to the inability to cultivate this bacterium in vitro, the most promising sources of antigens for diagnostics are recombinant proteins of T. pallidum. Evaluation of the analytical value of certain T. pallidum proteins is the approach to improve sensitivity, specificity, and reproducibility of syphilis serological tests, including possibilities of differential diagnosis of various forms of the disease. OBJECTIVE: The aim of the research was to evaluate the analytical values (sensitivity and specificity) of recombinant protein Tp0965 of T. pallidum as a candidate antigen for serological diagnosis of syphilis. METHODS: tp0965 gene was cloned into the expression vector pET28a and the construct was used for the transformation of E. coli BL-21 (DE3) cells and further expression and purification of the recombinant protein. The collected protein was used as T. pallidum antigen for serum analysis (ELISA) of groups of patients with various forms of syphilis (n=84) and the group of healthy donors (n = 25). RESULTS: High frequency of positive ELISA results was shown with serum of patients with syphilis, compared to the group of healthy donors. The sensitivity of serological reactions using recombinant protein Tp0965 was 98.8%, specificity--87.5%. The highest sensitivity (100%) was detected in the groups of patients with primary, secondary and early latent syphilis while in the group of patients with late latent syphilis it decreased to 95.2%. CONCLUSIONS: We concluded that due to its specificity T. pallidum recombinant protein Tp0965 can be used as a novel perspective antigen for development of syphilis serological diagnostic assays (for primary and early latent forms).

Expression of Neuropeptides, Neurotrophins, and Neurotransmitters in the Skin of Patients with Atopic Dermatitis and Psoriasis.

Expression of neural marker PGP9.5, amphiregulin, semaphorin-3A, calcitonin gene-related peptide and its receptor, nerve growth factor and its receptor, substance P and its receptor, and expression frequency were analyzed in biopsy specimens from patients with atopic dermatitis and psoriasis and healthy volunteers by immunohistochemical method. Nerve fibers penetrated into the epidermis more frequently, and amphiregulin expression was significantly higher in patients with atopic dermatitis and psoriasis comparing to the control. Expression of semaphorin-3A in patients with atopic dermatitis was less frequent than in the control. These findings reflect the involvement of amphiregulin and semaphorin-3A in the improvement of skin innervations and penetration of nerve fibers into the epidermis, and due of which these proteins mediate the development of itch.

[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.