Microbiological assessment of the effectiveness of standard therapy in atopic dermatitis

Background. Atopic dermatitis is an inflammatory skin disease characterized by recurrent lesions and intense pruritus. Nowadays there is a stepwise approach to the treatment of atopic dermatitis, which is defined by disease intensity and complications such as secondary skin infections. However, the current management of atopic dermatitis may not always lead to the expected outcome due to not only immune dysregulation of both adaptive and innate immunity but also imbalance of the skin microbiome. Aims. The aim of the study was to evaluate changes in the composition of the skin microbiome in both lesional and non-lesional skin in patients with atopic dermatitis during standard treatment. Materials and methods. Twenty patients with atopic dermatitis and twenty six healthy controls over 18 years old were included into the study. All microbiome samples were obtained from lesional and non-lesional skin sites of atopic dermatitis patients before and after therapy. Whereas samples from healthy controls were taken once from a flexor surface of the elbow. Species identification of clinical isolates were identified using MALDI Biotyper Sirius (Bruker Daltonics). Results. At baseline, the prevalence of S. aureus colonization among patients with atopic dermatitis was 34.20% in lesional skin and 32.50% in non-lesional skin. After treatment, there was a significant decrease in the prevalence of S. aureus carriage in both lesional and non-lesional skin areas (р 0.05). However, no significant difference was observed in the proportion of all other staphylococci (р 0.1). Interestingly, S. aureus was not found in healthy controls. Conclusions. The results of the study demonstrated the effectiveness of standard therapy for managing patients with atopic dermatitis as it had a positive impact on the skin microbial community and showed a decrease in S. aureus proportion after the treatment

Innate immune factor gene expression profiles in patients with atopic dermatitis

Atopic dermatitis is a multifactorial genetically determined inflammatory skin disease characterized by itching, chronically relapsing dermatitis, age-related features of localization and morphology of lesions. The pathogenesis of atopic dermatitis is complex and includes epigenetic alterations, involved in the genomic adaptation, immune response reactions and dysfunction of the epithelial barrier that together trigger the development of atopic dermatitis. The aim of this study is to detect the expression level for IL4, IL13, IL33, TLR2, TLR9 genes in the biological materials of atopic patients.The targeted genes for further expression evaluation were selected according to our previous findings on genome-wide methylation study. We detected the cascades with the differentially methylated genes that are most likely to take place in atopic dermatitis. Thus, we investigated expression levels for the IL4, IL13, IL33, TLR2, TLR9 genes in the skin, peripheral blood mononuclear cells and whole blood cells using RT-PCR on 55 pediatric patients and 26 healthy volunteers, and on 50 adult patients. Statistical analysis was performed with the use of Kruskal-Wallis H test and Mann-Whitney U test. Targeted expression analysis revealed that in the skin samples the expression of TLR9 and IL4 was 12 times significantly lower (p < 0.0001, p < 0.0005) in the lesional skin; and there was a 6-fold decrease in case of TLR2 (p < 0.01). The results for blood mononuclear cells differed and expression levels for most of the assessed targets were significantly higher before treatment. We have also found out that those differences were strongly pronounced especially in an elder age group (12-18 y.o.). Studying the IL33 gene expression in the whole blood samples of adults revealed that its level was significantly higher in case of patients with moderate form of AD. Besides, we concluded that locally in the affected skin inflammatory immune response may dominate; in the mononuclear cells Th2 immune response apparently takes place. New insights on immunological markers and links among them may shed a light on atopic dermatitis pathogenic mechanisms. The detected molecules could play role as potential therapeutic targets and form a management approach for patients with atopic dermatitis

Minoxidil in trichological practice

Management of patients with alopecia is still a challenging task. Minoxidil is a pyrimidine derivative, a potent KATP channel opener, and has been shown to act as a vasodilating agent. Minoxidil enhances the synthesis of growth factors, not only VEGF (vascular endothelial growth factor), but also FGF (fibroblast growth factor), IGF-1 (insulin-like growth factor), which stimulate growth processes in the hair follicle and trigger the anagen phase. The topical minoxidil formulations have been proven to be particularly efficient to a greater extent in androgenic alopecia (AGA) and to a lesser extent in nasal alopecia (GA).The best hair regrowth has been detected in early stages of alopecia. It generally takes 4-6 months before results are visible, and 12 months to stabilize the hair growth, which is maintained by a regular use of minoxidil. The level of the sulfotransferase enzyme in the hair follicles may predict the outcome of topical minoxidil therapy. The topical use of minoxidil does not cause any systemic side effects, and adverse effects in the form of skin reactions, irritation and dryness are short-term and rare. The article also presents the data of its own clinical experience in using minoxidil for the treatment of alopecia

New combined topical therapy for refractory rosacea

The main pathogenetic aspects of rosacea, various classifications of dermatosis and the modern clinical classification are presented. The symptoms of different forms of rosacea are described. Contradictory of modern scientific concepts of different researchers, approaches to etiology, pathogenesis and treatment has been noted. The main triggers for the manifestation of rosacea, which should be tak en into account to achieve the maximum effect in the treatment of the disease, are indicated. The first domestic positive experience of authors with 1 % ivermectin cream and 0.03 % tacrolimus ointment is presented. The high efficacy and very good tolerability of this combination, compared to monotherapy of 1 % ivermectin cream and traditional therapy of metronidazole cream and azelaic acid gel as well as the absence of any side effects in patients with moderate to severe rosacea are shown

Unspecified skin peripheral T-cell lymphoma: diagnostic difficulties

Peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS) have an aggressive, life-threatening course. 5‑year survival rate is less than 20 %, which may be due to not timely diagnosis. PTCL-NOS can histologically and immunophenotypically mimic other T-cell lymphomas of the skin, including mycosis fungoides. In this connection, the correct diagnosis is most often established in the late stages of the disease. We present a clinical case of PTCL-NOS misdiagnosed as mycosis fungoides

DEVELOPMENT OF AZATHIOPRINE-BASED TREATMENT REGIMEN FOR PATIENTS WITH STEROID RESISTANT PEMPHIGUS BASED ON ASSESSMENT OF MOLECULAR MECHANISMS AT THE POST-RECEPTOR LEVEL

Background: Autoimmune pemphigus is the most severe skin and mucous membranes disorders with production of IgG autoantibodies to desmogleins 1 and 3. Administration of systemic corticosteroids may help to abrogate active signs of pemphigus. However, some patients do not give an adequate response to systemic glucocorticosteroid monotherapy, as well as to their combination with immune suppressants and cytostatic agents. Aim: To develop an azathioprine-based treatment regimen for patients with autoimmune pemphigus resistant to steroids. Materials and methods: At the first step of development of a treatment regimen for patients with steroid-resistant pemphigus we analyzed retrospectively a clinical database on 23 patients who had been treated from 2000 to 2014 and whose treatment regimen included azathioprine, in addition to systemic glucocorticosteroids. At the second step, from 2013 to 2015, we assessed and treated with the azathioprine-based regimen 24 patients with autoimmune pemphigus, 14  of them being steroid resistant and 10, steroid sensitive (control group). To assess molecular mechanisms of steroid resistance at the post-receptor level (effect of prednisolone on incorporation of ³Н-uridine into lymphocyte mRNA, changes of intracellular levels of nuclear transcription factor NF-κB) we used a  real-time polymerase chain reaction, radioisotope method and liquid scintillation radiometry. Results: At the first step, we developed an azathioprine-based treatment regimen for patients with steroid resistant autoimmune pemphigus. Initial dose of azathioprine was 150 mg daily. After achievement of response, the dose was decreased to 100 mg daily. When the dose of systemic glucocorticosteroids was decreased to 20  mg daily, the dose of azathioprine was decreased to 50 mg daily, thereafter steroid resistant patients were taking azathioprine at a dose of 50 mg daily from 3  months to 2.5  years. Investigation of molecular mechanisms at the second step of the study showed that in 28%  of steroid resistant patients there was a decreased incorporation of ³Н-uridine into lymphocyte mRNA under prednisolone treatment with an increase in synthesis of total mRNA in lymphocytes (range, from 68.67 to 78.35%, р < 0.05). Compared to the control group, in all steroid resistant patients (n = 14), an increased NF-κB gene expression in lymphocytes was found (range, from 65.39 to 86.17%  and from 88.8  to 98.61%, respectively, р < 0.05). The combination therapy with systemic glucocorticosteroids and azathioprine in steroid resistant patients resulted in a  decrease in intracellular NF-κB gene expression (p < 0.01), which underlies the steroid-sparing effect of azathioprine. Conclusion: We demonstrated a statistically significant suppression of NF-κB gene expression in the cases of its high baseline levels before combination therapy in patients with autoimmune pemphigus. The steroid-sparing effect of azathioprine allows for its effective use in steroid-resistant pemphigus

Association of single nucleotide polymorphisms of <i>TLR2</i>, <i>TLR4</i> and <i>TLR9</i> with atopic dermatitis

Toll-like receptors (TLRs) are the most studied among all Pattern Recognition Receptors, the main function of which is to initiate innate immune response by recognizing pathogen-associated molecular patterns of various microorganisms on the skin surface. TLR-mediated recognition plays an important role in linking innate and adaptive immunity that ultimately leads to the production of key cytokines, chemokines and antimicrobial peptides. Today, there is growing interest in research on single nucleotide polymorphisms (SNPs) in TLR genes and its influence on susceptibility to inflammatory disease, including atopic dermatitis. The aim of the research was to study the association of the rs5743708 gene polymorphism in the TLR2 gene, the rs4986791 gene polymorphism in the TLR4 gene and the rs352140 gene polymorphism in the TLR9 gene with the risk of developing severe cases of AD. A total of 100 patients with AD were included in the study (38 male and 62 female). The age range was from 18 to 65 years old. All participants were divided into 2 groups according to the SCORAD index (SCORing Atopic Dermatitis). The control group included 72 volunteers over 18 years old. The results of our study showed a statistically significant difference between the moderate AD group and healthy controls in the rs352140 gene polymorphism in the TLR9 gene (Figure 1). The frequency of the GG genotype of SNP rs352140 in TLR9 was 0.169 in the AD group versus 0.329 in the control group (p &lt; 0.05; OR = 0.42; 95% CI = 0.18-0.97).In conclusion, the results of our study showed that the TLR9 rs352140 gene polymorphism may be linked to an increased risk of atopic dermatitis. Moreover, it was found that the GG genotype of SNP rs352140 in TLR9 can be used as a predictor of the risk of developing moderate AD

THE USE OF CONFOCAL LASER SCANNING MICROSCOPY IN THE DIAGNOSTICS OF BULLOUS PEMPHIGOID OF LEVER

Diagnostics of autoimmune bullous dermatoses, including bullous pemphigoid of Lever that is characterized by a large variety of clinical manifestations and in many cases by severe course and high mortality, remains one of the most complicated problems in dermatology. High diagnostic error rates are to be explained not only by variability of the forms of bullous pemphigoid of Lever, but also by insufficient accuracy of existing diagnostic methods, as well as by the complexity of their implementation. Non-invasive diagnostic methods have an undoubted advantage because they allow for prompt results of the assessment and therefore to fasten the initiation of therapy and to avoid damage to the skin, which is extremely important for patients with an autoimmune bullous dermatosis. The most accurate among non-invasive methods is a confocal laser scanning microscopy. The article describes the main features of the method and its potential application for the diagnosis of bullous pemphigoid of Lever with two clinical cases

The new differential diagnostic test for the lichenoid drug eruption

The differential diagnosis between lichenoid drug eruption (LDE) and lichen planus (LP) is difficult due to similar clinical and histological signs, but important for treatment and prognosis. The purpose of this study was to propose the new diagnosis method for differentiate LDE from lichen planus. During 2015-2018, 20 patients with confirmed LDE, 13 patients with LP and 134 controls were examined and treated at the Lenoblcenter. All enrolled patients were underwent the injection of 0.5 mL of the 2% lidocaine solution by insulin syringe into the papule with following histological examination. The formation of a blister (bulla) at the site of injection was considered a positive test result. Among LDE 18/20 patients were found positive for developing blister (bulla), 2 results were questionable. In 12 of 13 LP patents, bulla on the site of injection was not identified, the result of 1 patient was non-specific. All control patients were negative for the proposed test. The histological sections showed that the bulla has corresponded to the separation of the epidermis from the dermis. Intracutaneous injection of 0.5 mL of lidocaine into the papule is an easy highly specific and sensitive method to differentiate LDE from LP. This article is protected by copyright. All rights reserved