Transcriptional Activity of Some Genes Involved in Apoptosis in Patients with Vulvar Lichen Sclerosus.

Lichen sclerosus of the vulva is a common, but poorly studied disease. We assessed the level of transcriptional activity of APAF1, BAX, BCL2, BIRC5, CCND1, DAPK1, MCL1, and MYC genes encoding products that control apoptosis in the samples of tissues affected by vulvar lichen sclerosus and adjacent control tissues (n=24). Analysis of transcriptional activity was performed by real-time PCR using specific primers and SYBR Green intercalating dye. After the total group was divided by the presence of the concomitant gynecological diseases, a significant increase in the transcriptional activity of the CCND1 gene was revealed in patients with concomitant uterine fibroids. This may indicate the possible role of the activation of mitosis during tumor initiation.

IL-6 Gene Expression as a Marker of Pathological State in Psoriasis and Psoriatic Arthritis.

The expression of the IL-6 gene in mononuclear blood cells of 45 patients with psoriatic arthritis and 31 patients with plaque psoriasis was studied for possible differential diagnosis of the pathologies. The expression level of IL-6 in psoriatic arthritis and psoriasis surpassed that in healthy controls by 192 and 147 times, respectively. Significant differences in the gene expression were revealed between the patients with psoriatic arthritis and mild psoriasis. The level of IL-6 in patients with severe psoriasis approached that in patients with psoriatic arthritis. High level of IL-6 gene expression can be a marker of possible joint damage in patients with psoriasis and a signal for revising the therapeutic approach in a particular patient.

Genetic Markers of Therapeutic Efficacy of Methotrexate in Patients with Psoriasis.

We studied the effect of C677T and A1298C polymorphisms of the MTHFR gene and 2R/3R polymorphisms of the TYMS gene on the sensitivity to methotrexate in patients with psoriasis (n=139). It was shown that genotype 3R/3R TYMS (OR 8.86, 95%CI 2.00-39.22) and complex genotypes MTHFR1298:A;TYMS:3R (OR 8.20, 95%CI 2.36-28.48) and MTHFR677:C;TYMS:3R (OR 5.40, 95%CI 1.95-14.94) were associated with sensitivity to methotrexate, while genotype 2R/2R TYMS (OR 8.20, 95%CI 2.36-28.48) and complex genotypes MTHFR1298:C;MTHFR677:T;TYMS:2R (OR 0.18, 95%CI 0.06-0.56) and MTHFR1298:C;MTHFR677:T (OR 0.23, 95%CI 0.09-0.59) were associated with resistance to methotrexate. The results can be used for preventive assessment of the effectiveness of methotrexate treatment in patients with psoriasis.

LC-MS/MS analysis of lesional and normally looking psoriatic skin reveals significant changes in protein metabolism and RNA processing.

BACKGROUND: Plaque psoriasis is a chronic autoimmune disorder characterized by the development of red scaly plaques. To date psoriasis lesional skin transcriptome has been extensively studied, whereas only few proteomic studies of psoriatic skin are available. AIM: The aim of this study was to compare protein expression patterns of lesional and normally looking skin of psoriasis patients with skin of the healthy volunteers, reveal differentially expressed proteins and identify changes in cell metabolism caused by the disease. METHODS: Skin samples of normally looking and lesional skin donated by psoriasis patients (n = 5) and samples of healthy skin donated by volunteers (n = 5) were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After protein identification and data processing, the set of differentially expressed proteins was subjected to protein ontology analysis to characterize changes in biological processes, cell components and molecular functions in the patients' skin compared to skin of the healthy volunteers. The expression of selected differentially expressed proteins was validated by ELISA and immunohistochemistry. RESULTS: The performed analysis identified 405 and 59 differentially expressed proteins in lesional and normally looking psoriatic skin compared to healthy control. In normally looking skin of the patients, we discovered decreased expression of KNG1, APOE, HRG, THBS1 and PLG. Presumably, these changes were needed to protect the epidermis from spontaneous activation of kallikrein-kinin system and delay the following development of inflammatory response. In lesional skin, we identified several large groups of proteins with coordinated expression. Mainly, these proteins were involved in different aspects of protein and RNA metabolism, namely ATP synthesis and consumption; intracellular trafficking of membrane-bound vesicles, pre-RNA processing, translation, chaperoning and degradation in proteasomes/immunoproteasomes. CONCLUSION: Our findings explain the molecular basis of metabolic changes caused by disease in skin lesions, such as faster cell turnover and higher metabolic rate. They also indicate on downregulation of kallikrein-kinin system in normally looking skin of the patients that would be needed to delay exacerbation of the disease. Data are available via ProteomeXchange with identifier PXD021673.

Association of the NOS3 Gene Polymorphisms with the Risk of Atopic Dermatitis in Children.

We studied association of polymorphic markers Glu298Asp (rs1799983), C774T (rs1549758), and T786C (rs2070744) of the NOS3 gene with the risk of atopic dermatitis. It was found that T786C polymorphic marker of the NOS3 gene is associated with lower risk of erythematosquamous lichenoid atopic dermatitis. A significant association between the homozygous CC genotype in locus 786 of the NOS3 gene and the development of erythematosquamous atopic dermatitis with lichenification was revealed. The homozygous CC genotype can be considered as a risk factor of erythematosquamous atopic dermatitis with lichenification.

The role of regulatory RNAs (miRNAs) in asthma.

INTRODUCTION: Recently, a great deal of attention has been paid to the investigation of regulatory functions of microRNA. Currently, many different mechanisms involved in the pathogenesis of asthma are known, but the whole picture of pathogenesis has not yet been studied. CONCLUSIONS: MicroRNAs play an important role in the regulation of many cellular processes. Undoubtedly, these regulatory molecules are involved in the pathogenesis of asthma, and therefore can be potential targets for treatment.

[Genetically predetermined limitation in the use of HaCaT cells that affects their ability to serve as an experimental model of psoriasis].

Proinflammatory cytokines TNF, IFNG and ILl7 play an important role in eruption of psoriasis. The activation of epidermal keratinocytes with the named cytokines alters their terminal differentiation program and causes their hyperproliferation in the diseased skin. HaCaT cells, which are immortalized human keratinocytes, are often used as a cellular model of psoriasis. The aim of this study was to evaluate changes in gene expression and the proliferation rates in cultured HaCaT cells treated with TNF, IFNG and IL17. We found that HaCaT cells decrease their proliferation rate in response to either IL17 or a combination TNF and IF-NG. The analysis of microarray data discovered a group of 12 genes, which were downregulated in HaCaT after treatments with the named cytokines and upregulated in psoriatic lesional skin. Eight genes were important for DNA replication and they also contributed to two larger networks that regulated cell progression through the cell cycle. We conclude that HaCaT cells have a sufficient limitation as a cellular model of psoriasis due to their treatment with proinflammatory cytokines, namely TNF, IFNG and IL17 does not increase their proliferation rate. Thus, the studies of psoriasis based on HaCaT cells as an experimental model shall take in account this important phenomenon.

Pharmacological control of receptor of advanced glycation end-products and its biological effects in psoriasis.

Receptor for advanced glycation end-products is implicated in a development of chronic inflammatory response. Aim of this paper is to provide a review on commercial and experimental medicines that can interfere with RAGE and signaling through RAGE. We searched three bibliographical databases (PubMed, Web of Science and MEDLINE) for the publications from 2005 to March 2012 and identified 5 major groups of agents that can interfere with RAGE biological effects. In the first part of this paper, we discuss AGE crosslink breakers. These chemicals destroy advanced glycation end products (AGEs) that are crosslinked to the extracellular matrix proteins and can interact with RAGE as ligands. Then, we describe two non-conventional agents SAGEs and KIOM-79 that abolish certain biological effects of RAGE and have a strong anti-inflammatory potential. In the third part, we evaluate the inhibitors of the signaling cascades that underlie RAGE. Particularly, we discuss two groups of kinase inhibitors tyrphostins and the inhibitors of JAK kinases. Considering RAGE as a potential master regulator of processes that are crucial for the pathogenesis of psoriasis, we propose that these medicins may help in controlling the disease by abolishing the chronic inflammation in skin lesions.

[Expression of bioinformatically identified genes in skin of psoriasis patients].

Gene expression analysis for EPHA2 (EPH receptor A2), EPHB2 (EPH receptor B2), S100A9 (S100 calcium binding protein A9), PBEF(nicotinamide phosphoribosyltransferase), LILRB2 (leukocyte immunoglobulin-like receptor, subfamily B (with TM and ITIM domains), member 2), PLAUR (plasminogen activator, urokinase receptor), LTB (lymphotoxin beta (TNF superfamily, member 3)), WNT5A (wingless-type MMTV integration site family, member 5A) has been conducted using real-time polymerase chain reaction in specimens affected by psoriasis versus visually intact skin in 18 patients. It was revealed that the expression of the nine examined genes was upregulated in the affected by psoriasis compared to visually intact skin specimens. The highest expression was observed for S100A9, S100AS, PBEF, WNT5A2, and EPHB2 genes. S100A9 and S100A8 gene expression in the affected by psoriasis skin was 100-fold higher versus visually intact skin while PBEF, WNT5A, and EPHB2 gene expression was upregulated more than five-fold. We suggested that the high expression of these genes might be associated with the state of the pathological process in psoriasis. Moreover, the transcriptional activity of these genes might serve a molecular indicator of the efficacy of treatment in psoriasis.

[Role of receptor for advanced glycation end-products in pathgenesis of psoriasis].

This review summarizes the existing knowledge regarding the role of receptor for advanced glycation end products which is a key participant of the inflammatory process, in pathogenesis of psoriasis. By interacting with multiple ligands and activating several signaling mechanisms, receptor for advanced glycation end products regulates gene expression via a group transcription factors, that includes NFKB and AP1. According to the published data the expression of receptor for advanced glycation end products in both immune cells and their targets, a high stability of this receptor in complexes with ligands as well as a positive feedback loop, upregulating the expression of its certain ligands, suggest receptor for advanced glycation end products as a possible principal factor that makes the inflammatory response in psoriasis sustainable. Considering receptor for advanced glycation end products as a potential master regulator of several processes that play a crucial role in development of psoriatic plaques, we believe that further experimental studies are needed to elucidate how exactly this receptor converts a transient inflammatory reaction to a sustainable inflammatory response. These studies are also needed for the development of novel medications that target receptor for advanced glycation end products and signaling mechanisms that this receptor activates.

Three-dimensional model of mouse epidermis for experimental studies of psoriasis.

Three-dimensional models of skin and epidermis imitate the structure of real tissues and provide accurate information about certain skin conditions, such as psoriasis. A three-dimensional model of mouse epidermis was generated from the epidermal keratinocytes of newborn mice and treated with cytokines. The aim of this study was to evaluate this model as an experimental model of psoriasis and to assess the changes occurring in its structure and gene expression after the exposure to proinflammatory cytokines. Treatment of the three-dimensional model with either interleukin 17 or a combination of tumor necrosis factor and interferon γ was shown to produce morphological changes, which were similar to acanthosis in psoriatic skin. The observed changes in gene expression of metalloproteinases and certain psoriasis biomarkers, such as mki67, krt16 and fosl1, were similar to the changes in patients' skin. Notably, changes caused by interleukin 17 were less evident than those caused by the combination of interferon γ and tumor necrosis factor. On the contrary, HaCaT cells exhibited no significant changes in the expression of fosl1 and had decreased levels of mki67 after being treated with a combination of TNF and IFNG. Moreover, treatment with IL17 had no significant effect on krt16 and mki67 expression and even reduced the fosl1 levels. The findings suggest that artificially generated three-dimensional models of murine skin can be used to study psoriasis.

[Expression of genes for metalloproteinases (MMP-1, MMP-2, MMP-9, and MMP-12) associated with psoriasis].

Using real-time polymerase chain reaction (RT-PCR), we measured mRNA amounts of matrix metalloproteinases (MMPs): MMP-1, MMP-2, MMP-9, and MMP-12 genes in psoriatic lesions and unaffected skin of the same patients. We observed significant (about 15-fold) increase in the expression level of matrix metalloproteinase MMP-1 and MMP-12 genes associated with psoriasis. The results of our studies of MMP gene expression in cultured primary human keratinocytes treated with interleukin (IL-17) have shown upregulation of MMP gene expression both in cultured keratinocytes and in psoriatic skin lesions. Therefore, upregulation of MMP genes in the skin affected by psoriasis could result from IL-17 effects on skin cells.

Comparative study of the expression of ATF-3 and ATF-4 genes in vessels involved into atherosclerosis process and in psoriatic skin.

Expression of ATF-3 and ATF-4 genes was examined quantitatively by real-time PCR and changes in the expression of these genes in atherosclerotic lesions and in psoriatic skin were demonstrated. It was found that concomitant pathologies do not affect the expression of these genes. Opposite changes in the expression of ATF-3 and ATF-4 genes in atherosclerotic and psoriatic samples were revealed and a hypothesis was put forward that this parameter could be a criterion of pathological process in both diseases.

Effects of expression of transcriptional factor AP-1 FOSL1 gene on psoriatic process.

We performed quantitative analysis of FOSL1 gene expression in lesional psoriatic skin. The expression of this gene in lesional psoriatic skin was significantly increased compared to that in unaffected areas. Enhanced FOSL1 expression significantly correlated with high psoriasis area and severity index (PASI). High level of FOSL1 gene expression was proposed to be a marker of pathological process activity in psoriasis.

[Expression of the FOSL1 gene in psoriasis and atherosclerosis].

Psoriatic and atherosclerotic plaques were examined using the real-time polymerase chain reaction. Expression of the FOSL1 gene proved to substantially increase in both psoriatic lesions of the skin and atherosclerotic lesions of vessels as compared with nonlesion samples.

Study of Molecular Mechanisms Involved in the Pathogenesis of Immune-Mediated Inflammatory Diseases, using Psoriasis As a Model.

Psoriasis was used as a model to analyze the pathogenetic pathways of immune-mediated inflammatory diseases, and the results of bioinformatic, molecular-genetic and proteomic studies are provided. Cell mechanisms, common for the pathogenesis of psoriasis, as well as Crohn's disease, are identified. New approaches for immune-mediated diseases are discussed.

[Treatment tactics of patients with pyloroduodenal stenoses].

Overall 172 patients aged 29 to 87 years with stenosis of stomach pyloric part and duodenum were studied. The complications of gastric or duodenal ulcer (119), cancer of stomach (49) and pancreas (4) were the cause of stenosis. All the patients were divided into 3 groups: 1st group consisted of 43 patients who has undergone urgent surgery due to stenosis with bleedings and perforations without additional protein-energetic treatment before operation; 2nd group - 65 patients who were prepared preoperatively with complete parenteral nutrition; 3rd group - enteral tube feeding with balanced mixtures before surgery (64 patients). The best results were achieved in the patients with preoperative enteral tube feeding; it allowed to reduce lethality from 25.5 and 16.3% at 1st and 2nd group to 0% ant 3rd group.

[The role of non-invasive diagnostics of pulmonary embolism in the optimization of treatment].

177 patients with pulmonary embolism (PE) were examined to study the information value of transthoracal echoCG in early diagnostics of the disease, the possibility of noninvasive evaluation of its severity, and possibility of determining indications for thrombolytic therapy (TLT) on basis of such evaluation. The results show that the sensitivity of such a sign as elevation of pulmonary artery pressure (PAP) by 30 mmHg, is 95%, provided that the patient does not have severe or moderate concomitant diseases cousing PAP elevation (postinfarction cardiosclerosis, chronic obstructive lung disease, valvular diseases etc.). The specificity of this sign, vs the control group, was 5%. Thus, elevation of PAP in patients with clinical manifestations of PE and without concomitant pathology, is a highly informative symptom of acute PE. The sensitivity of the parameter of right ventricular (RV) dilatation was 85%, of paradoxial ventricular septum motion--54%. In patients with PE of various degree of severity and concomitant diseases, high sensitivity of the above-listed sonographic signs of RVoverload goes together with their low specificity, which is not higher than 25%. This does not allow evaluating the degree of embolism and determining indications for TLT in these patients on basis of echoCG data. The authors of the article have developed an algorithm of diagnostic and therapeutic measures in patients with suspected PE.

[The calculation and regulation of dust exposure doses to decrease the level of occupational diseases of dust etiology]. / Raschet i regulirovanie pylevykh ékspozitsionnykh doz s tsel'iu snizheniia urovnia professional'nykh zabolevanii pylevoi étiologii.

One of the real ways to reduce occupational diseases caused by dust is to cut the duration of contact with dust. The studies helped to suggest a method calculating occupational risk correlated to dust content of the air. The authors evaluated the duration of contact with dust so as to reduce occupational risk to the minimum. Those measures appeared to lower but not eliminate the occupational risk.