Previous studies have not completely elucidated the role of the histaminergic system in the pathogenesis of psoriasis. This study aimed to evaluate the effects of adalimumab and cyclosporine A on the expression of histaminergic system-related genes and miRNAs regulating these genes in bacterial lipopolysaccharide A (LPS)-stimulated human keratinocyte (HaCaT) cells. HaCaT cells were treated with 1 µg/mL LPS for 8 h, followed by treatment with 8 µg/mL adalimumab or 100 ng/mL cyclosporine A for 2, 8, or 24 h. Untreated cells served as controls. The cells were subjected to ribonucleic acid (RNA) extraction and microarray, quantitative real-time polymerase chain reaction, and enzyme-linked immunosorbent assay analyses. Statistical analysis was performed using the Statistica 13.0 PL (StatSoft, Cracow, Poland) and the Transcriptome Analysis Console programs (Affymetrix, Santa Clara, CA, USA) ( p < 0.05). The differential expression of the following two miRNAs was not affected in LPS-stimulated cells upon treatment with cyclosporine A or adalimumab: hsa-miR-583 (downregulated expression), involved in the regulation of histamine receptor 1 – HRH1 (overexpression); has-miR-1275 (downregulated expression), involved in the regulation of histamine receptor 1 – HRH3 (overexpression) and Solute carrier family 22 member 3 – SLC23A2 (downregulated expression)). Adalimumab and cyclosporine A modulated the histaminergic system in HaCaT cells in vitro . However, further studies are needed to elucidate the underlying mechanisms. Abbreviations: (-) – downregulated in comparison to the control, (+) – overexpression in comparison to the control, ACTB - β-actine, ADA - Adenosine deaminase, ADCYAP1 - Adenylate Cyclase Activating Polypeptide 1, BMP - bone morphogenetic protein, bp - base pair, cAMP - adenosine 3’ 5’-cyclic monophosphate, CBX7 - Chromobox protein homolog 7, cDNA - double-stranded complementary DNA, CSA - cyclosporine A DAG – diacylglycerol, DIAPH - Diaphanous related formin 1, DNMT - DNA methyltransferases, DRD2 - Dopamine receptor D2, EDN1 - Endothelin 1, EDNRA - Endothelin receptor type A, ELISA - Enzyme-linked immunosorbent assay, EZH2 - Enhancer of zeste homolog 2, FC - fold change, GABRB1 - Gamma-aminobutyric acid (GABA) A receptor, alpha 1, GABRB2 - Gamma-aminobutyric acid (GABA) A receptor, alpha 2, GABRB3 - Gamma-aminobutyric acid (GABA) A receptor, alpha 3, HaCaT - Human adult, low-calcium, high-temperature keratinocytes, HIS - Human Histamine, HLAs - human leukocyte antigens, HNMT - Histamine N-methyltransferase, HNMT - Histamine N-Methyltransferase, HRH1 – histamine receptor 1, HRH2 – histamine receptor 2, HRH3 – histamine receptor 3, HRH4 – histamine receptor 4, HTR6 - 5-Hydroxytryptamine Receptor 6, IGF1 - Insulin-like growth factor 1, IL10 -interleukin 10, IL12 -interleukin 12, IL...

Background: Increased levels of phosphorylated ERK and p38 MAPK proteins have been observed in psoriatic skin biopsies compared to controls, which may be associated with an impaired expression pattern of dual activity protein phosphatase (DUSP). Objective: The purpose of this study was to assess changes in the expression profile of mRNA DUSP 1-7 and miRNA regulating their expression in human keratinocyte cells (HaCaT) had exposed to the liposaccharide A (LPS). Methods: HaCaT was exposed to 1 µg/ml LPS and next adalimumab by 2,8,24h compared to untreated cells. The microarray method was used to analyze expression pattern of mRNAs, miRNAs, and ELISA to evaluate changes in the level of the proteins. RTqPCR was used to validate the microarray data. Transcriptome Analysis Console and Statistica Software 13 PL were used in statistical analysis (p<0.05). Results: The highest changes in expression was observed for DUSP2 (FC +11.12) and DUSP5 (FC +5.53) in HaCaT culture after 2 hours exposition on adalimumab. It was observed that miR-1275 (FC -2.39) and miR-34a (FC +6.52) might regulate level of DUSP2, and miR-27a (FC +3.55), miR-27b (FC +2.87) are involved in DUSP5 expression. Conclusion: The results obtained suggest that DUSP2 and DUSP5 may be considered as complementary molecular markers in the diagnosis and monitoring of the effectiveness of psoriasis therapy. It was confirmed that hsa-miR-34a, hsa-miR-1275, hsa-miR-3188, hsa-miR-382, hsa-miR-27a, hsa-miR-27b, hsa-miR-16 have the highest influence on the expression pattern of DUSP1-7.

The purpose of this study was to evaluate the relationship between human papillomavirus (HPV16/18), Epstein–Barr virus (EBV), and human cytomegalovirus (HCMV) infections and the occurrence of ovarian cancer in 48 women, of whom 36 underwent surgery and chemotherapy (group A), 12 in whom surgery was sufficient (group B), and 60 with endometroid endometrial cancer stage G1-G3 (group C), compared to patients in whom the uterus and its appendages were removed for nononcological reasons (control group). The detection of HPV, EBV, and HCMV in tumor tissue and normal tissue was performed using the real-time polymerase chain reaction (RT-PCR) technique. A statistically significantly higher risk of endometrial cancer was noted in patients infected only with HCMV (OR > 1; p < 0.05). In contrast, a significantly higher risk of ovarian cancer in group A was associated with HPV16, HPV18, and EBV (OR > 1; p < 0.05); a significantly higher risk of ovarian cancer in group B was associated with HPV18 and HMCV (OR > 1; p < 0.05). The obtained results suggest that HCMV infection is associated with the development of a stage of ovarian cancer when treatment can be completed with surgery alone. Meanwhile, EBV appears to be responsible for the development of ovarian cancer in more advanced stages.

Introduction: Cyclosporin A (CsA) treats moderate to severe psoriasis vulgaris. Psoriasis is a chronic inflammatory disease in which hyperproliferation of keratinocytes occurs. One of the most relevant signaling cascades in the development of psoriasis is the mitogen-activated protein kinase (MAPK) signaling pathway. It has been observed that dual-specificity phosphatases (DUSPs) dephosphorylate signaling molecules, such as MAPKs. background: Cyclosporin A (CsA) is used to treat moderate to severe psoriasis vulgaris. Psoriasis is a chronic inflammatory disease in which hyperproliferation of keratinocytes is noted. One of the most relevant signaling cascades in the development of psoriasis is the mitogen-activated protein kinase (MAPK) signaling pathway Dual-specificity phosphatases (DUSPs) dephosphorylate signaling molecules such as MAPKs. Aims: Aims: This study aims to determine changes in the expression pattern of Dual Activity Protein Phosphatase (DUSP1-7) and micro RNAs (miRNAs), potentially regulating their expression in the human adult, low-calcium, high-temperature keratinocytes cell line (HaCaT) cultures exposed to lipopolysaccharide A (LPS)-induced inflammation, followed by CsA. Methods: HaCaT cell line was exposed for 8 hours to 1 μg/mL LPS and then to 100 ng/mL CsA for 2, 8, and 24 hours compared to cultures not exposed to LPS and the drug. The molecular analysis included determining the DUSP1-7 expression and the miRNAs potentially regulating it using an expression microarray technique. An enzyme-linked immunosorbent assay (ELISA) was also performed to assess the concentration of DUSP1-7 in the culture medium. Statistical evaluation was performed assuming a statistical significance threshold (p) of < 0.05. method: Human keratinocyte (HaCaT) cell line was exposed for 8 hours to 1 μg/mL LPS and then to 100 ng/mL CsA for 2, 8, and 24 hours compared to cultures not exposed to LPS and the drug. The molecular analysis included determining DUSP1-7 expression and the miRNAs potentially regulating its expression by expression microarray technique. An Enzyme-Linked immunosorbent assay (ELISA) was also performed to assess the concentration of DUSP1-7 in the culture medium. Statistical evaluation was performed assuming a statistical significance threshold (p) &lt; 0.05. Results: Statistically significant differences were found in the expression of DUSP1-7 mRNAs and the miRNAs that regulate their expression. The most significant changes in expression were observed for DUSP1 and DUSP5, with the differences being most pronounced during the eight-hour incubation period of the cells, with the drug predictive analysis showing that miR-34 potentially regulates the expression of DUSP1-4,7, miR-1275: DUSP2, mir-3188:DUSP4, miR-382: DUSP4, miR-27a and miR-27b: DUSP5,6 and miR-16: DUSP7. No expression of DUSP1-7 was demonstrated at the protein level in CsA-exposed cultures. Conclusion: Our evaluation of the efficacy of CsA therapy on an in vitro model of HaCaT indicates that treatment with this drug is effective, resulting in changes in the expression of DUSP1-7 and, potentially, the miRNAs that regulate their expression. We also confirmed that the different expression pattern of mRNA and protein encoded by a given transcript is not only due to the regulatory role of miRNAs but also the lack of synchronization between transcription and translation processes. conclusion: Our evaluation of the efficacy of CsA therapy on an in vitro model of HaCaT indicates that treatment with this drug is effective, resulting in the observed changes in the expression of DUSP1-7 and, potentially, the miRNAs that regulate their expression. We also confirmed that the different expression pattern of mRNA and protein encoded by a given transcript is not only due to the regulatory role of miRNAs but also to the lack of synchronization between transcription and translation processes.