Lipoprotein(a) is associated with DNA damage in patients with heterozygous familial hypercholesterolemia.

Heterozygous familial hypercholesterolemia (HeFH) is a common autosomal-dominant inherited disorder associated with atherosclerotic cardiovascular disease (ASCVD). HeFH subjects have a higher lipoprotein(a), i.e. Lp(a), concentration than the general population. Patients with FH are exposed to elevated levels of LDL from birth and ox-LDL may induce other oxidation pathways. The aim of the study was to determine the levels of markers of oxidative stress and DNA damage in patients with HeFH and describe the effect of Lp(a) on the resulting damage. Higher DNA damage was identified in patients with HeFH compared to the normolipidemic ones, and ASCVD was associated with greater damage. Oxidative stress markers were elevated in HeFH patients; however, only ox-LDL was higher in the ASCVD group and its level correlated with DNA damage. A positive correlation was found between DNA damage and Lp(a) concentration in the HeFH patients. Higher levels of Lp(a) were associated with greater DNA damage, especially in patients with HeFH and ASCVD. In HeFH patients, the optimal Lp(a) cut-off point associated with ASCVD is > 23.45 nmol/L, i.e. much lower than for the general population; however this cut-off point needs validation in a larger group of HeFH patients.

The Role of Mast Cells in the Induction and Maintenance of Inflammation in Selected Skin Diseases.

Under physiological conditions, skin mast cells play an important role as guardians that quickly react to stimuli that disturb homeostasis. These cells efficiently support, fight infection, and heal the injured tissue. The substances secreted by mast cells allow for communication inside the body, including the immune, nervous, and blood systems. Pathologically non-cancerous mast cells participate in allergic processes but also may promote the development of autoinflammatory or neoplastic disease. In this article, we review the current literature regarding the role of mast cells in autoinflammatory, allergic, neoplastic skin disease, as well as the importance of these cells in systemic diseases with a pronounced course with skin symptoms.

Glyphosate disturbs various epigenetic processes in vitro and in vivo - A mini review.

Glyphosate in the concentrations corresponding to environmental or occupational exposure has been shown to induce epigenetic changes potentially involved in carcinogenesis. This substance (1) changes the global methylation in various cell types and organisms and is responsible for the methylation of different promoters of individual genes, such as TP53 and P21 in human PBMCs, (2) decreases H3K27me3 methylation and H3 acetylation and increases H3K9 methylation and H4 acetylation in rats, (3) increases the expression of P16, P21, CCND1 in human PBMCs, and the expression of EGR1, JUN, FOS, and MYC in HEK293 cells, but decreases TP53 expression in human PBMCs, (4) changes the expression of genes DNMT1, HDAC3, TET1, TET2, TET3 involved in chromatin architecture, e.g. in fish Japanese medaka, (5) alters the expression of various small, single-stranded, non-coding RNA molecules engaged in post-transcriptional regulation of gene expression, such as miRNA 182-5p in MCF10A cells, miR-30 and miR-10 in mammalian stem cells, as well as several dozen of murine miRNAs. Epigenetic changes caused by glyphosate can persist over time and can be passed on to the offsprings in the next generation; in the third generation they can result in some disorders development, such as prostate disease or obesity. Some epigenetic mechanisms have indicated a potential risk of breast cancer development in human as a result of the exposure to glyphosate. It should be emphasized that the majority of reported epigenetic changes have not yet been associated with the final metabolic effects, which may depend on many other factors.

New possible pharmacological targets for statins and ezetimibe.

Statin therapy is the gold standard in the treatment of dyslipidemia. Understanding the mechanisms of action of these drugs provides an opportunity to define new therapeutic goals for pharmacotherapy in patients with atherosclerotic lesions. The present review indicates the existence of previously unknown therapeutic targets for statins, such as Krüppel-like Factor 2 (KLF-2), Cystathionine γ lyase (CSE) and the microRNA regulating eNOS activity and synthesis; nuclear PXR receptor and EB transcription factor regulating Inflammasome NLRP3 activity; the Dickkopf-related protein 1 (DKK-1), which inhibits the WNT signalling pathway; the peroxisome proliferator-activated receptor (PPAR-γ) in vascular smooth muscle cells (VSMCs), which regulates the cell cycle, and the ERK5-Nrf2 pathway, which reduces the level of harmful advanced glycation end-products (AGE) in VSMCs during diabetic vasculopathy. Importantly, our review includes a number of promising discoveries, specifically those related to the effects of miR-221, miR-222 and miR-27b on the structure, synthesis and activity of eNOS, such as microRNA-based therapies, which offer promise in future targeted therapies. In contrast to numerous experiments confirming the pleiotropic effect of statins, there is still insufficient evidence on the pleiotropic effect of ezetimibe, which goes beyond its basic inhibitory effect on intestinal cholesterol absorption. However, recent studies indicate that this effect is limited to inhibiting macrophage migration, decreasing VCAM-1 expression and reducing the levels of reactive oxygen species. Defining new therapeutic goals for pharmacotherapy in patients with atherosclerotic lesions and ensuring effective treatment of dyslipidemia and its associated cardiovascular complications requires a thorough understanding of both the mechanisms of action of these drugs and of atherosclerosis itself.

The selected epigenetic effects of aminomethylphosphonic acid, a primary metabolite of glyphosate on human peripheral blood mononuclear cells (in vitro).

Aminomethylphosphonic acid (AMPA) is a primary metabolite of glyphosate and amino-polyphosphonate. We have determined the effect of AMPA on selected epigenetic parameters and major cell cycle drivers in human peripheral blood mononuclear cells (PBMCs). The cells were incubated with AMPA at 0.5, 10 and 250 µM for 24 h. The performed analysis included: global DNA methylation by colorimetric measurement of 5-methylcytosine in DNA, methylation in the promoter regions of selected tumor suppressor genes (P16, P21, TP53) and proto-oncogenes (BCL2, CCND1) as well as the expression profile of the indicated genes by Real-Time PCR assays. The obtained results have revealed significant reduction of global DNA methylation level in PBMCs exposed to AMPA. Investigated xenobiotic changed methylation pattern of the P21 and TP53 suppressor gene promoters, but in case of other analyzed genes: P16, BCL2 and CCND1 no statistically significant changes have been noted. Gene profiling have shown that AMPA only changed the expression of CCND1. Summing up, our results have revealed a small potential disturbance in methylation processes and the absence of changes in expression of tested tumor suppressor genes (P16, P21, TP53) and protooncogenes (BCL2) in human PBMCs exposed to AMPA.

Glyphosate affects methylation in the promoter regions of selected tumor suppressors as well as expression of major cell cycle and apoptosis drivers in PBMCs (in vitro study).

We have determined the effect of glyphosate on selected epigenetic parameters and major cell cycle drivers in human peripheral blood mononuclear cells (PBMCs). The cells were incubated with glyphosate at 0.5, 10 and 100 µM. The analysis included: global DNA methylation, methylation in the promoter regions of tumor suppressor genes (P16, P21, TP53) and proto-oncogenes (BCL2, CCND1) by the Real-Time PCR and the expression profile of the indicated genes by Real-Time PCR. The obtained results have revealed significant reduction of global DNA methylation level in PBMCs exposed to glyphosate. Tested compound changed methylation pattern of the P21 and TP53 suppressor gene promoters, but in case of other analyzed genes: P16, BCL2 and CCND1 we did not identify any statistically significant changes. Gene profiling showed that glyphosate changed the expression of genes involved in the regulation of cell cycle and apoptosis. Glyphosate decreased expression of P16 and TP53 as well as an increase in the expression of BCl2, CCND1 and P21. Summing up, our results have shown a potential disturbance in methylation processes and gene expression in human PBMCs exposed to glyphosate, but the observed changes do not prejudge about the final metabolic effects, which are depended on many other factors.

The effect of YAP expression in tumor cells and tumor stroma on the prognosis of patients with squamous cell carcinoma of the oral cavity floor and oral surface of the tongue.

Classic prognostic factors, such as clinical advancement of the disease and histological grade of the tumor, continue to have a decisive role in the selection of therapeutic strategy in patients with carcinoma of the oral cavity floor and oral surface of the tongue (OCC). YAP1/Yes-associated protein 1 (YAP) and transcriptional co-activator with PDZ-binding motif, WWTR1 (TAZ) proteins, appear to be promising markers that may be used to develop personalized therapies. The aim of the present study was to analyze the associations between the levels of YAP, TAZ and tyrosine-protein phosphatase non-receptor type 14 (PTPN14) and to determine whether the increased expression of YAP and TAZ had an effect on tumor cell proliferation, as determined by minichromosome maintenance 7, DNA replication licensing factor 7 expression. Their prognostic value was also assessed. In total, 127 patients who underwent radical surgery and were subjected to adjuvant radiation therapy due to squamous cell OCC were enrolled in the present study. The results demonstrated an evident effect as YAP expression increased in cancer-associated fibroblasts, which induced unfavorable prognosis in patients. In addition, a positive association between proliferation in cancer cells and YAP expression in stromal cells was observed. A lack of YAP expression in the cytoplasm of tumor cells was a factor for poor prognosis with regard to disease-free survival and disease specific survival. No statistically significant correlations between YAP and TAZ expression and PTPN14 expression were identified, nor was a correlation between cell proliferation and the presence of YAP and TAZ in tumor cells observed. The results indicated that YAP expression levels may support the development of personalized therapies for patients.

The mechanism of DNA damage induced by Roundup 360 PLUS, glyphosate and AMPA in human peripheral blood mononuclear cells - genotoxic risk assessement.

Glyphosate is the most heavily applied among pesticides in the world, and thus human exposure to this substance continues to increase. WHO changed classification of glyphosate to probably cancerogenic to humans, thus there is urgent need to assess in detail genotoxic mechanism of its action. We have assessed the effect of glyphosate, its formulation (Roundup 360 PLUS) and its main metabolite (aminomethylphosphonic acid, AMPA) in the concentration range from 1 to 1000 µM on DNA damage in human peripheral blood mononuclear cells (PBMCs). The cells were incubated for 24 h. The compounds studied and formulation induced DNA single and double strand-breaks and caused purines and pyrimidines oxidation. None of compounds examined was capable of creating adducts with DNA, while those substances increased ROS (including •OH) level in PBMCs. Roundup 360 PLUS caused damage to DNA even at 5 µM, while glyphosate and particularly AMPA induced DNA lesions from the concentration of 250 µM and 500 µM, respectively. DNA damage induced by glyphosate and its derivatives increased in order: AMPA, glyphosate, Roundup 360 PLUS. We may conclude that observed changes were not associated with direct interaction of xenobiotics studied with DNA, but the most probably they occurred through ROS-mediated effects.