Deregulated PI3K/AKT/mTOR signalling commonly exists in glioblastoma, making this axis an attractive target for therapeutic manipulation. Given that activation of PI3K/AKT/mTOR promotes tumour ...growth, metastasis, and resistance to anticancer therapies, mTOR inhibitors show promise in the treatment of cancer. The aim of this study was to investigate the underlying mechanism of novel dual PI3K/mTOR inhibitor, Apitolisib (GDC-0980), in A-172 and U-118-MG GBM tumour cell line suppression. It has been demonstrated that GDC-0980 induces time- and dose-dependent cytotoxicity and apoptosis in investigated glioma cell lines. In our study, the strongest induction of apoptosis was exhibited in the A-172 line after 48 h of incubation with 20 µM GDC-0980, where we observed 46.47% of apoptotic cells. In conclusion, we first discovered that dual PI3K/mTOR blockade by GDC-0980 markedly suppressed survival of human GBM cells and induced apoptosis, independent of the ER stress-mediated DR5 activation. We suggest that GDC-0980, by exerting an inhibitory effect on PERK expression, may thus block its inhibitory effect on protein synthesis, leading to intensification of translation, and this may result in an increase in apoptosis. On the other hand, CHOP stimulates protein synthesis and increases apoptosis. These findings suggest that GDC-0980 may be a candidate for further evaluation as a chemotherapeutic agent for anti-GBM therapy.
Psoriasis is associated with increased production of reactive oxygen species which leads to oxidative stress. As antioxidants can provide protection, the aim of this study was to evaluate the effects ...of cannabidiol (CBD) on neutrophil extracellular trap (NET) formation in psoriatic and healthy neutrophils. Important markers of NETosis were measured in healthy and psoriatic neutrophils after incubation with CBD, lipopolysaccharide (LPS), and LPS + CBD). The percentage of neutrophils undergoing NETosis and the level of NETosis markers (cfDNA, MPO, elastase) were higher in the neutrophils and blood plasma of psoriatic patients, compared to controls. After LPS treatment, all of the markers of NETosis, except elastase, and p47 and citrullinated histones, were increased in samples from healthy subjects and psoriasis patients. CBD reduced the concentrations of NETosis markers. This led to a reduction in NETosis, which was more pronounced in psoriatic neutrophils and neutrophils treated with LPS in both psoriatic and healthy participants. These results suggest that psoriatic patients neutrophils are at a higher risk of NETosis both in vitro and in vivo. CBD reduces NETosis, mainly in psoriatic neutrophils, possibly due to its antioxidant properties. The anti-NET properties of CBD suggest the positive effect of CBD in the treatment of autoimmune diseases.
Breast cancer has a multifactorial etiology. One of the supposed and novel mechanisms is an alteration of circadian gene expression. Circadian genes play a crucial role in many physiological ...processes. These processes, such as genomic stability, DNA repair mechanism and apoptosis, are frequently disrupted in breast tumors. To assess the significance of circadian gene expression in breast cancer, we carried out an analysis of CLOCK, BMAL1, NPAS2, PER1, PER2, PER3 and CRY1, CRY2, TIMELESS, CSNK1E expression by the use of the quantitative Real-Time PCR technique in tumor tissue and non-tumor adjacent normal tissue sampled from 107 women with a newly diagnosed disease. The obtained data were compared to the clinical and histopathological features. PER1, PER2, PER3, CRY2 were found to be significantly down-expressed, while CLOCK, TIMELESS were over-expressed in the studied tumor samples compared to the non-tumor samples. Only gene expression of CRY1 was significantly down-regulated with progression according to the TNM classification. We found significantly decreased expression of CRY2, PER1, PER2 genes in the ER/PR negative breast tumors compared to the ER/PR positive tumors. Additionally, expression of CRY2, NPAS2 genes had a decreased level in the poorly differentiated tumors in comparison with the well and moderately differentiated ones. Our results indicate that circadian gene expression is altered in breast cancer tissue, which confirms previous observations from various animal and in vitro studies.
This study was conducted to verify a hypothesis that immune cells are a target for the action of endocrine disrupting chemicals (EDCs) by investigating whether methylparaben (MeP) modulates human ...neutrophil functions. Neutrophils isolated from 15 donor samples were studied. Cells were incubated in the presence of increasing MeP concentrations (0.06, 0.8, 10, and 20 μM). The cytotoxic effect of MeP on neutrophils was evaluated by the MTT test. The ability of the neutrophils for chemotaxis, phagocytosis, NADPH oxidase activity, and superoxide anion generation was assessed in Boyden’s chamber, Park’s method with latex, the NBT test, and the cytochrome C reduction test, respectively. The total nitric oxide (NO) concentration was measured by the Griess reaction. There was no observable cytotoxic effect of MeP on human neutrophils. MeP (10 and 20 μM) exposure decreased neutrophilic ability for the tested functions, except for NO production. In neutrophils incubated with MeP (0.8 μM as well as 0.06 and 0.8 μM, respectively), we observed a decreased activity of NADPH oxidase as well as decreased generation of superoxide anion. Our results suggest a suppressive effect of MeP on the tested functions of human neutrophils, which confirms that immune cells are vulnerable to EDC action. Therefore, the disturbance of neutrophils’ oxygen-dependent phagocytic function as a result of exposure to environmental doses of MeP action could lead to impairment of innate immune responses in humans exposed to xenoestrogens.
Endocrine disrupting chemicals (EDCs) are added to food, cosmetics, plastic packages, and children's toys and have thus become an integral part of the human environment. In the last decade, there has ...been increasing interest in the effect of EDCs on human health, including their impact on the immune system. So far, researchers have proved that EDCs (e.g. bisphenols, phthalates, triclosan, phenols, propanil, tetrachlorodibenzo-p-dioxin, diethylstilbestrol, tributyltin (TBT), and parabens) affect the development, functions, and lifespan of immune cells (e.g., monocytes, neutrophils, mast cells, eosinophils, lymphocytes, dendritic cells, and natural killers).
In this review, we have summarized the current knowledge of the multivariable influence of EDCs on immune cells and underlined the novel approach to EDC studies, including dose-dependent effects and low-dose effects. We discuss critically the possible relationship between exposure to EDCs and immunity related diseases (e.g. allergy, asthma, diabetes, and lupus). Moreover, based on the literature, we construct a model of possible mechanisms of EDC action on immune cells at cellular, molecular, and epigenetic levels.
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•EDCs deregulate adaptive and innate immunity.•The immature immune system is more vulnerable to EDCs exposure than mature.•Molecular mechanism of EDCs' action in immune cells requires further studies.
Mesotrophic rich fens, that is, groundwater-fed mires, may be long-lasting, as well as transient ecosystems, displaced in time by poor fens, bogs, forests or eutrophic reeds. We hypothesized that fen ...stability is controlled by plant stress caused by waterlogging with calcium-rich and nutrient-poor groundwater, which limits expansion of hummock mosses, tussock sedges and trees. We analysed 32 European Holocene macrofossil profiles of rich fens using plant functional traits (PFTs) which indicate the level of plant stress in the environment: canopy height, clonal spread, diaspore mass, specific leaf area, leaf dry matter content, Ellenberg moisture value, hummock-forming ability, mycorrhizal status and plant functional groups. Six PFTs, which formed long-term significant trends during mire development, were compiled as rich fen stress indicator (RFSI). We found that RFSI values at the start of fen development were correlated with the thickness of subsequently accumulated rich fen peat. RFSI declined in fens approaching change into another mire type, regardless whether it was shifting into bog, forest or eutrophic reeds. RFSI remained comparatively high and stable in three rich fens, which have not terminated naturally until present times. By applying PFT analysis to macrofossil data, we demonstrated that fens may undergo a gradual autogenic process, which lowers the ecosystem’s resistance and enhances shifts to other mire types. Long-lasting rich fens, documented by deep peat deposits, are rare. Because autogenic processes tend to alleviate stress in fens, high levels of stress are needed at initial stages of rich fen development to enable its long persistence and continuous peat accumulation.
We have determined the effect of glyphosate and aminomethylphosphonic acid (AMPA) on expression of genes involved in chromatin architecture in human peripheral blood mononuclear cells (PBMCs). The ...cells were incubated with glyphosate and AMPA in the concentrations ranging from 0.5 to 100 μM and from 0.5, to 250 μM, respectively. The expression profile of the following genes by quantitative Real-Time PCR was evaluated: Genes involved in the DNA methylation (
,
) and DNA demethylation process (
) and those involved in chromatin remodeling: genes involved in the modification of histone methylation (
,
) and genes involved in the modification of histone deacetylation (
,
). Gene profiling showed that glyphosate changed the expression of
,
, and
, while AMPA changed the expression of
and
. The results also revealed that glyphosate at lower concentrations than AMPA upregulated the expression of the tested genes. Both compounds studied altered expression of genes, which are characteristic for the regulation of transcriptionally inactive chromatin. However, the unknown activity of many other proteins involved in chromatin structure regulation prevents to carry out an unambiguous evaluation of the effect of tested xenobiotics on the studied process. Undoubtedly, we have observed that glyphosate and AMPA affect epigenetic processes that regulate chromatin architecture.
Pregnancy loss is a multidisciplinary problem which concerns researchers from the fields of medicine, epidemiology, psychology, and public health. The primary objective of the present study was to ...explain the potential role of neutrophil extracellular traps (NETs) in the process of spontaneous miscarriage. Enzyme-linked immunosorbent assay to assess the levels of biomarkers of NETs in the serum of examined women was conducted. Furthermore, levels of nitric oxide (NO) and late markers of its action were measured in serum samples. Analyses results demonstrated the existence of NETs in the placental tissue of women who miscarried as well as a simultaneous increase in the levels of myeloperoxidase and pentraxin 3. This clearly confirms the participation of NETs in the course of pregnancy loss. Women who have had a miscarriage but did not show the presence of NETs in their placenta exhibited the highest contents of NO, nitrotyrosine, and malondialdehyde suggesting a different pathway leading to pregnancy loss associated with disturbed oxidative-antioxidative processes. Although study results demonstrate new aspects associated with the formation of NETs they are not, however, sufficient to unambiguously determine the role of NETs in the course of miscarriage.
Objectives: The study aimed to evaluate the impact of aging on the formation of neutrophil extracellular traps (NETs). The impaired formation of NETs is the cause of an abnormal innate immune ...response. Material and Methods: The study included a total of 45 healthy male subjects of different age groups. Whole blood was collected from the subjects, and the concentration of myeloperoxidase (MPO), the main biocidal protein in NETs, was determined in serum using ELISA. The serum levels of circulating free DNA (cfDNA), which are the structural basis of NETs, were also measured by fluorescence. In addition, the white blood cell count was determined, whole blood smear was evaluated, and the neutrophil-lymphocyte ratio was calculated. The variations in the levels of NET biomarkers were analyzed in different age groups. Results: The low levels of MPO (243.70 ng/ml) and cfDNA (6.24 ng/100 microl) in boys indicated neutrophil insufficiency for NETosis in children. A progressive increase in the levels of MPO and cfDNA with age was observed among adolescents (420.91, p = 0.04; 13.55, p = 0.03, respectively), with the highest level noted in the healthy adult group (466.58, p = 0.01; 14.07, p = 0.01, respectively). The levels of the studied parameters were comparable in adolescents and young adults, which proved that the NETosis process was appropriate and suggested the attainment of neutrophil maturity for the release of NETs in adolescence. The levels of MPO and cfDNA were low in older men (225.46, p < 0.01; 5.19, p < 0.01, respectively) indicating impaired NET formation. Conclusions: Data on the generation of NETs in different age groups obtained in this study can allow a better understanding of the ontogenesis of the immune system in terms of the course of NETosis, and also indicate the need to support nonspecific responses in children and adults. Further research should be performed to determine the possibility of regulating the NETosis process. Key words: biomarkers, neutrophils, MPO, NETs, cfDNA, immunoaging
The study documents a process of ice lenses persistence under hummocks during spring snowmelt in a rich fen in Poland. Air temperature, as well as the temperature under vascular plant litter,
...Hamatocaulis vernicosus
mat,
Tomentypnum nitens
hummocks and
Sphagnum teres
hummocks, was measured from December to March using temperature data loggers. The Capital Asset Pricing Model was adapted to analyse relationships between air temperature and temperature under litter and mosses.
Sphagnum teres
best insulated the temperature of the upper peat layer from changing air temperatures and maintained frozen peat under its hummocks the longest (for about a month after the frost had stopped).
Tomentypnum nitens
insulated similarly effectively in winter, also maintaining the ice lenses under its hummocks, but less effectively in warmer spring, which may be due to differences in hummock structure between the two species. The observed phenomenon may soon disappear due to global warming, which may affect the fen functioning.
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