Cancer metastatic spread to serous cavity causes malignant pleural effusions (MPEs), indicating dismal prognosis. Tumor microenvironment can implement suppressive activity on host immune responses. Thus, we investigated the prevalence of Tregs and the relationship between them and TGF-βand IL-10 concentrations and measured expression ofFOXP3,CTLA-4,CD28, andGITRgenes, as well as protein expression of selected genes in benign effusions and MPEs. The percentage of Tregs was determined by means of multicolor flow cytometry system. TGF-βand IL-10 concentrations were measured using human TGF-β1 and IL-10 ELISA kit. Relative mRNA expression of studied genes was analyzed by real-time PCR. The frequency of Tregs was significantly higher in MPEs compared to benign effusions; however, the level of TGF-βand IL-10 in analyzed groups was comparable, and no correlation between concentrations of TGF-βand IL-10 and percentage of Tregs was observed. Relative mRNA expression of all the genes was higher in CD4⁺CD25⁺compared to CD4⁺CD25⁻cells. In CD4⁺CD25⁺cells from MPEs, relative mRNA expression ofFOXP3,CTLA-4, andCD28genes was significantly higher than in benign effusions; however, the level of CD4⁺CD25⁺CTLA-4⁺cells in analyzed groups showed no significant differences. We found numerous genes correlations in an entire CD4⁺CD25⁺cell subset and CD4⁺CD25⁺cells from MPEs. Enhanced suppressive activity of Tregs is observed in the microenvironment of MPEs. Understanding of relations between cellular and cytokine immunosuppressive factors in tumor microenvironment may determine success of anticancer response.

The treatment of non-small cell lung cancer (NSCLC) has recently evolved with the introduction of targeted therapy based on the use of tyrosine kinase inhibitors (TKIs) in patients with certain gene alterations, including EGFR, ALK, ROS1, BRAF, and MET genes. Molecular targeted therapy based on TKIs has improved clinical outcomes in a large number of NSCLC patients with advanced disease, enabling significantly longer progression-free survival (PFS). Liquid biopsy is an increasingly popular diagnostic tool for treating TKI-based NSCLC. The studies presented in this article show that detection and analysis based on liquid biopsy elements such as circulating tumor cells (CTCs), cell-free DNA (cfDNA), exosomes, and/or tumor-educated platelets (TEPs) can contribute to the appropriate selection and monitoring of targeted therapy in NSCLC patients as complementary to invasive tissue biopsy. The detection of these elements, combined with their molecular analysis (using, e.g., digital PCR (dPCR), next generation sequencing (NGS), shallow whole genome sequencing (sWGS)), enables the detection of mutations, which are required for the TKI treatment. Despite such promising results obtained by many research teams, it is still necessary to carry out prospective studies on a larger group of patients in order to validate these methods before their application in clinical practice.

Mammalian cumulus–oocyte complexes (COCs) reach full developmental capability during folliculogenesis and oogenesis. It is well recognized that only gametes achieving MII stage after in vivo or in vitro maturation (IVM) are successfully fertilized by a single spermatozoon. Although the process of oocyte nuclear and/or cytoplasmic maturation in pigs is well determined, there exist many differences that promote these processes in vivo and in vitro. Therefore, this study aimed to investigate the differences in RNA expression profiles between porcine oocytes before and after IVM using microarray and real-time quantitative polymerase chain reaction (RT-qPCR) assays. Experiments were performed on oocytes isolated from 55 pubertal crossbred Landrace gilts. The oocytes were analyzed both before and after IVM and only Brilliant Cresyl Blue (BCB)-positive gametes were used for subsequent microarray analysis (Affymetrix) and RT-qPCR analysis. The microarray assay, which measures expression of 12,258 transcripts, revealed 419 differentially expressed transcripts in porcine oocytes, from which 379 were downregulated and 40 were upregulated before IVM compared to those analyzed after IVM. After DAVID analysis, we found eight different transcripts, including IHH, BMP1, WWTR1, CHRDL1, KLF10, EIF2AK3, MMP14, and STC1. Their expression is related to the “bone development” ontology group and was further subjected to hierarchical clusterization. Using RT-qPCR analysis, we confirmed the results of the microarray assay, showing increased expression of the eight genes in oocytes before IVM compared to oocytes after maturation in vitro. It has been suggested that “bone development” belongs to one ontological group involving genes substantially upregulated in porcine oocytes before IVM. We suggest that the gamete mRNA expression profile before IVM may comprise stored transcripts, which are templates for protein biosynthesis following fertilization. We also hypothesize that these mRNAs may be a specific “fingerprint” of folliculogenesis and oogenesis in pigs.

Background: HSC constitute a small portion of liver cells, but fulfill significant functions. LX-2 cells, a human cell line of HSC, are a model for the study of HSC. HSC express Toll-like receptors (TLRs) and some of them participate in the promotion of liver fibrosis. Little is known about other TLRs expression and function of HSC. Aims of this study were to search the expression and function of all TLRs known in LX-2 cells. Material and methods: Cells were subjected to immunocytochemistry, cell RNA was searched for mRNA specific for respective TLRs. Cells were treated with anti-TLR1-10 antibodies and flow cytometry (FC). They were cultured with TLR ligands and assessed by FC for TLR expression. Supernatants were examined for cytokines content and cells for the percentage of those in a S-phase of cell cycle. Results: All TLRs, both, protein and mRNA were expressed in these cells. TLR ligands had negligible effect on TLR expression. Culture of LX-2 with ligands resulted in the secretion of some cytokines, but did not increase the number of cells in S-phase. Conclusions: To conclude, LX-2 cells express all known TLRs. Their activation has only modest effect on their function.