MONITOROWANIE HODOWLI KOMÓRKOWYCH W CZASIE RZECZYWISTYM PRZY ZASTOSOWANIU NIKLOWYCH KONDENSATORÓW GRZEBIENIOWYCH

The aim of the study was to present a method for assessing the condition of cell culture by measuring the impedance of cells cultured in the presence of nickel. For this purpose, an impedance measurement technique using nickel comb capacitors was used. The capacitor electrodes were made using a thin film magnetron sputtering. In the experimental part, the culture of cells of mouse fibroblasts on the prepared substrate was performed. The cell culture lasted 43 hours and showed that the presented technique allows it to be used to analyze the effect of nickel on cells.Celem pracy było przedstawienie metody oceny stanu hodowli komórkowej poprzez pomiar impedancji komórek hodowanych w obecności niklu. W tym celu zastosowano technikę pomiaru impedancji z wykorzystaniem niklowych kondensatorów grzebieniowych. Cienkowarstwowe elektrody kondensatora wykonano metodą rozpylania magnetronowego. W części eksperymentalnej przeprowadzono hodowlę komórek mysich fibroblastów na przygotowanym podłożu. Hodowla komórkowa trwała 43 godziny i wykazała, że przedstawiona technika mogłaby być zastosowana do analizy wpływu niklu na komórki

Three Pathways of Cancer Cachexia: Inflammation, Changes in Adipose Tissue and Loss of Muscle Mass—The Role of miRNAs

According to the World Health Organization, in 2018, cancers, along with over 18 million new cases and over 9.5 million deaths remained one of the main causes of mortality globally. Cancer-cachexia, also called wasting syndrome is a complex, multifactorial disorder characterized by progressive skeletal muscle mass loss, with or without adipose tissue atrophy. It is considered as a state of cancer-related malnutrition (CRM) accompanied by inflammation, that is irreversible despite the introduction of nutritional support. Indication of markers of pre-cachectic state seems to be urgently needed. Moreover, such markers have also potential to be used in the assessment of the effects of anti-cachexia treatment, and prognosis. miRNAs are non-coding RNA molecules that are about 20–30 nucleotides long. Single miRNA has the potential to control from few dozen to several hundred different genes. Despite the fact, that the number of miRNAs keep growing. we are making steady progress in establishing regulatory targets and their physiological levels. In this review we described the current knowledge on the impact of miRNAs on processes involved in cancer cachexia development: inflammation, adipose tissue remodelling, and loss of muscle mass both in animal models and the human cohorts. The available studies suggest that miRNAs, due to their properties, e.g., the possibility of regulating even hundreds of different genes, signalling pathways, and biological processes by one molecule, but also due their stability in biological material, the fact, that the change in their level reflects the disease status or the response to the applied treatment, they have great potential to be used as valuable biomarkers in the diagnosis, treatment, and prognosis of cancer cachexia

Monitoring vital functions of A-375 melanoma cell cultures via thin-film nickel capacitors

This article deals in the constantly developing branch of microelectronic devices used in various fields of medicine, i.e. diagnostics and treatment of previously incurable human diseases. A method for assessing and monitoring the vital functions of living cells by measuring cellular impedance in real-time using the ECIS® system and a commercial culture substrate is presented. The goal was to develop a substrate significantly less expensive than a commercial substrate that would be suitable for multiple uses and compatible with the ECIS® measurement station. Moreover, thanks to the use of a material with electrochemical properties other than the biocompatible material (gold or platinum) it is possible to observe the cells behavior with regard to the toxic agent. For this purpose, a culture substrate with nickel comb capacitors was used. To make the electrodes, a thin metal layer was sputtered on polycarbonate plates in the magnetron sputtering process. Prior to the next stages, technological masks were designed so as to fit in the ECIS® measuring station. Subsequently, the microelectronic processes of photolithography and etching the metal layer were performed. Finally, the wells were glued onto the culture medium with a biocompatible adhesive. The completed substrates were transferred to the Department of Human Physiology, Medical University of Lublin, for the culture test on A-375 human melanoma cells. The results of the experiment determined the usefulness of the device for monitoring cell culture vital functions by means of impedance measurement

TNF-α Induced Myotube Atrophy in C2C12 Cell Line Uncovers Putative Inflammatory-Related lncRNAs Mediating Muscle Wasting

Background: Muscle atrophy is a complex catabolic condition developing under different inflammatory-related systemic diseases resulting in wasting of muscle tissue. While the knowledge of the molecular background of muscle atrophy has developed in recent years, how the atrophic conditions affect the long non-coding RNA (lncRNAs) machinery and the exact participation of the latter in the mediation of muscle loss are still unknown. The purpose of the study was to assess how inflammatory condition developing under the tumor necrosis factor alpha (TNF-α) treatment affects the lncRNAs’ expression in a mouse skeletal muscle cell line. Materials and method: A C2C12 mouse myoblast cell line was treated with TNF-α to develop atrophy, and inflammatory-related lncRNAs mediating muscle loss were identified. Bioinformatics was used to validate and analyze the discovered lncRNAs. The differences in their expression under different TNF-α concentrations and treatment times were investigated. Results: Five lncRNAs were identified in a discovery set as atrophy related and then validated. Three lncRNAs, Gm4117, Ccdc41os1, and 5830418P13Rik, were selected as being significant for inflammatory-related myotube atrophy. Dynamics changes in the expression of lncRNAs depended on both TNF-α concentration and treatment time. Bioinformatics analysis revealed the mRNA and miRNA target for selected lncRNAs and their putative involvement in the molecular processes related to muscle atrophy. Conclusions: The inflammatory condition developing in the myotube under the TNF-α treatment affects the alteration of lncRNAs’ expression pattern. Experimental and bioinformatics testing suggested the prospective role of lncRNAs in the mediation of muscle loss under an inflammatory state

Can Dietary Actives Affect miRNAs and Alter the Course or Prevent Colorectal Cancer?

Colorectal cancer is a diet-related cancer. There is much research into the effects of nutrients on the prevention, modulation, and treatment of colorectal cancer. Researchers are trying to find a correlation between epidemiological observations indicating certain dietary components as the originator in the process of developing colorectal cancer, such as a diet rich in saturated animal fats, and dietary components that could eliminate the impact of harmful elements of the daily nutritional routine, i.e., substances such as polyunsaturated fatty acids, curcumin, or resveratrol. Nevertheless, it is very important to understand the mechanisms underlying how food works on cancer cells. In this case, microRNA (miRNA) seems to be a very significant research target. MiRNAs participate in many biological processes connected to carcinogenesis, progression, and metastasis. However, this is a field with development prospects ahead. In this paper, we review the most significant and well-studied food ingredients and their effects on various miRNAs involved in colorectal cancer

Characterization of Undiscovered miRNA Involved in Tumor Necrosis Factor Alpha-Induced Atrophy in Mouse Skeletal Muscle Cell Line

Muscular atrophy is a complex catabolic condition that develops due to several inflammatory-related disorders, resulting in muscle loss. Tumor necrosis factor alpha (TNF-α) is believed to be one of the leading factors that drive inflammatory response and its progression. Until now, the link between inflammation and muscle wasting has been thoroughly investigated, and the non-coding RNA machinery is a potential connection between the candidates. This study aimed to identify specific miRNAs for muscular atrophy induced by TNF-α in the C2C12 murine myotube model. The difference in expression of fourteen known miRNAs and two newly identified miRNAs was recorded by next-generation sequencing between normal muscle cells and treated myotubes. After validation, we confirmed the difference in the expression of one novel murine miRNA (nov-mmu-miRNA-1) under different TNF-α-inducing conditions. Functional bioinformatic analyses of nov-mmu-miRNA-1 revealed the potential association with inflammation and muscle atrophy. Our results suggest that nov-mmu-miRNA-1 may trigger inflammation and muscle wasting by the downregulation of LIN28A/B, an anti-inflammatory factor in the let-7 family. Therefore, TNF-α is involved in muscle atrophy through the modulation of the miRNA cellular machinery. Here, we describe for the first time and propose a mechanism for the newly discovered miRNA, nov-mmu-miRNA-1, which may regulate inflammation and promote muscle atrophy

In Vitro Comparison of Several Thrombus Removal Tools

Background: Although the routine use of thrombus aspiration is not recommended, the thrombectomy technique still might be considered for a selected population of patients. Therefore, the assessment of the effectiveness of commercially available thrombectomy devices is still clinically relevant. Aim: Here, we present an in vitro comparison of several different types of catheters that can be used for thrombus aspiration or removal. Methods: Through the removal of 6 h and 24 h human blood clots in an in vitro model, four catheters were compared: the Launcher, Pronto V4, Vasco+ and the stent-retriever Catchview. The aspiration efficacy was expressed as a percentage of the initial thrombus weight. The effectiveness of the patient’s aspiration was dependent on the time of thrombus formation and was significantly higher for a thrombus formed over 24 h (58.5 ± 26.5%) than for one formed over 6 h (48.0 ± 22.5%; p < 0.001). In the presented in vitro model, Pronto V4 and Launcher showed the highest efficiency. Conclusions: Large-bore aspiration catheters were found to be more effective than narrow-bore catheters or stent-retrievers in an in vitro model of thrombus removal. The thrombus aspiration efficacy increases with longer thrombus formation times

Biological Activity of an Epilobium angustifolium L. (Fireweed) Infusion after In Vitro Digestion

The biological activity of an in vitro digested infusion of Epilobium angustifolium (fireweed) was examined in a model system of intestinal epithelial and colon cancer tissues. The content of selected phenolic compounds in the digested aqueous extract of fireweed was determined using HPLC-ESI-QTOF-MS/MS. Biological activity was examined using the human colon adenocarcinoma cell lines HT-29 and CaCo-2 and the human colon epithelial cell line CCD 841 CoTr. Cytotoxicity was assessed by an MTT assay, a Neutral Red uptake assay, May-Gr&uuml;nwald-Giemsa staining, and a label-free Electric Cell-Substrate Impedance Sensing cytotoxicity assay. The effect of the infusion on the growth of selected intestinal bacteria was also examined. The extract inhibited the growth of intestinal cancer cells HT-29. This effect can be attributed to the activity of quercetin and kaempferol, which were the most abundant phenolic compounds found in the extract after in vitro digestion. The cytotoxicity of the fireweed infusion was dose-dependent. The highest decrease in proliferation (by almost 80%) compared to the control was observed in HT-29 line treated with the extract at a concentration of 250 &mu;g/mL. The fireweed infusion did not affect the growth of beneficial intestinal bacteria, but it did significantly inhibit E. coli. The cytotoxic effect of the fireweed extract indicates that it does not lose its biological activity after in vitro digestion. It can be concluded that the fireweed infusion has the potential to be used as a supporting agent in colon cancer therapy

Electric Cell-Substrate Impedance Sensing (ECIS) as a Convenient Tool to Assess the Potential of Low Molecular Fraction Derived from Medicinal Fungus Cerrena unicolor in Action on L929 and CT-26 Cell Lines

The increase in the incidence of cancer has contributed to the search for new therapeutic methods. In recent years, the use of preparations of natural origin from medical fungi has increased. One such active substance is the extracellular, low molecular active fraction obtained from the medicinal fungus Cerrena unicolor. This study aimed to monitor the pharmacokinetics of different concentrations of substances isolated from the medicinal fungus Cerrena unicolor (ex-LMS) using the ECIS technique. In the study, mouse L929 fibroblasts and colon cancer CT26 cell lines were treated with different concentrations of the active fractions obtained from Cerrena unicolor: C1 = 2.285 (&mu;g/mL); C2 = 22.85 (&mu;g/mL); and C3 = 228.5 (&mu;g/mL). This study demonstrated that the tested preparation from Cerrena unicolor had no considerable effect on the resistance, capacitance, and impedance of L929 fibroblast cells, which was an indicator of no significant effect on its physiological processes. At the same time, those parameters exhibited a decrease in colon cancer cell viability. Following our previous and current studies on Cerrena unicolor, ex-LMS extracts can be safely used in anticancer therapy or chemoprevention with no significant harmful effects on normal cells