The Impact of Mutations in the HvCPD and HvBRI1 Genes on the Physicochemical Properties of the Membranes from Barley Acclimated to Low/High Temperatures

1) Background: The study characterized barley mutants with brassinosteroid (BR) biosynthesis and signaling disturbances in terms of the physicochemical/structural properties of membranes to enrich the knowledge about the role of brassinosteroids for lipid metabolism and membrane functioning. (2) Methods: The Langmuir method was used to investigate the properties of the physicochemical membranes. Langmuir monolayers were formed from the lipid fractions isolated from the plants growing at 20 C and then acclimated at 5 C or 27 C. The fatty acid composition of the lipids was estimated using gas chromatography. (3) Results: The BR-biosynthesis and BR-signaling mutants of barley were characterized by a temperature-dependent altered molar percentage of fatty acids (from 14:0 to 20:1) in their galactolipid and phospholipid fractions in comparison to wild-type (WT). For example, the mutants had a lower molar percentage of 18:3 in the phospholipid (PL) fraction. The same regularity was observed at 5 C. It resulted in altered physicochemical parameters of the membranes (Alim, coll, Cs1). (4) Conclusions: BR may be involved in regulating fatty acid biosynthesis or their transport/incorporation into the cell membranes. Mutants had altered physicochemical parameters of their membranes, compared to the WT, which suggests that BR may have a multidirectional impact on the membrane-dependent physiological processes

Stress conditions affect the immunomodulatory potential of Candida albicans extracellular vesicles and their impact on cytokine release by THP-1 human macrophages

Human immune cells possess the ability to react complexly and effectively after contact with microbial virulence factors, including those transported in cell-derived structures of nanometer sizes termed extracellular vesicles (EVs). EVs are produced by organisms of all kingdoms, including fungi pathogenic to humans. In this work, the immunomodulatory properties of EVs produced under oxidative stress conditions or at host concentrations of $CO_{2}$ by the fungal pathogen Candida albicans were investigated. The interaction of EVs with human pro-monocytes of the U-937 cell line was established, and the most notable effect was attributed to oxidative stress-related EVs. The immunomodulatory potential of tested EVs against human THP-1 macrophages was verified using cytotoxicity assay, ROS-production assay, and the measurement of cytokine production. All fungal EVs tested did not show a significant cytotoxic effect on THP-1 cells, although a slight pro-oxidative impact was indicated for EVs released by C. albicans cells grown under oxidative stress. Furthermore, for all tested types of EVs, the pro-inflammatory properties related to increased IL-8 and TNF-$\alpha$ production and decreased IL-10 secretion were demonstrated, with the most significant effect observed for EVs released under oxidative stress conditions

Curcumin’s membrane localization and disruptive effects on cellular processes - insights from neuroblastoma, leukemic cells, and Langmuir monolayers

Abstract In therapies, curcumin is now commonly formulated in liposomal form, administered through injections or creams. This enhances its concentration at the cellular level compared to its natural form ingestion. Due to its hydrophobic nature, curcumin is situated in the lipid part of the membrane, thereby modifying its properties and influencing processes The aim of the research was to investigate whether the toxicity of specific concentrations of curcumin, assessed through biochemical tests for the SK-N-SH and H-60 cell lines, is related to structural changes in the membranes of these cells, caused by the localization of curcumin in their hydrophobic regions. Biochemical tests were performed using spectrophotometric methods. Langmuir technique were used to evaluate the interaction of the curcumin with the studied lipids. Direct introduction of curcumin into the membranes alters their physicochemical parameters. The extent of these changes depends on the initial properties of the membrane. In the conducted research, it has been demonstrated that curcumin may exhibit toxicity to human cells. The mechanism of this toxicity is related to its localization in cell membranes, leading to their dysfunction. The sensitivity of cells to curcumin presence depends on the saturation level of their membranes; the more rigid the membrane, the lower the concentration of curcumin causes its disruption

Assessment of the oxidative stress intensity and the integrity of cell membranes under the manganese nanoparticles toxicity in wheat seedlings

Abstract A response to manganese nanoparticles was studied in seedlings of two wheat cultivars and a model system of plant cell membranes. Nanoparticles at concentrations of 125 and 250 mg/ml were applied foliar. The application of NPs enhanced the content of Mn in plant cells, indicating its penetration through the leaf surface. The stressful effect in the plant cells was estimated based on changes in the activity of antioxidant enzymes, content of chlorophylls and starch. MnNPs evoked no significant changes in the leaf morphology, however, an increase in enzyme activity, starch accumulation, and a decrease in chlorophyll synthesis indicated the stress occurrence. Moreover, a rise in the electrokinetic potential of the chloroplast membrane surface and the reconstruction of their hydrophobic parts toward an increase in fatty acid saturation was found

Brassinosteroid-lipid membrane interaction under low and high temperature stress in model systems

Background: In earlier studies [1], we indicated that applying brassinosteroids (BRs) to lipids that had been isolated from plants altered the physicochemical properties of the monolayers. A continuation of these dependencies using the defined model lipid systems is presented in this paper. The influence of homocastasterone (HCS) and castasterone (CS) (BRs for which the increase in concentration were characteristic of plants grown at low temperatures) on the membrane properties of their polar and the hydrophobic parts were studied. Results: Changes in the electrokinetic potential indicate that both BRs decreased the negative charge of the surface, which is an important factor in modifying the contacts with the polar substances. This property of BRs has not yet been described. The studies of the interactions that occur in the hydrophobic part of the membrane were investigated using the EPR methods and Langmuir techniques. The physicochemical parameters of the lipid structure were determined, and the excess of Gibbs free energy was calculated. Conclusion: We conclude that examined BRs modify both the hydrophilic and hydrophobic properties of the membranes, but to a greater extent HCS. The consequence of these changes may be the attempt to maintain the stability of the membranes in stressful temperature conditions and/or to the possibility of adsorption of other substances on membranes surfaces. The change of plant metabolism towards increasing the amount of BR, mainly HCS (under cooling) may by an important factor for maintaining optimal structural properties of membranes and their functionality despite temperature changes

Exploring the effects of three-finger toxins from Naja ashei venom on neuronal and immunological cancer cell membranes

Abstract Three-finger proteins are the most abundant toxins in the venom of Naja ashei, a snake species from the Elapidae family. This research aimed to describe the effects of varying charges of these proteins, isolated from Naja ashei venom using SEC and IEX chromatography. The study examined how differently charged three-finger toxin fractions interact with and affect neuroblastoma (SK-N-SH) and promyeloblast (HL-60) cells, as well as model Langmuir membranes and liposomes designed to mimic cellular lipid composition. Findings revealed that protein surface charges significantly impact cell survival (MTT assay), membrane damage (lactate dehydrogenase release, malondialdehyde formation), and the structural and electrochemical properties of model membranes (Langmuir membranes and zeta potential for liposomes and cancer cell lines). Results indicated that SK-N-SH cells, characterized by a higher negative charge on their cell membranes, interacted more effectively with positively charged toxins than HL-60 cells. However, the mechanism of these electrostatic interactions is complex. The research demonstrated that electrostatic and mechanical membrane modifications induced by venom proteins can significantly affect cell metabolism. Additionally, the total charge of the membrane, influenced by polar lipid components and phospholipid saturation, plays a decisive role in toxin interaction

Effects of 3FTx Protein Fraction from Naja ashei Venom on the Model and Native Membranes: Recognition and Implications for the Mechanisms of Toxicity

Three-finger toxins are naturally occurring proteins in Elapidae snake venoms. Nowadays, they are gaining popularity because of their therapeutic potential. On the other hand, these proteins may cause undesirable reactions inside the body′s cells. A full assessment of the safety of Naja ashei venom components for human cell application is still unknown. The aim of the study was to determine the effect of the exogenous application of three-finger toxins on the cells of monocytes (U-937) and promyelocytes (HL-60), with particular emphasis on the modification of their membranes under the influence of various doses of 3FTx protein fraction (0–120 ng/mL). The fraction exhibiting the highest proportion of 3FTx proteins after size exclusion chromatography (SEC) separation was used in the experiments. The structural response of cell membranes was described on the basis of single-component and multi-component Langmuir monolayers that mimicked the native membranes. The results show that the mechanism of protein–lipid interactions depends on both the presence of lipid polar parts (especially zwitterionic type of lipids) and the degree of membrane saturation (the greatest-for unsaturated lipids). The biochemical indicators reflecting the tested cells (MDA, LDH, cell survival, induction of inflammation, LD50) proved the results that were obtained for the model

Cold Acclimation and Deacclimation of Winter Oilseed Rape, with Special Attention Being Paid to the Role of Brassinosteroids

Winter plants acclimate to frost mainly during the autumn months, through the process of cold acclimation. Global climate change is causing changes in weather patterns such as the occurrence of warmer periods during late autumn or in winter. An increase in temperature after cold acclimation can decrease frost tolerance, which is particularly dangerous for winter crops. The aim of this study was to investigate the role of brassinosteroids (BRs) and BR analogues as protective agents against the negative results of deacclimation. Plants were cold-acclimated (3 weeks, 4 °C) and deacclimated (1 week, 16/9 °C d/n). Deacclimation generally reversed the cold-induced changes in the level of the putative brassinosteroid receptor protein (BRI1), the expression of BR-induced COR, and the expression of SERK1, which is involved in BR signal transduction. The deacclimation-induced decrease in frost tolerance in oilseed rape could to some extent be limited by applying steroid regulators. The deacclimation in plants could be detected using non-invasive measurements such as leaf reflectance, chlorophyll a fluorescence, and gas exchange monitoring

Foliar spraying with amino acids and their chitosan nanocomposites as promising way to alleviate abiotic stress in iceberg lettuce grown at different temperatures

Abstract We analyzed the effects of foliar spraying with amino acids, chitosan (CHS) and nanocomposites (NCs) of chitosan with the amino acids proline, l-cysteine and glycine betaine (CHS-Pro NCs; CHS-Cys NCs, CHS-GB NCs, respectively) on the changes in the physiological and biochemical parameters of iceberg lettuce grown at the control temperature (20 °C) and under chilling conditions (4 °C). The physicochemical parameters of the phospholipid monolayers (PLs) extracted from plants showed the effects of the treatments on the properties of the monolayers, namely, the packing density and flexibility. We observed increased accumulation of proline at 4 °C, and differences in the concentrations of sugars in most of the analyzed variants were a consequence of the lowered temperature and/or the use of organic compounds. A temperature of 4 °C caused a significant increase in the l-ascorbic acid level compared with that at 20 °C. Differences were also found in glutathione (GSH) content depending on the temperature and treatment with the tested organic compounds. CHS NCs loaded with Pro and GB were effective at increasing the amount of phenols under stress temperature conditions. We noted that a significant increase in the antioxidant activity of plants at 4 °C occurred after priming with Cys, CHS-Cys NCs, Pro and CHS-Pro NCs, and the CHS nanocomposites were more effective in this respect. Both low-temperature stress and foliar spraying of lettuce with various organic compounds caused changes in the activity of antioxidant enzymes. Two forms of dismutase (SOD), iron superoxide dismutase (FeSOD) and copper/zinc superoxide dismutase (Cu/ZnSOD), were identified in extracts from the leaves of iceberg lettuce seedlings. The application of the tested organic compounds, alone or in combination with CHS, increased the amount of malondialdehyde (MDA) in plants grown under controlled temperature conditions. Chilling caused an increase in the content of MDA, but some organic compounds mitigated the impact of low temperature. Compared with that of plants subjected to 20 °C, the fresh weight of plants exposed to chilling decreased. However, the tested compounds caused a decrease in fresh weight at 4 °C compared with the corresponding control samples. An interesting exception was the use of Cys, for which the difference in the fresh weight of plants grown at 20 °C and 4 °C was not statistically significant. After Cys application, the dry weight of the chilled plants was greater than that of the chilled control plants but was also greater than that of the other treated plants in this group. To our knowledge, this is the first report demonstrating that engineered chitosan–amino acid nanocomposites could be applied as innovative protective agents to mitigate the effects of chilling stress in crop plants