Aggregation Of Liposomes Research Articles

A highly sensitive fluorescence biosensor for aflatoxins B1 detection based on polydiacetylene liposomes combined with exonuclease III-assisted recycling amplification.

Afluorescence biosensor for determination of aflatoxin B1 (AFB1)based on polydiacetylene (PDA) liposomes and exonuclease III (EXO III)-assisted recycling amplification was developed. The AFB1 aptamer partially hybridizes with complementary DNA (cDNA), which is released upon recognition of AFB1 by the aptamer. Subsequently, the cDNA hybridizes with hairpin H to form double-stranded DNA that undergoes digestion by EXO III, resulting in the cyclic release of cDNA and generation of capture DNA for further reaction. The capture DNA then hybridizes with probe modified on PDA liposomes, leading to aggregation of liposomes and subsequent fluorescence production. This strategy exhibited a limit of detection of 0.18 ng/mL within the linear range 1-100 ng/mL with adetermination coefficient > 0.99. The recovery ranged from 92.81 to 106.45%, with relative standard deviations (RSD) between 1.73 and 4.26%, for corn, brown rice, peanut butter, and wheatsamples. The stability, accuracy, and specificity of the method demonstrated the applicability for realsampleanalysis.

Differential effects of theasinensins and epigallocatechin-3-O-gallate on phospholipid bilayer structure and liposomal aggregation

(−)-Epigallocatechin-3-O-gallate (EGCg), the major catechin responsible for the health-enhancing and disease-preventive effects of green tea, is susceptible to auto-oxidation at physiological pH levels. However, whether the oxidized EGCg resulting from its oral consumption possesses any bioactive functions remains unclear. This study presents a differential analysis of intact and oxidized EGCg regarding their interactions with phosphatidylcholine liposomes, serving as a simple biomembrane model. In the presence of ascorbic acid, pre-oxidized EGCg induced liposomal aggregation in a dose-dependent manner, whereas intact EGCg did not. Toxicity evaluation using calcein-loaded liposomes revealed that liposomal aggregation is associated with minimal membrane damage. Through fractionation of the oxidized EGCg sample, the fraction containing theasinensins showed high liposomal aggregation activity. Overall, these results suggest that oxidatively condensed EGCg dimers may stimulate various cells by altering the plasma membrane in a manner different from that of EGCg monomers.

Technique for assessing the astringency of persimmon fruit by measuring the liposome aggregation

Technique for assessing the astringency of persimmon fruit by measuring the liposome aggregation

Annexin A5 stabilizes matrix vesicle-biomimetic lipid membranes: unravelling a new role of annexins in calcification

Matrix vesicles are a special class of extracellular vesicles thought to actively contribute to both physiologic and pathologic mineralization. Proteomic studies have shown that matrix vesicles possess high amounts of annexin A5, suggesting that the protein might have multiple roles at the sites of calcification. Currently, Annexin A5 is thought to promote the nucleation of apatitic minerals close to the inner leaflet of the matrix vesicles’ membrane enriched in phosphatidylserine and Ca2+. Herein, we aimed at unravelling a possible additional role of annexin A5 by investigating the ability of annexin A5 to adsorb on matrix-vesicle biomimetic liposomes and Langmuir monolayers made of dipalmitoylphosphatidylserine (DPPS) and dipalmitoylphosphatidylcholine (DPPC) in the absence and in the presence of Ca2+. Differential scanning calorimetry and dynamic light scattering measurements showed that Ca2+ at concentrations in the 0.5–2.0 mM range induced the aggregation of liposomes probably due to the formation of DPPS-enriched domains. However, annexin A5 avoided the aggregation of liposomes at Ca2+ concentrations lower than 1.0 mM. Surface pressure versus surface area isotherms showed that the adsorption of annexin A5 on the monolayers made of a mixture of DPPC and DPPS led to a reduction in the area of excess compared to the theoretical values, which confirmed that the protein favored attractive interactions among the membrane lipids. The stabilization of the lipid membranes by annexin A5 was also validated by recording the changes with time of the surface pressure. Finally, fluorescence microscopy images of lipid monolayers revealed the formation of spherical lipid-condensed domains that became unshaped and larger in the presence of annexin A5. Our data support the model that annexin A5 in matrix vesicles is recruited at the membrane sites enriched in phosphatidylserine and Ca2+ not only to contribute to the intraluminal mineral formation but also to stabilize the vesicles’ membrane and prevent its premature rupture.

Liposomal aggregates sustain the release of rapamycin and protect cartilage from friction

Liposomes show promise as biolubricants for damaged cartilage, but their small size results in low joint and cartilage retention. We developed a zinc ion-based liposomal drug delivery system for local osteoarthritis therapy, focusing on sustained release and tribological protection from phospholipid lubrication properties. Our strategy involved inducing aggregation of negatively charged liposomes with zinc ions to extend rapamycin (RAPA) release and improve cartilage lubrication. Liposomal aggregation occurred within 10 min and was irreversible, facilitating excess cation removal. The aggregates extended RAPA release beyond free liposomes and displayed irregular morphology influenced by RAPA. At nearly 100 µm, the aggregates were large enough to exceed the previously reported size threshold for increased joint retention. Tribological assessment on silicon surfaces and ex vivo porcine cartilage revealed the system's excellent protective ability against friction at both nano- and macro-scales. Moreover, RAPA was shown to attenuate the fibrotic response in human OA synovial fibroblasts. Our findings suggest the zinc ion-based liposomal drug delivery system has potential to enhance OA therapy through extended release and cartilage tribological protection, while also illustrating the impact of a hydrophobic drug like RAPA on liposome aggregation and morphology.

Influence of Lipidation Pattern of the KR12 Fragment of Peptide LL-37 on Its Antibacterial and Hemolytic Activities

Contemporary medicine has been confronted by multidrug resistance. Therefore, new antibiotics are sought to alleviate the problem. In this study, we estimated the effect of the positioning and extent of lipidation (mainly octanoic acid residue) in the KR12-NH2 molecule on antibacterial and hemolytic activities. The effect of the conjugation of benzoic acid derivatives (C6H5-X-COOH, where X: CH2, CH2-CH2, CH=CH, C≡C, and CH2-CH2-CH2) with the N-terminal part of KR12-NH2 on biological activity was also studied. All analogs were tested against planktonic cells of ESKAPE bacteria and reference strains of Staphylococcus aureus. The effect of lipidation site on the helicity of the KR12-NH2 analogs was studied using CD spectroscopy. The ability of the selected peptides to induce the aggregation of POPG liposomes was evaluated with DLS measurements. We demonstrated that both the site and extent of peptide lipidation play an essential role in the bacterial specificity of the lipopeptides. Most of the C8α-KR12-NH2 (II) analogs that were more hydrophobic than the parent compound were also more hemolytic. A similar relationship was also found between the α-helical structure content in POPC and hemolytic activity. It is worth emphasizing that in our study, the highest selectivity against S. aureus strains with an SI value of at least 21.11 exhibited peptide XII obtained by the conjugation of the octanoic acid with the N-terminus of retro-KR12-NH2. All lipidated analogs with the highest net charge (+5) were the most selective toward pathogens. Therefore, the overall charge of KR12-NH2 analogs plays pivotal role in their biological activity.

Co-delivery of gemcitabine and salinomycin in PEGylated liposomes for enhanced anticancer efficacy against colorectal cancer

Colorectal cancer remains one of the major causes of morbidity and mortality in both developed and emerging countries. Cancer stem cells (CSCs) are a subpopulation of cells within the tumor mass harboring stem cell characteristics, considered responsible for tumor initiation, growth, relapse, and treatment failure. Lately, it has become clear that both CSCs and non-CSCs have to be eliminated for the successful eradication of cancer. Drug delivery systems have been extensively employed to enhance drug efficacy. In this study, salinomycin (SAL), a selective anti-CSC drug, and gemcitabine (GEM), a conventional anticancer drug, were co-loaded in liposomes and tested for optimal therapeutic efficacy. We employed the Design of Experiments approach to develop and optimize a liposomal delivery system for GEM and SAL. The antiproliferative effect of the liposomes was evaluated in SW-620 human colorectal cancer cells. The GEM and SAL-loaded liposomes exhibited adequate size, polydispersity, zeta potential, and drug content. The in vitro release study showed a sustained release of GEM and SAL from the liposomes over 72 h. Moreover, no sign of liposome aggregation was seen over 1 month and in a biological medium (FBS). The in vitro cytotoxic effects of the co-loaded liposomes were superior to that of single GEM either in free or liposomal form. The combination therapy using GEM and SAL co-loaded in liposomes could be a promising strategy for tackling colorectal cancer.

Proliposome-Based Nanostrategies: Challenges and Development as Drug Delivery Systems.

Many attempts have been made to the refinement of liposomal stability. In 1986, Payne et al. developed the approach of proliposomes to derelict the physicochemical instability confronted in some liposome suspensions, i.e., fusion, aggregation, hydrolysis, and oxidation. This review attempts to cover different aspects of proliposomes along with their types and preparation methods. The review is also focused on the scope of proliposomes as a nano-based drug delivery system and subsequent applications. An attempt has been made to cover all the facets of proliposomes, from their composition to clinical trials. The extensive scientific data from proliposomes provide substantial shreds of evidence for its huge delivery potential.

Nano-visualization of the in vitro antiviral activity of black tea based on production area using a liposome-based virus membrane model.

Black tea extracts (BTEs) from four different production areas showed a higher aggregation strength for phosphatidylcholine-based liposomes containing cholesterol used as a viral membrane model. Furthermore, the anti-influenza A virus (IAV) activity of each BTE in vitro demonstrated that although Sri Lanka, Kenya, and Assam had higher anti-IAV activities, Darjeeling had a lower anti-IAV activity, showing a correlation between each BTE and the liposome aggregation strength. Moreover, the antiviral activity strength of BTEs was consistent with the antioxidant activity strength of BTEs, suggesting that the component(s) in black tea that exhibits antioxidant activity would also be the component(s) that accounts for its antiviral activity. Thus, our results propose that BTEs exert their antiviral effects by binding not only hemagglutinin and neuraminidase but also viral membranes directly, especially "cholesterol-rich lipid rafts" and affect the membrane structure, causing the virus to aggregate, thereby inhibiting infection of the host cells.

Structuring and de-structuring of nanovectors from algal lipids. Part 1: physico-chemical characterization

Lipid nanocarriers are among the most employed systems for drug delivery purposes in several research and industrial sectors, since their favorable properties ensure broad applicability. The design and characterization of these nanosystems are of paramount importance to obtain controlled outcome, since the supramolecular structure and molecular interactions deeply impact the functionality of the resulting aggregates. The choice of the most appropriate formulation for the target of interest relies on in-depth physico-chemical characterization in order to optimize stability, loading rates and sustained release. Several supramolecular architectures suited for carrier development can be obtained from lipid building blocks, by varying lipid composition and packing parameter. In particular, cubosome and liposome aggregates are often used as drug vectors thanks to their high cargo capability and biocompatibility. Moreover, the possibility to employ lipids from natural sources i.e. biomasses to prepare nanosystems makes them especially attractive. In this work, two aggregate types were characterized and compared as drug vectors for poorly water-soluble antioxidants, particularly curcumin and two adjuvants (i.e. tocopherol and piperine). The nanovectors were obtained by extracting lipids from algal biomasses with different lipid composition, and characterized by advanced structural (DLS, SAXS, Cryo-TEM) techniques, spectroscopy (NMR) and calorimetry (ITC). Finally, the structural stability of both aggregate types was evaluated.

Size-dependent placental retention effect of liposomes in ICR pregnant mice: Potential superiority in placenta-derived disease therapy

The great challenge in developing safe medications for placenta-derived diseases is to reduce or eliminate fetal drug exposure while still providing the necessary therapeutic effect. Rapid advances in nanotechnology have brought opportunities for the therapy of placenta-derived disease through accumulating the drug in the placenta while reducing its placental penetration. Among various nanocarriers, liposomes are regarded as an ideal type of carrier for placental drug delivery due to their biosafety and biodegradability. However, their placental retention effect with different particle sizes has not been studied. This research aimed to explore a suitable size of liposomes for placenta drug delivery. Cy 5 dye was chosen as a model molecule for tracing the distribution of three different-sized liposomes (∼80 nm, 200 nm, and 500 nm) in ICR pregnant mice. The stability, cytotoxicity, and cellular uptake study of Cy 5-loaded liposomes were performed. The in vivo fluorescence studies on ICR pregnant mice suggested that the particle size of liposomes was positively correlated with the degree of liposome aggregation in the placenta. The ratio of fluorescence in the placenta and fetus section (P/F value) was proposed to evaluate the placental retention effect of different-sized liposomes. The results showed that the liposomes with 500 nm had the highest P/F value and thus exhibited the strongest placental retention effect and the weakest placental penetration ability. Moreover, liquid chromatography-mass spectrometry analysis confirmed the reliability of the fluorescence section analysis in exploring the placental retention effect of nanovehicles. In general, this study introduced a simple and intuitive method to evaluate the placental retention effect of nanoplatforms and defined a suitable size of liposomes for placenta-derived disease drug delivery.

Interactions of (−)-epigallocatechin-3-gallate with model lipid membranes

(−)-Epigallocatechin-3-gallate (EGCG) is a flavonoid known for its good antioxidant potential and health benefits. It is one of the most intriguing flavonoids, especially because of its specific interactions with model lipid membranes. It was noticed that EGCG might form EGCG rich domains/rafts at certain compositions of lipid membranes. In this article, we investigate whether EGCG forms EGCG rich domains when incorporated in 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) liposomes. Our results show that EGCG decreases lipid ordering parameter in ordered membranes and increases it in the case of disordered ones. Also, incorporation of EGCG does not affect the zeta-potential and shape of the liposomes, but it can induce aggregation of liposomes. Our study also demonstrates that liposomes with incorporated EGCG are highly protected against UV-light induced oxidation.

Lipid headgroup and side chain architecture determine manganese-induced dose dependent membrane rigidification and liposome size increase.

Metal ion-membrane interactions have gained appreciable attention over the years resulting in increasing investigations into the mode of action of toxic and essential metals. More work has focused on essential ions like Ca or Mg and toxic metals like Cd and Pb, whereas this study investigates the effects of the abundant essential trace metal manganese with model lipid systems by screening zwitterionic and anionic glycerophospholipids. Despite its essentiality, deleterious impact towards cell survival is known under Mn stress. The fluorescent dyes Laurdan and diphenylhexatriene were used to assess changes in membrane fluidity both in the head group and hydrophobic core region of the membrane, respectively. Mn-rigidified membranes composed of the anionic phospholipids, phosphatidic acid, phosphatidylglycerol, cardiolipin, and phosphatidylserine. Strong binding resulted in large shifts of the phase transition temperature. The increase was in the order phosphatidylserine > phosphatidylglycerol > cardiolipin, and in all cases, saturated analogues > mono-unsaturated forms. Dynamic light scattering measurements revealed that Mn caused extensive aggregation of liposomes composed of saturated analogues of phosphatidic acid and phosphatidylserine, whilst the mono-unsaturated analogue had significant membrane swelling. Increased membrane rigidity may interfere with permeability of ions and small molecules, possibly disrupting cellular homeostasis. Moreover, liposome size changes could indicate fusion, which could also be detrimental to cellular transport. Overall, this study provided further understanding into the effects of Mn with biomembranes, whereby the altered membrane properties are consequential to the proper structural and signalling functions of membrane lipids.

Inhibition of cryoaggregation of phospholipid liposomes by an Arabidopsis intrinsically disordered dehydrin and its K-segment

Dehydrin is an intrinsically disordered protein involved in the cold tolerance of plants. Although dehydrins have been thought to protect biomembranes under cold conditions, the underlying protective mechanism has not been confirmed. Here we report that Arabidopsis dehydrin AtHIRD11 inhibited the aggregation of phospholipid liposomes after freezing and thawing. AtHIRD11 showed significantly greater cryoaggregation-prevention activity than cryoprotective agents such as trehalose, proline, and polyethylene glycols. Amino acid sequence segmentation analysis indicated that the K-segment of AtHIRD11 inhibited the cryoaggregation of phosphatidylcholine (PC) liposomes but other segments did not. This showed that K-segments conserved in all dehydrins were likely to be the cryoprotective sites of dehydrins. Amino acid replacement for a typical K-segment (TypK for short) sequence demonstrated that both hydrophobic and charged amino acids were required for the cryoaggregation-prevention activity of PC liposomes. The amino acid shuffling of TypK remarkably reduced cryoprotective activity. Although TypK did not bind to PC liposomes in solution, the addition of liposomes reduced its disordered content under crowded conditions. Together, these results suggested that dehydrins protected biomembranes via conserved K-segments whose sequences were optimized for cryoprotective activities.

Stability of bioactive compounds in liposomes after pasteurisation and storage of functional chocolate milk

SummaryIn the present study, liposomes with bioactive compounds (BC), vitamin E (VE) and vitamin C (VC), were characterised after pasteurisation and storage in chocolate milk. Liposomes were obtained using soy phosphatidylcholine (SPC), SPC with stearic acid (SA) or calcium stearate (CaS). Oxidative stability, size, shape and membrane packing were studied after pasteurisation and storage for 28 days at 4 °C. Oxidative stability was also studied after pasteurisation in extreme conditions: 6 months, 25 °C and light. The oxidative level determined in liposomes is much under the permitted limits, generating good quality food. SPC maintained stability in size, shape and membrane packing for 28 days at 4 °C. SPC:SA and SPC:CaS evidenced a decrease in liposomal aggregation and a higher membrane packing for 28 days at 4 °C, which favoured stability. These results were optimum because commercial functional milk must have a shelf life of 7 days at 4 °C. Pasteurised liposomes were incorporated in chocolate milk to study the sensory (after storage for 7 days at 4 °C) and microbiological stability. None of the liposomes contributed to the microbial load. SPC:SA presented high sensorial stability in the milk, being a potentially suitable medium to incorporate BC generating functional food.

Photoinduced aggregation of liposome modified with DNA containing ultrafast DNA photo‐cross‐linker

AbstractBACKGROUNDIn this study, we demonstrated the aggregation of liposome using ultrafast DNA photo‐cross‐linking with 3‐cyanovinylcarbazole nucleoside (CNVK). Liposomes are not only used in model experiments of cell membranes, but also are attracting attention as a carrier for drug delivery. The surface of liposomes can be modified with biomolecules such as peptides and DNA. So far, several studies have been conducted to control liposome aggregation by DNA, and it was found that oligoDNA (ODN) containing CNVK can photo‐cross‐link to pyrimidine bases in complementary strand using photoirradiation for a few seconds. Therefore, this DNA photo‐cross‐linking procedure was adapted for liposome aggregation.RESULTBased on results of confocal laser scanning microscopy, it was confirmed that the liposomes were aggregated by photoirradiation. Also, from the results of denaturing PAGE, it was confirmed that the DNA fixed on the liposome surface and the linker connecting the liposomes were photo‐cross‐linked. In addition, the energy of photoirradiation can be used to extract the translated part, so it is expected that liposomes can be prepared according to each degree of aggregation.CONCLUSIONLinker DNA containing CNVK can induce liposome aggregation upon photoirradiation, and the proportion of liposomes aggregated can be varied by changing the irradiation energy (intensity). © 2021 Society of Chemical Industry (SCI).

Coating function and stabilizing effects of surface layer protein from Lactobacillus acidophilus ATCC 4356 on liposomes

Surface layer proteins (SLPs) are crystalline arrays in the outermost layer of cell envelope in many archaea and bacteria. SLPs subunits have the ability to reassemble on the surface of lipid layers. In this work, the SLP from Lactobacillus acidophilus ATCC 4356 was extracted and reassembled on the surface of positively charged liposomes composed of dipalmitoyl phosphatidylcholine, cholesterol and octadecylamine. Zeta potentials and particle size were determined to describe the adsorption process of SLP on liposomes. The liposomes completely coated with SLP were observed by transmission electron microscope. To investigate the stabilizing effects of SLP on liposomes, carboxyfluorescein (CF) was encapsulated and its leakage was determined as an evaluation index. The results showed that the L. acidophilus ATCC 4356 SLP significantly (P < 0.05) increased the stability of the liposomes in the course of thermal challenge. Furthermore, SLP was able to reduce the aggregation of liposomes in serum. Storage stability of liposomes was performed at 25 °C, 4 °C and −20 °C for 90 days. And the SLP-coated liposomes released less CF than the control liposomes during storage at the three evaluated temperatures. Our findings extended the application field of Lactobacillus SLPs and introduced a novel nanocarrier system with good chemical stability.

Interactions of natural deep eutectic solvents (NADES) with artificial and natural membranes

Natural deep eutectic solvents (NADES) are a third type of liquids, apart from water and lipids. NADES show promise as drug carriers in dermal formulations or formulations for local administration in the oral cavity due to their low toxicity, tunability and biodegradability as well as their solubilizing and stabilizing properties. However, a thorough investigation of their effects in a biological system is needed. The present study aimed to investigate the physicochemical properties of six selected NADES and dilutions thereof. Further, the suitability of NADES as vehicles in topical preparations was evaluated through the application of liposome model membranes and ex vivo pig skin models. The effects of NADES on the membrane stability of egg-phosphatidylcholine (Egg-PC) liposomes and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine 16:0 PC (DPPC) liposomes were investigated by physical stability studies, membrane permeability studies, calorimetric analysis and visual representation by transmission electron microscopy (TEM). The liposomes appeared in most cases physically stable for 24 h in NADES, although there were changes in size and morphology most likely related to increased osmolality. Some NADES were also able to disaggregate formed DPPC liposome aggregates. The ex vivo permeation studies indicated a slightly reduced permeability of chloramphenicol through the pig skin after NADES application, which most likely was caused by hydrogen bonding between the NADES and proteins in the skin based on FT-IR-results. Altogether, our findings suggest that NADES are promising drug carriers in new drug delivery systems for topical applications.

A single-liposome assay that enables temperature-dependent measurement of proton permeability of extremophile-inspired lipid membranes

Thermoacidophilic archaea are able to survive in low pH environments by maintaining large proton gradients across their cell membranes. The unique structure of the unipolar and bipolar lipids that compose these membranes, with branched isoprene chains and ether linkages, may contribute to their reduced permeability to protons at elevated temperature. Here we present a proton permeability assay that interrogates hundreds of surface-tethered single liposomes in parallel using total internal reflection fluorescence (TIRF) microscopy to determine a proton permeability value for each liposome. The large number of liposomes that can be individually interrogated by this microscopy platform allow for the collection and statistical analysis of large quantities of data within a single experiment, as opposed to the single, averaged result produced by a bulk fluorimetry assay. In addition, tethering individual liposomes to a surface prevents liposome aggregation in free solution, which is especially prevalent when performing studies at elevated temperatures. Using a temperature-controlled chamber, we demonstrated a decreased temperature-dependence of proton permeability for bipolar archaea-mimetic membranes in comparison to unipolar eukaryote-mimetic membranes. Eyring-Polanyi analysis revealed a high entropic cost associated with proton flux across the archaea-mimetic membranes. Furthermore, analysis of the distribution of proton permeability values within populations of liposomes revealed a positive correlation between size and permeability in the eukaryote-mimetic liposomes that was absent in the archaea-mimetic liposomes.

The degree and position of phosphorylation determine the impact of toxic and trace metals on phosphoinositide containing model membranes

This work assessed effects of metal binding on membrane fluidity, liposome size, and lateral organization in biomimetic membranes composed of 1 mol% of selected phosphorylated phosphoinositides in each system. Representative examples of phosphoinositide phosphate, bisphosphate and triphosphate were investigated. These include phosphatidylinositol-(4,5)-bisphosphate, an important signaling lipid constituting a minor component in plasma membranes whereas phosphatidylinositol-(4,5)-bisphosphate clusters support the propagation of secondary messengers in numerous signaling pathways. The high negative charge of phosphoinositides facilitates electrostatic interactions with metals. Lipids are increasingly identified as toxicological targets for divalent metals, which potentially alter lipid packing and domain formation.Exposure to heavy metals, such as lead and cadmium or elevated levels of essential metals, like cobalt, nickel, and manganese, implicated with various toxic effects were investigated. Phosphatidylinositol-(4)-phosphate and phosphatidylinositol-(3,4,5)-triphosphate containing membranes are rigidified by lead, cobalt, and manganese whilst cadmium and nickel enhanced fluidity of membranes containing phosphatidylinositol-(4,5)-bisphosphate. Only cobalt induced liposome aggregation. All metals enhanced lipid clustering in phosphatidylinositol-(3,4,5)-triphosphate systems, cobalt in phosphatidylinositol-(4,5)-bisphosphate systems, while all metals showed limited changes in lateral film organization in phosphatidylinositol-(4)-phosphate matrices. These observed changes are relevant from the biophysical perspective as interference with the spatiotemporal formation of intricate domains composed of important signaling lipids may contribute to metal toxicity.