Lipid Phase Research Articles

Iron-Induced Lipid Oxidation Alters Membrane Mechanics Favoring Permeabilization.

Ferroptosis is a form of regulated necrosis characterized by the iron-dependent accumulation of lipid peroxides in cell membranes. However, how lipid oxidation via iron-mediated Fenton reactions affects the biophysical properties of cellular membranes and how these changes contribute to the opening of plasma membrane pores are major questions in the field. Here, we characterized the dynamics of membrane alterations during lipid oxidation induced onsite by Fenton reactions in chemically defined in vitro model membrane systems. We find that lipid vesicle permeabilization kinetically correlates with the appearance of malondialdehyde (MDA), a product of lipid oxidation. Iron-induced lipid oxidation also alters the lateral organization of supported lipid bilayers (SLBs) with lipid phase coexistence in a time-dependent manner, reducing the lipid phase mismatch and the circularity of liquid ordered domains, which indicates a decrease in line tension at the phase boundaries. Further analysis of oxidized SLBs by force spectroscopy reveals a significant decrease in the average membrane breakthrough force upon oxidation, resulting from changes in lipid bilayer organization that make it more susceptible to permeabilization. Our findings suggest that lipid oxidation via iron-mediated Fenton-like reactions induces strong changes in membrane lipid interactions and mechanical properties leading to reduced line tension in the permeabilized state of the bilayer, which promotes membrane pore formation.

Development and Optimization of Transferosomal Gel for Efficient Topical Delivery of Berberine Hydrochloride

Background: Berberine is an isoquinoline alkaloid with potent anti-inflammatory effects. However, its therapeutic efficacy is often restricted by its poor solubility, absorption, and permeability, especially in topical applications. Transferosomes are elastic vesicular carriers with high skin permeability values and retention, making them suitable for encapsulating hydrophilic and lipophilic actives. Objective: The objective of this research was to develop a transferosome-based topical gel formulation of Berberine hydrochloride (BER) to improve its skin permeability and anti-inflammatory efficacy. Method: The thin film hydration method was used to formulate the BER transferosomes. The effects of independent variables, amount of BER in lipid phase (X1), and lipid (Phospholipon 90G) to surfactant ratio (X2) on BER entrapment and vesicle size were studied using face-centered central composite design. The characterization was performed using differential scanning calorimetry, transmission electron microscopy, and X-ray diffraction. The optimized batch (F5) was incorporated in Carbopol gel and further investigated for viscosity, in vitro and ex-vivo diffusion, skin retention by tape stripping, and in-vivo anti-inflammatory efficiency. Results: The formulation optimized with 50 mg of drug and a 5:1 lipid-to-surfactant ratio (F5) demonstrated higher drug entrapment efficiency (72.11%) and lower vesicle size (77.9 nm). TEM validated the spherical vesicle morphology, whereas DSC and XRD analysis confirmed the molecular entrapment of BER within the phospholipid vesicles. The transferosomal gel demonstrated improved BER diffusion (0.63 mg/cm2) confirmed by in vitro and ex-vivo diffusion experiments that revealed a 6-fold increase in flux and permeability coefficient (0.1053 mg. cm-2.h-1). The drug release from transferosome gel was non-Fickian in nature (n = 0.6575), indicating an integration of diffusion and erosion processes. Furthermore, BER transferosomal gel displayed substantial anti- inflammatory activity in rats (p < 0.001). result: The independent variables exerted a substantial impact on drug entrapment and transferosome vesicle size. The optimized batch (F5) exhibited 72.11 % of BER entrapment and 77.9 nm vesicle size. DSC studies indicated molecular dispersion of BER in phospholipon G. The change in BER crystalline state was indicated by the XRD study. In vitro, diffusion study of Transferosome gel formulation indicated the sustained release by a non-fickian mechanism. Ex vivo skin permeation revealed significant improvement in BER permeation (p< 0.05). Further, the tape-stripping method revealed higher BER skin permeability. In-vivo anti-inflammatory efficacy of BER transferosome gel was significantly higher as compared to plain BER gel. Conclusion: The findings demonstrated the potential of transferosomal gel as a promising approach for efficient drug delivery and therapeutic efficacy.

Evaluating the Impact of the Membrane Thickness on the Function of the Intramembrane Protease GlpG

Cellular membranes exhibit a huge diversity of lipids and membrane proteins that differ in their properties and chemical structure. Cells organize these molecules into distinct membrane compartments characterized by specific lipid profiles and hydrophobic thicknesses of the respective domains. If a hydrophobic mismatch occurs between a membrane protein and the surrounding lipids, there can be functional consequences like reduced protein activity. This phenomenon has been extensively studied for single-pass transmembrane proteins, rhodopsin, and small polypeptides such as gramicidin. Here we investigate the E. coli rhomboid intramembrane protease GlpG as a model to systematically explore the impact of membrane thickness on GlpG activity. We used fully saturated dilauroylpohosphatidylcholine (DLPC) and dimyristoylphosphatidylcholine (DMPC) model lipids and altered membrane thickness by varying the cholesterol content. Physical membrane parameters were determined by 2H and 31P NMR spectroscopy and correlated with GlpG activity measurements in the respective host membranes. Differences in bulk and annular lipids as well as alterations in protein structure in the respective host membranes were determined using molecular dynamics simulations. Our findings indicate that GlpG can influence the membrane thickness in DLPC/cholesterol membranes but not in DMPC/cholesterol membranes. Moreover, we observe that GlpG protease activity is reduced in DLPC membranes at low cholesterol content, which was not observed for DMPC. While a change in GlpG activity can already be due to smallest differences in the lipid environment, potentially enabling allosteric regulation of intramembrane proteolysis, there is no overall correlation to cholesterol-mediated lipid bilayer organization and phase behavior. Additional factors such as the influence of cholesterol on membrane bending rigidity and curvature energy need to be considered. In conclusion, the functionality of α-helical membrane proteins such as GlpG relies not only on hydrophobic matching but also on other membrane properties, specific lipid interaction and the composition of the annular layer.

Microscopic water dispersion and hydrogen-bonding structures in margarine spreads with Raman hyperspectral imaging and machine learning

Margarine, a water-in-oil (W/O) emulsion, offers advantages such as lower costs in comparison to similar products, but large amounts of saturated fats pose health risks. Reduction of saturated fat content is difficult and often leads to “oil-off,” i.e., the seepage of liquid oil from the mixture, resulting in undesirable appearance and texture. Investigations into the phenomenon have often focused on morphology at the water-oil interfaces, and this work establishes Raman imaging as a powerful application for observing microscopic morphologies of W/O emulsions. We analyze morphologies of 5 distinct margarine spreads that differ in manufacturing date, formulation, and manufacturing process. More robust H-bonding in the oil phase of the emulsions co-occurred with smaller amounts of oil-off, suggesting that H-bonding interactions between emulsifier molecules, water, and crystallized fats in the lipid phase of the W/O emulsions results in an emulsion that is less susceptible to the production of oil-off.

Improved Transdermal Delivery of Anti-hypertensive Drug Loaded Nanostructured Lipid Carriers: Statistical Design, Optimization, Depiction and Pharmacokinetic Assessment

Background: The vasoselective calcium-channel blocker lercanidipine hydrochloride (LCH) is poorly absorbed orally (only 10% bioavailability) owing to its low solubility and hepatic metabolism. Because of the LCH's poor solubility and permeability, bioavailability is low and very variable, stable aqueous liquid formulations are challenging to create, and a uniform distribution of the medication is almost impossible to produce. Objective: The purpose of this research was to see whether an approach involving the development of nanostructured lipid carriers (NLCs) might be used to create an effective, innovative oral formulation of LCH. The efficacy of several synthetic and natural liquid lipids was compared using a hot homogenization-ultrasonication strategy. Methods: Following initial improvements with hot homogenization and ultrasonication, the LCHloaded NLCs formulation was fine-tuned by Box-Behnken statistical analysis. The optimal LCHNLCs composition includes the lipid phase (2-4% w/v) of stearic acid and oleic acid, the surfactants poloxamer 188 (1%) and Tween 80(1%), and other ingredients. Results: The optimized NLCs formulation was found to have mean vesicle sizes of 128.72 ± 1.59 nm, polydispersity indices of 0.169 ± 0.06, zeta potentials of -36.81 ± 0.42 mV, and entrapment efficiencies of 79.84 ± 0.11%. The optimized NLCs formulation released much more LCH (88.74 ± 4.62) than the LCH-suspension (36.84 ± 0.37%) in in-vitro drug release experiments lasting up to 24 hours. Ex vivo studies on the ability of LCH-NLCs to pass through the gut showed that drug permeation was much better than it was with plain LCH-solution. The in vivo pharmacodynamic analysis demonstrated that, compared to conventional LCH-suspension, NLCs released LCH more slowly and steadily over a longer time period. Conclusion: These findings provide additional evidence that NLCs have great promise as a drug delivery technology for the treatment of hypertension, just as they show promise as a controlled release formulation for the treatment of LCH.

Cubosomes hydrogel containing silver based organometallic compounds for enhanced wound healing of burns: In vitro and in vivo studies

The aim of this study is to investigate the treatment of burn wounds and bacterial infections, specifically focusing on silver(I) N-heterocyclic carbene (Ag-NHC) loaded into cubosomes. Ag-NHC is an organometallic complex of silver (Ag) and can be considered an active pharmaceutical ingredient for the topical treatment of burns. A novel nanoparticle system, cubosome dispersions, was formulated using high pressure homogenization methods with different concentrations of lipid phase glyceryl monooleate (GMO), surfactants such as poloxamer 407 (F-127), and polyvinyl alcohol (PVA). The optimized formulation (U6) was characterized by in vitro drug release profile (90.54 ± 1.17 %), particle size (126.7 ± 0.98 nm), zeta potential (−21.6 ± 6.83 mV), entrapment efficiency (89.30 ± 1.46 %), and morphology as observed by transmission electron microscopy (TEM). The U6 formulation was incorporated into a hydrogel containing Carbopol 940 to form a cubosomal hydrogel (cubogel). The cubogel was characterized by the in vitro release of Ag-NHC (92 %), entrapment efficiency (>90 %), pH (5.3), viscosity (1.2 cP), spreadability, and TEM. From the in vitro drug release pattern, it was concluded that the U6 formulation showed slow and sustained drug release over 24 h, and demonstrated superior burn healing compared to marketed products. The in vivo wound healing study revealed that the cubogel demonstrated a superior burn healing rate compared to commercially available products (Quench® and Clotrimazole®). The in vivo histopathological results showed that the prepared cubogel was effective in treating second-degree burns, with no side effects or skin irritation, compared to commercial products. Additionally, Ag-NHC-loaded cubogel facilitated sustained release for treating burn wounds without any bacterial infections. The current findings provide a strong basis for the initiating phase 0 clinal trials, highlighting significant potential for the further research to benefit the pharmaceutical industry.

Niosomes: A promising approach for targeted drug delivery

Niosomes, also known as nonionic surfactant vesicles, are small lamellar structures that are created by combining nonionic surfactants from the alkyl or dialkyl polyglycerol ether category with cholesterol, and then hydrating them in water-based solutions. These are vesicle systems resembling liposomes that can be utilized as carriers for drugs that are both amphiphilic and lipophilic. The production process of niosomes is derived from liposome technology. The fundamental manufacturing method remains unchanged, wherein the lipid phase is hydrated by the aqueous phase. The lipid phase can consist of either a pure surfactant or a combination of surfactant and cholesterol. Niosomes effectively tackle the challenges associated with medication insolubility, instability, inadequate bioavailability, and fast degradation. The amphiphilic character of niosomes, which combines both hydrophilic and lipophilic properties, enhances their capacity to encapsulate medicines that are either hydrophilic or lipophilic. Cholesterol is frequently utilized as one of the ingredients. Preserving the stiffness of the niosome structure. The present article discusses the essential elements of niosomes, including their structural constituents, methods of manufacturing, and their uses in different disorders.

Environmentally Friendly Microemulsions of Essential Oils of Artemisia annua and Salvia fruticosa to Protect Crops against Fusarium verticillioides.

Essential oils (EOs) are reported to be natural pesticides, but their use to protect crops is very limited due to EOs' high instability and great volatility. Nanovectors represent a very smart alternative, and in this study, EOs from Artemisia annua (AEO) and Salvia fruticosa (SEO) were formulated into microemulsions and tested against Fusarium verticillioides. The EOs were extracted by steam distillation and analyzed by GC-MS. The main constituents of AEO were camphor, artemisia ketone, and 1,8-cineole; the main constituents of SEO were 1,8-cineole, camphor, α-pinene, and β-pinene. Artemisia ketone and 1,8-cineole were used to calculate the recovery and chemical stability of the microemulsions. The microemulsions were loaded with 10 mg/mL of EOs, and the recoveries were 99.8% and 99.6% for AEO and SEO, respectively. The sizes of the lipid phases were 255.3 ± 0.6 nm and 323.7 ± 2.3 nm for the AEO and SEO microemulsions, respectively. Activity against F. verticillioides was tested using amphotericin B as the positive control. F. verticillioides was very susceptible to both EOs. When loaded in the microemulsions, AEO and SEO remained very active at a dose of 1.4 and 1.2 mg, with a 99.99% reduction of F. verticillioides. The findings suggest AEO and SEO microemulsions are suitable carriers for the protection of crops against F. verticillioides.

Insights into the phase behavior at interfaces using vibrational sum frequency generation spectroscopy.

Phase separation is ubiquitous at the interface between two distinct phases. Physical transformation during phase separation often plays a crucial role in many important mechanisms, such as lipid phase separation, which is fundamental for transport through biological membranes. Phase separation can be complex, involving changes in the physical state and the reorganization of molecular structures, influencing the behavior and function of materials and biological systems. Surface-sensitive vibrational sum frequency generation (VSFG) spectroscopy provides a powerful tool for investigating these interfacial processes. As a non-linear optical technique, VSFG spectroscopy is sensitive to changes in molecular orientation and interactions at interfaces, making it an ideal method for studying phase separation processes. Here, we review the molecular interaction mechanisms underlying phase separation. We also explore the application of VSFG spectroscopy in studying phase separation processes at different interfaces. In particular, we focus on oil-water interfaces, which are relevant in environmental and industrial contexts; polymer and lipid surfaces, important for materials science and biological membranes; and intrinsically disordered protein systems, which play key roles in cellular function and disease.

Sheet-on-sheet fixed target data collection devices for serial crystallography at synchrotron and XFEL sources

Serial crystallography (SX) efficiently distributes over many crystals the radiation dose absorbed during diffraction data acquisition, enabling structure determination of samples at ambient temperature. SX relies on the rapid and reliable replacement of X-ray-exposed crystals with fresh crystals at a rate commensurate with the data acquisition rate. `Solid supports', also known as `fixed targets' or `chips', offer one approach. These are microscopically thin solid panes into or onto which crystals are deposited to be individually interrogated by an X-ray beam. Solid supports are generally patterned using photolithography methods to produce a regular array of features that trap single crystals. A simpler and less expensive alternative is to merely sandwich the microcrystals between two unpatterned X-ray-transparent polymer sheets. Known as sheet-on-sheet (SOS) chips, these offer significantly more versatility. SOS chips place no constraint on the size or size distribution of the microcrystals or their growth conditions. Crystals ranging from true nanocrystals up to microcrystals can be investigated, as can crystals grown in media ranging from low viscosity (aqueous solution) up to high viscosity (such as lipidic cubic phase). Here, we describe our two SOS devices. The first is a compact and lightweight version designed specifically for synchrotron use. It incorporates a standard SPINE-type magnetic base for mounting on a conventional macromolecular crystallography goniometer. The second and larger chip is intended for both X-ray free-electron laser and synchrotron use and is fully compatible with the fast-scanning XY-raster stages developed for data collection with patterned chips.

Double (W1/O/W2) emulsions prepared with natural vegetable oils and varied inner glucose concentration: Microstructure, rheology and stability

The objective was to study the microstructure, rheology and stability of double (W1/O/W2) emulsions prepared with Tween 80 as hydrophilic emulsifier and coconut, palm or sunflower oil. The effects of glucose concentration in inner aqueous phase and crystallization of lipid phase were analyzed. Aggregation of oil globules by partial coalescence was observed in emulsions prepared with coconut or palm oil because of the crystallization of the lipid phase. The increase of glucose concentration increased the amount of inner water by osmotic swelling and produced a higher partial coalescence degree due to a higher collision frequency between oil globules. Consequently, higher elastic and viscous modules were observed at higher glucose concentration. Fat crystals in lipid phase allowed a partial retention of inner water in the absence of glucose, but they also favored the release of inner water due to partial coalescence. Creaming stability was enhanced by the presence of glucose, because of the formation of a three-dimensional network of aggregated oil globules in emulsions with coconut or palm oil, and a higher packing and global density of oil globules in systems with sunflower oil. The obtained results contribute to the understanding of partial coalescence in W1/O/W2 emulsions, including its effects on inner water release and creaming stability. This work could be relevant for the design and stabilization of vegetable, lipid-reduced food emulsions prepared with natural oils.

Entangling roles of cholesterol-dependent interaction and cholesterol-mediated lipid phase heterogeneity in regulating listeriolysin O pore-formation.

Cholesterol-dependent cytolysins (CDCs) are the distinct class of β-barrel pore-forming toxins (β-PFTs) that attack eukaryotic cell membranes, and form large, oligomeric, transmembrane β-barrel pores. Listeriolysin O (LLO) is a prominent member in the CDC family. As documented for the other CDCs, membrane cholesterol is essential for the pore-forming functionality of LLO. However, it remains obscure how exactly cholesterol facilitates its pore formation. Here, we show that cholesterol promotes both membrane-binding and oligomerization of LLO. We demonstrate cholesterol not only facilitates membrane-binding, it also enhances the saturation threshold of LLO-membrane association, and alteration of the cholesterol-recognition motif in the LLO mutant (LLOT515G-L516G) compromises its pore-forming efficacy. Interestingly, such defect of LLOT515G-L516G could be rescued in the presence of higher membrane cholesterol levels, suggesting cholesterol can augment the pore-forming efficacy of LLO even in the absence of a direct toxin-cholesterol interaction. Furthermore, we find the membrane-binding and pore-forming abilities of LLOT515G-L516G, but not those of LLO, correlate with the cholesterol-dependent rigidity/ordering of the membrane lipid bilayer. Our data further suggest that the line tension derived from the lipid phase heterogeneity of the cholesterol-containing membranes could play a pivotal role in LLO function, particularly in the absence of cholesterol binding. Therefore, in addition to its receptor-like role, we conclude cholesterol can further facilitate the pore-forming, membrane-damaging functionality of LLO by asserting the optimal physicochemical environment in membranes. To the best of our knowledge, this aspect of the cholesterol-mediated regulation of the CDC mode of action has not been appreciated thus far.

Probing Deuteration-Induced Phase Separation in Supported Lipid Monolayers using Hyperspectral TERS Imaging.

In this study, we investigate the impact of deuteration on the formation of phase-separated domains in supported lipid monolayers using hyperspectral Tip-Enhanced Raman Spectroscopy (TERS) imaging. The intricate organization of biological membranes plays a crucial role in cellular functions. Various factors that influence domain formation have been identified in previous studies such as lipid tail length and cholesterol concentration. Deuterium labeling of lipids has proven useful for probing cellular structures and dynamics, but its impact on lipid phase separation remains underexplored. By examining 1:1 mixed monolayers of dipalmitoylphosphatidylcholine (DPPC) and deuterated DPPC on Au(111) surfaces, we reveal partial segregation of domains rich in deuterated and nondeuterated lipids. This study addresses a gap in knowledge by examining the impact of deuteration on lipid tail behavior, offering new insights into how even subtle structural modifications can influence phase behavior. Furthermore, it demonstrates that TERS can be a powerful, nondestructive, and label-free nanoanalytical tool for analyzing lipid membranes and advance the field of membrane biophysics.

The effects of plant flavones on the membrane boundary potential and lipid packing stress

Here we have revealed the effects of different plant flavones on the physicochemical properties of model lipid membranes. We have demonstrated that baicalein increases the boundary potential of membranes composed of phosphatidylcholine, while wogonin does not affect it. Other flavones tested reduce membrane boundary potential, with this ability increasing among scutellarein, chrysin, apigenin, morin, fisetin, and luteolin. Molecular dynamics simulations demonstrate connection of alteration in boundary potential with the preferential orientation of intrinsic flavone dipole moments in membranes. We have also shown that flavones reduce the melting point of phosphatidylcholine, and this ability increases in the series of luteolin, morin, wogonin, scutellarein, apigenin, baicalein, chrysin, and fisetin. The introduction of baicalein, chrysin and fisetin also leads to a significant decrease in the sharpness of the lipid phase transition. We have hypothesized that the localization of flavones in the glycerol backbone or in the C1-C8 methylene region of lipid hydrocarbon chains leads to an increase in the area per lipid and, as a consequence, to an expansion of the lipid melting peak. Replacement of neutral phosphatidylcholine with negatively charged phosphatidylserine affects the membrane-modifying activity of flavones which given the externalization of phosphatidylserine on the surface of cancer cells may be crucial in the flavone anticancer effects.

The Influence of Processing Using Conventional and Hybrid Methods on the Composition, Polysaccharide Profiles and Selected Properties of Wheat Flour Enriched with Baking Enzymes.

In this study, a developed wheat flour blend (F), consisting of a high content of non-starch polysaccharides, was fortified with cellulase (C) and a cellulase-xylanase complex (CX) and then processed via conventional and hybrid treatment methods. Dry heating (T), hydrothermal treatment (H) and extrusion processing (E) were applied without or with enzyme addition as hybrid treatments. Proximate composition and polysaccharide profiles selected techno-functional and structural properties of modified wheat flours, were analyzed. Conventional and hybrid treatments induced changes in polysaccharide fraction compositions (especially the arabinoxylans) and the rheology of modified flour. Dry heating caused an inconsiderable effect on flour composition but reduced its baking value, mainly by reducing the elasticity of the dough and worsening the strain hardening index, from 49.27% (F) to 44.83% (TF) and from 1.66 (F) to 1.48 (TF), respectively. The enzymes added improved the rheological properties and baking strength, enhancing the quality of gluten proteins. Hydrothermal enzyme-assisted treatment increased flour viscosity by 14-26% and improved the dough stability by 12-21%; however, the use of steam negatively affected the protein structure, weakening dough stretchiness and elasticity. Extrusion, especially enzyme-assisted, significantly increased the hydration properties by 55-67% but lowered dough stability, fat content and initial gelatinization temperature due to the changes in the starch, mostly induced by the hybrid enzymatic-extrusion treatment. The structure of extruded flours was different from that obtained for other treatments where the peak intensity at 20° was the highest, suggesting the presence of amorphous phases of amylose and lipids. The results can be helpful in the selection of processing conditions so as to obtain flour products with specific techno-functional properties.

Lipid Selectivity of Membrane Action of the Fragments of Fusion Peptides of Marburg and Ebola Viruses.

The life cycle of Ebola and Marburg viruses includes a step of the virion envelope fusion with the cell membrane. Here, we analyzed whether the fusion of liposome membranes under the action of fragments of fusion peptides of Ebola and Marburg viruses depends on the composition of lipid vesicles. A fluorescence assay and electron microscopy were used to quantify the fusogenic activity of the virus fusion peptides and to identify the lipid determinants affecting membrane merging. Differential scanning calorimetry of lipid phase transitions revealed alterations in the physical properties of the lipid matrix produced by virus fusion peptides. Additionally, we found that plant polyphenols, quercetin, and myricetin inhibited vesicle fusion induced by the Marburg virus fusion peptide.

High-throughput Approaches to Detect Cytokines and Chemokines Secretion by Both Autophagy and Endosome Vesicle Fusion of HBMEC by Knocking Down the NLRP2 Gene

Background: Given the rapid increase in serious infectious and inflammatory diseases in the general population worldwide, it is vital to advance our understanding of the process of infection and inflammation. Objective: These illnesses can be treated by diverse cytokines, chemokines, and chemical compounds, but how to find and identify effective protein drugs is unknown. In this study, large–scale analytical approaches, such as high–throughput chips with RNA-Seq that construct the protein regulatory networks, were established. Methods: NLRP2:shRNA (1–3) and GFP: Control lentivirus were fabricated to infect Human Brain Microvascular Endothelial Cells (HBMEC) to knock down NLRP2 gene that not only possesses biochemical activity but also bio-mechanical properties. Once HBMEC was loaded on the 18mmX18mm circular soft cover slip, by knocking down NLRP2 gene processing and adding puromycin, a large number of cytokines were secreted to activate both autophagy and endosome vesicle signaling by KEGG pathway analysis, which was successfully detected by both ELISA approach and QAH-Neu-1 chip. However, no secretion from GFP: Control was reported. Results: It was demonstrated that the NLRP2 gene was highly responsive to cytokines. The protein regulatory network from RNA–Seq platform demonstrated that the secretion by knocking down the NLRP2 gene was highly correlated with autophagy, endosome, multivesicular body, phosphatidylinositol, and necrosis signalling pathway. Furthermore, most of the cytokines expressed were found to be specific for intracellular vesicle–dependent secretion, leading to obvious cell swelling and shedding, membrane protein dotting to nucleation, and actin dynamics. Interestingly, it was also found that autophagy, together with endosome signalling, was collectively activated to boost the secretion to cause a “cytokine storm”, which led to lipid phase separation. Conclusion: This study proposed high throughput approaches centered on the NLRP2 gene network for many severe diseases, providing novel insight into biological pathways influencing infection and inflammation (e.g., COVID-19/E.coli). They can be modulated as potential therapeutic targets and used as biomarkers in the diagnosis and treatment of many diseases to promote human health. This increases our interest in developing new leadless-peptides at the genomics and proteomics levels to obtain cytokines and chemokines for micro-array constructions (VirD‒cytokines, VirD‒enzymes), antibody and vaccine development for infections, diabetes, cardiovascular disease, atherosclerosis, non–alcoholic fatty liver, auto-immune, neurodegenerative disorder, and even cancer-related disease therapy. They have the most valuable applications in molecular diagnosis, protein marker discovery, and bio-therapy.

Mechanism of two styryl BODIPYs as fluorescent probes and protective agents in lipid bilayers against aqueous ClO.

Two styryl BODIPY derivatives, BOH and BOE, with different hydrophilic properties, were investigated for their reaction mechanisms in lipid bilayers against aqueous ClO-, by both experimental and theoretical methods. Density functional theory (DFT) calculations confirmed their identical conformations in solution. Fluorescence spectra and high-resolution mass spectra corroborated the central vinyl group as a common antioxidation moiety against ClO- oxidation. In giant unilamellar vesicles (GUVs), distinct reaction kinetics with ClO- suggested that BOE provided superior protective effects compared to BOH on lipids. Molecular dynamics simulations indicated that the lipophilic octyloxy group in BOE led to its deeper localization within the lipid phase, bringing it closer to the corresponding lipid target group. This study establishes the two styryl BODIPYs as promising fluorescent probes for detecting aqueous ClO- in lipid-water polyphasic systems.

Bisphenol F and Bisphenol S in a Complex Biomembrane: Comparison with Bisphenol A.

Bisphenols are a group of endocrine-disrupting chemicals used worldwide for the production of plastics and resins. Bisphenol A (BPA), the main bisphenol, exhibits many unwanted effects. BPA has, currently, been replaced with bisphenol F (BPF) and bisphenol S (BPS) in many applications in the hope that these molecules have a lesser effect on metabolism than BPA. Since bisphenols tend to partition into the lipid phase, their place of choice would be the cellular membrane. In this paper, I carried out molecular dynamics simulations to compare the localization and interactions of BPA, BPF, and BPS in a complex membrane. This study suggests that bisphenols tend to be placed at the membrane interface, they have no preferred orientation inside the membrane, they can be in the monomer or aggregated state, and they affect the biophysical properties of the membrane lipids. The properties of bisphenols can be attributed, at least in part, to their membranotropic effects and to the modulation of the biophysical membrane properties. The data support that both BPF and BPS, behaving in the same way in the membrane as BPA and with the same capacity to accumulate in the biological membrane, are not safe alternatives to BPA.

An in vitro study for reducing the cytotoxicity and dose dumping risk of remdesivir via entrapment in nanostructured lipid carriers

The aim of this study was to synthesize and evaluate nanostructured lipid carriers (NLCs) loaded with Remdesivir (RDV) to control its side effects in COVID-19 patients. Due to the low solubility and short half-life of RDV in the blood, an injectable formulation was prepared using sulphobutylether-beta-cyclodextrin. However, it can accumulate in the kidney and cause renal impairment. NLCs improve the parenteral delivery of hydrophobic drugs such as RDV by increasing drug solubility and bioavailability. For the synthesis of RDV-NLCs, the aqueous phase containing Tween 80 was injected into the lipid phase under rapid stirring and was sonicated. The experimental conditions were optimized using Box-Behnken design and Design Expert software. The optimum formulation contained a total lipid of 2.13%, a total surfactant of 1%, and a hot bath time of 71 min. The optimum formulation showed particle size, polydispersity index, zeta potential, and entrapment efficiency values of 151.0 ± 1.7 nm (from 149.1 to 152.1), 0.4 ± 0.1 (from 0.3 to 0.5), −43.8 ± 1.2 mV (from −42.4 to −44.7), and 81.34 ± 1.57% (from 79.52 to 82.33%), respectively. RDV-NLCs showed acceptable stability for 30 days at 25 ℃ and were compatible with commonly used intravenous infusion fluids for 48 h. FE-SEM images of RDV-NLC showed spherical particles with a mean diameter of 207 nm. The NLC-RDV formulation showed a sustained release of RDV with a low risk of dose-dumping, minimizing potential side effects. In addition, RDV in the form of RDV-NLC causes less cytotoxicity to healthy normal kidney cells, which is expected to reduce renal impairment in COVID-19 patients.