Lipid Phase Research Articles

NANOSTRUCTURED LIPID CARRIERS AS A PROMISING DERMAL DELIVERY PLATFORM FOR ST. JOHN'S WORT EXTRACT: PRELIMINARY STUDIES

urpose: Nanostructured lipid carriers (NLC) can improve the stability of various phytochemicals, so this research aimed to develop and employ such delivery systems for Hypericum perforatum extract containing the light- and oxygen-sensitive phloroglucinol hyperforin. Materials and methods: By varying the processing parameters and the solid and liquid lipids used, different NLC models were obtained via emulsification, followed by high-shear homogenization and ultrasonication. After characterization of the elaborated nanocarriers, those that exhibited optimal properties were loaded with St. John's wort extract by incorporating it in the lipid phase before emulsification and also studied. Results: The NLC models developed by prolonged ultrasonication demonstrated favorable characteristics regarding particle size, polydispersity index (PDI), and zeta potential (ZP). The physically stable during long-term storage nanosystems, which also showed a reduced degree of crystallinity of the solid lipid included, were loaded with St. John's wort extract. Of the carriers so-obtained, those developed by ultrasonication at ambient temperatures can be described as acceptably uniform systems (PDI from 0.23±0.01 to 0.27±0.01), comprising particles with dimensions below 200 nm, favorable ZP values (ζ > |30 mV|), and superior entrapment efficacy (EE) greater than 85%. Conclusion: In this research, different stable NLC dispersions were successfully developed. After the inclusion of the St. John's wort extract, the model possessing the highest EE (87.77±0.64%) was chosen as a carrier to conduct further studies evaluating its wound-healing potential.

Assessment of Purity, Functionality, Stability, and Lipid Composition of Cyclofos-nAChR-Detergent Complexes from Torpedo californica Using Lipid Matrix and Macroscopic Electrophysiology

The main objective of the present study was to find detergents that can maintain the functionality and stability of the Torpedo californica nicotinic acetylcholine receptor (Tc-nAChR). We examined the functionality, stability, and purity analysis of affinity-purified Tc-nAChR solubilized in detergents from the Cyclofos (CF) family [cyclofoscholine 4 (CF-4), cyclofoscholine 6 (CF-6), and cyclofloscholine 7 (CF-7)]. The functionality of the CF-Tc-nAChR-detergent complex (DC) was evaluated using the Two Electrode Voltage Clamp (TEVC) method. To assess stability, we used the florescence recovery after photobleaching (FRAP) in Lipidic Cubic Phase (LCP) methodology. We also performed a lipidomic analysis using Ultra-Performance Liquid Chromatography (UPLC) coupled to electrospray ionization mass spectrometry (ESI–MS/MS) to evaluate the lipid composition of the CF-Tc-nAChR-DCs. The CF-4-Tc-nAChR-DC displayed a robust macroscopic current (− 200 ± 60 nA); however, the CF-6-Tc-nAChR-DC and CF-7-Tc-nAChR-DC displayed significant reductions in the macroscopic currents. The CF-6-Tc-nAChR and CF-4-Tc-nAChR displayed higher fractional florescence recovery. Addition of cholesterol produced a mild enhancement of the mobile fraction on the CF-6-Tc-nAChR. The lipidomic analysis revealed that the CF-7-Tc-nAChR-DC displayed substantial delipidation, consistent with the lack of stability and functional response of this complex. Although the CF-6-nAChR-DC complex retained the largest amount of lipids, it showed a loss of six lipid species [SM(d16:1/18:0); PC(18:2/14:1); PC(14:0/18:1); PC(16:0/18:1); PC(20:5/20:4), and PC(20:4/20:5)] that are present in the CF-4-nAChR-DC. Overall, the CF-4-nAChR displayed robust functionality, significant stability, and the best purity among the three CF detergents; therefore, CF-4 is a suitable candidate to prepare Tc-nAChR crystals for structural studies.Graphical abstract

Coupling of protein condensates to ordered lipid domains determines functional membrane organization.

During T cell activation, the transmembrane adaptor protein LAT (linker for activation of T cells) forms biomolecular condensates with Grb2 and Sos1, facilitating signaling. LAT has also been associated with cholesterol-rich condensed lipid domains; However, the potential coupling between protein condensation and lipid phase separation and its role in organizing T cell signaling were unknown. Here, we report that LAT/Grb2/Sos1 condensates reconstituted on model membranes can induce and template lipid domains, indicating strong coupling between lipid- and protein-based phase separation. Correspondingly, activation of T cells induces cytoplasmic protein condensates that associate with and stabilize raft-like membrane domains. Inversely, lipid domains nucleate and stabilize LAT protein condensates in both reconstituted and living systems. This coupling of lipid and protein assembly is functionally important, as uncoupling of lipid domains from cytoplasmic protein condensates abrogates T cell activation. Thus, thermodynamic coupling between protein condensates and ordered lipid domains regulates the functional organization of living membranes.

Cholesterol and melatonin regulated membrane fluidity does not affect the membrane breakage triggered by amyloid-beta peptide

We have studied by means of small angle neutron scattering and diffraction, and molecular dynamics simulations the effect of lipid membrane fluidity on the amyloid-beta peptide interactions with the membrane. These interactions have been discovered previously to trigger the reorganization of model membranes between unilamellar vesicles and planar membranes (bicelle-like structures) during the lipid phase transition. The morphology changes were taking place in rigid membranes prepared of fully saturated lipids and were proposed to play a role in the onset of amyloid related disorders. We show in this study that the replacement of fully saturated lipids by more fluid mono-unsaturated lipids eliminates the mentioned morphology changes, most likely due to the absence of phase transition within the temperature range investigated. We have therefore controlled the membrane rigidity also while ensuring the presence of membrane phase transition within the biologically relevant temperatures. It was done by the addition of melatonin and/or cholesterol to the initial membranes made of saturated lipids. Small angle neutron scattering experiments performed over a range of cholesterol and melatonin concentrations show their distinctive effects on the local membrane structure only. The cholesterol for example affects the membrane curvature such that spontaneously formed unilamellar vesicles are of much larger sizes than those formed by the neat lipid membranes or membranes with melatonin added. The temperature dependent experiments, however, reveal no influence on the previously discovered membrane breakage whether cholesterol or melatonin have been added.

Comment on lipid emulsion for xenobiotic overdose

We read with interest the assessment of Drs Fettiplace and Weinberg concerning antidotal use of intravenous lipid emulsion.1 While the authors suggest that lipophilicity of the toxicant is not likely to be a useful predictor of clinical benefit of intravenous lipid emulsion in a poisoned patient, French et al. convincingly disagree.2 Further, if the mechanism of action of intravenous lipid emulsion includes the lipid sink or lipid shuttle, it is likely that lipophilicity (measured by the octanol/water partition coefficient or log P) would be one explanatory variable for apparent effect (or lack thereof) after lipid emulsion. An example is organophosphate poisoning, which continues to be a major cause of poisoning morbidity and mortality, with estimates of 3 000 000 organophosphate poisonings and 300 000 organophosphate poisoning deaths worldwide.3, 4 A recent case report described successful lipid emulsion treatment of poisoning by chlorpyrifos (log P = 4.96).5 A pilot study of intravenous lipid emulsion in organophosphate poisoning in India indicated that lipid emulsion may be a safe therapeutic intervention in organophosphate poisoning, but the study lacked the power needed to evaluate for efficacy.6 The same research group recently published an open-label, randomized control trial of lipid emulsion for organophosphate poisoning. They again found lipid emulsion to be safe, but they did not detect benefit.7 Regarding the 2019 pilot study,6 Mullins et al. pointed out that the identified organophosphates varied widely in lipophilicity and recommended stratifying patients by the ingested organophosphate.8 In both the pilot study and the randomized controlled trial,6, 7 at least half of the patients had ingested unknown organophosphates (Table 1), so inferences about outcomes are difficult. In both studies, the most frequently identified organophosphate was chlorpyrifos (log P = 4.96), and the second most frequently identified organophosphate was dichlorvos (log P = 1.43). If log P does predict efficacy as previously suggested,2 chlorpyrifos should achieve concentrations in the lipid phase over 2000 times greater than dichlorvos would reach. It is likely that the unknown category includes chlorpyrifos and dichlorvos. If an outcome difference exists, the dichlorvos cases in the unknown category would probably obscure the possibly better outcomes among the chlorpyrifos cases. A new randomized, controlled trial of intravenous lipid emulsion for organophosphate poisoning is launching in Egypt.9 We recommend that these researchers report the log P values for all identified organophosphates and include subgroup analysis by organophosphate. Knowing both the toxicant and the treatment assignment for each organophosphate-poisoned patient is necessary to understand the results more clearly.

Application of sodium caseinate as protein based coating/stabilizing agent in development and optimization of vitamin D3-loaded nanostructured lipid carriers (NLCs)

Sodium caseinate is a food-grade protein that has several functional properties, such as acting as an emulsifying and stabilizing agent in a colloidal delivery system. Nanostructured lipid carriers (NLCs) have been proven to be highly effective carriers for encapsulating bioactive compounds. In this research, several novel food-grade NLC systems based on sodium caseinate (as a protein surfactant), anhydrous milk fat (as a solid lipid), and vitamin E acetate (as a liquid oil) were developed as an encapsulant for vitamin D3. The D-optimal combined design was employed to optimize the effects of lipid phase ratios and protein concentrations on particle size and loading efficiency. The optimized formulation obtained from the predicted model was composed of 1% w/v caseinate protein and a 73.23/26.78 solid lipid/liquid oil ratio with a desirability function of 0.983. This optimum formulation showed the smallest particle size at 120.16 nm with a narrow size distribution of 0.208 and an encapsulation efficiency of 96.14%. The cytotoxicity study showed that the anticancer effect of the vitamin was improved after incorporation into the NLC structure. The current study suggests that this composition of biopolymeric surfactant and applied lipid matrix could be an effective, health-promoting, and promising carrier system for developing food-grade bioactive delivery systems.

Unusual phosphatidylcholine lipid phase behavior in the ionic liquid ethylammonium nitrate

HypothesisThe forces that govern lipid self-assembly ionic liquids are similar to water, but their different balance can result in unexpected behaviour. ExperimentsThe self-assembly behaviour and phase equilibria of two phospholipids, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), in the most common protic ionic liquid, ethylammonium nitrate (EAN) have been investigated as function of composition and temperature by small- and wide-angle X-ray scattering (SAXS/WAXS) and small-angle neutron scattering (SANS). FindingsBoth lipids form unusual self-assembly structures and show complex and unexpected phase behaviour unlike that seen in water; DSPC undergoes a gel Lβ to crystalline Lc phase transition on warming, while POPC forms worm-like micelles L1 upon dilution. This surprising phase behaviour is attributed to the large size of the EAN ions that solvate the lipid headgroup compared to water changing amphiphile packing. Weaker H-bonding between EAN and lipid headgroups also contributes. These results provide new insight for the design of lipid based nanostructured materials in ionic liquids with atypical properties.

Novel Approach Self Nanoemulsifying Drug Delivery System: A Review

The self-nanoemulsifying drug delivery system is solitarily the most frequent dosage form for administering drug molecules towards the target region in the pharmaceutical industry. It has gotten a lot of interest in the current scenario because of its budding applications in various industries. In the pharmaceutical business, self-nanoemulsifying drug delivery systems have been used to distribute medications via oral, topical, and parenteral routes. SNEDDS are submicron-sized emulsions being researched as drug carriers to improve therapeutic agent delivery. Due to their tiny droplets, elevated solubility ability, broad interfacial region, reduced consistency, transparent or semi-transparent emergence, & remarkable kinetic constancy, self-nanoemulsifying medicines are meant for an extensive choice of application. Through a combination of a water-insoluble lipid phase with a high-stress aqueous phase, these emulsions may be manufactured in huge quantities quickly. These are created using surfactants, co-surfactants, and co-solvents. In BCS class II and IV drugs, the self-nanoemulsifying drug delivery system is useful in favor of increasing the water solubility of water-soluble pharmaceuticals. In this review, various aspects of the self-nanoemulsifying drug delivery system are highlighted, including method of preparation, characterization techniques such as particle size, zeta potential, polydispersity, drug content, advantages, disadvantages, and a particular importance on various applications of SNEDDS in different areas, such as cancer treatment, drug targeting, and as a vehicle for transdermal drug delivery.

Rheological properties and microstructure of thermodynamically stable microemulsions as factors influencing the release rate of liposoluble vitamins

Microemulsions (ME) are stable nano-sized emulsions that constitute one of the most effective ways to incorporate lipophilic active compounds into water-based food matrices. Great stability, colloidal domain droplets, and optical isotropy are some of the advantages of ME. The present work analyzed the influence of rheological behavior and microstructural characteristics of food microemulsions on the release rate of lipophilic vitamins E and D. A fluid ME was compared to a ME with a higher dispersed phase content and a bicontinuous structure (gel-ME). Additionally, carboxymethyl cellulose (CMC) was added to fluid ME (3.5 and 10 g/100 g), obtaining systems with different apparent viscosities and microstructure. The percentage of released vitamins was determined by HPLC, and mathematically modeled. All ME were characterized using TEM microscopy FTIR, rheological and viscoelastic analysis. Fluid and gel-ME reached higher percentages of vitamin release in a very fast manner, while systems containing CMC showed a matrix-driven nature of the release. Those systems with similar zero-shear viscosity and different microstructure exhibited significantly different viscoelastic behavior. Microemulsions with CMC exhibited a viscoelastic solid type behavior with G’ > G’’. Mechanical spectra were satisfactorily fitted with the Generalized Maxwell model and relaxation time spectra were determined. In the quiescent state, gel-ME exhibited a higher plateau modulus than those containing large amount of thickener in the continuous phase. The relaxation time behavior of the structure could explain the kinetic release of liposoluble vitamins from the inner lipid phase of the microemulsions.

Tuning Targeted Liposome Avidity to Cells via Lipid Phase Separation.

The addition of both cell-targeting moieties and polyethylene glycol (PEG) to nanoparticle (NP) drug delivery systems is a standard approach to improve the biodistribution, specificity, and uptake of therapeutic cargo. The spatial presentation of these molecules affects avidity of the NP to target cells in part through an interplay between the local ligand concentration and the steric hindrance imposed by PEG molecules. Here, we show that lipid phase separation in nanoparticles can modulate liposome avidity by changing the proximity of PEG and targeting protein molecules on a nanoparticle surface. Using lipid-anchored nickel-nitrilotriacetic acid (Ni-NTA) as a model ligand, we demonstrate that the attachment of lipid anchored Ni-NTA and PEG molecules to distinct lipid domains in nanoparticles can enhance liposome binding to cancer cells by increasing ligand clustering and reducing steric hindrance. We then use this technique to enhance the binding of RGD-modified liposomes, which can bind to integrins overexpressed on many cancer cells. These results demonstrate the potential of lipid phase separation to modulate the spatial presentation of targeting and shielding molecules on lipid nanocarriers, offering a powerful tool to enhance the efficacy of NP drug delivery systems.

Moisturizing at a molecular level - The basis of Corneocare.

This review covers the last 20 years of research we and our collaborators have conducted on ethnic differences in facial skin moisturization placed in historical context with previous research. We have focussed particularly on the biochemical and cellular gradients of the stratum corneum (SC) with the aim of discovering new skin moisturization and SC maturation mechanisms, identifying new technologies and/or providing conceptual innovations for ingredients that will improve our understanding and treatment of dry skin. Specifically, we discuss gradients for corneodesmosomes and proteases, corneocyte phenotype-inducing enzymes, filaggrin and natural moisturizing factor (NMF), and barrier lipids. These gradients are interdependent and influence greatly corneocyte maturation. The interrelationship between corneodesmolysis and the covalent attachment of ω-hydroxy ceramides and ω-hydroxy fatty acids to the corneocyte protein envelope forming the corneocyte lipid envelope is especially relevant in our new understanding of mechanisms leading to dry skin. This process is initiated by a linoleoyl-ω-acyl ceramide transforming enzyme cascade including 12R lipoxygenase (12R-LOX), epidermal lipoxygenase-3 (eLOX3), epoxide hydrolase 3 (EPHX3), short-chain dehydrogenase/reductase family 9C member 7 (SDR9C7), ceramidase and transglutaminase 1. Our research has opened the opportunity of using novel treatment systems for dry skin based on lipids, humectants, niacinamide and inhibitors of the plasminogen system. It is clear that skin moisturization is a more complex mechanism than simple skin hydration.

Impact of dispersion media and carrier type on spray-dried proliposome powder formulations loaded with beclomethasone dipropionate for their pulmonary drug delivery via a next generation impactor.

Drug delivery via aerosolization for localized and systemic effect is a non-invasive approach to achieving pulmonary targeting. The aim of this study was to prepare spray-dried proliposome (SDP) powder formulations to produce carrier particles for superior aerosolization performance, assessed via a next generation impactor (NGI) in combination with a dry powder inhaler. SDP powder formulations (F1-F10) were prepared using a spray dryer, employing five different types of lactose carriers (Lactose monohydrate (LMH), lactose microfine (LMF), lactose 003, lactose 220 and lactose 300) and two different dispersion media. The first dispersion medium was comprised of water and ethanol (50:50% v/v ratio), and the second dispersion medium comprised wholly of ethanol (100%). In the first dispersion medium, the lipid phase (consisting of Soya phosphatidylcholine (SPC as phospholipid) and Beclomethasone dipropionate (BDP; model drug) were dissolved in ethanol and the lactose carrier in water, followed by spray drying. Whereas in second dispersion medium, the lipid phase and lactose carrier were dispersed in ethanol only, post spray drying. SDP powder formulations (F1-F5) possessed significantly smaller particles (2.89 ± 1.24-4.48 ± 1.20 μm), when compared to SDP F6-F10 formulations (10.63 ± 3.71-19.27 ± 4.98 μm), irrespective of lactose carrier type via SEM (scanning electron microscopy). Crystallinity of the F6-F10 and amorphicity of F1-F15 formulations were confirmed by XRD (X-ray diffraction). Differences in size and crystallinity were further reflected in production yield, where significantly higher production yield was obtained for F1-F5 (74.87 ± 4.28-87.32 ± 2.42%) then F6-F10 formulations (40.08 ± 5.714-54.98 ± 5.82%), irrespective of carrier type. Negligible differences were noted in terms of entrapment efficiency, when comparing F1-F5 SDP formulations (94.67 ± 8.41-96.35 ± 7.93) to F6-F10 formulations (78.16 ± 9.35-82.95 ± 9.62). Moreover, formulations F1-F5 demonstrated significantly higher fine particle fraction (FPF), fine particle dose (FPD) and respirable fraction (RF) (on average of 30.35%, 890.12 μg and 85.90%) when compared to counterpart SDP powder formulations (F6-F10). This study has demonstrated that when a combination of water and ethanol was employed as dispersion medium (formulations F1-F5), superior formulation properties for pulmonary drug delivery were observed, irrespective of carrier type employed.

Capsaicin encapsulated in W/O/W double emulsions fabricated via ethanol-induced pectin gelling: Improvement of bioaccessibility and reduction of irritation

Capsaicin is a water-insoluble bioactive component with several beneficial physiological functions. However, the widespread application of this hydrophobic phytochemical is limited by its low water-solubility, intense irritation, and poor bioaccessibility. These challenges can be overcome by entrapping capsaicin within the internal water phase of water-in-oil-in-water (W/O/W) double emulsions via using ethanol to induce pectin gelling. In this study, ethanol was used both to dissolve capsaicin and to promote pectin gelation, thereby forming capsaicin-loaded pectin hydrogels that were used as the internal water phase of the double emulsions. Pectin addition improved the physical stability of the emulsions and led to a high encapsulation efficiency of capsaicin (>70 % after 7d storage). After simulated oral and gastric digestion, capsaicin-loaded double emulsions maintained their compartmentalized structure, avoiding capsaicin leakage in the month and stomach. The double emulsions were digested in the small intestine, thereby releasing the capsaicin. Capsaicin bioaccessibility was significantly enhanced after encapsulation, which was attributed to mixed micelle formation by the digested lipid phase. Furthermore, encapsulation of capsaicin within the double emulsions reduced the irritation in the gastrointestinal tissues of mice. This kind of double emulsion may have great potential for the development of more palatable capsaicin-loaded functional food products.

Baking using oleogels

Baking using oleogels

Composition-based statistical model for predicting CO2 solubility in modified atmosphere packaging application

Carbon dioxide (CO 2 ) is an important gas used in modified atmosphere packaging of non-respiring foods where it solubilizes into the aqueous and lipid phases of food and exerts an antimicrobial effect. Prediction of CO2 solubility within food is thus of paramount importance to anticipate its benefit on food preservation. In the present study, machine learning algorithms were applied on a set of 362 values of CO 2 solubilities collected from the scientific literature to tentatively predict the solubility as a function of food composition (water, protein, fat and salt content) and temperature. The best option kept was a random forest algorithm that was used to predict CO 2 solubility in four food case studies (ham, salmon, cheese and pâté) that were further used as input parameters in the MAP’ OPT tool, predicting the evolution of headspace gas composition. Predicted CO 2 solubilities used as input parameters succeeded in representing the CO 2 headspace dynamic as a function of time in the four case studies.

Fluorescent Probes cis- and trans-Parinaric Acids in Fluid and Gel Lipid Bilayers: A Molecular Dynamics Study.

Fluorescence probes are indispensable tools in biochemical and biophysical membrane studies. Most of them possess extrinsic fluorophores, which often constitute a source of uncertainty and potential perturbation to the host system. In this regard, the few available intrinsically fluorescent membrane probes acquire increased importance. Among them, cis- and trans-parinaric acids (c-PnA and t-PnA, respectively) stand out as probes of membrane order and dynamics. These two compounds are long-chained fatty acids, differing solely in the configurations of two double bonds of their conjugated tetraene fluorophore. In this work, we employed all-atom and coarse-grained molecular dynamics simulations to study the behavior of c-PnA and t-PnA in lipid bilayers of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), representative of the liquid disordered and solid ordered lipid phases, respectively. All-atom simulations indicate that the two probes show similar location and orientation in the simulated systems, with the carboxylate facing the water/lipid interface and the tail spanning the membrane leaflet. The two probes establish interactions with the solvent and lipids to a similar degree in POPC. However, the almost linear t-PnA molecules have tighter lipid packing around them, especially in DPPC, where they also interact more with positively charged lipid choline groups. Probably for these reasons, while both probes show similar partition (assessed from computed free energy profiles across bilayers) to POPC, t-PnA clearly partitions more extensively than c-PnA to the gel phase. t-PnA also displays more hindered fluorophore rotation, especially in DPPC. Our results agree very well with experimental fluorescence data from the literature and allow deeper understanding of the behavior of these two reporters of membrane organization.

Correction of rhodopsin serial crystallography diffraction intensities for a lattice-translocation defect.

Rhodopsin is a G-protein-coupled receptor that detects light and initiates the intracellular signalling cascades that underpin vertebrate vision. Light sensitivity is achieved by covalent linkage to 11-cis retinal, which isomerizes upon photo-absorption. Serial femtosecond crystallography data collected from rhodopsin microcrystals grown in the lipidic cubic phase were used to solve the room-temperature structure of the receptor. Although the diffraction data showed high completeness and good consistency to 1.8 Å resolution, prominent electron-density features remained unaccounted for throughout the unit cell after model building and refinement. A deeper analysis of the diffraction intensities uncovered the presence of a lattice-translocation defect (LTD) within the crystals. The procedure followed to correct the diffraction intensities for this pathology enabled the building of an improved resting-state model. The correction was essential to both confidently model the structure of the unilluminated state and interpret the light-activated data collected after photo-excitation of the crystals. It is expected that similar cases of LTD will be observed in other serial crystallography experiments and that correction will be required in a variety of systems.

Seed Storage Physiology of Lophomyrtus and Neomyrtus, Two Threatened Myrtaceae Genera Endemic to New Zealand.

There is no published information on the seed germination or seed storage physiology of Lophomyrtus bullata, Lophomyrtus obcordata, and Neomyrtus pedunculata. This lack of information is hampering conservation efforts of these critically endangered species. This study investigated the seed morphology, seed germination requirements, and long-term seed storage methods for all three species. The impact of desiccation, desiccation and freezing, as well as desiccation plus storage at 5 °C, -18 °C, and -196 °C on seed viability (germination) and seedling vigour was assessed. Fatty acid profiles were compared between L. obcordata and L. bullata. Variability in storage behaviour between the three species was investigated through differential scanning calorimetry (DSC) by comparing thermal properties of lipids. L. obcordata seed were desiccation-tolerant and viability was retained when desiccated seed was stored for 24 months at 5 °C. L. bullata seed was both desiccation- and freezing-sensitive, while N. pedunculata was desiccation-sensitive. DSC analysis revealed that lipid crystallisation in L. bullata occurred between -18 °C and -49 °C and between -23 °C and -52 °C in L. obcordata and N. pedunculata. It is postulated that the metastable lipid phase, which coincides with the conventional seed banking temperature (i.e., storing seeds at -20 ± 4 °C and 15 ± 3% RH), could cause the seeds to age more rapidly through lipid peroxidation. Seeds of L. bullata, L. obcordata and N. pedunculata are best stored outside of their lipid metastable temperature ranges.

A robust approach for MicroED sample preparation of lipidic cubic phase embedded membrane protein crystals

Crystallizing G protein-coupled receptors (GPCRs) in lipidic cubic phase (LCP) often yields crystals suited for the cryogenic electron microscopy (cryoEM) method microcrystal electron diffraction (MicroED). However, sample preparation is challenging. Embedded crystals cannot be targeted topologically. Here, we use an integrated fluorescence light microscope (iFLM) inside of a focused ion beam and scanning electron microscope (FIB-SEM) to identify fluorescently labeled GPCR crystals. Crystals are targeted using the iFLM and LCP is milled using a plasma focused ion beam (pFIB). The optimal ion source for preparing biological lamellae is identified using standard crystals of proteinase K. Lamellae prepared using either argon or xenon produced the highest quality data and structures. MicroED data are collected from the milled lamellae and the structures are determined. This study outlines a robust approach to identify and mill membrane protein crystals for MicroED and demonstrates plasma ion-beam milling is a powerful tool for preparing biological lamellae.

Food resources in food system technology: Bifunctional food system technology based on pickering emulsions

The development of domestically generated food additives based on eco-technologies with specified functionaltechnological and bioactive qualities is now a significant focus of contemporary study. Pickering emulsions are more stable both physically and chemically, making them a highly promising method of introducing biologically active chemicals into the body. The purpose of this research was to determine if ultrasound could be used to create a bioactive colloidal Pickering emulsion system using flax cellulose or brown algal polysaccharides as a stabilizer. Pickering emulsions stabilized with fucoidan, sodium alginate, or flax cellulose, based on a lipid fraction of sunflower oil and cinnamon oil blend, or a lipid fraction of linseed oil, were applied to the subjects, who were then subjected to ultrasound treatment at 630W/l for 16 min (4 min on, 3 min off) at a controlled temperature of 50°C. The results obtained demonstrate the effectiveness of using ultrasonic exposure to create bioactive Pickering emulsion colloidal systems. Fucoidan stabilizes pickering emulsion, with the highest AOA values (5.120±0.005 DPPH, %) and a rise in AOA values in linseed oil-based emulsion samples from 30% to 100% in comparison to emulsion samples based on a blend of lipid phases. The viscoelastic characteristics of Pickering emulsions with various oil fractions vary dramatically for the integrated structure-forming components.