Lipid Phase Research Articles

Evaluation of the synergistic effect of plant-based components on the stability of curcuminoid emulsion

In this study, the effect of matrix compounds from natural curcuminoid resources on the stability of curcuminoids and emulsions thereof was evaluated. Curcuminoid emulsions were prepared curcuminoid rich sources (curcuminoid extract, an aqueous turmeric concentrate and turmeric powder) with medium-chain triglyceride oil as lipid phase, lecithin, and pectin as emulsifiers. The curcuminoid emulsions were exposed to light in the visible wavelength range (300 nm–800 nm) at the specific energy input of 0.47 kW/m2 for 7 days and to the temperature of 4 °C, 25 °C, 40 °C for 49 days. The total curcuminoid retention (TC), droplet size (DS) change, instability index (InI), and yellowness reduction (YR) was observed during the storage time. The half-life of curcuminoids in emulsions was increased to 21 h, while the half-life of free curcuminoids was 1.3 h in the light exposure test. The co-compounds from the curcuminoid sources contributed to the emulsion stability by increasing the viscosity. In the thermal exposure test, the matrix compound system retained more than 93% curcuminoids after 49 days of storage at 40 °C, whereas the phase separation increased significantly. However, the TC reduction was independent of the InI change and droplet agglomeration. The YR depended on the TC and the amount of co-components in the emulsion.

Antioxidant and Neuroprotective Activity of Vitamin E Homologues: In Vitro Study.

Here we present comparative data on the inhibition of lipid peroxidation by a variety of tocochromanols in liposomes. We also show for the first time the potential neuroprotective role of all the vitamin E homologues investigated on the neuronally differentiated human neuroblastoma SH-SY5Y cell line. α-Tocopherol had nearly no effect in the inhibition of lipid peroxidation, while β-, γ-, and δ-tocopherols inhibited the reaction completely when it was initiated in a lipid phase. Similar effects were observed for tocotrienol homologues. Moreover, in this respect plastochromanol-8 was as effective as β-, γ-, and δ-tocochromanols. When the prenyllipids were investigated in a 1,1-diphenyl-2-picrylhydrazyl (DPPH) test and incorporated into different lipid carriers, the radical oxidation was most pronounced in liposomes, followed by mixed micelles and the micellar system. When the reaction of tocochromanols was examined in niosomes, the oxidation was most pronounced for α-tocopherol and plastochromanol-8, followed by α-tocotrienol. Next, using retinoic acid-differentiated SH-SY5Y cells, we tested the protective effects of the compounds investigated on hydrogen peroxide (H2O2)-induced cell damage. We showed that tocotrienols were more active than tocopherols in the oxidative stress model. Plastochromanol-8 had a strong inhibitory effect on H2O2-induced lactate dehydrogenase (LDH) release and H2O2-induced decrease in cell viability. The water-soluble α-tocopherol phosphate had neuroprotective effects at all the concentrations analyzed. The results clearly indicate that structural differences between vitamin E homologues reflect their different biological activity and indicate their potential application in pharmacological treatments for neurodegenerative diseases. In this respect, the application of optimal tocochromanol-carrying structures might be critical.

Librational Dynamics of Spin-Labeled Membranes at Cryogenic Temperatures From Echo-Detected ED-EPR Spectra.

Methods of electron spin echo of pulse electron paramagnetic resonance (EPR) spectroscopy are increasingly employed to investigate biophysical properties of nitroxide-labeled biosystems at cryogenic temperatures. Two-pulse echo-detected ED-spectra have proven to be valuable tools to describe the librational dynamics in the low-temperature phases of both lipids and proteins in membranes. The motional parameter, , given by the product of the mean-square angular amplitude, , and the rotational correlation time, , of the motion, is readily determined from the nitroxide ED-spectra as well as from the W-relaxation rate curves. An independent evaluation of is obtained from the motionally averaged 14N-hyperfine splitting separation in the continuous wave cw-EPR spectra. Finally, the rotational correlation time can be estimated by combining ED- and cw-EPR data. In this mini-review, results on the librational dynamics in model and natural membranes are illustrated.

Lycorine impedes 7,12-dimethylbenz(a)anthracene exposed hamster oral carcinogenesis through P13K/Akt and NF-κB inhibition

Abstract Objectives Oncogenic signaling pathways that are activated abnormally play a key activity in tumor initiation and development. This research aimed to examine the preventive efficiency of lycorine in the buccal pouch hamster tumor model based on its capacity to target phosphoinositide 3-kinase (PI3K)/Akt and nuclear factor-kappa B (NF-κB) signaling cascades. Methods The induction of oral tumor in male golden Syrian hamsters was done by 7,12-dimethylbenz [a] anthracene (DMBA) painting on the left buccal pouch thrice a week for 10 weeks. The chemopreventive effect of lycorine (20 mg/kg b.w.) was assessed by treating orally for 14 weeks of the experimental period. The biochemical endpoints such as lipid peroxidation (LPO), antioxidants, and phase I and II detoxification agents were analyzed. Results The treatment of lycorine to DMBA-induced hamsters drastically suppressed tumor incidence and tumor size and reverted the levels of the biochemical indicator. Moreover, lycorine significantly downregulated the p53, Cyclooxygenase 2 (cox-2), and P13K/Akt signaling and inhibited the phosphorylation of NF-κB and nuclear factor-kappa-B-inhibitor alpha (Iκ-Bα) in DMBA-induced hamsters. Conclusions The oral administration of lycorine effectively inhibited tumor cell proliferation, restored the antioxidant, LPO, and detoxification enzymes, and inhibited NF-κB signaling in oral tumorigenesis. Thus, the use of lycorine after a proper clinical trial could be effective for oral tumorigenesis treatment.

Long acting injectables for therapeutic proteins

Biotherapeutic development presents a myriad of challenges in relation to delivery, in particular for protein therapeutics. Protein delivery is complicated due to hydrophilicity, size, rate of degradation in vivo, low permeation through biological barriers, pH and temperature sensitivity, as well as the need to conserve its quaternary structure to retain function. To preserve therapeutic levels in vivo, proteins require frequent administration due to their short half-lives. Formulation strategies combining proteins with lipid carriers for parenteral administration show potential for improving bioavailability, while preserving protein activity and bypassing the mucosal barriers of the body. Encapsulating protein in long acting injectable delivery systems can improve therapeutic indices by prolonging and controlling protein release and reducing the need for repeat interventions. Two lyotropic crystal forming lipids, monoolein and phytantriol, have been formulated to produce lipidic cubic phases and assessed for their use as long acting protein eluting injectables. Three soluble proteins, cytochrome c, glyceraldehyde-3-phosphate dehydrogenase and aldehyde dehydrogenase and one membrane protein, cytochrome c oxidase, were incorporated into bulk cubic phase formulations of each lipid system to comparatively assess protein release kinetics. The activity of the soluble proteins was measured upon release from a phytantriol bulk cubic phase and phytantriol cubosomes, produced using a liquid precursor method.

Imidazo[1,5-a]pyridine-Based Fluorescent Probes: A Photophysical Investigation in Liposome Models.

Imidazo[1,5-a]pyridine is a stable scaffold, widely used for the development of emissive compounds in many application fields (e.g., optoelectronics, coordination chemistry, sensors, chemical biology). Their compact shape along with remarkable photophysical properties make them suitable candidates as cell membrane probes. The study of the membrane dynamics, hydration, and fluidity is of importance to monitor the cellular health and to explore crucial biochemical pathways. In this context, five imidazo[1,5-a]pyridine-based fluorophores were synthesized according to a one-pot cyclization between an aromatic ketone and benzaldehyde in the presence of ammonium acetate and acetic acid. The photophysical features of prepared compounds were investigated in several organic solvents and probes 2–4 exhibited the greatest solvatochromic behavior, resulting in a higher suitability as membrane probes. Their interaction with liposomes as artificial membrane model was tested showing a successful intercalation of the probes in the lipid bilayer. Kinetic experiments were carried out and the lipidic phase influence on the photophysical features was evaluated through temperature-dependent experiments. The results herein reported encourage further investigations on the use of imidazo[1,5-a]pyridine scaffold as fluorescent membrane probes.

Shea Butter Potentiates the Anti-Bacterial Activity of Fusidic Acid Incorporated into Solid Lipid Nanoparticle.

Fusidic acid (FA) is an efficient anti-bacterial drug proven to be efficient against a wide range of bacteria. Nevertheless, the main restriction in its formulation is the limited solubility. To avoid such an obstacle, the drug is incorporated into the lipid core of the nanolipid formulation. Consequently, the present study was an attempt to formulate nanolipid preparation, mainly, solid lipid nanoparticle (SLN) integrating FA. FA-SLN was prepared using shea butter as a lipid phase owing to its reported anti-bacterial activity. Different FA-SLNs were fabricated using the central composite design (CCD) approach. The optimized formula was selected and integrated into a hydrogel base to be efficiently used topically. FA-SLN-hydrogel was evaluated for its character, morphology, in vitro release and stability. The formula was examined for irritation reaction and finally evaluated for its anti-bacterial performance. The optimized formula showed particle size 283.83 nm and entrapment 73.057%. The formulated FA-SLN-hydrogel displayed pH 6.2, viscosity 15,610 cP, spreadability 51.1 mm and in vitro release 64.6% following 180 min. FA-SLN-hydrogel showed good stability for three months at different conditions (room temperature and refrigerator). It exhibited no irritation reaction on the treated rats. Eventually, shea butter displayed a noteworthy effect against bacterial growth that improved the effect of FA. This would indicate prospective anti-bacterial activity of FA when combined with shea butter in SLN formulation as a promising nanocarrier.

Fluidity-Guided Assembly of Au@Pt on Liposomes as a Catalase-Powered Nanomotor for Effective Cell Uptake in Cancer Cells and Plant Leaves.

The fluidity of the liposomes is essential to nanoparticle-membrane interactions. We herein report a liposomal nanomotor system by controlling the self-assembly behavior of gold core-platinum shell nanoparticles (Au@Pt) on liposomes. Au@Pt can aggregate immediately on fluid-phase dioleoyl-sn-glycero-3-phosphocholine (DOPC) liposomes, forming an uneven distribution. By control of the lipid phase and fluidity, either using pure 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) above its phase transition temperature or adding cholesterol as an adjuvant to DPPC lipids, we precisely control the assembly of Au@Pt on liposomes. Au@Pt maintained high catalase-like activity on the liposomal surface, promoting the decomposition of H2O2 and the movement of the liposomal nanomotors. Finally, we demonstrate that liposomal nanomotors are biocompatible and they can speed up the cellular uptake in mammalian HepG2 cancer cells and Nicotiana tabacum (Nb) plant leaves. This liposomal nanomotor system is expected to be further investigated in biomedicine and plant nanotechnology.

Abstract 5327: Characterization of a green, temperature activated nano formulation that drives the mechanism of doxorubicin toxicity from apoptosis to ferroptosis

Abstract Introduction: Nano formulations continue to be a very attractive venue for the development of “smart” drug delivery systems. Emerging green (organic) formulations do not require harsh chemical solvents and are easy to prepare. Solid Core Lipid Formulations: The lipid phase (l.p.) contained a solid fatty acid (lauric acid) and liquid lipid (oleic acid) with or without Doxorubicin. 55° surfactant containing an emulsifying agent (Brij-58) mixed with nonionic surfactant (Span 80) was dropwise added. The emulsion was cooled to form temperature-activated nanoparticles (TAN). Before usage, the emulsion was activated by heating to 41°C. Experiments: B16F10 melanoma, U87-MR glioblastoma, MDA-MB-231 and MCF-7 breast cancer cell lines were analyzed on a variety of quantitative Imaging platforms. Results iCyte (CompuCyte): Laser based instrument for obtaining quantitative fluorescence and light scatter measurements. We obtained single endpoint dosage response curves to compare the TAN with free Dox, and found the TANs were effective at 4-6 times lower equivalent Dox concentrations. Holomonitor (Phase Holographic Imaging): The HM4 resides in tissue culture incubators and allows long-term medium resolution holographic imaging. It offersa complete image processing library including routines for proliferation and cell tracking. We devised methods for 4-dimensional imaging (X pos. vs. Y pos. vs Time, vs. Density). Thus, we are able to monitor the temporal effects of the compounds. Cell Explorer (Nanolive): The Cell Explorer has a a rotating mirror that directs the laser in an orbit around the sample to obtain super-resolution 3D tomographs. It enables long-term time-lapse imaging via stage-top environmental chamber. It is well suited for quantifying sub cellular components such as mitochondria and lipids. HT-2 (Tomocube): The HT-2 is a label-free ultra-high resolution holotomography system complemented by fluorescence capabilities. Combined RI and fluorescence images give the highest information content of any of the platforms. Discussion: In 2012, Dixon et al described ferroptosis as a distinct new form of cell death through the iron dependent accumulation of oxidatively damaged phospholipids. They state that a diagnostic feature is mitochondrial shrinkage and collapse. We confirmed this in our tomographic imaging images. Other features that were detected and consistent with ferroptosis are loss of nuclear contents, a thickening of the nuclear membrane, and peri-nuclear mitochondrial accumulation. In 48-hour HM4 plots control cells show numerous mitotic events. Dox treated cells show cellular enlargement but no mitotic events. In the TAN population at about 20 hours, there is sudden death occurring in the entire population. Conclusion: Our experiments suggest a high potency of the newly developed TANs. Ferroptosis is the most likely mechanism of action. Citation Format: Farzana Parveen, Nina Filipczak, Asadullah Madni, Vladimir P. Torchilin, Ed Luther. Characterization of a green, temperature activated nano formulation that drives the mechanism of doxorubicin toxicity from apoptosis to ferroptosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5327.

Short-Chained Alcohols Make Membrane Surfaces Conducive for Melittin Action: Implication for the Physiological Role of Alcohols in Cells.

Alcohols are a part of cellular metabolism, but their physiological roles are not well understood. We investigated the effects of short-chain alcohols on Daphnia pulex and model membranes mimicking the lipid composition of eukaryotic inner mitochondrial membranes. We also studied the synergistic effects of alcohols with the bee venom membrane-active peptide, melittin, which is structurally similar to endogenous membrane-active peptides. The alcohols, from ethanol to octanol, gradually decreased the heart rate and the mitochondrial ATP synthesis of daphnia; in contrast, in combination with melittin, which exerted no sizeable effect, they gradually increased both the heart rate and the ATP synthesis. Lipid packing and the order parameter of oriented films, monitored by EPR spectroscopy of the spin-labeled probe 5-doxylstrearic acid, revealed gradual alcohol-assisted bilayer to non-bilayer transitions in the presence of melittin; further, while the alcohols decreased, in combination with melittin they increased the order parameter of the film, which is attributed to the alcohol-facilitated association of melittin with the membrane. A 1H-NMR spectroscopy of the liposomes confirmed the enhanced induction of a non-bilayer lipid phase that formed around the melittin, without the permeabilization of the liposomal membrane. Our data suggest that short-chain alcohols, in combination with endogenous peptides, regulate protein functions via modulating the lipid polymorphism of membranes.

Peppermint essential oil-loaded solid lipid nanoparticle in gelatin coating: Characterization and antibacterial activity against foodborne pathogen inoculated on rainbow trout (Oncorhynchus mykiss) fillet during refrigerated storage.

The present study was conducted to determine the characterization and antibacterial activity of peppermint essential oil-loaded solid lipid nanoparticle (PEO-SLN) and its impact on the quality of trout fillet stored at 4 ± 1°C for 12 days. The SLNs were prepared through a bath sonication technique. PEO-SLNs contained 0.2% (w/v) PEO in 2% of lipid phase glycerol monostearate (GMS) and tween 80 (1% w/v) used as a surfactant in the aqueous phase. The characterization parameter of PEO-SLN was evaluated, and the antibacterial activity of PEO-SLNs was conducted under in vitro conditions. Trout samples were analyzed for inoculated Pseudomonas aeruginosa, Listeria monocytogenes, and Escherichia coli O157:H7 during refrigerated storage. The mean particle size of PEO-SLNs was 154.83 ± 1.21nm with a polydispersity index (PDI) of 0.35 ± 0.01 and zeta potential was about -24.16 ± 0.51mV. The results indicated that PEO-SLN had higher antibacterial activity than the free form of PEO and also when used in combination with gelatin coating (gel + PEO-SLN) had a significant effect on preventing microbial growth in trout fillets (p<0.05). The most decreasing rate of P. aeruginosa (1.92 log CFU/g), E. coli O157:H7 (0.71 log CFU/g), and L. monocytogenes count (1.69 log CFU/g) was seen in gel + PEO-SLN. These findings illustrated that PEO-SLNs could potentially be utilized in the food industry to increase the shelf life of fish fillets.

Membrane Protein Activity Induces Specific Molecular Changes in Nanodiscs Monitored by FTIR Difference Spectroscopy.

It is well known that lipids neighboring integral membrane proteins directly influence their function. The opposite effect is true as well, as membrane proteins undergo structural changes after activation and thus perturb the lipidic environment. Here, we studied the interaction between these molecular machines and the lipid bilayer by observing changes in the lipid vibrational bands via FTIR spectroscopy. Membrane proteins with different functionalities have been reconstituted into lipid nanodiscs: Microbial rhodopsins that act as light-activated ion pumps (the proton pumps NsXeR and UmRh1, and the chloride pump NmHR) or as sensors (NpSRII), as well as the electron-driven cytochrome c oxidase RsCcO. The effects of the structural changes on the surrounding lipid phase are compared to mechanically induced lateral tension exerted by the light-activatable lipid analogue AzoPC. With the help of isotopologues, we show that the ν(C = O) ester band of the glycerol backbone reports on changes in the lipids’ collective state induced by mechanical changes in the transmembrane proteins. The perturbation of the nanodisc lipids seems to involve their phase and/or packing state. 13C-labeling of the scaffold protein shows that its structure also responds to the mechanical expansion of the lipid bilayer.

Effect of alkyl chain length on the antioxidant activity of alkylresorcinol homologs in low-moisture crackers

Alkylresorcinols (ARs) are a naturally occurring homologous series of phenolipids extracted from rye bran which have previously shown antioxidant activity in lipid-containing foods. This study extends previous work to consider their use in low-moisture foods using crackers as a model system. ARs (153 μmol) were capable of inhibiting lipid oxidation reactions based on the delayed generation of lipid hydroperoxides and headspace hexanal compared to a control treatment and were more effective than α-tocopherol which displayed prooxidant behavior. The antioxidant activity of ARs was observed to increase as a function of increasing alkyl chain length, with optimum activity at an alkyl chain length of C23:0; however, ARs were lost at a rate independent of alkyl chain length. A mixture of ARs displayed comparable antioxidant activity to the C23:0 and C25:0 homologs, all of which exerted stronger activity than butylated hydroxytoluene providing an effective alternative to synthetic antioxidants. These findings suggest that ARs are effective antioxidants in low-moisture foods likely because of their hydrophobicity, which allowed them to localize in the lipid phase, the purported site of lipid oxidation in the model cracker system.

Absorption of Phosphonium Cations and Dications into a Hydrated POPC Phospholipid Bilayer: A Computational Study.

Molecular dynamics (MD) based on an empirical force field is applied to investigate the effect of phosphonium cations ([P6,6,6,6]+) and geminal dications ([DxC10]2+) inserted at T = 300 K into the hydration layer separating planar POPC phospholipid bilayers. Up to high concentration, nearly every added cation and dication becomes absorbed into the lipid phase. Absorption takes place during several microseconds and is virtually irreversible. The neutralizing counterions ([Cl]−, in the present simulation) remain dissolved in water, giving origin to the charge separation and the strong electrostatic double layer at the water/lipid interface. Incorporation of cations and dications changes the properties of the lipid bilayer such as diffusion, viscosity, and the electrostatic pattern. At high ionic concentration, the bilayer acquires a long-wavelength standing undulation, corresponding to a change of phase from fluid planar to ripple. All these changes are potentially able to affect processes relevant in the context of cell biology. The major difference between cations and dications concerns the kinetics of absorption, which takes place nearly two times faster in the [P6,6,6,6]+ case, and for [DxC10]2+ dications displays a marked separation into two-stages, corresponding to the easy absorption of the first phosphonium head of the dication and the somewhat more activated absorption of the second phosphonium head of each dication.

Location of the TEMPO moiety of TEMPO-PC in phosphatidylcholine bilayers is membrane phase dependent

The (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO) moiety tethered to the headgroup of phosphatidylcholine (PC) lipid is employed in spin labeling electron paramagnetic resonance spectroscopy to probe the water dynamics near lipid bilayer interfaces. Due to its amphiphilic character, however, TEMPO spin label could partition between aqueous and lipid phases, and may even be stabilized in the lipid phase. Accurate assessment of the TEMPO-PC configuration in bilayer membranes is essential for correctly interpreting the data from measurements. Here, we carry out all-atom molecular dynamics (MD) simulations of TEMPO-PC probe in single-component lipid bilayers at varying temperatures, using two standard MD force fields. We find that, for a dipalmitoylphosphatidylcholine (DPPC) membrane whose gel-to-fluid lipid phase transition occurs at 314 K, while the TEMPO spin label is stabilized above the bilayer interface in the gel phase, there is a preferential location of TEMPO below the membrane interface in the fluid phase. For bilayers made of unsaturated lipids, 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), which adopt the fluid phase at ambient temperature, TEMPO is unequivocally stabilized inside the bilayers. Our finding of membrane phase-dependent positioning of the TEMPO moiety highlights the importance of assessing the packing order and fluidity of lipids under a given measurement condition.

Saponins as Natural Emulsifiers for Nanoemulsions.

The awareness of sustainability approaches has focused attention on replacing synthetic emulsifiers with natural alternatives when formulating nanoemulsions. In this context, a comprehensive review of the different types of saponins being successfully used to form and stabilize nanoemulsions is presented, highlighting the most common natural sources and biosynthetic routes. Processes for their extraction and purification are also reviewed altogether with the recent advances for their characterization. Concerning the preparation of the nanoemulsions containing saponins, the focus has been initially given to screening methods, lipid phase used, and production procedures, but their characterization and delivery systems explored are also discussed. Most experimental outcomes showed that the saponins present high performance, but the challenges associated with the saponins' broader application, mainly the standardization for industrial use, are identified. Future perspectives report, among others, the emerging biotechnological processes and the use of byproducts in a circular economy context.

Lipidic cubic phase serial femtosecond crystallography structure of a photosynthetic reaction centre.

Serial crystallography is a rapidly growing method that can yield structural insights from microcrystals that were previously considered to be too small to beuseful in conventional X-ray crystallography. Here, conditions for growing microcrystals of the photosynthetic reaction centre of Blastochloris viridis within a lipidic cubic phase (LCP) crystallization matrix that employ a seeding protocol utilizing detergent-grown crystals with a different crystal packing are described. LCP microcrystals diffracted to 2.25 Å resolution when exposed to XFEL radiation, which is an improvement of 0.15 Å over previous microcrystal forms. Ubiquinone was incorporated into the LCP crystallization media and the resulting electron density within the mobile QB pocket is comparable to that ofother cofactors within the structure. As such, LCP microcrystallization conditions will facilitate time-resolved diffraction studies of electron-transfer reactions to the mobile quinone, potentially allowing the observation of structural changes associated with the two electron-transfer reactions leading to complete reduction of the ubiquinone ligand.

Comparison of structural properties of cyclosporin A and its analogue alisporivir and their effects on mitochondrial bioenergetics and membrane behavior

The paper considers the effect of the MPT pore inhibitor cyclosporin A (CsA) and its non-immunosuppressive analogue alisporivir (Ali) on the functioning of rat skeletal muscle mitochondria. We have shown that both agents at a standard in vitro concentration of 1 μM increase the calcium capacity of organelles and have no effect on the parameters of oxidative phosphorylation. However, an increase in their concentration to 5 μM leads to the suppression of oxygen consumption by mitochondria, which is more pronounced in the case of Ali. This effect is accompanied by a decrease in the membrane potential of organelles and, apparently, is based on the inhibition of electron transport along the mitochondrial respiratory chain due to limited mobility of coenzyme Q. We have noted that both agents do not affect the production of hydrogen peroxide by isolated mitochondria. NMR spectroscopy and molecular dynamics simulation did not reveal significant differences in the structure and backbone flexibility of CsA and Ali. Both agents decrease the overall fluidity of the membrane of DPPC liposomes, inducing an increase in laurdan generalized polarization parameter. A similar effect was also found in the case of mitochondrial membranes. We suggested that these effects of CsA and Ali, associated with their lipophilic nature and the ability to accumulate in the lipid phase of membranes, may cause a decrease in the efficiency of electron transport in the respiratory chain of mitochondria and suppression of the bioenergetics of these organelles.

Total ice content and lipid saturation determine adipose tissue cryolipolysis by injection of ice-slurry.

Cryolipolysis uses tissue cooling to solidify lipids, preferentially damaging lipid-rich cells. Topical cooling is popular for the reduction of local subcutaneous fat. Injection of biocompatible ice-slurry is a recently introduced alternative. We developed and verified a quantitative model that simulates the heat exchange and phase changes involved, offering insights into ice-slurry injection for treating subcutaneous fat. Finite element method was used to model the spatial and temporal progression of heat transfer between adipose tissue and injected ice-slurry, estimating dose-response relationships between properties of the slurry and size of tissue affected by cryolipolysis. Phase changes of both slurry and adipose tissue lipids were considered. An in vivo swine model was used to validate the numerical solutions. Oils with different lipid compositions were exposed to ice-slurry in vitro to evaluate the effects of lipid freezing temperature. Microscopy and nuclear magnetic resonance (NMR) were performed to detect lipid phase changes. A ball of granular ice was deposited at the injection site in subcutaneous fat. Total injected ice content determines both the effective cooling region of tissue, and the duration of tissue cooling. Water's high latent heat of fusion enables tissue cooling long after slurry injection. Slurry temperature affects the rate of tissue cooling. In swine, when 30 ml slurry injection at -3.5°C was compared to 15 ml slurry injection at -4.8°C (both with the same total ice content), the latter led to almost twice faster tissue cooling. NMR showed a large decrease in diffusion upon lipid crystallization; saturated lipids with higher freezing temperatures were more susceptible to solidification after ice-slurry injection. Total injected ice content determines both the volume of tissue treated by cryolipolysis and the cooling duration after slurry injection, while slurry temperature affects the cooling rate. Lipid saturation, which varies with diet and anatomic location, also has an important influence.

Fluorescence Studies of Nicotinic Acetylcholine Receptor and Its Associated Lipid Milieu: The Influence of Erwin London's Methodological Approaches.

Erwin London dedicated considerable effort to understanding lipid interactions with membrane-resident proteins and how these interactions shaped the formation and maintenance of lipid phases and domains. In this endeavor, he developed ad hoc techniques that greatly contributed to advancements in the field. We have employed and/or modified/extended some of his methodological approaches and applied them to investigate lipid interaction with the nicotinic acetylcholine receptor (nAChR) protein, the paradigm member of the superfamily of rapid pentameric ligand-gated ion channels (pLGIC). Our experimental systems ranged from purified receptor protein reconstituted into synthetic lipid membranes having known effects on receptor function, to cellular systems subjected to modification of their lipid content, e.g., varying cholesterol levels. We have often employed fluorescence techniques, including fluorescence quenching of diphenylhexatriene (DPH) extrinsic fluorescence and of nAChR intrinsic fluorescence by nitroxide spin-labeled phospholipids, DPH anisotropy, excimer formation of pyrene-phosphatidylcholine, and Förster resonance energy transfer (FRET) from the protein moiety to the extrinsic probes Laurdan, DPH, or pyrene-phospholipid to characterize various biophysical properties of lipid-receptor interactions. Some of these strategies are revisited in this review. Special attention is devoted to the anionic phospholipid phosphatidic acid (PA), which stabilizes the functional resting form of the nAChR. The receptor protein was shown to organize its PA-containing immediate microenvironment into microdomains with high lateral packing density and rigidity. PA and cholesterol appear to compete for the same binding sites on the nAChR protein.