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Lecithinized brain-derived neurotrophic factor promotes the differentiation of embryonic stem cells in vitro and in vivo.

The addition of lecithin molecules to brain-derived neurotrophic factor (BDNF) has been reported to markedly enhance its pharmacological effect in vivo. In the current study, we show that lecithinized BDNF (PC-BDNF) has a higher affinity than BDNF for neural precursor cells. Although BDNF only slightly increased the expression of the genes for Mash-1, p35, 68 kDa neurofilament, and TrkB receptor, PC-BDNF caused a significant increase in their expression. PC-BDNF also increased the level of neurofilament protein and dramatically increased TrkB mRNA gene expression, which was followed by a sustained activation of the p42/p44 extracellular-regulated kinases. Finally, transplantation of PC-BDNF-treated cells was more effective than BDNF-treated cells at improving impaired motor function caused by spinal cord injury. These findings showed that PC-BDNF has a better potential than BDNF for promoting neural differentiation, partly due to a higher cellular affinity. Furthermore, PC-BDNF-treated cells could be useful for transplantation therapy for central nervous system injuries.

Animals↗

A synthetic alternative to natural lecithins with antimicrobial properties.

Two soft biocompatible cationic surfactants from the amino acid arginine, 1,2-dilauroyl-3-acetylarginyl-rac-glycerol (1212RAc) and 1,2-dimirystoyl-3-acetylarginyl-rac-glycerol (1414RAc), were prepared. Their physicochemical properties show that they can be classified as multifunctional surfactants with self-aggregation behaviour comparable to that of short-chain lecithins. The two surfactants can simultaneously stabilise water-in-oil (W/O) droplets and oil-in-water (O/W) droplets, forming multiple emulsions. They have antimicrobial activity similar to that of conventional cationic surfactants and are as harmless as amphoteric betaines. These surfactants constitute an interesting alternative to the diglycerides and lecithins in formulations that require antimicrobial properties.

Anti-Bacterial Agents↗

Transdermal permeation of tetracaine hydrochloride by lecithin microemulsion: in vivo.

In vivo transdermal permeation of tetracaine hydrochloride encapsulated in lecithin water-in-oil and oil-in-water microemulsion was studied. The effect of the composition of the lecithin microemulsion on analgesic response of tetracaine hydrochloride was evaluated on Wistar rats by tail flick method. To find out the toxicity of lecithin/n-propanol/isopropyl myristate/water/tetracaine hydrochloride microemulsion histopathological and irritation response were measured in Swiss mice. Time course studies were also conducted for the biochemical response of microemulsion by measuring catalase, glutathione and lipid peroxidation levels of the treated mice skin. The analgesic response was found to be dependent on the drug concentration and composition of the systems. The histopathological, irritation and biochemical findings reveal that lecithin/n-propanol/isopropyl myristate/water/tetracaine hydrochloride microemulsion is a safe carrier for transdermal drug delivery systems. Confocal laser scanning microscopy observation indicated that sweat gland and hair follicle also provided the path for transdermal permeation of lecithin/n-propanol/isopropyl myristate/water microemulsion.

1-Propanol↗

Iontophoresis of lecithin vesicles of cyclosporin A.

Site-specific immunosuppression with topical cyclosporin A (CsA) has broad clinical implications in the treatment of skin disorders like psoriasis, pyoderma gangrenosum, lichen planus, cutaneous graft-versus-host disease and contact hypersensitivity and the temporary treatment of skin allografts on burn wounds. However, like any other peptide drug, its skin delivery is hindered by the barrier property of stratum corneum and the physicochemical properties of CsA. We have attempted to deliver CsA across human cadaver epidermis in vitro using colloidal systems like microemulsion and lecithin vesicles and iontophoresis. Although, passive diffusion did not result in permeation of quantifiable amounts of CsA, anodal iontophoresis of the negatively charged colloidal systems facilitated the permeation. Electroosmosis and compromised epidermis might have contributed to the higher skin flux. Lecithin vesicles were better than microemulsion for the iontophoretic delivery of CsA and appear to have potential in site-specific immunosuppression.

Cyclosporine↗

Physicochemical investigation of acrylamide solubilization in sodium bis(2-ethylhexyl)sulfosuccinate and lecithin reversed micelles.

The state of acrylamide confined within dry sodium bis(2-ethylhexyl)sulfosuccinate (AOT) and lecithin reversed micelles dispersed in CCl(4) has been investigated by FTIR and (1)H NMR spectroscopy. Measurements have been performed at 25 degrees C as a function of the acrylamide-to-surfactant molar ratio (R) at a fixed surfactant concentration (0.1 mol kg(-1)). The analysis of experimental data, corroborated by the results of SAXS measurements, is consistent with the hypothesis that acrylamide is quite uniformly distributed among reversed micelles mainly located in proximity to the surfactant head-group region and that its presence induces significant unidimensional growth of micellar aggregates. Moreover, the confinement of acrylamide within reversed micelles involves some changes of the typical H-bonded structure of pure solid acrylamide attributable to the establishment of system-specific acrylamide/surfactant head group interactions. Preliminary experiments showed that, by exposure to X-rays, the polymerization of acrylamide can be induced in the confined space of dry AOT and lecithin reversed micelles.

Acrylamide↗

Lecithinized superoxide dismutase (PC-SOD) improved spinal cord injury-induced motor dysfunction through suppression of oxidative stress and enhancement of neurotrophic factor production.

PC-SOD (lecithinized superoxide dismutase) is a derivative of human Cu, Zn-SOD conjugated with 4 molecules of lecithin, yet having the enzyme activity of scavenging superoxide anion (O2-). Intravenous administration of PC-SOD promoted the recovery from spinal cord injury (SCI)-induced motor dysfunction in a dose-dependent manner in rat model, when evaluated by BBB (Basso Beattie Bresnahan) score. Even when given at 24 h after SCI, PC-SOD (1 mg/kg) significantly improved motor dysfunction. Distribution study demonstrated that PC-SOD gradually accumulated to the injured site. Enzyme-linked immunoassay revealed that PC-SOD prevented quantitative loss of neurons, astrocytes, and oligodendrocytes. PC-SOD inhibited SCI-induced oxidative stress, such as the decrease of free sulfhydryl residue, acetylcholine esterase activity, and the increase of lipid peroxidation. PC-SOD increased the production of neuroprotective factors. HIF-1alpha gene expression increased following SCI, and PC-SOD further increased it. In conclusion, PC-SOD gradually accumulated and retained at the damaged site to scavenge excessive O2-, and suppressed neuronal death through reducing oxidative stress, increasing neuroprotective factor production and HIF-1alpha gene expression.

Animals↗

Lecithin translational diffusion studied by pulsed nuclear magnetic resonance.

The translational diffusion coefficient of egg yolk and dilauroyl lecithin in optically isotropic phases containing sodium cholate has been measured using the pulsed NMR magnetic field gradient method. After a correction for geometrical factors the measured diffusion coefficient is found to agree well with previous determinations in phospholipid systems. The experimental data imply that the cubic mesophase of the lecithin-sodium cholate-water system contains continuous lipid aggregates. A possible model of the arrangement of the different amphiphile molecules in the cubic phase is discussed.

Diffusion↗

Magnetic anisotropy of egg lecithin membranes.

Magnetic realignment and rotational diffusion of cylindrical egg lecithin vesicles were measured under a phase contrast microscope. The anisotropy of magnetic susceptibility times membrane thickness was calculated from the data for several thin-walled vesicles. The resulting values were assigned to discrete numbers of bilayers. The difference between the susceptibilities parallel and perpendicular to the long axes of the lecithin molecules is deduced to be X parallel - X perpendicular = -(0.28 +/- 0.02) . 10(-8) cgs at 23 degrees C, if a bilayer thickness of 60 A is assumed.

Magnetics↗

Phase behavior of ganglioside-lecithin mixtures. Relation to dispersion of gangliosides in membranes.

Ganglioside GM1 and mixed brain gangliosides were mixed with 1-stearoyl-2-oleoyl lecithin (SOPC) and examined by differential scanning calorimetry as a function of ganglioside content and temperature. Low mole fractions of ganglioside GM1 and of mixed brain gangliosides are shown to be miscible with SOPC in the gel phase up to X = 0.3, with the possible exception of a small region of immiscibility for the mixed brain gangliosides system centered around X = 0.05. Above X = 0.3, the low-temperature phases demix into a (gel) phase of composition X = 0.3 and a (micellar) phase of composition X = 1.0. Above the endothermic phase transition temperature, no phase boundaries are discerned. It is pointed out that phase structures need to be determined in each domain delineated in the phase diagrams, and that cylindrical phases may exist at higher temperatures and intermediate compositions. The effects of addition of wheat germ agglutinin, which binds to ganglioside GM1, on a ganglioside GM1-SOPC mixture (X = 0.5), are described and interpreted in terms of partial demixing of ganglioside and lecithin. Behavior of the ganglioside-SOPC system is discussed with respect to the kinetics of cholera toxin action in lymphocytes, as well as to other physiological roles of gangliosides in membranes.

Calorimetry, Differential Scanning↗

Thermoelasticity of large lecithin bilayer vesicles.

Micromechanical experiments on large lecithin bilayer vesicles as a function of temperature have demonstrated an essential feature of bilayer vesicles as closed systems: the bilayer can exist in a tension-free state (within the limits of experimental resolution, i.e., less than 10(-2) dyn/cm). Furthermore, because of the fixed internal volume, there is a critical temperature at which the vesicle becomes a tension-free sphere. Below this temperature, thermoelastic tension builds up in the membrane and the vesicle's internal pressure increases while the surface area remains constant. Above this temperature, the vesicle's surface area increases while the tension and internal pressure are negligible. Without mechanical support, the vesicles fragment into small vesicles because they have insufficient surface rigidity. In the upper temperature range we have measured the increase of surface area with temperature. These data established the thermal area expansivity to be 2.4 X 10(-3)/degrees C. At constant temperature, we used either pipet aspiration with suction pressures up to 10(4) dyn/cm2 or compression against a flat surface with forces up to 10(-2) dyn to produce area dilation of the vesicle surface on the order of 1%. The rate of increase of membrane tension with area dilation was calculated, which established the elastic area compressibility modulus to be 140 dyn/cm. The tension limit that produced lysis was observed to be 3-4 dyn/cm (equivalent to 2-3% area increase). The product of the elastic area compressibility modulus, the thermal area expansivity, and the temperature gives the reversible heat of expansion at constant temperature for the bilayer. This value is 100 ergs/cm2 at 25 degrees C, or approximately 5 kcal/mol of lecithin. Similarly, the product of the thermal area expansivity multiplied by the area compressibility modulus determines the rate of increase of thermoelastic tension with decrease in temperature when the area is held constant, i.e., -0.34 dyn/cm/degrees C.

Elasticity↗

Structure of gel phase saturated lecithin bilayers: temperature and chain length dependence.

Systematic low-angle and wide-angle x-ray scattering studies have been performed on fully hydrated unoriented multilamamellar vesicles of saturated lecithins with even chain lengths N = 16, 18, 20, 22, and 24 as a function of temperature T in the normal gel (L beta') phase. For all N, the area per chain Ac increases linearly with T with an average slope dAc/dT = 0.027 A2/degree C, and the lamellar D-spacings also increase linearly with an average slope dD/dT = 0.040 A/degree C. At the same T, longer chain length lecithins have more densely packed chains, i.e., smaller Ac's, than shorter chain lengths. The chain packing of longer chain lengths is found to be more distorted from hexagonal packing than that of smaller N, and the distortion epsilon of all N approaches the same value at the respective transition temperatures. The thermal volume expansion of these lipids is accounted for by the expansion in the hydrocarbon chain region. Electron density profiles are constructed using four orders of low-angle lamellar peaks. These show that most of the increase in D with increasing T is due to thickening of the bilayers that is consistent with a decrease in tilt angle theta and with little change in water spacing with either T or N. Because of the opposing effects of temperature on area per chain Ac and tilt angle 0, the area expansivity alpha A is quite small. A qualitative theoretical model based on competing head and chain interactions accounts for our results.

Gels↗

Two histidine residues are essential for catalysis by lecithin retinol acyl transferase.

Lecithin retinol acyl transferase (LRAT) is a novel membrane bound enzyme that catalyzes the formation of retinyl esters from vitamin A and lecithin. The enzyme is both essential for vision and for the general mobilization of vitamin A. The sequence of LRAT defines it as a novel enzyme unrelated to any other protein of known function. LRAT possesses a catalytically essential active site cysteine residue. The enzyme also contains six histidine residues. It is shown here that two of these residues (H57 and H163) are essential for catalysis. A mechanistic hypothesis is presented to account for these observations.

Acyltransferases↗

Dietary sitostanol reduces plaque formation but not lecithin cholesterol acyl transferase activity in rabbits.

The effects of graded amounts of dietary sitostanol (0.01, 0.2 and 0.8% (w/w)) were examined on plasma lipid-profile, coronary artery plaque development and lecithin:cholesterol acyl transferase activity in male New Zealand White rabbits given semi-purified diets for 10 weeks. All diets provided < 10% energy in the form of fat and contained 0.5% (w/w) cholesterol (C). Rabbits fed the semi-purified diet with 0.8% (w/w) (0.64 g/day) sitostanol had lower plasma total cholesterol (TC) (p = 0.006) (15.2 +/- 4.80 mmol/l) and very low-density lipoprotein-cholesterol (VLDL-C) (p = 0.007) (6.31 +/- 3.11 mmol/l) levels compared to the atherogenic control group (n = 6) (29.6 +/- 5.52 and 17.16 +/- 7.43 mmol/l, respectively). Dietary sitostanol at 0.8% (w/w) depressed plaque accretion in coronary arteries (p = 0.0014) and ascending aorta (p = 0.0004) compared with the atherogenic control, 0.01 and 0.2% (w/w) sitostanol-fed groups. No differences (p = 0.24) in the activity of lecithin:cholesterol acyl transferase (LCAT) were observed across groups, although plasma cholesterol fractional esterification rate was higher (p = 0.004) in the 0.8% (w/w) sitostanol fed animals compared with the atherogenic control. Significant negative correlations were demonstrated between sitostanol intake and plasma TC, LDL-C and VLDL-C levels. Hepatic campesterol levels were correlated (r = 0.3, p = 0.03) with plasma but not hepatic TC concentrations. These results demonstrate that dietary sitostanol at a concentration of 0.8% (w/w) or 0.64 g/day lowered plasma cholesterol levels and depressed atherosclerosis development in rabbits, but did not alter LCAT activity.

Animals↗

In vitro and in vivo comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of bovine semen.

Semen diluents containing egg yolk as a cryoprotectant may pose hygienic risks and are difficult to standardize. Although a new generation of semen diluents free of animal ingredients is available, egg yolk-containing extenders are still widely used for cryopreserving semen. We compared the effects of using different extenders on bovine sperm function in vitro and on fertility in vivo. A soy lecithin extender (SL; AndroMed) and an egg yolk-containing (TRIS-EY) extender were tested. No differences (P>0.05) were detected between the two extenders for sperm-zona pellucida binding capacity (HZI=115+/-13). Assessment of the inducibility of the acrosome reaction with progesterone showed no differences (P>0.05) between extenders for live acrosome-reacted sperm (15+/-2.36 and 14.42+/-2.02%, respectively, for SL and TRIS-EY). However, post-thaw sperm motility was significantly lower (P<0.05) when semen was extended in the TRIS-EY diluent. Field trials revealed that nonreturn rates of SL-extended semen showed significantly higher insemination success (P<0.0001) compared with the nonreturn rates for the TRIS-EY extender (70.45 and 67.85%, respectively). We suggest that consistent with quality standards that should be required for cryoprotectant media and because of the superior quality of the egg yolk-free extender, a defined soybean lecithin-containing diluter might be the better choice as a semen extender in the future.

Acrosome↗

Solution and adsorption behaviour of lecithin surfactants in CFC suspensions: a light scattering study in aerosol propellants.

The behaviour of the lecithin Epikuron 200 was studied in solution in a chlorofluorocarbon (CFC) aerosol propellant (P113) using light scattering to determine the micellar structure as a function of temperature. At room temperature Epikuron 200 formed micelles with a radius of approximately 25 nm. The radius increased with decreasing temperature until a phase boundary was reached at 15-16 degrees C, below which the micellar radius increased rapidly to 150-250 nm. This was accompanied by a large increase in the viscosity and turbidity of the solution. We consider that this behaviour is due to the formation of large wormlike micelles similar to those reported for lecithin in cyclohexane. The adsorption of Epikuron 200 on salbutamol sulphate was measured over this temperature range. The adsorbed amount of Epikuron 200 increased with decreasing temperature, and the hemimicelle concentration decreased rapidly at a temperature corresponding to the threshold for the formation of wormlike micelles. It is likely that these structures do form in CFC-MDI systems (particularly during cold-fill formulation), and that the structure of the adsorbed surfactant layer changes at low temperature. This may have implications for the formulation of stable aerosol suspensions.

Adsorption↗

Study of partition of nitrazepam in bile salt micelles and the role of lecithin.

The effect of trihydroxy (sodium cholate and sodium glycocholate) and dihydroxy (sodium deoxycholate and sodium glycodeoxycholate) bile salt micelles on the spectrophotometric properties and on the solubility of nitrazepam in aqueous solution, at 25.0 degrees C and at ionic strength 0.1 M in sodium chloride, has been assessed. From the results obtained it was possible to calculate the partition coefficients (Kp) of nitrazepam between aqueous and micellar phases. The partition coefficients of nitrazepam have also been determined in mixed micelles of cholate or deoxycholate with lecithin (egg yolk phosphatidylcholine), which were used as a model of the gastrointestinal tract. Drug partition was found to depend on the bile acid (number of hydroxyl groups and conjugation with glycine), and our data indicate further that addition of lecithin to bile salt micelles decreases the values of the partition coefficients in the mixed micelles at physiological pH.

Bile Acids and Salts↗

Determination of lecithin drug specialties and diet integrators; by means of first or second derivative enzymatic-spectrophotometric analysis.

The possibility has been investigated of applying derivative analysis to a classical enzymatic-spectrophotometric method for lecithin determination for the purpose of developing an analytical direct method that does not require long pretreatment of the test sample even in the case of turbid samples. Several samples of drugs and food integrators containing lecithin were thus tested using both the standard and the derivative method and then comparing the results obtained. The RSD% values of measurements on real (food or drugs) samples were always <2.5. Using a first derivative spectrophotometric method, average recovery was always between 102 and 105%.

Alcohol Oxidoreductases↗

Photoreactions of psoralens with lecithins.

The formation of cyclobutane (cb) photoadducts of psoralen with a model lecithin has been shown. The adducts are formed both in ethanol solution and in micellar suspension in water. In spite of their sensitivity to various factors such as light, temperature, air, etc., they are isolated by thin-layer chromatography (TLC) and high-pressure liquid chromatography (HPLC) and characterized by mass spectrometry, NMR and UV absorption spectroscopy. The NMR analysis indicates the similarity of isomeric forms of cb adducts in lecithin to those formed with free oleic acid.

Ficusin↗