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Dissolution rate studies of cholesterol monohydrate in bile acid-lecithin solutions using the rotatingdisk method.

A physical model approach was used to investigate cholesterol gallstone dissolution kinetics in simulated bile. Critical experimental and theoretical investigations simulating in vivo conditions showed that, in the bile acid-lecithin solutions, there is a significant interfacial barrier for both cholesterol gallstone and cholesterol monohydrate pellet dissolution. In the present study, the rotating-disk dissolution method and the accompanying Levich theory were applied to assess the contributions of the diffusion convection mass transfer resistance and of the interfacial barrier to the overall kinetics. Cholesterol dissolution rates in bile acid solutions were about 2-20 times slower than diffusion-controlled rates depending upon the degree of agitation. As found in previous studies, these rates in the presence of sufficient concentrations of dissolution accelerators approached the theoretical diffusion-convection-controlled rates. To account for the much slower dissolution rates in bile acid-lecithin solutions, two possible kinetic interpretations were investigated. The first is based upon slow crystal-micellar solution interfacial kinetics, and the second is based upon a slow rate of cholesterol solubilization in the aqueous diffusion layer. For the latter, an analytical mathematical solution was obtained.

Benzoates↗

Dissolution kinetics of cholesterol in simulated bile I: Influence of bile acid type and concentration, bile acid-lecithin ration, and added electrolyte.

A physical model approach was utilized to investigate cholesterol monohydrate dissolution kinetics in simulated bile. The static pellet method and the Berthoud theory were employed to assess the contributions of the diffusion-convection mass transfer resistance and those of the interfacial resistance to the overall kinetics. For almost all situations studied, the interfacial resistance was the dominant rate-determining factor. The effects of four bile acids and their concentrations, the bile acid-lecithin ratio, and the added electrolytes and their concentrations on the interfacial resistance were examined. The results were correlated with those obtained with human bile samples, and the indications were that the kinetics of cholesterol dissolution in bile may be explainable on the basis of the principal bile acids, lecithin, and the electrolytes in the bile.

Bile↗

Interaction between erythrocytes from various animals and emulsions stabilized with various lecithins.

The degree of hemolysis caused by the interaction between erythrocytes from various animals and emulsions stabilized with various lecithins was evaluated as a measure of the safety of emulsions for drug carriers. The stability of the emulsions was estimated using the gradient of the slope derived from the direct linear correlation between the percentage hemolysis and the phosphatidylcholine (PC) content of the erythrocyte membrane. When members of the egg lecithin (EPC) series were used as emulsifiers of emulsions, the percentage hemolysis increased as the PC content of the erythrocyte membrane increased and as the sphingomyelin (SM) content of the erythrocyte membrane decreased. Lysolecithin, a contaminant present in the emulsifying agent of emulsions, did not have any significant influence on the hemolysis of erythrocytes. These experimental findings show that the hemolysis caused by interaction between emulsions and erythrocytes was dependent on the PC content of both the emulsifying agent used and the erythrocyte membrane, and that the SM present in the erythrocyte membrane was an essential component for the stability of erythrocytes against emulsion-induced hemolysis.

Animals↗

Effect of interfacial pressure on the binding and phospholipase A2 activity of recombinant human lecithin-cholesterol acyltransferase.

We examined the effect of surface pressure on the interfacial binding and phospholipase A2 activity of lecithin-cholesterol acyltransferase. The enzyme bound to phosphatidylcholine monolayers with an apparent dissociation constant of 1.5 nM was excluded from the interface at pressures > 29 mN/m and exhibited maximal phospholipase activity at pressures between 29-28 mN/m. These data suggest that lipoprotein surface pressure may regulate lecithin-cholesterol acyltransferase activity in vivo.

Humans↗

The influence of paracetamol on the hepatic biosynthesis of lecithin.

In paracetamol induced liver injury changes in the phospholipid components of microsomal membranes are usual. Direct incubation of paracetamol with subcellular fractions of human liver specimens did not alter the biosynthesis of lecithin. Preincubation of liver slices with paracetamol led to an increased synthesis of lecithin. The presence of intact cells and the integrity of hepatocellular function seem to be a prerequisite for the poisoning effects in vitro.

Acetaminophen↗

Influence of phytogenic surfactants (quillaya saponin and soya lecithin) on bio-elimination of phenanthrene and fluoranthene by three bacteria.

The influence of two phytogenic surfactants on the elimination of polycyclic aromatic hydrocarbons (PAH) was studied in shaken-batch cultures of three soil bacteria under axenic conditions. At sufficiently high concentrations, quillaya saponin and soybean lecithin solubilized phenanthrene or fluoranthene efficiently. However, complete solubilization of the PAH by lecithin only doubled the maximal rate of elimination of the two PAH compounds by Pseudomonas 0259, strain MKm (Rhizomonas ?) and Mycobacterium EMI 2. By contrast, quillaya saponin did not improve PAH bioavailability, and in strain MKm it caused significant growth lags above 2.5 g/l. Simultaneously with the elimination of the PAH the bacteria utilized the surfactants as substrates for growth. Intermediate formation of PAH metabolites was noted. The results suggest that some phytogenic surfactants might improve PAH bioavailability in rhizospheres.

Bacteria↗

Mixtures of lecithin with polymerizable derivatives of cholesterol. A monolayer film balance study.

One of the best investigated binary lipid mixtures is the lecithin-cholesterol system. We show here that it is possible to modify the cholesterol in such a way that it can be polymerized without changing its behaviour in mixtures with lecithin. The polymerizable derivatives exhibit a very similar phase diagram in the mixture with dipalmitoylphosphatidylcholine as the cholesterol itself. This is demonstrated by filmbalance measurements.

1,2-Dipalmitoylphosphatidylcholine↗

Tetrahydroaminoacridine and lecithin in the treatment of Alzheimer's disease. Effect on cognition, functioning in daily life, behavioural disturbances and burden experienced by the carers.

In 12 ambulant patients with a clinical diagnosis of probable Alzheimer's disease, a 12-week, double-blind, placebo-controlled study with 100 mg/day tetrahydroaminoacridine (THA) and 10 g/day lecithin is reported. The aim of the study was to find whether treatment would result in an improvement of cognition, of functioning in daily life, decrease of behavioural disturbances, and decrease in burden experienced by the carers. Two of the six THA-treated patients demonstrated an increase on cognitive test scores with a moderate increase in the other outcome measurements. There was, however, no difference between the two groups in any outcome measurement after treatment. In addition, we found a reversible rise of liver transaminases in 4 of 6 patients in the treated group. This pilot study is too small to draw definite conclusions on the use of THA alone or in combination with lecithin. Our results suggest, however, that semi-structured interviews with the carers may be of value in the evaluation of treatment effects in patients with Alzheimer's disease.

Aged↗

Orientation and rotational freedom of fluorescent probes in lecithin bilayers.

The fluorescence polarization properties of lecithin bilayers stained with 2,6-MANS and 1,8 ANS under applied potential steps have been studied. The fluorescence signal components of both dyes were found to have different sign and relative amplitude, suggesting that 1,8-ANS and 2,6-MANS behave differently when bound to black lipid membranes. In order to determine the location and the extent of rotational brownian motions of the bound chromophores, the experimental data were analyzed by using a simplified physico-mathematical model. According to it 2,6-MANS appears to have a ratio rho/tau higher than 1.8-ANS (rho being the rotational relaxation of in plane rotations and tau the lifetime of the excited singlet state of the bound molecules), suggesting that the former chromophore is more tightly held inside the bilayers. Furthermore, 2,6-MANS is found to possess the absorption and emission oscillators more closely oriented to the normal of membrane surface, while 1,8-ANS has both oscillators almost near the plane of the bilayers. The results furnish also a fair estimate of the random molecular motion own by the phospholipid molecules at room temperature. The comparison of the present data with those obtained from squid axon membranes confirms the validity of the proposed physical model, yielding a rough estimate of the axon membrane-area covered by integral protein macromolecules. These preliminary results derived from lecithin model membranes suggest that fluorescence polarization techniques can provide valuable informations if applied to study the macromolecular organization of in vitro reconstituted membranes.

Animals↗

[A rapid method for determination of oil in lecithins (author's transl)].

A rapid method for routine determination of oil in commercial lecithins and similar phospholipid samples has been worked out. This based on the observation that under the described experimental conditions 1 g of phospholipids is completely adsorbed on 45 g silica gel which has previously been disactivated by the addition of 15% water. The lecithin sample is filtered in ethereal solution through a column filled with disactivated silica gel which is subsequently washed with a little ether. The combined ether eluates are evaporated to dryness invacuo and the residue weighed. This residue is the oil part of the sample and is constituted of mono-, di-, and triglycerides, free fatty acids, sterols and their derivatives. The duration for one such determination is about 20 min.

Chromatography, Thin Layer↗

Pigment containing lipid vesicles. I. Preparation and characterization of chlorophyll a-lecithin vesicles.

Vesicles obtained by sonication of chlorophyll a-lecithin mixtures dispersed in anaqueous medium closely resemble the well-characterized vesicles similarly prepared from pure lipids. They are bounded by one spherical lipid bilayer which contains the chlorophyll a. Appropriate conditions for sonication prevent substantial degradation of the membrane constituents. Up to one chlorophyll a molecule per 55 lecithins can be incorporated into membranes. The average Stokes' radius of the vesicles determined by analytical sieve chromatography is 102 +/- 5 A and independent of the chloropyll a content. The membrane is visible in the electron-microscope when the vesicles are treated with osmium tetroxide prior to negative staining. The osmium fixation is, however, not strong enough to allow for a preparation of the vesicles for thin sectioning (dehydration, embedding in epoxide).

Chlorophyll↗

Influence of enzymatic phospholipid cleavage on the permeability of the erythrocyte membrane: III. Discrimination between the causal role of split products and of lecithin removal.

Cleavage of 55% of the lecithin in intact human erythrocytes by phospholipase A2 (bee venom) markedly inhibits the mediated transport of L-lactate (via the monocarboxylate carrier) and of L-arabinose (via the monosaccharide carrier), while the major anion exchange system (probed by oxalate) and diffusion via the lipid domain (probed by erythritol) remain essentially unaltered. the causal role of the split products, unsaturated fatty acids and saturated lysolecithin, and of lecithin removal were now studied by sequential extraction of split products with serum albumin and by their controlled insertion into normal membranes. Careful choice of the albumin-to-cell ration allowed the extraction of more than 95% or the fatty acids and up to 80% of the lysolecithin without hemolysis. Extraction of fatty acids abolished inhibition of lactate and arabinose transfer, but induced inhibition of anion exchange and translipid permeation. Subsequent extraction of lysolecithin produced no further effects except on lactate transfer, which was inhibited. Exogenous oleic and linoleic acid, at intramembrane concentrations equal to those produced by phospholipase A2, inhibit lactate and arabinose transfer, while accelerating oxalate and erythritol movements, in agreement with effects of endogenous fatty acids. Exogenous lysolecithin inhibits all mediated transfer processes but does not alter translipid permeation. This pattern differs from that obtained for endogenous lysolecithin. The action of exogenous lysolecithin can be suppressed by loading of the cells with cholesterol. Insertion of exogenous lysolecithin into cells depleted of endogenous lysolecithin does not restore the functional state before depletion, indicating that exogenous and endogenous lysolecithin may act differently.

Biological Transport↗

Changes in relative fatty acid composition of serum lecithin and cholesterol ester after treatment with two gonane progestins administered alone and in combination with ethinyl estradiol.

Twenty women, oophorectomized as part of treatment for cervical carcinoma of the cervix participated in the study. Ten of them were given desogestrel (DG) 150 micrograms/day for 3 weeks followed by DG + 30 micrograms ethinyl estradiol (EE) for 6 weeks, and finally EE alone for 3 weeks. The other 10 women were given 150 micrograms levonorgestrel (NORG) and EE in a similar way. Before treatment and after each period of treatment the relative fatty acid composition of serum lecithin and serum cholesterol ester were assessed by gas-liquid chromatography. In both groups ethinyl estradiol, when administered alone, increased arachidonic acid in serum lecithin. This increase was not observed on the DG + EE or the NORG + EE combination indicating that both progestins counteracted this estrogenic effect.

Adult↗

The fatty acid composition of serum lecithin after pregnancy complicated by pre-eclampsia.

Using gas-liquid chromatography the relative fatty acid composition of serum lecithin was measured in 22 women, during and after pregnancies complicated by pre-eclampsia, and the results were compared with those in 21 "normal" puerperal women. The increased values of 22:6 in the linolenic acid series found during pregnancy in women who had pre-eclampsia were also noted in the puerperium and the 22:6 value fell to normal more slowly. This might be due to an enhanced activity of the deacylation-reacylation cycle in this group of patients. All other fatty acids showed a rapid return to normal with values comparable to those of "normal" puerperal women 1 week after delivery. The differences in essential fatty acid distribution between normal and pre-eclamptic women did not seem to have a dietary cause. The prostaglandin precursors (arachidonic and dihomo-gamma-linolenic acids) were not reduced during pre-eclampsia or in the puerperium. Neither breast-feeding nor treatment of hypertension with hydralazine seemed to influence the fatty composition of serum lecithin.

Adult↗

Lecithin:cholesterol acyltransferase activity in hypercholesterolemic subjects and in hypercholesterolemic subjects treated with clofibrate.

The lecithin:cholesterol acyl transfer reaction in the plasma of hypercholesterolemic subjects and of hypercholesterolemic subjects treated with clofibrate was studied. An increased enzyme activity was found in the first group of patients, while lecithin:cholesterol acyl transfer activity tended to normalize in the second group. This increased enzyme activity might be a defense mechanism against the accumulation of cholesterol in the arterial wall.

Acyltransferases↗

Identification of vasopressor phospholipid in crude soybean lecithin.

The vasopressor phospholipid in crude soybean lecithin was isolated by column chromatography on Sephadex LH-20. It represented 0.1% of crude soybean lecithin. The isolated phospholipid was identified to be lysophosphatidic acid by gas chromatography-mass spectrometry analysis of TMS-deacylated product and acetolysis product. Nuclear magnetic resonance analysis favored the 1-monoacyl isomer over the 2-isomer. By enzymic determination with L-3-glycerophosphate dehydrogenase, the isolated phospholipid was identified as 1-monoacyl-L-3-glycerophosphate. Gas chromatographic examination revealed that it was composed of a large percentage of unsaturated fatty acids, especially linoleic acid. The activity of isolated lysophosphatidic acid was slightly less than that of synthetic 1-linoleoyl-L-3-glycerophosphate.

Animals↗

Formation of complexes between lecithin and apovitellenin I, an avian egg-yolk apoprotein.

In a study of lipid-protein interactions in egg yolk, it was found that L-alpha-dipalmitoyl lecithin gave two distinct noncovalent complexes (A and B) with apovitellenin I, an apoprotein in the major yolk lipoprotein. Interaction took place under widely varied conditions, and yolk lecithin gave similar complexes. Complex A, which was formed within minutes, consisted of round particles of about 9 nm diameter. Complex B, which was formed more slowly, consisted of larger particles, possibly resembling curved discs, with diameter of 30-40 nm. The preparation and some properties of these complexes are described. It is suggested that they may be suitable for an extensive study of phospholipid-protein interactions in yolk.

Animals↗

Positional specificity of trans fatty acids in fetal lecithin.

Differences in the positional incorporation of 9-trans[1-(14)C] octadecenoic (elaidic) and 9-trans,12-trans[1-(14)C] octadecadienoic (linoelaidic) acids in fetal lecithin of rats were demonstrated. On the 20th day of gestation, a 14C-labeled albumin complex of elaidic or linoelaidic acid was injected into the jugular vein of pregnant rats. For comparative purposes, 9-cis[1-(14)C] octadecenoic (oleic) or 9-cis,12-cis[1-(14)C] octadecadienoic (linoleic acid) was injected into the maternal circulation of rats. Animals were killed 6 hr later. Distribution of label in total lipids and phospholipids (PL) of fetal issue was measured by TLC. Irrespective of the label, the highest percentage of total radioactivity was associated with PL-59 to 67%. Within PL, the major portion of radioactivity was found in choline phosphoglycerides (CPG)-53 to 67%, and in ethanolamine phosphoglycerides (EPG)-18 to 33%. While linoelaidic acid was predominantly esterified in the 2-position of CPG, elaidic acid was nearly equally distributed between positions 1 and 2 of lecithin. Distribution of radioactivity within fatty acid methyl esters (FAME) of CPG measured by radio-GLC suggested that oleic and possibly linoleic acids may be converted to nervonic and arachidonic acid, respectively, in the rat by the 20th day of gestation. Following injection of elaidate, radioacivity of FAME was distributed between palmitate and elaidic acid indicating that rat fetal tissue may metabolize elaidic acid via beta-oxidation. In contrast, following injection of linoelaidate, radioactivity of FAME was primarily associated with tt-18:2, suggesting little biotransformation to other fatty acids by fetal tissues.

Animals↗