Role of Endogenous Ingredients in Meibum and Film Structures on Stability of the Tear Film Lipid Layer against Lateral Compression.

The property and structure of spread films of meibum extruded from rabbit eyelids and its fractions were investigated using the Langmuir film balance technique and Brewster angle microscopy in order to understand the influence of endogenous ingredients in meibum on the structure and stability of the tear film lipid layer against mechanical stimulus. Surface pressure (?)?film area ( A) isotherms for meibum were measured upon repetitive high-speed compression?expansion cycles and were found almost identical to each other with very small hysteresis, indicating the high stability of the meibum film. Brewster angle microcopy observation implied the spontaneous formation of condensed-phase network structures which consist primarily of wax esters and cholesteryl esters as nonpolar ingredients, coexisting with a monolayer phase of polar lipids two-dimensionally confined by the networks, which were spontaneously formed in the meibum film. The networks were gathered densely and deformed when the film was laterally compressed with barriers of Langmuir trough, but returned to the dispersed networks when expanded. The influence of temperature and salts dissolved in an aqueous subphase was also investigated. The results indicated that the temperature change (20 and 35 ?C) induced a difference of surface pressure at the same film areas in rather compressed films, and the presence of salts in the subphase expanded the films. However, the features of isotherm and surface morphology of the film, including their reversibility, were maintained. Phospholipid-removed meibum also formed a stable film, but slight changes were found in the hysteresis and film morphology compared to those in the meibum film. In contrast, in a film of phospholipid- and cholesterol-removed meibum, three-dimensional aggregates grew upon the first compression and not redispersed by the subsequent expansion, giving noticeable hysteresis between the isotherms. It is considered that high deformability upon compression and resilience upon expansion of the networks as well as reversible collapse and spreading property of the confined monolayer phase would hold the stability of the meibum film against repeated compressions.

Effect of Mineral-Oil Addition on Film Stability of Polar Lipid Constituents Derived from Meibum.

We studied the effects of mineral oil (MO) on the properties and structure of a spread monolayer of polar lipid constituents in meibum, by performing cyclic lateral compression-expansion experiments using a Langmuir trough. A meibum sample without nonpolar lipids (meibumΔnonpolar-lipid) was prepared by removing the nonpolar lipids from biological meibum extruded from rabbit eyelids and spread on a water surface for measuring the cyclic surface pressure (π)-film area (A) isotherms with in situ observation of the film morphology using a Brewster angle microscope. The meibumΔnonpolar-lipid formed a homogeneous fluid monolayer and underwent collapse upon compression. The π-A isotherm shifted to a smaller area upon repeating the compression-expansion cycles. These observations contrasted those obtained for meibum previously, which may have resulted from the absence of nonpolar lipids. The recovery of the film stability against the lateral compression-expansion cycles was analyzed by adding MO as a nonpolar compound to the film system. A spread film of 1:1 mixture (by weight) could recover the high reversibility of the π-A isotherms during the repeated compression and expansion processes.

A thin layer of sucrose octasulfate protects the oesophageal mucosal epithelium in reflux oesophagitis.

Sucralfate is effective for the treatment of gastric and duodenal ulcers owing to its protective gel-forming ability. However, the mechanism by which sucralfate protects the oesophageal mucosa against reflux oesophagitis has not been clarified. We aimed to investigate the mechanisms of action of sucralfate and sucrose octasulfate (SOS), a component of sucralfate. SOS and sucralfate were administered to oesophagitis-induced rats, and the ulcer lesion size was macroscopically examined and scored. Effective pepsin activity in the gastric juices obtained from the animal model was evaluated by a casein digestion test. Sucralfate and SOS improved the pathology scores in a dose-dependent manner, whereas gastric juice pepsin activity was not impaired by therapeutic doses of SOS. As SOS lacks the ability to form a thick gel layer by polymerisation, we examined the distribution of SOS in the mucosal lumen by imaging mass spectrometry (IMS) to determine whether SOS directly adheres to the mucosal surface. A clear homogeneous thin-layer SOS film (>100 µm thick) was visualized on the oesophageal mucosal surface. Moreover, this SOS film formation was enhanced at ulcer lesion sites. Taken together, SOS appears to protect oesophageal mucosa against reflux oesophagitis via thin-layer formation on the mucosal surface.

Membrane permeation of giant unilamellar vesicles and corneal epithelial cells with lipophilic vitamin nanoemulsions.

Nanoemulsions of a lipophilic vitamin, retinol palmitate (vitamin A; VA), have a therapeutic effect on corneal damage. The nanoemulsion based on a triblock-type polymer surfactant with polyoxyethylene and polypropylene, EO100PO70EO100 (EOPO) showed superior efficacy, as compared with a nanoemulsion based on polyoxyethylene (60) hydrogenated castor oil (HCO). We studied the mechanism of VA nanoemulsions related to efficacy from the viewpoint of the interaction with plasma membrane-mimicking giant unilamellar vesicles (GUVs) and the plasma membrane permeation in corneal epithelial cells. When nanoemulsions and GUVs doped with fluorescent compounds were mixed each other, and observed by confocal laser microscopy, EOPO nanoemulsions induced endocytic morphological changes like strings and vesicles of the bilayer drawn inside a GUV by budding. Judging by isothermal titration calorimetry and ζ potential measurements, the EOPO nanoemulsions seemed to have stronger hydrophobic interactions with the lipid bilayer because of lower coverage of the core interface. Next, when the nanoemulsions prepared with a pyrene derivative of retinol (VApyr) were applied to corneal epithelial cells, the EOPO nanoemulsions greatly permeated the cells and gathered around the cell nucleus, as compared with HCO nanoemulsions. Furthermore, according to the three-dimensional images of the cell, it was found that the vesicles that absorbed nanoemulsions formed from the plasma membrane as real endocytosis, and were transported to the area around the nucleus. Consequently, it is likely that EOPO nanoemulsions entered the cell by membrane-mediated transport, delivering VA to the cell nucleus effectively and enhancing the effects of VA.

DHA-PC and PSD-95 decrease after loss of synaptophysin and before neuronal loss in patients with Alzheimer's disease.

Alzheimer's disease (AD) is a progressive neurodegenerative disease that is characterized by senile plaques, neurofibrillary tangles, synaptic disruption, and neuronal loss. Several studies have demonstrated decreases of docosahexaenoic acid-containing phosphatidylcholines (DHA-PCs) in the AD brain. In this study, we used matrix-assisted laser desorption/ionization imaging mass spectrometry in postmortem AD brain to show that PC molecular species containing stearate and DHA, namely PC(18:0/22:6), was selectively depleted in the gray matter of patients with AD. Moreover, in the brain regions with marked amyloid ß (Aß) deposition, the magnitude of the PC(18:0/22:6) reduction significantly correlated with disease duration. Furthermore, at the molecular level, this depletion was associated with reduced levels of the postsynaptic protein PSD-95 but not the presynaptic protein synaptophysin. Interestingly, this reduction in PC(18:0/22:6) levels did not correlate with the degrees of Aß deposition and neuronal loss in AD. The analysis of the correlations of key factors and disease duration showed that their effects on the disease time course were arranged in order as Aß deposition, presynaptic disruption, postsynaptic disruption coupled with PC(18:0/22:6) reduction, and neuronal loss.