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Inhibition of pulmonary surfactant by oleic acid: mechanisms and characteristics.

The inhibitory effects of oleic acid (OA) on the surface activity of pulmonary surfactant were characterized by use of the oscillating bubble surfactometer, the Wilhelmy balance, and excised rat lungs. Oscillating bubble studies showed that OA prevented lavaged calf surfactant [0.5 mM phospholipid (PL)] from lowering surface tension below 15 mN/m at or above a molar ratio of OA/PL = 0.5. In contrast to inhibition of surfactant by plasma proteins, increasing the surfactant concentration did not eliminate inhibition by oleic acid, which occurred at OA/PL greater than 0.67 on the oscillating bubble even at surfactant concentrations of 1.5 and 12 mM PL. Studies of surfactant adsorption showed that preformed films of OA had little effect on the adsorption of pulmonary surfactant. Wilhelmy balance studies showed that OA did interfere with the ability of spread films of surfactant to reach low surface tensions during dynamic compression. Further balance experiments with binary films of OA and dipalmitoyl phosphatidylcholine showed that these compounds were miscible in surface films. Together these findings suggested that OA inhibited pulmonary surfactant activity by disrupting the rigid interfacial film responsible for the generation of very low surface tension during dynamic compression. Mechanical studies in excised rat lungs showed that instillation of OA gave altered deflation pressure-volume characteristics with decreased quasi-static compliance, indicating disruption of pulmonary surfactant function in situ. This alteration of mechanics occurred without major changes in the composition of lavaged PLs or in the tissue compliance of the lungs defined by mechanical measurements during inflation-deflation with saline.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Production and characterization of O/W emulsions containing cationic droplets stabilized by lecithin-chitosan membranes.

Oil-in-water emulsions containing cationic droplets stabilized by lecithin-chitosan membranes were produced using a two-stage process. A primary emulsion was prepared by homogenizing 5 wt % corn oil with 95 wt % aqueous solution (1 wt % lecithin, 100 mM acetic acid, pH 3.0) using a high-pressure valve homogenizer. This emulsion was diluted with aqueous chitosan solutions to form secondary emulsions with varying compositions: 1 wt % corn oil, 0.2 wt % lecithin, 100 mM acetic acid, and 0-0.04 wt % chitosan (pH 3.0). The particle size distribution, particle charge, and creaming stability of the primary and secondary emulsions were measured. The electrical charge on the droplets increased from -49 to +54 mV as the chitosan concentration was increased from 0 to 0.04 wt %, which indicated that chitosan adsorbed to the droplet surfaces. The mean particle diameter of the emulsions increased dramatically and the emulsions became unstable to creaming when the chitosan concentration exceeded 0.008 wt %, which was attributed to charge neutralization and bridging flocculation effects. Sonication, blending, or homogenization could be used to disrupt flocs formed in secondary emulsions containing droplets with high positive charges, leading to the production of emulsions with relatively small particle diameters (approximately 1 microm). These emulsions had good stability to droplet aggregation at low pH (< or =5) and ionic strengths (<500 mM). The interfacial engineering technology utilized in this study could lead to the creation of food emulsions with improved stability to environmental stresses.

Chitin↗

Poly(ethylene carbonate) microspheres: manufacturing process and internal structure characterization.

The granulocyte-macrophage colony stimulating factor (GM-CSF), a water-soluble cytokine, was encapsulated in poly(ethylene carbonate) microspheres (MS) by a double emulsion w(1)/o/w(2) solvent evaporation method. Poly(ethylene carbonate) is a new polymer of high molecular weight (MW) and forms polymer matrices that are exclusively surface bioerodible. In the frame of this study, the influence of the polymer molecular weight and the polymer concentration in the organic phase on the physico-chemical characteristics of the microspheres were investigated. Ninety percent of the microspheres had a diameter ranging between 4 and 136 microm, with a mean value of 30 microm. The encapsulation ratios ranged from 2.22 to 2.51% (w/w) depending on the molecular weight of the polymer corresponding to an encapsulation efficiency of 70 to 100%, respectively. Independent of the polymer molecular weight used, the in vitro drug release was very low, ranging from 5.61 to less than 1% of the total encapsulated GM-CSF amount. Scanning electron microscopy (SEM) analysis showed microparticles with spherical shapes and smooth surfaces containing a few small globules. The inner structure of the microspheres appeared to consist of a polymeric matrix surrounding numerous globules. These globules have different sizes, shape and distribution in the polymeric matrix, depending on the concentration of the polymer solution and on the polymer molecular weight. In addition, it was demonstrated that the GM-CSF lowered the interfacial tension between the GM-CSF aqueous solution and the methylene chloride organic phase. The active critical concentration was as low as 0.008 mg/ml. It was therefore suggested that this particular behavior contributed to the stabilization of the primary emulsion during the formation of the microspheres, leading to rather high encapsulation efficiency.

Cell Division↗

A Newtonian rheological model for the interface of microbubble contrast agents.

A quantitative model of the dynamics of an encapsulated microbubble contrast agent will be a valuable tool in contrast ultrasound (US). Such a model must have predictive ability for widely varying frequencies and pressure amplitudes. We have developed a new model for contrast agents, and successfully investigated its applicability for a wide range of operating parameters. The encapsulation is modeled as a complex interface of an infinitesimal thickness. A Newtonian rheology with surface viscosities and interfacial tension is assumed for the interface, and a modified Rayleigh-Plesset equation is derived. The rheological parameters (surface tension and surface dilatational viscosity) for a number of contrast agents (Albunex, Optison and Quantison) are determined by matching the linearized model dynamics with experimentally obtained attenuation data. The model behavior for Optison (surface tension 0.9 N/m and surface dilatational viscosity 0.08 msP) was investigated in detail. Specifically, we have carried out a detailed interrogation of the model, fitted in the linear regime, for its nonlinear prediction. In contrast to existing models, the new model is found to capture the characteristic subharmonic emission of Optison observed by. A detailed parametric study of the bubble behavior was executed using the ratio of scattering to attenuation (STAR). It shows that the encapsulation drastically reduces the influence of resonance frequency on scattering cross-section, suggesting possible means of improvement in imaging at off-resonant frequencies. The predictive capability of the present model indicates that it can be used for characterizing different agents and designing new ones.

Albumins↗

Structure and supramolecular architecture of membrane channel-forming peptides.

Peptides gathering together to induce channels in lipid bilayers may be classified in several categories according to the spatial structures involved. For example, gramicidin A forms intramolecular tubes, alamethicin, bundles of helical rods with intermolecular pores, porins (being proteins, properly speaking) are rich in beta-sheets that may form barrels, whereas cyclic peptides might stack together resulting in the formation of pores. The chemical structure of these compounds is now well characterized. The transmembrane electrical signals that they transmit are also typical of the particular supramolecular configurations (or architecture). Investigations in this field are thus relevant to structure-function relationship studies due to the availability of natural or synthetic analogues allowing the measurement of the influence of physico-chemical parameters upon the energy profiles of the pores. Consequently, questions such as the existence and probabilities of conductance substrates, their voltage-dependence and their ion or molecular selectivity can be tackled. Today, the loosest aspect of these studies lies in the actual molecular conformations and architecture in the membranes of the peptide aggregates, the knowledge of which remains imprecise, even 'at rest' in the best-studied cases. This review attempts to point out still unresolved questions and to propose some plausible approaches concerning, for example: 1) the configurations of the molecular aggregates responsible for ion transfer; 2) the mechanisms for channel-opening and closing (gating); 3) the eventual cooperative phenomena between channels, via the bilayer or interfacial components. Possible applications of these structures will be tentatively outlined.

Alamethicin↗

The proteins of the surfactant system.

The structural and functional integrity of pulmonary surfactant depends on several specific proteins. Two of these, SP-A and SP-D, are large and water-soluble, while SP-B and SP-C are small and very hydrophobic. SP-A is an 18-mer of 26 kDa polypeptide chains and contains N-linked oligosaccharides. Structurally, it can be characterized as a collagen/lectin hybrid. Together with SP-B, SP-A is required for conversion of secreted endogenous surfactant to tubular myelin in the alveolar lining. It also regulates surfactant secretion and reuptake of surfactant lipids by type II cells; these functions are probably receptor mediated. SP-D, a 12-mer of 39 kDa polypeptide chains, is a collagenous glycoprotein with structural similarities to C-type lectins. Both SP-A and SP-D stimulate alveolar macrophages. SP-B is a 79-residue polypeptide that contains three intrachain disulphide bridges. It exists mainly as a homodimer, which is strongly positively charged and may selectively remove anionic and unsaturated lipid species from the alveolar surface film, thereby increasing surface pressure. SP-C is a mainly alpha-helical, extraordinarily hydrophobic polypeptide containing 35 amino acid residues and covalently linked palmitoyl groups. Its alpha-helical portion is inserted into surfactant lipid bilayers. SP-C accelerates the adsorption of lipid bilayers to an interfacial monolayer. In babies with respiratory distress syndrome, the clinical response to treatment with surfactant containing SP-B and SP-C is much faster than in babies treated with protein-free synthetic surfactant. We speculate that, in the near future, surfactant preparations based on recombinant hydrophobic proteins will be available for clinical use.

Glycoproteins↗

High-resolution electron microscope observation of interface microstructure of a cast Al-Mg-Si-Bi-Pb(6262)/Al2O3p composite.

High-resolution electron microscopy was employed to characterize the interface structure of a cast Al-Mg-Si-Bi-Pb aluminium(6262)-based composite reinforced by alpha alumina particles with a trace of beta alumina in order to investigate the behaviour of alloying elements in cast composites. Except for a few primary Mg2Si particles, few reaction products were detected at the interface of Al/alpha-Al2O3 due to the unfavourable reaction kinetics during the squeeze-casting process. The Mg2Si particle has an orientation relationship with alpha-Al2O3 of [011]Mg2Si//[1210]alpha-Al2O3 (111)Mg2Si//(0006)alpha-Al2O3. A significant amount of MgAl2O4 was found on the surface of the beta-Al2O3 particles, which is in contrast to the small degree of reaction found on alpha-Al2O3 particles. MgAl2O4 and beta-Al2O3 particles have the following orientation relationship: [011]MgAl2O4//[1210]beta-Al2O3 (111) MgAl2O4//(0006)beta-Al2O3. The similar crystal structure of beta-Al2O3 to MgAl2O4 favours MgAl2O4 nucleation and growth on the surface of beta-Al2O3. Interfacial energy minimization dominates the atomic structure of the interface with the result that close packed planes and directions in the Al2O3 reinforcement and reaction products are parallel to the interfaces. Bi and Pb were found in the form of metallic nanometre particles between Al2O3 particles, or between the MgAl2O4 and Al2O3 particles, or in the open channels of beta-Al2O3 filled by the Al matrix.

Journal Article↗

Differential Localization of Carbohydrate Epitopes in Plant Cell Walls in the Presence and Absence of Arbuscular Mycorrhizal Fungi.

Two monoclonal antibodies (McAbs) generated against rhamnogalacturonan I and characterized as specific for a terminal [alpha]-(1->2)-linked fucosyl-containing epitope (CCRC-M1) and for an arabinosylated [beta]-(1,6)-galactan epitope (CCRC-M7) were used in immunogold experiments to determine the distribution of the epitopes in four plants. Allium porrum, Zea mays, Trifolium repens, and Nicotiana tabacum plants were chosen as representatives of monocots and dicots with different wall structures. Analyses were performed on root tissues in the presence and absence of arbuscular mycorrhizal fungi. A differential localization of the two cell wall epitopes was found between tissues and between species: for example, in leek, CCRC-M1 labeled epidermal and hypodermal cells, whereas CCRC-M7 labeled cortical cells only. Clover walls were labeled by both McAbs, whereas maize and tobacco were only labeled by CCRC-M7. In the presence of the arbuscular mycorrhizal fungi, labeling was additionally found in an apoplastic compartment typical of the symbiosis (the interface) occurring around the intracellular hyphae. Epitopes binding both McAbs were found in the interfacial material, and their distribution mirrored the pattern found in the host cell wall. These findings demonstrate that the composition of the interface zone in a fungus-plant symbiosis reflects the composition of the wall of the host cell.

Journal Article↗

Synthesis and thermotropic characterization of a homologous series of racemic beta-D-glucosyl dialkylglycerols.

The phase behaviour of aqueous dispersions of a series of synthetic 1,2-di-O-alkyl-3-O-(beta-D-glucosyl)-rac-glycerols with both odd and even hydrocarbon chain lengths was studied by differential scanning calorimetry and low angle X-ray diffraction (XRD). Thermograms of these lipids show a single, strongly energetic phase transition, which was shown to correspond to either a lamellar gel/liquid crystalline (L(beta)/L(alpha)) phase transition (short chain compounds, n < or =14 carbon atoms) or a lamellar gel/inverted hexagonal (L(beta)/H(II)) phase transition (longer chain compounds, n > or =15 carbon atoms) by XRD. The shorter chain compounds may exhibit additional transitions at higher temperatures, which have been identified as lamellar/nonlamellar phase transitions by XRD. The nature of these nonlamellar phases and the number of associated intermediate transitions can be seen to vary with chain length. The thermotropic phase properties of these lipids are generally similar to those reported for the corresponding 1,2-sn-diacyl alpha- and beta-D-glucosyl counterparts, as well as the recently published 1, 2-dialkyl-3-O-(beta-D-glycosyl)-sn-glycerols. However, the racemic lipids studied here show no evidence of the complex patterns of gel phase polymorphism exhibited by the above mentioned compounds. This suggests that the chirality of the glycerol molecule, by virtue of its position in the interfacial region, may significantly alter the phase properties of a lipid, perhaps by controlling the relative positions of hydrogen bond donors and acceptors in the polar region of the membrane.

Calorimetry, Differential Scanning↗

Interactions of anesthetics with the water-hexane interface. A molecular dynamics study.

The free energy profiles characterizing the transfer of nine solutes across the liquid-vapor interfaces of water and hexane and across the water-hexane interface were calculated from molecular dynamics simulations. Among the solutes were n-butane and three of its halogenated derivatives, as well as three halogenated cyclobutanes. The two remaining molecules, dichlorodifluoromethane and 1,2-dichloroperfluoroethane, belong to series of halo-substituted methanes and ethanes, described in previous studies (J. Chem. Phys. 1996, 104, 3760; Chem. Phys. 1996, 204, 337). Each series of molecules contains structurally similar compounds that differ greatly in anesthetic potency. The accuracy of the simulations was tested by comparing the calculated and the experimental free energies of solvation of all nine compounds in water and in hexane. In addition. the calculated and the measured surface excess concentrations of n-butane at the water liquid-vapor interface were compared. In all cases, good agreement with experimental results was found. At the water-hexane interface, the free energy profiles for polar molecules exhibited significant interfacial minima, whereas the profiles for nonpolar molecules did not. The existence of these minima was interpreted in terms of a balance between the free energy contribution arising from solute-solvent interactions and the work to form a cavity that accommodates the solute. These two contributions change monotonically, but oppositely, across the interface. The interfacial solubilities of the solutes, obtained from the free energy profiles, correlate very well with their anesthetic potencies. This is the case even when the Meyer-Overton hypothesis, which predicts a correlation between anesthetic potency and solubility in oil, fails.

Algorithms↗

Identifying and mapping surface amorphous domains.

PURPOSE: Undesirable amorphous material generation during formulation is implicated in a growing number of pharmaceutical problems. Due to the importance of interfacial properties in many drug delivery systems, it seems that surface amorphous material is particularly significant. Consequently, this study investigates a range of methods capable of detecting and mapping surface amorphous material. METHODS: A micron-sized localized surface domain of amorphous sorbitol is generated using a novel localized heating method. The domain is subsequently investigated using atomic force microscopy (AFM) imaging, nanomechanical measurements, and Raman microscopy 3-D profiling. RESULTS: AFM phase and height images reveal nanoscale-order variations within both crystalline and amorphous sorbitol domains. Nanomechanical measurements are able to quantitatively distinguish the amorphous and crystalline domains through local Young's modulus measurements. Raman microscopy also distinguishes the amorphous and crystalline sorbitol through variations in peak width. This is shown to allow mapping of the 3-D distribution of the amorphous phase and is hence complementary to the more surface sensitive AFM measurements. CONCLUSIONS: AFM and Raman microscopy map the distribution of amorphous material at the surface of a sorbitol crystal with submicron spatial resolution, demonstrating surface analysis methods for characterizing semicrystalline solids generated during pharmaceutical processing.

Chemistry, Pharmaceutical↗

The role of autoimmune phenomena in the pathogenesis of cataract.

Investigations were carried out to clarify the role of autoimmune phenomena in the pathogenesis of cataract in the adult human lens. Studies were carried out to determine the presence of serum antibodies to lens protein in patients with senile cataract, in patients with diabetes mellitus with and without cataract, and in healthy adult controls using the interfacial test and the gel-diffusion technique. Non-specific antibodies were removed by adsorption of sera with homogenized rat liver. A high proportion of healthy adults was found to have anti-lens protein antibodies (44.4% by the gel-diffusion method). In contrast, patients with cataract and diabetic patients with no cataract demonstrated double this incidence (82% and 80%), while all diabetic patients with cataract showed the presence of antibodies (P = 0.0002). The possible causes for the development of lens antibodies in normal healthy humans are discussed. Also, the causes for the higher incidence of lens antibodies in patients with cataract and in diabetic subjects with no clinical evidence of cataract are considered in relation to cataract formation. Homogenates of cataractous lenses when investigated revealed the presence of both IgG and IgM immunoglobulins, the former probably to a greater extent. Fluorescent microscopy on cryosections of senile and diabetic cataractous lenses revealed the presence of immunoglobulins within the lens. The antigen in the immune complexes isolated from homogenized cataractous lenses was characterized by the SDS-polyacrylamide gel electrophoresis method. A single band was consistently obtained and the molecular weight of the protein was estimated to be between 35,000 and 40,000. The strong possibility of auto-antibodies to lens protein being of aetiological significance in the pathogenesis of cataract is discussed.

Adult↗

Synthesis, characterization, and black lipid membrane studies of [7-L-alanine] gramicidin A.

With a view to study the relevance of side-chain orientation in the transport of cations through a gramicidin transmembrane channel and to identify an analogue with favorable characteristics, [L-Ala7] gramicidin A was synthesized, purified, verified, and characterized by high-performance liquid chromatography, by carbon-13 and proton magnetic resonance spectra, and by circular dichroism spectra in methanol. Complete incorporation as the channel state was achieved when packaged in lysolecithin-containing lipid bilayers. The single-channel conductance data in diphytanoyllecithin/n-decane membranes are presented along with those of synthetic gramicidin A (GA). [L-Ala7] GA exhibits the highest most probable single-channel conductance so far reported for an analogue occurring at 28 pS as compared to 21 pS for GA under similar conditions. Also, a dramatic reduction in the dispersity of conducting states is observed with about 76% of the events falling in a narrow 1.75-pS conductance window as compared to about 31% of the events for GA under identical conditions. Thus, with the above characteristics, [L-Ala7]GA appears to be a very good candidate for a thorough study of ionic mechanism. The present results indicate that elements intrinsic to the channel proper are rate-limiting for GA and that there is no interfacial polarization or diffusion-controlled association at 1 M KCl and a 100-mV applied potential.

Chromatography, High Pressure Liquid↗

The selenoenzyme phospholipid hydroperoxide glutathione peroxidase.

The reduction of membrane-bound hydroperoxides is a major factor acting against lipid peroxidation in living systems. This paper presents the characterization of the previously described 'peroxidation-inhibiting protein' as a 'phospholipid hydroperoxide glutathione peroxidase'. The enzyme is a monomer of 23 kDa (SDS-polyacrylamide gel electrophoresis). It contains one gatom Se/22 000 g protein. Se is in the selenol form, as indicated by the inactivation experiments in the presence of iodoacetate under reducing conditions. The glutathione peroxidase activity is essentially the same on different phospholipids enzymatically hydroperoxidized by the use of soybean lipoxidase (EC 1.13.11.12) in the presence of deoxycholate. The kinetic data are compatible with a tert-uni ping-pong mechanism, as in the case of the 'classical' glutathione peroxidase (EC 1.11.1.9). The second-order rate constants (K1) for the reaction of the enzyme with the hydroperoxide substrates indicate that, while H2O2 is reduced faster by the glutathione peroxidase, linoleic acid hydroperoxide is reduced faster by the present enzyme. Moreover, the phospholipid hydroperoxides are reduced only by the latter. The dramatic stimulation exerted by Triton X-100 on the reduction of the phospholipid hydroperoxides suggests that this enzyme has an 'interfacial' character. The similarity of amino acid composition, Se content and kinetic mechanism, relative to the difference in substrate specificity, indicates that the two enzymes 'classical' glutathione peroxidase and phospholipid hydroperoxide glutathione peroxidase are in some way related. The latter is apparently specialized for lipophylic, interfacial substrates.

Animals↗

Biomaterial coatings by stepwise deposition of silk fibroin.

A completely aqueous, stepwise deposition process with Bombyx mori silk fibroin for the assembly of nanoscale thin film coatings is reported the first time. The focus of this work was to develop an understanding of the control of this deposition process and to characterize the films formed from a physicochemical perspective. The deposition process was monitored by UV spectrophotometry and research quartz crystal microbalance. Both absorbance and film thickness correlated linearly with the number of silk fibroin layers deposited, analogous to multilayered materials fabricated from conventional polyelectrolytes. The polymer adsorption process was stable and reproducible, with control of a single layer thickness ranging from a few to tens of nanometers, determined by the concentrations of silk fibroin, salt concentration in the dipping solution, and method of rinsing. The driving force for the assembly of silk fibroin onto the substrate was primarily hydrophobic interactions, while some electrostatic interactions were also involved. The difference with this approach from traditional polyelectrolyte layer-by-layer techniques is that an intervening drying step is used to control the structure and stability of the self-assembled silk fibroin. The assembled films were stable under physiological conditions and supported human bone marrow stem cell adhesion, growth, and differentiation. This approach offers new options to engineer biomaterial coatings as well as bulk materials with control of both interfacial properties conducive to specific cellular or tissue responses and the potential to entrap and deliver labile molecules or other components due to the all-aqueous process described.

Bone Marrow Cells↗

Characterization of polyethylene crystallization from an oriented melt by molecular dynamics simulation.

Molecular dynamics is used to characterize the process of crystallization for a united atom model of polyethylene. An oriented melt is produced by uniaxial deformation under constant load, followed by quenching below the melting temperature at zero load. The development of crystallinity is monitored simultaneously using molecular-based order parameters for density, energy, and orientation. For crystallization temperatures ranging from 325 to 375 K, these simulations clearly show the hallmarks of crystal nucleation and growth. We can identify multiple nucleation events, lamellar growth up to the limit imposed by periodic boundaries of the simulation cell, and lamellar thickening. We observe a competition between the rate of nucleation, which results in multiple crystallites, the rate of chain extension, which results in thicker lamellae, and the rate of chain conformational relaxation, which is manifested in lower degrees of residual order in the noncrystalline portion of the simulation. The temperature dependence of lamellar thickness is in accord with experimental data. At the higher temperatures, tilted chain lamellae are observed to form with lamellar interfaces corresponding approximately to the [201] facet, indicative of the influence of interfacial energy.

Journal Article↗

Effects of enamel matrix proteins on tissue formation along the roots of human teeth.

OBJECTIVE: Enamel matrix-derived proteins (EMD) are thought to trigger the formation of acellular extrinsic fibre cementum (AEFC), while other reports indicate that EMD may have osteogenic potential. The aim of the present study was to characterize the tissues developing on the root surface following application of EMD. METHODS: Twelve human periodontitis-affected teeth, scheduled for extraction, were treated with EMD. Two to 6 weeks later, the teeth were extracted, demineralized and processed for embedding in acrylic and epoxy resins. New tissue formation was analysed by light and transmission electron microscopy. RESULTS: New tissue formation on the root was observed in the notch and on both scaled and unscaled root surfaces distant of the notch area in six defects. The newly formed tissues on the root were thick, collagenous, devoid of extrinsic fibres, and had an irregular surface contour. The presence of electron-dense, organic material in the collagenous matrix indicated at least partial mineralization. Embedded cells were numerous and the cells on the matrix surface were very large in size. Abundant rough endoplasmic reticulum and a prominent Golgi complex were evident. The presence of a split between the treated root surfaces and the newly formed tissue was a common observation, as was the presence of bacteria and host cells in the interfacial gap. CONCLUSION: Following treatment with EMD, a bone-like tissue resembling cellular intrinsic fibre cementum may develop on the root surfaces, instead of AEFC. Furthermore, EMD may both induce de novo formation of a mineralized connective tissue on scaled root surfaces and stimulate matrix deposition on old native cementum. Interfacial bonding appeared to be weak after 6 weeks of healing.

Cementogenesis↗

Lung surfactant gelation induced by epithelial cells exposed to air pollution or oxidative stress.

Lung surfactant lowers surface tension and adjusts interfacial rheology to facilitate breathing. A novel instrument, the interfacial stress rheometer (ISR), uses an oscillating magnetic needle to measure the shear viscosity and elasticity of a surfactant monolayer at the air-water interface. The ISR reveals that calf lung surfactant, Infasurf, exhibits remarkable fluidity, even when exposed to air pollution residual oil fly ash (ROFA), hydrogen peroxide (H2O2), or conditioned media from resting A549 alveolar epithelial cells (AEC). However, when Infasurf is exposed to a subphase of the soluble fraction of ROFA- or H2O2-treated AEC conditioned media, there is a prominent increase in surfactant elasticity and viscosity, representing two-dimensional gelation. Surfactant gelation is decreased when ROFA-AEC are pretreated with inhibitors of cellular reactive oxygen species (ROS), or with a mitochondrial anion channel inhibitor, as well as when A549-rho0 cells that lack mitochondrial DNA and functional electron transport are investigated. These results implicate both mitochondrial and nonmitochondrial ROS generation in ROFA-AEC-induced surfactant gelation. A549 cells treated with H2O2 demonstrate a dose-dependent increase in lung surfactant gelation. The ISR is a unique and sensitive instrument to characterize surfactant gelation induced by oxidatively stressed AEC.

Air Pollutants↗