Microvascular remodelling in chronic airway inflammation in mice.

1. Chronic inflammation is associated with blood vessel remodelling, including vessel proliferation and enlargement, and changes in vessel phenotype. We sought to characterize these changes in chronic airway inflammation and to determine whether corticosteroids that inhibit inflammation, such as dexamethasone, can also reduce microvascular remodelling. 2. Chronic airway inflammation was induced in C3H mice by infection with Mycoplasmapulmonis and the tracheal vessels treatment also decreased the immunoreactivity for P-selectin and the number of adherent leucocytes (595 +/- 203 vs 2,024 +/- 393 cells/ mm2 in treated and non-treated infected mice, respectively). 6. We conclude that microvascular enlargement and changes in vessel phenotype are features of some types of chronic inflammation and, furthermore, that dexamethasone reverses the microvascular enlargement, changes in vessel phenotype and leucocyte influx associated with chronic inflammatory airway disease.

Openings between defective endothelial cells explain tumor vessel leakiness.

Leakiness of blood vessels in tumors may contribute to disease progression and is key to certain forms of cancer therapy, but the structural basis of the leakiness is unclear. We sought to determine whether endothelial gaps or transcellular holes, similar to those found in leaky vessels in inflammation, could explain the leakiness of tumor vessels. Blood vessels in MCa-IV mouse mammary carcinomas, which are known to be unusually leaky (functional pore size 1.2-2 microm), were compared to vessels in three less leaky tumors and normal mammary glands. Vessels were identified by their binding of intravascularly injected fluorescent cationic liposomes and Lycopersicon esculentum lectin and by CD31 (PECAM) immunoreactivity. The luminal surface of vessels in all four tumors had a defective endothelial monolayer as revealed by scanning electron microscopy. In MCa-IV tumors, 14% of the vessel surface was lined by poorly connected, overlapping cells. The most superficial lining cells, like endothelial cells, had CD31 immunoreactivity and fenestrae with diaphragms, but they had a branched phenotype with cytoplasmic projections as long as 50 microm. Some branched cells were separated by intercellular openings (mean diameter 1.7 microm; range, 0.3-4.7 microm). Transcellular holes (mean diameter 0.6 microm) were also present but were only 8% as numerous as intercellular openings. Some CD31-positive cells protruded into the vessel lumen; others sprouted into perivascular tumor tissue. Tumors in RIP-Tag2 mice had, in addition, tumor cell-lined lakes of extravasated erythrocytes. We conclude that some tumor vessels have a defective cellular lining composed of disorganized, loosely connected, branched, overlapping or sprouting endothelial cells. Openings between these cells contribute to tumor vessel leakiness and may permit access of macromolecular therapeutic agents to tumor cells.

Lidar equations for turbid media with pulse stretching.

Lidar equations for a system with multiple-scattering beam spreading and pulse stretching are developed from an analytical model for the beam spread function. The resulting lidar equations are transparent to the physics and with some simple approximations for system transfer functions become mathematically simple engineering models for system studies. Application to and comparison with a variety of lidar applications in ocean environments (turbidity and bathymetry) and clouds (aerosol scattering) are presented. These examples provide model validation. The lidar model developed represents a significant extension beyond historical lidar models that exclude pulse stretching. Their mathematical simplicity should foster use in a broader class of problems involving light propagation in turbid media.

Cationic liposomes target angiogenic endothelial cells in tumors and chronic inflammation in mice.

This study sought to determine whether angiogenic blood vessels in disease models preferentially bind and internalize cationic liposomes injected intravenously. Angiogenesis was examined in pancreatic islet cell tumors of RIP-Tag2 transgenic mice and chronic airway inflammation in Mycoplasma pulmonis-infected C3H/HeNCr mice. For comparison, physiological angiogenesis was examined in normal mouse ovaries. We found that endothelial cells in all models avidly bound and internalized fluorescently labeled cationic liposomes (1,2-dioleoyl-3-trimethylammonium-propane [DOTAP]/cholesterol or dimethyldioctadecyl ammonium bromide [DDAB]/cholesterol) or liposome-DNA complexes. Confocal microscopic measurements showed that angiogenic endothelial cells averaged 15-33-fold more uptake than corresponding normal endothelial cells. Cationic liposome-DNA complexes were also avidly taken up, but anionic, neutral, or sterically stabilized neutral liposomes were not. Electron microscopic analysis showed that 32% of gold-labeled liposomes associated with tumor endothelial cells were adherent to the luminal surface, 53% were internalized into endosomes and multivesicular bodies, and 15% were extravascular 20 min after injection. Our findings indicate that angiogenic endothelial cells in these models avidly bind and internalize cationic liposomes and liposome-DNA complexes but not other types of liposomes. This preferential uptake raises the possibility of using cationic liposomes to target diagnostic or therapeutic agents selectively to angiogenic blood vessels in tumors and sites of chronic inflammation.

Beam spread function with time dispersion.

An analytical model for the beam spread function with time dispersion is modeled and validated against Monte Carlo calculations. The model, which is structured on simple statistical concepts and relies on only first and second moments for displacement, angle, and multipath time, is suitable for describing pulsed laser radiation propagation in nonconservative scattering media out to tens of scattering lengths. Numerical examples for marine environments are used to show its robustness and versatility.

Organ-specific endothelial cell uptake of cationic liposome-DNA complexes in mice.

This study identified the organ and cellular distribution of cationic liposome-DNA complexes injected intravenously into CD-1 mice for gene delivery. DOTIM-cholesterol liposomes were labeled with the fluorescent dye CM-Dil and complexed with plasmid DNA encoding the chloramphenicol acetyltransferase reporter gene. The distribution of the complexes was examined in 29 organs and tissues by fluorescence, confocal, and electron microscopy from 5 min to 24 h after injection. The complexes formed clusters in blood, which were cleared within 20 min. Complexes visible by fluorescence microscopy were taken up by endothelial cells, leukocytes, and macrophages and did not leave the vasculature except in the spleen. At 5 min, the complexes formed a patchy coating on the endothelial surface, but by 4 h, they were internalized into endosomes and lysosomes in organ- and vessel-specific patterns. Uptake by capillary endothelial cells was greatest in the lung, ovary, and anterior pituitary, less in muscle and the heart, and nearly absent in the brain and pancreatic islets. In lymph nodes and intestinal Peyer's patches, the uptake was sparse in capillaries but abundant in high endothelial venules. In the liver and spleen, most of the uptake was in Kupffer cells and macrophages. Measurements of chloramphenicol acetyltransferase reporter gene expression were generally consistent with the pattern of uptake by endothelial cells. The uptake and gene expression were accompanied by a decrease in circulating leukocytes and platelets. Overall, our results showed that the complexes were internalized by endothelial cells in organ- and vessel-specific patterns that did not match any previously identified properties of the microvasculature. The unusual distribution of endothelial cell uptake may be explained by a heterogeneously distributed membrane receptor for which the complexes are ligands.

Dentinal bonding agents versus glass-ionomer cements.

The long-term bonding of dental material to dentin remains an area of great controversy and the results of in vitro testing do not always reflect those found in vivo. The clinician is faced with a large number of dentinal bonding agents that have had limited testing in vivo and are frequently replaced before any long-term clinical testing has been completed. Glass-ionomer cements, although having a longer history of good adhesion to dentin, are not suitable for use in high-stress-bearing areas. The selection of materials for specific clinical situations has become more and more difficult. This paper gave a personal view of the history and evolution of both resin bonding agents and glass-ionomer cements and their potential in clinical use.

Effects of ocean waves on airborne lidar imaging.

The effects of ocean waves on lidar imaging of submerged objects are investigated. Two significant consequences of wave focusing or defocusing are quantified: (a) intensification of near-surface backscatter in which the mean return is increased relative to that for a flat interface, and (b) spatial-temporal modulations of the backscattered return. For the former, mean returns can be as much as 50% larger than flat surface returns at shallow depth. For the latter, the strong modulations induced by wave motion present a dominant clutter field that significantly affects the imaging of shallow objects. Both effects are compensated at greater depths by beam spreading caused by multiple scattering, which diminishes the intensity of the wave focusing.

RGD-containing peptides inhibit adhesion of 293 cells transfected with GpIIb/IIIa to fibrinogen: comparison to inhibition of platelet aggregation.

Cyclic RGD-containing peptides caused a dose-dependent inhibition of binding of human embryonic kidney cells transfected with recombinant GpIIb/IIIa (r293 clone B) to human fibrinogen coated on to non-tissue culture plates. The inhibitory activity, IC50, of a panel of seventeen RGD-containing peptides ranged from 0.12 to 89.2 microM. These IC50 values correlated with those determined by the inhibition of platelet aggregation (r = 0.99). Even though there was a correlation, there were differences between the platelet aggregation and the bioadhesion assay. The binding of r293 clone B to fibrinogen was not increased by ADP suggesting that GpIIb/IIIa expressed on the surface of r293 clone B cells may be in the 'activated' form. Moreover, preincubation of r293 clone B cells with a monoclonal antibody (mAb) specific for GpIIIa (4B12) resulted in a dose-dependent decrease of binding to fibrinogen while a mAb specific for GPIIb (2D2) had no effect. Neither of these mAbs inhibited platelet aggregation. The binding of r293 clone B cells to fibrinogen required Ca2+ or Mg2+. This cell-based bioadhesion method can provide a tool for screening potential GpIIb/IIIa antagonists and investigating the interaction of GpIIb/IIIa and fibrinogen not possible with platelet aggregation.

A preliminary report on the effect of storage in water on the properties of commercial light-cured glass-ionomer cements.

Two commercially available light-curable glass-ionomer cements, Vitrebond and XR-Ionomer, have been studied and their compressive strengths measured following storage under wet and dry conditions for varying lengths of time up to 3 months. The strongest cements were those stored in air and allowed to age. On the other hand, cements that were stored in water were found to become progressively weaker with time. Their failure mode was different from that of cements stored in air in that specimens became barrel-shaped as they were loaded and exhibited considerable plastic deformation prior to fracturing. By contrast, air-stored specimens behaved as predominantly brittle materials, the specimens essentially maintaining their integrity up to the point of catastrophic failure. Both of these findings indicate that the properties of these particular light-cured cements change markedly on exposure to moisture, a fact which is of clinical significance.

The clinical use of glass-ionomer cements.

The use of glass-ionomer cements in clinical dentistry has expanded greatly over the last decade. Their use in treating early carious or erosion lesions has been investigated widely and established techniques include fissure filling, restoration of erosion lesions without cavity preparation, and the internal or tunnel restoration. Because of their adhesion to moist tooth structure, biologic compatibility, and fluoride release, increasing use also has been made of their anticariogenic properties in treating geriatric patients. Glass-ionomers have proved very successful as dentin substitutes for attaching composites to enamel without involving risk of pulpal damage in the deeper cavity. The deficiencies of glass-ionomer cements are well known, including lack of toughness, early water sensitivity, low abrasion resistance, and porosity, leading to poor surface polish. Solving these problems is formidable because inherently the strength of these cements is related to their water content. The clinician should be aware of these deficiencies and stay within the parameters of the techniques outlined in this article. In particular, clinical success depends on early protection of the cement from hydration or dehydration and the current use of light-cured bonding agents largely has solved this problem. The future probably lies in using laminate techniques in which materials that attach to dentin and form a biologic seal can be covered by tougher and harder enamel veneers, thus mimicking the structure of the tooth. It is possible that future materials will be developed on the lines of these polyelectrolyte cements in which higher molecular weight polymers are used in conjunction with polymers that contain photoinitiators to effect light curing and toughen the matrix. In addition, the possibility of developing laboratory-cured glass-ionomer inlays in which porosity can be reduced and tougher polymers used should be considered.