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Biomedical subjects

J W McLean

Publications and source records attributed to J W McLean.

At least 19 recordsLinked to original sources
PubMed2000

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.

Animals↗
PubMed1998

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.

Animals↗
PubMed1997

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.

Animals↗
PubMed1996

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.

Dental Bonding↗
PubMed1993

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.

Adenosine Diphosphate↗
PubMed1992

Molecular cloning of a cDNA encoding the glycoprotein of hen oviduct microsomal signal peptidase.

Detergent-solubilized hen oviduct signal peptidase has been characterized previously as an apparent complex of a 19 kDa protein and a 23 kDa glycoprotein (GP23) [Baker & Lively (1987) Biochemistry 26, 8561-8567]. A cDNA clone encoding GP23 from a chicken oviduct lambda gt11 cDNA library has now been characterized. The cDNA encodes a protein of 180 amino acid residues with a single site for asparagine-linked glycosylation that has been directly identified by amino acid sequence analysis of a tryptic-digest peptide containing the glycosylated site. Immunoblot analysis reveals cross-reactivity with a dog pancreas protein. Comparison of the deduced amino acid sequence of GP23 with the 22/23 kDa glycoprotein of dog microsomal signal peptidase [Shelness, Kanwar & Blobel (1988) J. Biol. Chem. 263, 17063-17070], one of five proteins associated with this enzyme, reveals that the amino acid sequences are 90% identical. Thus the signal peptidase glycoprotein is as highly conserved as the sequences of cytochromes c and b from these same species and is likely to be found in a similar form in many, if not all, vertebrate species. The data also show conclusively that the dog and avian signal peptidases have at least one protein subunit in common.

Amino Acid Sequence↗
PubMed1992

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.

Dental Restoration, Permanent↗
PubMed1992

Clinical applications of glass-ionomer cements.

The use of glass-ionomer cements in clinical dentistry is now well established. They have a number of unique properties, including adhesion to moist tooth structure, biological compatibility, and anticariogenic properties due to their fluoride release. Their use in treating early carious or erosion lesions has been widely investigated. Established techniques include fissure filling and sealing, restoration of class 5 erosion lesions without cavity preparation, and the internal occlusal fossa or tunnel restoration. The "sandwich" technique using glass-ionomer cements as "dentin substitutes" has enabled composite restorations to be used with greater safety where pulpal damage may occur. The future probably lies in using a laminate technique where materials that attach to dentin and form a biological seal can be covered by tougher and harder enamel veneers, thus mimicking the structure of the tooth. 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, since 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 paper. In particular, clinical success depends upon early protection of the cement from hydration or dehydration, and the current use of light-cured bonding agents has largely solved this problem.

Cermet Cements↗
PubMed1991

Cloning and expression of a divergent integrin subunit beta 8.

Rabbit and human cDNA clones have been identified that encode a novel integrin beta subunit. The sequences that encode this subunit, which has been designated as beta 8, were isolated initially from rabbit placental cDNA libraries using an oligonucleotide probe derived from a highly conserved region of integrin beta subunit sequences. The rabbit clone was used to isolate human beta 8 cDNA clones from human placental and MG-63 osteosarcoma cell libraries. The putative beta 8 polypeptides, which comprise 769 and 768 residues in human and rabbit, respectively, show a high degree of inter-species conservation (approximately 90% identity). In contrast, beta 8 is distinct from the other integrin beta subunits. At the amino acid level human beta 8 ranges from 31 to 37% identity with human beta 1-7. The domain structure of beta 8 is typical of the integrin beta subunits. Human beta 8 has a 42-residue N-terminal signal peptide, a large extracellular domain (approximately 639 residues) that contains four cysteine-rich repeats, a transmembrane domain (approximately 30 residues), and a C-terminal cytoplasmic domain (approximately 58 residues). There are several structural features that are unique to the beta 8 polypeptide, as compared with the other integrin beta subunits. Six of the 56 cysteine residues that are conserved within the extracellular domains of beta 1, beta 2, beta 3, beta 5, beta 6, and the beta subunit from Drosophila are absent in the beta 8 polypeptide. Also, the cytoplasmic domain of the beta 8 subunit shares no homology with the cytoplasmic regions of any of the other integrin beta subunits. Northern analysis demonstrated an approximately 8-kilobase beta 8 mRNA in rabbit placenta, kidney, brain, ovary, and uterus. PCR analysis revealed that beta 8 mRNA is also present in several transformed human cell lines. The beta 8 polypeptide has been transiently expressed in 293 human embryonic kidney cells. A polyclonal antipeptide antibody specific for beta 8 and a polyclonal antibody that recognizes alpha v epitopes were used to show that beta 8 can complex with the endogenous alpha v subunit in 293 cells and that the resulting integrin is expressed as a cell surface complex.

Amino Acid Sequence↗
PubMed1991

The clinical use of glass-ionomer cements--future and current developments.

Although the glass-ionomer cements were developed in the 1970s, it is only recently that their clinical use has expanded internationally. Improvements in formulation have resulted in better mechanical strengths, increased translucency and, more importantly, faster setting times. Their use as 'dentine substitutes' has expanded the versatility of laminate restorations in anterior and posterior teeth and the introduction of the silver-cermet ionomers provided materials with increased abrasion resistance. Glass-ionomer cements have a number of unique properties, including adhesion to moist tooth structure, biological compatibility, and anticariogenic properties due to their fluoride release. One of the major roles is the treatment of early carious lesions using microcavity preparation. However, like all materials, glass-ionomer cements have their weaknesses, the prime ones being lack of toughness, early water sensitivity, and porosity leading to poor surface polish. Future research should concentrate on correcting these deficiencies.

Cermet Cements↗