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Biomedical subjects

J Leighton

Publications and source records attributed to J Leighton.

83 records · Page 5Linked to original sources

Collagen-coated cellulose sponge: three dimensional matrix for tissue culture of Walker tumor 256.

Three-dimensional growth of large populations of cells in vitro has been observed in the interstices of a matrix consisting of collagen-coated cellu lose sponge. The growth of Walker tumor 256 in this composite matrix is com pared with that found in a matrix composed of either cellulose sponge alone or collagen sponge alone. The composite matrix is superior to either one. Collagen coated cellulose sponge may provide a simple tool for the study of social interaction of cells in the formation of organized elementary tissue structures.

Animals↗

Increased numbers of annulate lamellae in myocardium of chick embryos incubated at abnormal temperatures.

Annulate lamellae have been observed in the myocardium of 18-day-old chick embryos maintained at the normal temperature of 100 degrees F and at 90 degrees F during the last week of incubation. An increased number of annulate lamellae was observed in heart muscle of embryos incubated at 90 degrees F. This is probably caused by a persistent production of these organelles, since annulate lamellae are present in greater frequency than in 11-day-old embryos incubated at 100 degrees F. In the hypertrophic hearts of 18-day-old embryos incubated at 90 degrees F, the annulate lamellae were associated with a net increase of protein content and an elevated concentration of myocardial glycogen. It is suggested that the increased number of annulate lamellae is a sequela of reduced environmental temperature during incubation.

Animals↗

Interactions between newly formed endothelial channels and carcinoma cells in plasma clot culture.

Bovine capillary endothelial cells (BCEC), cultured in suspension on a rotary shaker, formed aggregates ranging from 50 to 300 micron in diameter. In plasma clot these aggregates sprouted in multiple directions and gave rise to vascular channels. Aggregates of the squamous cell carcinoma line of rat bladder NBT-II-81, cultured in plasma clot, formed solid spheroids that grew slowly by expansion. When cultured together with BCEC, however, NBT-II-81 infiltrated the plasma clot extensively. The tumor cells, after establishing contacts with the vascular channels, spread into the fibrin meshwork using the subendothelial space as their path of propagation. Endothelial cells that were separated from the surrounding matrix by invading tumor cells degenerated, leaving behind channels lined only by neoplastic epithelium. The adhesive properties of the subendothelial matrix were studied by seeding NBT-II-81 cells on dishes coated with the extracellular matrix produced by BCEC. Tumor cells attached readily and in large numbers to dishes coated with the subendothelial matrix. In contrast they attached poorly to dishes coated with fibrin. We conclude that the spread of carcinoma cells into plasma clot is markedly enhanced by endothelial channels, developed in the absence of blood flow. The production of a highly adhesive extracellular matrix by the capillary endothelium during angiogenesis may represent an important element in the preferential growth of the tumor along the vascular route.

Animals↗

Structural biology of epithelial tissue in histophysiologic gradient culture.

Epithelial cells proliferate, forming organized tissues, when positioned in the lumen of a thin-walled, transparent, elongated cylindrical, cystlike culture chamber. The closed chamber, 2.5 mm in diameter and 25 mm long, bathed in medium, incubated with continuous gentle agitation, enables the inoculum to exchange metabolites including oxygen by diffusion across the thin, nylon filament-reinforced collagen membrane wall of the chamber. After periods of culture of a week or more, using inocula derived from urothelium, the inner surface of the cystic chamber is lined by a stratified epithelium. Proliferation of cells is seen in the basal zone, which is attached to the collagen substrate. The development of the model is briefly described. Some of the applications of the procedure are illustrated using cell lines, chick embryo tissues, and clinical tissues. Implications of the procedure are considered for studying categories of tissue biology, e.g., problems of aging, neoplasia, and toxicology.

Animals↗

Human mammary cancer cell lines and other epithelial cells cultured as organoid tissue in lenticular pouches of reinforced collagen membranes.

A system has been developed for the culture of cells that provides conditions favoring the formation of tissues comparable to conditions existing in nature. The culture chamber is a lens-shaped pouch composed of two thin-walled, reinforced, waffled collagen membranes facing each other. The chamber is immersed in medium in a closed transparent container and incubated on a rocker. On histologic study, after days to weeks in culture, human mammary cancer cell lines BT-20, MCF-7, MDA-231, MDA-468, and T47D grow in the chamber as distinctive structured epithelial tissue. Dog kidney cell line MDCK grows as a papillary adenocarcinoma and rat bladder cancer line NBT-II as an epidermoid carcinoma; cells from clinical effusion tumors produce distinct tissue. Changes in histologic phenotype may be driven by molecular changes at the level of the genome. Resulting alteration of the biochemical functions essential for the integrity of specific durable tissue organization should alter or reset the pattern of tissue organization and of biological behavior, including malignancy and response to cytotoxic chemicals. Lenticular pouch culture promises to be an effective tool for exploring the molecular changes associated with histogenesis and malignancy.

Adenocarcinoma, Papillary↗

Recovery of cyclic antidepressants with gastric lavage.

The role of lavage fluid volume in recovery of cyclic antidepressant with gastric lavage was determined in 13 patients treated for moderate to severe cyclic antidepressant intoxication. An orogastric tube was placed, gastric contents aspirated, and gastric lavage performed with 60 to 180 mL aliquots of tap water or 0.9% NaCl for irrigation. A volume of 12.8 +/- 4.4 liters of lavage fluid was instilled, and 97.6 +/- 6.9% of this volume was recovered. The recovered aspirate and lavage fluid were inspected for the presence of particulate matter and saved in 4 to 5 liter collections. Determination of cyclic antidepressant and metabolite concentrations was performed using gas chromatography-mass spectrometry, and the amount of cyclic antidepressant recovered was calculated. The mean total amount recovered was 110 mg and ranged from 2.4 mg to 342 mg. Of the total amount recovered, 88% +/- 13% was recovered in the first 4 to 5 liters of lavage fluid. An estimate of the dose ingested was available in 7 patients, with 8.7% (0.4% to 21.7%) of the estimated dose recovered by lavage. There were no cases of significant drug recovery in clear lavage fluid. We conclude that if gastric lavage is undertaken in cyclic antidepressant intoxications, it should utilize 5 liters initially and then continue only if particulate matter is seen.

Adolescent↗

Expression and intracellular localisation of odorant receptors in mammalian cell lines using Semliki Forest virus vectors.

Odorant receptors are members of the G protein-coupled receptor superfamily. They are expressed on the surface of cilia of olfactory neurons, where they bind ligand (odorant). Studies of the molecular mechanisms of olfaction are complicated by the extremely large number of receptor genes, and difficulties in pairing a particular mammalian receptor to a specific odorant ligand in vivo. Here we report expression and localisation studies of two rat odorant receptor genes (17 and OR5), and C. elegans odr-10, using the Semliki Forest virus (SFV) system. All receptors were epitope-tagged at the N- or C-terminus in order to facilitate their detection in infected cells, and determine the localisation and membrane-orientation of recombinant proteins. The immortalised mouse olfactory neuronal cell line OLF 442, rat cortical and striatal primary neuron cultures, and the baby hamster kidney (BHK) cells, were infected and tested. Immunofluorescence and confocal microscopy studies performed on permeabilised, non-permeabilised and native cells revealed that in BHK cells the rat receptors 17 and OR5 were not targeted to the plasma membrane and remained in the endoplasmic reticulum. In contrast, in the mouse olfactory cell line OLF 442 both rat receptors were correctly inserted into the plasma membrane. Similar results were obtained using primary neurons, indicating that like mature neurons, the immortalised OLF 442 cells are capable of providing for correct odorant receptor processing and targeting.

Animals↗