A morphometric study quantifying the mucus content of proliferating colonic epithelial cells.

We report a quantification of the maximum mucus accumulation in proliferating rat colonic epithelial cells. The proliferative potential was determined by radioautographic study of one-hour pulse exposures to tritiated thymidine, mucous content was determined by Periodic-acid Schiff (PAS) staining. We examined 55 labeled mucous cells in 0.5- to 1-micrometer serial sections. The maximum thecal and nuclear profiles of these cells were photographed and their surface areas were determined utilizing a coordinate sensor. The data were expressed as a theca-to-nucleus (T/N) ratio. The maximum (T/N) ratio for a labeled mucous cell was 3.0. We performed a similar analysis on 22 unlabeled mucous cells from upper crypt regions and surface epithelium to derive the range of (T/N) ratios for terminally differentiated mature mucous cells. The range of (T/N) ratios from these cells was from 4.8 to 16.4. Our study shows that proliferative potential of mucous cells is determined by the interrelationship between mucus accumulation and nuclear size.     

Solubility characteristics of Micrococcus lysodeikticus membrane components in detergents and chaotropic salts analyzed by immunoelectrophoresis

In order to evaluate the effectiveness and selectivity of various reagents in the solubilization of bacterial membranes, membranes of Micrococcus lysodeikticus were treated with detergents and chaotropic agents. The composition of the extracts so obtained was analyzed by rocket and two-dimensional immunoelectrophoretic techniques. Recovery of succinate-, malate-, and reduced nicotinamide adenine dinucleotide- (NADH) dehydrogenases, ATPase, succinylated lipomannan and cytochromes in the extracts was measured. Treatment with a variety of non-denaturing detergents produced extracts that were generally qualitatively uniform although quantitative differences were observed. The degree of extraction of various components was correlated with the hydrophile-lipophile balance. Several chaotropic agents were also evaluated as reagents for membrane solubilization. These agents were less effective in extraction of bulk protein, but produced extracts enriched in some membrane components.     

Analysis of ligand-receptor binding by the difference method

A new method has been proposed for analysis of experimental data on ligand-receptor binding at equilibrium. This method makes it possible to detect heterogeneity of a receptor system in cases where the contribution of the high-affinity site to total binding is rather small and the problem of graphic discrimination of a model cannot be solved unambiguously by other methods. The difference method permits us to exclude experiments on measuring nonspecific binding. A computer program for analysis of ligand-receptor binding has been worked out in which the difference method and traditional methods of binding isotherm analysis are realized. Numerical modeling has shown that the best strategy in experimental data processing is the treatment of total binding isotherms by both the difference method and regression analysis, including the nonspecific binding constant as one of the regression parameters.