In vitro adherence of radioactively labeled Escherichia coli in normal and cystitis-prone females.

Numerous investigators report data obtained using an in vitro quantitative assay for measuring bacterial adherence to epithelial cells. We found this assay to contain significant sources of error in the large variation in number of bacteria bound per cell and in the dependence on the investigator's visual counting of bacteria bound per cell. In the modified assay described here, we eliminated the need for visual counting of bacteria by incorporating the use of radioactively labeled Escherichia coli. This allowed quantitation of bacterial adherence to as many as 50,000 vaginal cells, whereas the visual counting system limits the determination to perhaps 50 cells. We feel that the use of radioactively labeled bacteria in place of the visual counting system increases the validity and sensitivity of this assay. Using the modified method, we found no statistically significant differences among values for adherence of E. coli type 04 to the vaginal cells of control and cystitis-prone women at either pH 6.4 or 4.0.     

Mating ability in laboratory-adapted and field-derivedDrosophila melanogaster: The stress of domestication

Mating ability differences between flies of different alcohol dehydrogenase (Adh) genotypes have been assessed in the temperature range 15 to 29°C for laboratory-adapted and field-derivedDrosophila melanogaster. Significant differences amongAdh genotypes were detected principally for the laboratory-adapted strains due to departures from random mating associated with heterozygote superiority at the relatively extreme temperature of 29°C, although mating ability differences could not be attributed directly to theAdh locus. The difference between the laboratory and the field populations can be explained by the effects of genetic back-ground manifested in the form of fitness differences, being enhanced for the laboratory-adapted flies as a consequence of the stress of laboratory culture. In contrast with larval survival and development time, laboratory and field flies do not differe appreciably in their overall abilities to obtain mates, which indicates that mating ability is a direct fitness character not greatly affected by laboratory culture. It follows that direct fitness traits are the least amenable to change under domestication.     

UNILATERAL BRAIN DAMAGE AND BILATERAL SKIN CONDUCTANCE LEVELS IN HUMANS

Left and right, palmar and dorsal skin conductance levels (SCLs) were obtained from hospital controls, left hemisphere lesion Ss, right hemisphere lesion Ss, and diffuse or bilateral lesion Ss during several experimental conditions involving rest, passive auditory stimulation, motor reactions, and simple “perception”. The unilateral lesion groups generally displayed significantly higher palmar SCLs on the side contralateral to their lesion. Such “laterality” was not demonstrated in dorsal recordings or in the hospital controls or diffuse lesion group. These unilateral lesion groups had higher palmar SCLs during passive stimulation than during rest, motor, or perception phases. Results were discussed in terms of possible neural mechanisms underlying the phenomena.     

Oligopeptidases of brush border membranes of rat small intestinal mucosal cells

1. The properties of peptidases located in the brush borders of rat small intestinal mucosal cells have been investigated using a new method for the assay of peptidase activity. In this method amino acids produced by hydrolysis of peptides are oxidized by ophidian L-amino acid oxidase and the hydrogen peroxide produced during reoxidation of the reduced enzyme is estimated spectrophotometrically.2. 10-20% of the total cellular peptidase activity and 50% of the leucylnaphthylamidase (LNAase) activity were found to be tightly bound to the brush borders and to possess different substrate specificity from the supernatant peptidase activity.3. Evidence from kinetic and competition studies indicates the presence of more than one peptidase in the brush borders. The peptidases exhibit pH optima of 8.0-8.5, are inhibited by EDTA, but are not usually activated by divalent metal ions. The brush border peptidases hydrolyse di- and tripeptides, some oligopeptides, leucinamide and leucyl-beta-naphthylamine. On the basis of the Michaelis constants, these substrates differ in their affinities for the enzymes.4. It is proposed that the brush border peptidases serve an analogous function in the terminal stages of protein digestion to that of the disaccharidases in the case of carbohydrate digestion and absorption.     

Distinct patterns of tyrosine phosphorylation during the life cycle of Trypanosoma brucei.

Regulation of tyrosine phosphorylation is a critical element in controlling growth and differentiation in higher eukaryotes. We have determined that the protozoan Trypanosoma brucei, which diverged early in the eukaryotic lineage, possesses multiple proteins which react with a specific anti-phosphotyrosine antiserum. Anti-phosphotyrosine immunoprecipitates of [32P]orthophosphate-labeled cells were shown to contain phosphotyrosine by two-dimensional electrophoresis. Western analysis of cells from different stages of the life cycle demonstrates the appearance of tyrosine-phosphorylated proteins at 40-42 kDa during the transition from slender to stumpy blood-forms. Growth of procyclic form cells in orthovanadate resulted in increased levels of specific tyrosine-phosphorylated proteins. The demonstration of phosphotyrosine-containing proteins in T. brucei and their differential regulation during the life cycle suggests that tyrosine kinases and phosphatases may play an important role in the biology of primitive protozoa.