Bacterial killing and inhibition of inner membrane activity by C5b-9 complexes as a function of the sequential addition of C9 to C5b-8 sites

The assembly of the C5b-9 complex on the outer membrane of C-sensitive cells of Escherichia coli results in a rapid inhibition of inner membrane function and ultimately a loss of cell viability. Cells bearing C5b-8 sites suffer no deleterious effects; however, the addition of C9 results in a rapid inhibition of inner membrane function and cell death. An attempt was made to examine the relationship between the toxic effects of the C5b-9 complex and the number of C9 molecules per C5b-8 site. Cells bearing C5b-8 sites were exposed to excess C9 at 0 degrees C and washed three times at 4 degrees C. The number of C9 molecules bound to each cell was equivalent to the number of C5b-8 sites present on each cell, and no additional C9 molecules could be bound when the cells were maintained at 4 degrees C. These cells were then incubated at 37 degrees C for 3 min and returned to 0 degrees C, a technique which exposed additional C9-binding sites equivalent to the number of C9 molecules previously bound to the cells. This technique was repeated and demonstrated that the sequential build-up of a C5b-9 site with two C9 molecules per C5b-8 site was capable of inhibiting both inner membrane function (respiration and amino acid transport) and cell viability. Three C9 molecules per complex had effects that approached the inhibitory effects of complexes formed in the presence of excess C9.     

Identification of disulfide-linked transforming growth factor-beta 1-specific binding proteins in rat glomeruli

We have identified two distinct classes of transforming growth factor-beta (TGF-beta)-binding proteins by affinity labeling rat glomeruli with 125I-TGF-beta 1 and 125I-TGF-beta 2. The first type consists of a group of proteins that bind TGF-beta 1 but do not bind TGF-beta 2. When 125I-TGF-beta 1 affinity-labeled glomeruli were separated under nonreducing conditions, four prominent bands with Mr values of 320,000, 260,000, 170,000, and 90,000 were observed. Following reduction, the 320,000 and 170,000 bands yielded only a 100,000 band, the 260,000 complex yielded bands of 200,000, 100,000, and 85,000, and the 90,000 band migrated with an Mr of 85,000. Binding of 125I-TGF-beta 1 to these proteins was unaffected by the addition of as much as a 1,000-fold excess of TGF-beta 2. The second type of glomerular TGF-beta-binding protein consists of Mr 160,000-200,000 and 280,000 proteins that bind both TGF-beta 1 and beta 2. Digestion of these affinity-labeled proteins with heparitinase and chondroitinase resulted in a decrease of approximately 40,000 in their apparent molecular weights. Glomerular TGF-beta 1-binding proteins are distinct from previously described TGF-beta-binding proteins in their specificity for TGF-beta 1 and their formation of disulfide-linked multimers. The TGF-beta 1/beta 2-binding proteins share some properties of the previously described type III TGF-beta receptor.     

A comparison of the energy budgets of three species of Pogonomyrmex harvester ants (Hymenoptera: Formicidae)

Summary This investigation compares the energy budgets of three species of Pogonomyrmex harvester ants along an altitudinal transect in southern California. All three demonstrate similar seasonal patterns, with high foraging activity and high respiratory costs in mid-summer and little or no activity during winter. Respiration, predominantely metabolism of the workers, was estimated to account for 84–92% of the energy assimilated by the nests. The remaining 8–16% of the energy was invested in the production of new individuals. However, total production was not correlated with food input. It is suggested that worker care of the brood may be the most important determinant of brood production, and thus food may not be a direct limiting resource in harvester ants.A higher percentage of production is invested in workers than in reproductives in all three species. The species usually partition similar proportions of energy between the production of males and females, but since females are larger, more males are produced. Although the species are in different habitats and have very different numbers of workers per nest, the numbers of sexuals produced per nest are similar. The sex-ratio appears to be ecologically determined. Nests that were provided with additional food invested more energy in the production of males. Control nests, nests which had food removed and older nests invested more in the production of females or invested equally in the production of the two sexes.     

Segregating random amplified polymorphic DNAs (RAPDs) in Betula alleghaniensis

Summary Molecular markers are currently being developed for Betula alleghaniensis Britton using random amplified polymorphic DNA (RAPD). Arbitrarily designed 11-mer primers were tested on three intraspecific controlled crosses for which more than 15 full-sibs were available. Using two of these primers, we were able to genetically characterize a total of nine polymorphic RAPD markers. Segregation of these markers was consistent with a biparental diploid mode of inheritance, and all appeared dominant. RAPDs were valuable in detecting contaminants and, therefore, in assessing the validity of controlled crosses. Limitations of the technique are discussed in relation to the determination of parental genotypes and construction of linkage maps for hardwood species.     

Genetic engineering of trees: progress and new horizons

Genetic engineering of trees to improve productivity, wood quality, and resistance to biotic and abiotic stresses has been the primary goal of the forest biotechnology community for decades. We review the extensive progress in these areas and their current status with respect to commercial applications. Examples include novel methods for lignin modification, solutions for long-standing problems related to pathogen resistance, modifications to flowering onset and fertility, and drought and freeze tolerance. There have been numerous successful greenhouse and field demonstrations of genetically engineered trees, but commercial application has been severely limited by social and technical considerations. Key social factors are costly and uncertain regulatory hurdles and sweeping market barriers in the form of forest certification systems that disallow genetically modified trees. These factors limit and, in many cases, preclude field research and commercial adoption. Another challenge is the high cost and uncertainty in transformation efficiency that is needed to apply genetic engineering and gene editing methods to most species and genotypes of commercial importance. Recent advances in developmental gene-based transformation systems and gene editing, if combined with regulatory and certification system reform, could provide the foundation for genetic engineering to become a significant tool for coping with the increasing environmental and biological stresses on planted and wild forests.     

Inhibition of neuromuscular transmission in the guinea-pig saphenous artery by atriopeptin II

In the guinea-pig saphenous artery, stimulation of perivascular nerves elicited contraction and two types of synaptic potentials: the excitatory junction potential and the slow depolarization. The synaptic potentials were inhibited by atriopeptin II but not by sodium nitroprusside. Exogenous noradrenaline induced membrane depolarization and contraction, and both sodium nitroprusside and atriopeptin II inhibited the contraction but not the depolarization. These results suggest that atriopeptin II has an inhibitory effect both presynaptically at the nerve terminals and postsynaptically on the vascular smooth muscle cells.     

Quantitative analysis of cyclic AMP waves mediating aggregation in Dictyostelium discoideum

We have previously reported the detection of cAMP waves within monolayers of aggregating Dictyostelium discoideum cells (K. J. Tomchik and P.N. Devreotes, 1981, Science 212, 443-446). The computer-assisted analysis presented here of the fluorographic images of the cAMP waves reveals (1) all the waves have a consistent width and height; (2) cAMP concentrations within centers of concentric aggregation territories oscillate periodically while at spiral centers the concentration builds up to a plateau value within 2 mm; (3) cells within the region of intersection of two oppositely directed cAMP waves are stimulated to produce more cAMP than those responding to a single wave; (4) cells start to move when the cAMP level begins to increase and cease movement when the peak cAMP concentration reaches the cell.