Analysis of the proteins,glycoproteins and glycosaminoglycans of fibroblast adhesions to substratum

The focal adhesion preparations which remain attached to a glass substratum when fibroblast bodies are removed by a gentle stream of buffer have been analysed by gel electrophoresis coupled with other selective methods of analysis. The results are consistent with the presence of three classes of macromolecular components. (i) Muscle and associated proteins amongst which actin was abundant with significant amounts of tropomyosin, some myosin and traces of α-actinin. Some vimentin was present but no vinculin. We detected a major new protein component, as yet unidentified, with a molecular weight in the region of 50 000–55 000 which is not desmin or tubulin and could have an important function at the focal adhesion. (ii) Glycoproteins which are a specialised subset of those in the whole plasma membrane and included a family which bind ricin and therefore contain β-galactose end groups, together with a series having carbohydrate chains which bound neither ricin nor concanavalin A. The relative proportion of ricin-binding glycoproteins compared to concanavalin A-binding glycoproteins was higher than in whole plasma membranes. (iii) Glycosaminoglycans, with hyaluronate identified as the major component by column chromatography and its susceptibility to Streptomyces hyaluronidase.     

Microtubule affinity chromatography: a new technique for isolating microtubule-binding proteins from rat pancreas.

We have developed an affinity chromatography method for the isolation of microtubule-associated proteins (MAPs) from soluble cytoplasmic extracts of rat pancreas. Among the ten proteins which copurify with pancreas tubulin on a colchicine derivatives-affinity chromatography, three polypeptides of respectively 58, 55 and 48 kDa strongly bind to the microtubule affinity column. To begin to characterize these proteins, we have generated polyclonal antibodies against tau polypeptides from brains of immature chicken or rat. As judged by immunoblots, the three polypeptides seem to be immunologically related to the tau proteins previously localized in brain.     

Interactions between M proteins of Streptococcus pyogenes and glycosaminoglycans promote bacterial adhesion to host cells.

Several microbial pathogens have been reported to interact with glycosaminoglycans (GAGs) on cell surfaces and in the extracellular matrix. Here we demonstrate that M protein, a major surface-expressed virulence factor of the human bacterial pathogen, Streptococcus pyogenes, mediates binding to various forms of GAGs. Hence, S. pyogenes strains expressing a large number of different types of M proteins bound to dermatan sulfate (DS), highly sulfated fractions of heparan sulfate (HS) and heparin, whereas strains deficient in M protein surface expression failed to interact with these GAGs. Soluble M protein bound DS directly and could also inhibit the interaction between DS and S. pyogenes. Experiments with M protein fragments and with streptococci expressing deletion constructs of M protein, showed that determinants located in the NH2-terminal part as well as in the C-repeat region of the streptococcal proteins are required for full binding to GAGs. Treatment with ABC-chondroitinase and HS lyase that specifically remove DS and HS chains from cell surfaces, resulted in significantly reduced adhesion of S. pyogenes bacteria to human epithelial cells and skin fibroblasts. Together with the finding that exogenous DS and HS could inhibit streptococcal adhesion, these data suggest that GAGs function as receptors in M protein-mediated adhesion of S. pyogenes.     

Characterization of glycosaminoglycans from human normal and scoliotic nasal cartilage with particular reference to dermatan sulfate.

The composition and the distribution of glycosaminoglycans (GAGs) present in normal human nasal cartilage (HNNC), were examined and compared with those in human scoliotic nasal cartilage (HSNC). In both tissues, hyaluronan (HA), keratan sulfate (KS) and the galactosaminoglycans (GalAGs)--chondroitin sulfate (CS) and dermatan sulfate (DS)--were identified. The overall GAG content in HSNC was approx. 30% higher than the HNNC. Particularly, a 114% increase in HA, and 46% and 86% in KS and DS, respectively, was recorded. CS was the main type of GAG in both tissues with no significant compositional difference. GalAG chains in HSNC exhibited an altered disaccharide composition which was associated with significant increases of non-sulfated and 6-sulfated disaccharides. DS, which was identified and quantitated for the first time in HNNC and HSNC, contained low amounts of iduronic acid (IdoA), 18% and 28% respectively. In contrast to other tissues, where IdoA residues are organized in long IdoA rich repeats, the IdoA residues of DS in human nasal cartilage seemed to be randomly distributed along the chain. DS chains in HSNC were of larger average molecular size than those from HNNC. These results clearly indicate the GAG content and pattern in both HNNC and HSNC and demonstrate that scoliosis of nasal septum cartilage is related to quantitative and structural modifications at the GAG level.     

Studies on the Golgi apparatus by electron microscopy with particular reference to Aoyama's technique

1. Aoyama's silver impregnation method for the Golgi apparatus has been used on exocrine cells of the pancreas of the mouse and studied by electron microscopy in order to determine as precisely as possible where the silver is deposited. Similar cells have also been fixed in buffered osmium tetroxide solution and compared with cells treated by the silver technique. 2. Examination of the Aoyama preparations usually revealed a light deposition of silver in the cytoplasm (hyaloplasm or matrix) and a heavy deposition of silver around a series of closely apposed vacuoles. The heavy deposition of silver was regarded as revealing the chromophilic region of the Golgi apparatus while the vacuoles were identified as the chromophobic component. 3. Comparison of the silver preparations with those fixed in buffered osmium tetroxide solution showed that the silver was primarily deposited in the region of the Golgi membranes.     

Limb bud chondrogenesis in cell culture, with particular reference to serum concentration in the culture medium

When limb bud mesodermal cells of stages 23–24 chick embryos were plated at low cell density (2 × 10⁵ cells/cm²) and cultured in medium containing 10% fetal calf serum (FCS) (serum-rich medium), all cells became fibroblastic and no chondrocyte differentiation occurred in the culture. However, when cells of the same origin were cultured in a medium containing only 0.1% FCS (serum-poor medium), almost all the cells formed aggregates which developed further to form cartilage nodules. The loss of chondrogenic activity in serum-rich medium culture was irreversible: cultivation of the limb bud cells in serum-rich medium for 12 h abolished chondrogenic activity completely and these cells could not resume activity on re-cultivation in serum-poor medium. Calf, horse and chick serum at a concentration of 10% also induced the loss of chondrogenic activity in low cell density culture. Failure of chondrogenesis in serum-rich medium culture seemed to be due to the commitment of bipotential limb bud mesodermal cells to fibroblastic cells rather than to selective detachment of pre-committed chondroblasts.     

Nuba agriculture and ethnobotany,with particular reference to sesame and sorghum

There is a remarkably high level of variation within cultivated sesame and sorghum in the Nuba Mountains of Sudan although the region is relatively small. The Nuba people are geographically isolated and culturally diverse in religion, language, material inventory, agricultural practices and in their rituals involving crop plants, and this contributes to the diversity in their cultivars. Nuba crop husbandry is sophisticated and high levels of genetic diversity are maintained by deliberate selection of crop varieties that are well adapted to each of the microenvironments of the region and best suited for different economic uses.     

Curvature factor and membrane solubilization, with particular reference to membrane rafts

The composition of membrane rafts (cholesterol/sphingolipid-rich domains) cannot be fully deduced from the analysis of a detergent-resistant membrane fraction after solubilization in Triton X-100 at 4°C. It is hypothesized that the membrane curvature-dependent lateral distribution of membrane components affects their solubilization. The stomatocytogenic, Triton X-100, cannot effectively solubilize membrane components, especially with regard to the outward membrane curvature.     

Analyses of ribosomal proteins from mycobacteria with particular reference to the β antigen

Abstract The proteins of the ribosomal subunits of Mycobacterium phlei were analysed by sodium dodecylsulphate (SDS)-polyacrylamide gel electrophoresis (PAGE) and by 2-dimensional PAGE. These techniques revealed that both the size and the charge of the mycobacterial ribosomal proteins were quite different from those of Escherichia coli . The divergent chemical properties of the mycobacterial ribosomes might be related to other exceptional properties of mycobacteria, e.g., their slow growth. An antigen designated β was furthermore revealed in both 30S and 50S subunits of M. phlei and Mycobacterium bovis BCG. 2 Proteins from each subunit migrated in a similar way in SDS-PAGE, being thus prime candidates as carriers of β-activity.     

Axoplasmic proteins of the squid giant nerve fiber with particular reference to the fibrous protein

1. Axoplasm of squid giant nerve fibers is examined with the ultracentrifuge and electrophoresis apparatus and several distinct components demonstrated. 2. One of these components, a protein called axon filaments, is isolated by fractional extraction followed by differential ultracentrifugation and redissolving in glycine solution. Axon filaments are monodisperse by ultracentrifugation. Their physical chemical properties have been studied. 3. The existence of a reversible transformation of axon filaments into a particle of lower molecular weight and lower asymmetry has been demonstrated.     

Evolutionary ecology of colonial reef-organisms, with particular reference to corals

Sedentary reef-organisms such as sponges, colonial coelenterates, bryozoans and compound ascidians produce repeated modules (aquiferous systems, polyps, zooids) as they grow. Modular construction alleviates constraints on biomass imposed by mechanical and energetic factors that are functions of the surface area to volume ratio. Colonies thus may grow large whilst preserving optimal modular dimensions. Among corals, optimal polyp size is smaller in the more autotrophic than in the more heterotrophic species. Modular construction allows flexibility of growth form, which can adapt to factors such as water currents, silting, light intensity and proximity of competitors. Modular colonies have great regenerative capacities, even separated fragments may survive and grow into new colonies. All fragments from a parental colony are genetically identical and large branching corals frequently undergo clonal propagation through fragmentation during storms. Soft corals can also fragment endogenously. By spreading the risk of mortality among independent units, the generation and dispersal of fragments lessens the likelihood of clonal extinction. In spite of their ability to propagate asexually, most benthic colonial animals also reproduce asexually. The selective advantages of the genetic diversity among sexually produced offspring seem not to be linked with dispersal, but probably lie in the biological interactions with competitors, predators and pathogens in the parental habitat. Age at first sexual maturity and the proportional investment of resources in sexual reproduction are related to colonial survivorship. Small branching corals on reef flats grow quickly, attain sexual maturity within 1–4 years, planulate extensively, but reach only small sizes before dying. Massive corals are longer lived and have the opposite characteristics of growth and reproduction. Most sessile reef organisms compete for space, food or light. Faster growers can potentially outcompete slower growers, but are often prevented from doing so by several forms of aggression from competitors and by the damage inflicted by storms. Competitive interactions among sedentary organisms on coral reefs are unlikely to be linear or deterministic, and so the co-existence of diverse species is possible.     

Individual versus social complexity, with particular reference to ant colonies

Insect societies colonies of ants, bees, wasps and termites--vary enormously in their social complexity. Social complexity is a broadly used term that encompasses many individual and colony-level traits and characteristics such as colony size, polymorphism and foraging strategy. A number of earlier studies have considered the relationships among various correlates of social complexity in insect societies; in this review, we build upon those studies by proposing additional correlates and show how all correlates can be integrated in a common explanatory framework. The various correlates are divided among four broad categories (sections). Under 'polyphenism' we consider the differences among individuals, in particular focusing upon 'caste' and specialization of individuals. This is followed by a section on 'totipotency' in which we consider the autonomy and subjugation of individuals. Under this heading we consider various aspects such as intracolony conflict, worker reproductive potential and physiological or morphological restrictions which limit individuals' capacities to perform a range of tasks or functions. A section entitled 'organization of work' considers a variety of aspects, e.g. the ability to tackle group, team or partitioned tasks, foraging strategies and colony reliability and efficiency. A final section, 'communication and functional integration', considers how individual activity is coordinated to produce an integrated and adaptive colony. Within each section we use illustrative examples drawn from the social insect literature (mostly from ants, for which there is the best data) to illustrate concepts or trends and make a number of predictions concerning how a particular trait is expected to correlate with other aspects of social complexity. Within each section we also expand the scope of the arguments to consider these relationships in a much broader sense of'sociality' by drawing parallels with other 'social' entities such as multicellular individuals, which can be understood as 'societies' of cells. The aim is to draw out any parallels and common causal relationships among the correlates. Two themes run through the study. The first is the role of colony size as an important factor affecting social complexity. The second is the complexity of individual workers in relation to the complexity of the colony. Consequently, this is an ideal opportunity to test a previously proposed hypothesis that 'individuals of highly social ant species are less complex than individuals from simple ant species' in light of numerous social correlates. Our findings support this hypothesis. In summary, we conclude that, in general, complex societies are characterized by large colony size, worker polymorphism, strong behavioural specialization and loss of totipotency in its workers, low individual complexity, decentralized colony control and high system redundancy, low individual competence, a high degree of worker cooperation wher tackling tasks, group foraging strategies, high tempo, multi-chambered tailor-made nests, high functional integration, relatively greater use of cues and modulatory signals to coordinate individuals and heterogeneous patterns of worker-worker interaction.