Calcium-activated chloride channels in the retina

This review examines the function of calcium-activated chloride currents (I(Cl(Ca))) in the retina with an emphasis on their physiological role in photoreceptors. Although found in a variety of neurons and glial cells of the retina, I(Cl(Ca)) has been most prominently studied in cones, where it activates in response to depolarization-evoked Ca(2+) influx. The slow and complex gating kinetics of the chloride current have been considered to reflect the changing submembrane concentration of intracellular calcium. It is likely that the role of I(Cl(Ca)) is to stabilize the membrane potential of cones during synaptic activity and presynaptic Ca channel modulation. Several candidates in the molecular identification of the channel have been put forward but the issue remains unresolved.     

Bovine mammary gland UDP-GalNAc:GlcNAcbeta-R beta1-->4-N- acetylgalactosaminyltransferase is glycoprotein hormone nonspecific and shows interaction with alpha-lactalbumin

We have identified a novel N -acetylgalactosaminyltransferase activity in lactating bovine mammary gland membranes. Acceptor specificity studies and analysis of products obtained in vitro by 400 MHz1H-NMR spectroscopy revealed that the enzyme catalyses the transfer of N - acetylgalactosamine (GalNAc) from UDP-GalNAc to acceptor substrates carrying a terminal, beta-linked N -acetylglucosamine (GlcNAc) residue and establishes a beta1-->4-linkage forming a GalNAcbeta1-->4GlcNAc ( N, N '-diacetyllactosediamine, lacdiNAc) unit. Therefore, the enzyme can be identified as a UDP-GalNAc:GlcNAcbeta-R beta1-->4-N- acetylgalactosaminyltransferase (beta4-GalNAcT). This enzyme resembles invertebrate beta4-GalNAcT as well as mammalian beta4- galactosyltransferase (beta4-GalT) in acceptor specificity. It can, however, be clearly distinguished from the pituitary hormone-specific beta4-GalNAcT by its incapability of acting with an elevated activity on a glycoprotein substrate carrying a hormone-specific peptide motif. Furthermore, the GalNAcT activity appeared not to be due to a promiscuous action of a beta4-GalT as could be demonstrated by comparing the beta4-GalNAcT and beta4-GalT activities of the mammary gland, bovine colostrum, and purified beta4-GalT, by competition studies with UDP-GalNAc and UDP-Gal, and by use of an anti-beta4-GalT polyclonal inhibiting antibody. Interestingly, under conditions where mammalian beta4-GalT forms with alpha-lactalbumin (alpha-LA) the lactose synthase complex, the mammary gland beta4-GalNAcT was similarly induced by alpha-LA to act on Glc with an increased efficiency yielding the lactose analog GalNAcbeta1-->4Glc. This enzyme thus forms the second example of a mammalian glycosyltransferase the specificity of which can be modified by this milk protein. It is proposed that the mammary gland beta4-GalNAcT functions in the synthesis of lacdiNAc- based, complex-type glycans frequently occurring on bovine milk glycoproteins. The action of this enzyme is to be considered when aiming at the production of properly glycosylated protein biopharmaceuticals in the milk of transgenic dairy animals.      

Callus regeneration from Alnus incana protoplasts isolated from cell suspensions

Nagata and Takebe's (NT) medium, supllementedte with 2.5 μm 2,4-dichlorphenoxyacetic acid (2,4-D), induced development of friable calluses from leaves of axenic shoot cultures of Alnus incana. Fast-growing cell suspensions were established in the same medium without agar. Suspensions gave high yields of viable protoplasts after an overnight incubation in an enzyme mixture consisting of 1% (w/v) Onozuka R-10, 0.5% (w/v) Rhozyme HP-150, 0.03% (w/v) Macerase, CPW salts, and 13% (w/v) mannitol (pH 5.8). Protoplasts cultured on K8p medium underwent cell wall regeneration within 24 h. The optimum protoplast-derived colony formation and growth was obtained on the NT medium supplemented, as was the K8p medium, with glucose as the osmoticum, growth regulators, coconut milk and casein hydrolysate. Compared with other culture techniques, the agarose bead technique of Shillito et al. (Plant Cell Reports, 2 (1983) 244) improved cell division and colony formation frequency. Protoplast-derived macrocalluses grew under the same conditions as those used for leaf calluses.     

A covariotide model explains apparent phylogenetic structure of oxygenic photosynthetic lineages

The aims of the work were (1) to develop statistical tests to identify whether substitution takes place under a covariotide model in sequences used for phylogenetic inference and (2) to determine the influence of covariotide substitution on phylogenetic trees inferred for photosynthetic and other organisms. (Covariotide and covarion models are ones in which sites that are variable in some parts of the underlying tree are invariable in others and vice versa.) Two tests were developed. The first was a contingency test, and the second was an inequality test comparing the expected number of variable sites in two groups with the observed number. Application of these tests to 16S rDNA and tufA sequences from a range of nonphotosynthetic prokaryotes and oxygenic photosynthetic prokaryotes and eukaryotes suggests the occurrence of a covariotide mechanism. The degree of support for partitioning of taxa in reconstructed trees involving these organisms was determined in the presence or absence of sites showing particular substitution patterns. This analysis showed that the support for splits between (1) photosynthetic eukaryotes and prokaryotes and (2) photosynthetic and nonphotosynthetic organisms could be accounted for by patterns arising from covariotide substitution. We show that the additional problem of compositional bias in sequence data needs to be considered in the context of patterns of covariotide/covarion substitution. We argue that while covariotide or covarion substitution may give rise to phylogenetically informative patterns in sequence data, this may not always be so.      

The history of sensibilities: Of the standard of taste in mid-eighteenth century England and the circulation of smells in post-revolutionary France

Conclusion We have presented a somewhat different picture of the sensory profile of the mid-to late-eighteenth century than is found in the work of so renowned an historian as Michel Foucault.⁴⁹ According to Foucault, sight is the dominant (and dominating) sense of the modern ara; we live in a society of surveillance. In his account, there has been a steady progression in the power of the gaze to organize both knowledge and society since the Enlightenment. It was the reorganization of the space of the prison, hospital, and workplace in accordance with the principle of individualizing partitioning under the scrupulously classificatory eye of the master-disciplinarian that crystallized this tendency and laid the foundations for the scopic regime of contemporary Western society.⁵⁰ The evidence presented here concerning taste in England and smell in France suggests that Foucault's preoccupation with the visual may be misplaced. Rather than a steady progression or intensification of sight, we have glimpsed some of its vicissitudes relative to taste and smell. The question which this alternative picture raises is whether it was visuality that created the individuality of modern society,⁵¹ or merely cemented a change that was effected by other sensory means. Sight is eminently capable of structuring a field and of distributing objects or individuals within that field. But it is not as discriminating as taste or smell. We suggest that it was the enlistment of the latter senses, with their power to attract and to repel as well as to make distinctions, that helped catalyze the novel distribution of individuals into classes, as opposed to estates or ranks, that distinguishes the modern from the premodern era. Having done their work, the affective senses could recede again, leaving the job of policing the new boundaries to sight.David Howes is Assistant Professor of Anthropology at Concordia University, Montreal. Marc Lalonde is a Doctoral Candidate in the Department of Religion at Concordia University.     

Identification of a hemolysin from Actinobacillus pleuropneumoniae and characterization of its channel properties in planar phospholipid bilayers

A proteinaceous hemolysin secreted by strain 4074 of serotype 1 of Actinobacillus pleuropneumoniae was purified by diafiltration and ion exchange chromatographic techniques. The hemolytic activity is associated with a 107-kDa band as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and confirmed by Western blotting and immunoprecipitation. This hemolysin produces pores in membranes as demonstrated by osmotic protection studies using red blood cells and carbohydrate compounds of various molecular weights. These assays suggest a pore diameter in the order of 2 nm. Phospholipid bilayers composed of 1:1 w/w phosphotidylserine:phosphotidylethanolamine exposed to this toxin display discrete current flow events typical of transmembrane channels and consistent with the interpretation that this toxin acts by forming pores in phospholipid membranes. The linear relationship of current amplitude to holding potential when examined over the -60 to +60 mV range indicates that this pore has a constant mean single channel conductance level of 350-400 pS.     

Co-culture ofFrankia alni subsp.pommerii (strain ACN1 AG ) with birch (Betula papyrifera) protoplasts

The present study was undertaken to set up an experimental system in which barriers to infection of a non-host plant related to the presence of the cell wall, at the level of recognition and/or the necessity of penetrating the cell wall, might be bypassed. Co-cultures betweenFrankia alni subsp.pommerii (strain ACN1 ^AG ) andBetula papyrifera protoplasts were established. Betula protoplasts remained viable after 2 weeks with no substantial cell wall regeneration. Suppression of the wall barrier was not sufficient to allowFrankia infection under the conditions tested. The non-infectivity ofFrankia on Betula protoplasts may also reflect difficulties inherent to thein vitro environment, which might not permit duplication of infection mechanisms.     

Isozyme Variation among 40 Frankia Strains

Forty Frankia strains belonging to the Alnus and Elaeagnus host specificity groups and isolated from various plant species from different geographical areas were characterized by the electrophoretic separation of isozymes of eight enzymes. All the enzyme systems that were investigated showed large variation. Diaphorases and esterases gave multiple band patterns and confirmed the identification of specific Frankia strains. Less variability was observed with enzymes such as phosphoglucose isomerase, leucine aminopeptidase, and malate dehydrogenase, which allowed for the delineation of larger groups of Frankia strains. Cluster analysis, based on the pair-wise similarity coefficients calculated between strains, delineated three large, dissimilar groups of Frankia strains, although each of these groups contained a large amount of heterogeneity. However, numerous Frankia strains, mainly from the Alnus host specificity group, demonstrated a perfect homology for all the enzymes tested.