A rapid procedure for cell colony hybridization using DNA probes

A rapid, direct method for screening single cell-derived colonies or foci is described. The method allows the screening of a large number of colonies or foci by nitrocellulose filter hybridization using DNA probes. This technique simplifies current screening procedures and is a reliable, rapid, and sensitive method for the selection of cell clones containing a desired transfected gene.     

Cell adhesion molecule in the hexactinellid Aphrocallistes vastus

Abstract. The Hexactinellida sponge Aphrocallistes vastus contains a soluble aggregation factor (AF) whose purification has been described in this communication. It is characterized by a S°_20.w value of 37 and a buoyant density of 1.45 g/cm³. The AF is a glycoporteinaceous particle composed of three major protein species; no core structure could be visualized. In the presence of Ca²⁺, the AF causes secondary aggregation of single cells. The aggregation process is temperature, pH, and ionic strength independent within a broad range. Evidence is presented indicating that two (or more) AF molecules are required for the establishment of a stable cell: cell interaction. In contrast to the AFs from demosponges, the hexactinellid AF functions species-unspecifically.     

Population structure,dispersal and colonization history of the garden bumblebee <Emphasis Type="Italic">Bombus hortorum</Emphasis> in the Western Isles of Scotland

New methods of analysing genetic data provide powerful tools for quantifying dispersal patterns and reconstructing population histories. Here we examine the population structure of the bumblebee Bombus hortorum in a model island system, the Western Isles of Scotland, using microsatellite markers. Following declines in other species, B. hortorum is the only remaining long-tongued bumblebee species found in much of Europe, and thus it is of particular ecological importance. Our data suggest that populations of B. hortorum in western Scotland exist as distinct genetic clusters occupying groups of nearby islands. Population structuring was higher than for other bumblebee species which have previously been studied in this same island group (F_st = 0.16). Populations showed significant isolation by distance. This relationship was greatly improved by using circuit theory to allow dispersal rates to differ over different landscape features; as we would predict, sea appears to provide far higher resistance to dispersal than land. Incorporating bathymetry data improved the fit of the model further; populations separated by shallow seas are more genetically similar than those separated by deeper seas. We argue that this probably reflects events following the last ice age when the islands were first colonized by this bee species (8,500–5,000 ybp), when the sea levels were lower and islands separated by shallow channels would have been joined. In the absence of significant gene flow these genetic clusters appear to have since diverged over the following 5,000 years and arguably may now represent locally adapted races, some occurring on single islands.     

Quantification of viable biomass in biofilm reactors by extractable lipid phosphate

 Lipid phosphate, which is a measure of viable biomass, was determined using biofilm samples from three different laboratory-scale reactors. The analysis procedure proposed in the literature was modified and tested for suitability in experiments with biofilm reactors. The microbial contents of the biofilms studied are compared in three types of reactor. Received: 2 November 1994/Accepted: 23 January 1995     

Sodium chloride treatment of amphotericin B nephrotoxicity. Standard of care?

Amphotericin B is an effective therapeutic agent for most systemic or invasive mycoses, but its usefulness is limited by the frequent occurrence of nephrotoxicity. Given the high and increasing frequency of serious fungal infections, especially in immunocompromised patients, the importance of the morbidity caused by this toxicity is substantial. Salt loading may prevent and even reverse amphotericin B-induced azotemia by an unknown mechanism. A prospective, randomized, placebo-controlled trial in a relevant patient group would strengthen the support for this simple, safe therapy, but will not likely be carried out because of practical and ethical considerations. Thus, a few prospective and limited controlled human studies may be the only supportive evidence for using this therapy. Supplementing dietary sodium chloride intake with 150 mEq of sodium chloride daily intravenously or orally beginning when or before amphotericin B therapy is initiated will likely prevent much of the observed nephrotoxicity and should be carried out routinely.     

Role of different lymphoid tissues in the initiation and maintenance of DNA-raised antibody responses to the influenza virus H1 glycoprotein.

Antibody responses in mice immunized by a single gene gun inoculation of plasmid expressing the influenza virus H1 hemagglutinin and in mice immunized by a sublethal H1 influenza virus infection have been compared. Both immunizations raised long-lived serum responses that were associated with the localization of antibody-secreting cells (ASC) to the bone marrow. However, the kinetics of these responses were 4 to 8 weeks slower in the DNA-immunized than in the infection-primed mice. Following a gene gun booster, the presence of ASC in the inguinal lymph nodes, but not in other lymph nodes, revealed gene gun responses being initiated in the nodes that drain the skin target site. Both pre- and postchallenge, the DNA-immunized mice had 5- to 10-times-lower levels of antibody and ASC than the infection-primed mice.