Kinetics of excited states of pigment clusters in solubilized light-harvesting complex II: photon density-dependent fluorescence yield and transmittance.

Relative fluorescence yield, phi F, and transmittance, T, were measured in solubilized light-harvesting complex II (LHCII) as a function of photon density, Ip, of monochromatic 645-nm laser pulses (duration: approximately 2.5 ns). Special efforts were made in constructing an optical set-up that allows the accurate determination of the fluorescence from an area of constant Ip, phi F(Ip) starts to decline at approximately 10(14) and drops to values below 0.01% at maximum Ip (approximately 10(19) photons cm-2 pulse-1). T(Ip) decreases only slightly at photon densities of approximately 10(15) but increases steeply at values of > 10(17) photons cm-2 pulse-1. The interpretation of the phi F(Ip) data using the saturation limit of Mauzerall's multiple hit model leads to a unit size of about 10-15 chlorophyll molecules. One interpretation is to attribute this result to a very fast exciton-exciton annihilation of multiple excited states generated within this small domain. Alternatively, based on the assumption that delocalized cluster states within the monomeric/trimeric subunit of LHCII exist, the results can be consistently described by a kinetic model comprising ground, monoexcitonic, and biexcitonic states of clusters and a triplet state that is quenched by carotenoids in LHCII. Within the framework of this model the annihilation of multiple excitations is explained as ultrafast radiationless relaxation of higher excited cluster states. Comparative measurements in diluted acetonic Chl a solution are consistently described by the depletion of the ground state, taking the absorption cross section at the used wavelength.     

Binding of ionic ligands to polyelectrolytes.

Ionic ligands can bind to polyelectrolytes such as DNA or charged polysaccharides. We develop a Poisson-Boltzmann treatment to compute binding constants as a function of ligand charge and salt concentration in the limit of low ligand concentration. For flexible chain ligands, such as oligopeptides, we treat their conformations using lattice statistics. The theory predicts the salt dependence and binding free energies, of Mg(2+) ions to polynucleotides, of hexamine cobalt(III) to calf thymus DNA, of polyamines to T7 DNA, of oligolysines to poly(U) and poly(a), and of tripeptides to heparin, a charged polysaccharide. One parameter is required to obtain absolute binding constants, the distance of closest separation of the ligand to the polyion. Some, but not all, of the binding entropies and enthalpies are also predicted accurately by the model.     

Engineering the Drosophila Genome: Chromosome Rearrangements by Design

We show that site-specific recombination can be used to engineer chromosome rearrangements in Drosophila melanogaster. The FLP site-specific recombinase acts on chromosomal target sites located within specially constructed P elements to provide an easy screen for the recovery of rearrangements with breakpoints that can be chosen in advance. Paracentric and pericentric inversions are easily recovered when two elements lie in the same chromosome in opposite orientation. These inversions are readily reversible. Duplications and deficiencies can be recovered by recombination between two elements that lie in the same orientation on the same chromosome or on homologues. We observe that the frequency of recombination between FRTs at ectopic locations decreases as the distance that separates those FRTs increases. We also describe methods to determine the absolute orientation of these P elements within the chromosome. The ability to produce chromosome rearrangements precisely between preselected sites provides a powerful new tool for investigations into the relationships between chromosome arrangement, structure, and function.     

Spatial and environmental components of variation in the distribution patterns of subarctic plant species at Kevo, N Finland — a case study at the meso-scale level

This study presents a quantitative partitioning of the total variance in the patterns of occurrence of 231 vascular plant taxa in 362 1 × 1 km grids in the Kevo Nature Reserve into four independent components: purely spatial variation, spatially structured environmental variation, non-spatial environmental variation, and unexplained variation. This partitioning is done with (partial) constrained ordinations (canonical correspondence analysis) and associated Monte Carlo permutation tests. The numerical results suggest that most of the biological variance captured by the external explanatory variables is related to 'local' meso-scale environmental factors, as 12.6% of the variation in the species data is explained solely by the environmental variables. Part of the variance (6%) represents a spatially covarying environmental component, but only a very small part, ca 2%, is related to purely spatial variation. The amount of unexplained variation is very high (>75%). The results are compared and discussed in relation to the relative amounts of these four variance components at broader- and finer-scales and to the concepts of domains and transition zones of scales in biological patterning.     

Seasonal variation in the effects of hard substratum biofilming on settlement of marine invertebrate larvae

The effect of season on "biofilming";, as a cue for the settlement of marine invertebrate larvae, was investigated in a long-term field study during the years 1992-1994. The series of settlement experiments was conducted in a tidal rapid on the west coast of Scotland, and involved manipulations of artificial panels. Biofilming of substrata, whilst excluding larval settlement, was achieved by the enclosure of panels within tight-fitting (but removable) mesh screens so that the number of settlers on filmed and unfilmed substrata were counted in the initial absence of other incumbent post-larvae. Depending on larval species, the effects of biofilming were found to be either facilitatory or inhibitory. Significant within- and between-species seasonal differences in the settlement responses were detected, and a reversal of the effect of biofilming on larval settlement response, from inhibitory to facilitatory and vice versa, was noted with season in the case of some taxonomic groups and species (e.g. Tubulipora sp., Plagioecia sp., Electra pilosa (L.)). The present study emphasizes the need for extended field studies of larval responses to environmental cues, when the focus of interest is in drawing general inferences about naturally occurring behavioural patterns at settlement.     

Fluoranthene as a model toxicant in sediment studies with Chironomus riparius

The bioavailability of fluoranthene (as a model toxicant) in freshwater sediment tests with Chironomus riparius was investigated in relation to the partitioning of the chemical between water and sediment. Effects on survival and successful development of newly hatched larvae through to adult emergence were measured over a range of sediment concentrations. Radiolabelled fluoranthene was employed to enable sediment, aqueous and tissue concentrations to be determined. A significant effect on emergence was obtained at a measured sediment concentration of 80 mg kg^-1 (dry weight). The measured concentration in the overlying water was 88 g l^-1. Although effects at 170 mg kg^-1 (dry weight) were more severe, the measured concentration in the overlying water was similar (89 g l^-1), probably reflecting the approach towards the solubility limit. Bioaccumulation of fluoranthene, determined by analysis of emerging adults, was approximately proportional to the sediment concentration. Although the aqueous phase concentration appeared to be the principal factor controlling bioavailability, the increasing toxicity, when water concentrations had reached a plateau, suggested that other mechanisms may also be involved. Different sediment spiking procedures resulted in broadly similar partitioning of the chemical between the sediment and the overlying water. There were only minor differences in toxicity between the procedures, two of which allowed the carrier solvent used for spiking to be eliminated before addition of the test organisms. It was concluded that, for most purposes, all four procedures were equally acceptable, and by retaining a flexible approach, even for standardised regulatory tests, spiking procedures may be tailored to the physico-chemical properties of the test material.     

Lack of Staphylococcal Enterotoxin Production among Strains of Staphylococcus aureus from Bovine Mastitis in Denmark

Staphylococcal enterotoxins (SE’s) are a group of small exoproteins produced by some strains of Staphylococcus aureus. The SE’s, designated A to E according to their antigenic specificities, are important causes of food poisoning worldwide. Milk and dairy products are frequently associated with S. aureus enter-otoxin food poisoning, and it is supposed that infected milk from mastitic animals constitute the main source of enterotoxigenic S. aureus of animal origin (Bryon 1983, Gilmour & Harvey 1990, Bergdoll 1989). Indeed, S. aureus is the most common cause of bovine mastitis worldwide, and if mastitis strains produce SE this makes up an enormous reservoir of potential enterotoxin producers. The production of SE by S. aureus isolated from bovine mastitis have been investigated in several countries (Matsunaga et al. 1993, Kenny et al. 1993, Olson et al 1970, Orden et al. 1992, Olsvik et al. 1981, Adekeye 1980, Garcia et al. 1980, Abbar 1986, Harvey & Gilmour 1985). Since no studies have been performed on the prevalence of enterotoxigenic strains of S. aureus isolated from bovine mastitis in Denmark, a well characterized collection of S. aureus (Aarestrup et al. 1995) was investigated with respect to this property.     

Transmissibility of Bacterial Endosymbionts Between Isolates of Acanthamoeba Spp.

ABSTRACT. Experimental transmission of two bacterial endosymbionts to symbiont-free isolates of Acanthamoeba spp. was studied to determine specificity of the host-symbiont relationship. Both symbionts originated from amoebic isolates displaying an identical mitochondrial DNA Eco RI fingerprint (group AcUW II). Symbioses were readily established in one amoebic isolate which displayed a homologous mtDNA fingerprint (group AcUW II). Exposure of a heterologous amoebic isolate (group AcUW IV) to the two symbionts resulted in either cell death or encystation without the establishment of symbioses. While symbioses were established with an amoebic isolate from a second heterologous group (AcUW I), a unique membranous sheath appeared and persisted around one of the symbionts which did not exist in the original host. An isolate representing a third heterologous amoebic group (AcUW VI) was variable in its susceptibility with one symbiont unable to infect the host and the other becoming established only after an initial reaction in which trophozoites rounded-up and floated off the substrate. These studies suggest that a specific recognition system exists between particular isolates of Acanthamoeba and their symbionts, and that the appearance of a killer phenotype is related to contact between mismatched, though recognized, pairs.