Understorey vegetation change in a Picea mariana chronosequence

Eighteen black spruce (Picea mariana) stands, representing postfire ages of 26 to 120 yr, were surveyed for understorey vegetation and site/microsite characteristics at two spatial scales. This enabled comparison of within- versus among-stand compositional variation.Detrended correspondence analysis (DCA) ordination among the 18 stands revealed a complex age/moisture gradient. DCA ordination among 1 800 quadrats within the stands indicated a similar gradient with much compositional overlap. Quadrats were grouped, using two-way indicator species analysis (TWINSPAN), into 9 classes each representing a phase in understorey vegetation composition. These phases shifted in abundance from young to old stands with a high degree of concordance among replicates in the same age class. Understorey succession is strongly linked to the stages in tree growth, mortality and thinning coupled with the accumulation of site moisture.Abbreviations DCA Detrended Corrospondence Analysis     

Anonymous nuclear DNA markers in the American oyster and their implications for the heterozygote deficiency phenomenon in marine bivalves

A puzzling population-genetic phenomenon widely reported in allozyme surveys of marine bivalves is the occurrence of heterozygote deficits relative to Hardy-Weinberg expectations. Possible explanations for this pattern are categorized with respect to whether the effects should be confined to protein-level assays or are genomically pervasive and expected to be registered in both protein- and DNA-level assays. Anonymous nuclear DNA markers from the American oyster were employed to reexamine the phenomenon. In assays based on the polymerase chain reaction (PCR), two DNA-level processes were encountered that can lead to artifactual genotypic scorings: (a) differential amplification of alleles at a target locus and (b) amplification from multiple paralogous loci. We describe symptoms of these complications and prescribe methods that should generally help to ameliorate them. When artifactual scorings at two anonymous DNA loci in the American oyster were corrected, Hardy-Weinberg deviations registered in preliminary population assays decreased to nonsignificant values. Implications of these findings for the heterozygote-deficit phenomenon in marine bivalves, and for the general development and use of PCR-based assays, are discussed.      

Molecular genetic studies of Creutzfeldt-Jakob disease

Genetic study of over 200 cases of Creutzfeldt-Jakob disease (CJD), Gerstmann-Sträussler-Scheinker disease (GSS), fatal familial insomnia (FFI), and kuru have brought a reliable body of evidence that the familial forms of CJD and all known cases of GSS and FFI are linked to germline mutations in the coding region of the PRNP gene on chromosome 20, either point substitutions or expansion of the number of repeat units. No pathogenic mutations have so far been found in sporadic or infectious forms of CJD, although there are features of genetic predisposition in iatrogenic CJD and kuru. In FFI and familial CJD, clinically and pathologically distinct syndromes that are both linked to the 178^Asp→Asn substitution, phenotypic expression is dependent on a polymorphism at codon 129. Synthetic peptides homologous to several regions of PrP spontaneously form insoluble amyloid fibrils with unique morphological characteristics and polymerization tendencies. Peptides homologous to mutated regions of PrP exhibit enhanced fibrilogenic properties and, if mixed with the wild-type peptide, produce even more abundant and larger fibrous aggregates. A similar process in vivo may lead to amyloid accumulation and disease, and transmission of “baby fibrils” may induce disease in other hosts.     

Coastal-marine discontinuities and synergisms: implications for biodiversity conservation

Coastal-marine biodiversity conservation must focus increasingly at the level of the land- and seascape. Five cases illustrate discontinuities and synergisms and how system changes may take place. For Caribbean coral reefs, the result of overfishing and disease has been a shrinkage in the entire system, the effects of which may cascade through the coastal seascape. For Beringia, patterns of benthic diversity are best understood in a manner that matches the multiscale, integrated dynamics of weather, ice, marine mammal feeding, and community structure. In the case of US East Coast estuaries, oyster reefs may be keystone elements, with important effects on functional diversity. Large-scale coastal systems depend upon the connectivity of fresh and marine waters in the coastal zone, having implications for the apparent stochasticity of coastal fisheries. And, for a coastal barrier-lagoon site, a state change may be described in terms of a combination of succession, the attainment of a quasi-equilibrium state, and disturbance. A profound problem for conservation is that there is very little information about the relationship between species diversity and ecological function. Coastal-marine biodiversity conservation is best addressed at the level of the land- and seascape.     

Role of glycoprotein PE2 in formation and maturation of the Sindbis virus spike.

Sindbis virus envelope assembly is a multistep process resulting in the maturation of a rigid, highly ordered T=4 icosahedral protein lattice containing 80 spikes composed of trimers of E1-E2 heterodimers. Intramolecular disulfide bonds within E1 stabilize E1-E1 associations required for envelope formation and maintenance of the envelope's structural integrity. The structural integrity of the envelope protein lattice is resistant to reduction by dithiothreitol (DTT), indicating that E1 disulfides which stabilize structural domains become inaccessible to DTT at some point during virus maturation. The development of E1 resistance to DTT occurs prior to the completion of E1 folding and is temporally correlated with spike assembly in the endoplasmic reticulum. From these data we have predicted that in the final stages of spike assembly, E1 intramolecular disulfides, which stabilize the structural integrity of the envelope protein lattice, are buried within the spike and become inaccessible to the reductive activity of DTT. The spike is formed prior to the completion of E1 folding, and we have suggested that PE2 (the precursor to E2) may play a critical role in E1 folding after PE2-E1 oligomer formation has occurred. In this study we have investigated the role of PE2 in E1 folding, oligomer formation, and development of E1 resistance to both protease digestion and reduction by DTT by using a Sindbis virus replicon (SINrep/E1) which allows for the expression of E1 in the presence of truncated PE2. Through pulse-chase analysis of both Sindbis virus- and SINrep/E1-infected cells, we have determined that the folding of E1 into a trypsin-resistant conformation and into its most compact and stable form is not dependent upon association of E1 with PE2. However, E1 association with PE2 is required for oligomer formation, the export of E1 from the endoplasmic reticulum, and E1 acquisition of resistance to DTT.     

Quantitative video sampling of coral reef benthos: large-scale application

The feasibility of reliably estimating percent cover of coral reef benthos by video techniques is examined. Video belt transects were recorded within study areas on Davies and John Brewer Reefs in the central Great Barrier Reef during September 1988. Two years later in September 1990, the study area at Davies Reef was resampled concurrently by video and line intercept transects. Percent cover data of major coral growth forms and non-biotic physionomic attributes were extracted from video footage by scoring the identity of items located at even or random spaced points along the transect. A cost-benefit analysis which compared increases in precision with increases in sampling effort suggested that the optimum regime for analyzing 200 m long video transects was five subsamples of 110 random points or one sample of 550. This regime resolved both spatial variability at large and smaller spatial scales (between study areas and among transects) and temporal change (within a single study area over a two year period) for total live coral and individual growth forms. The strengths of the technique lie in its cost-savings in filed expenses, and in the production of a permanent visual record. The limitations of the technique lie in reduced taxonomic resolution when compared with hands on field techniques. The results suggest that for broad taxonomic categories of coral reef benthos reliable estimates of relative abundance can be obtained by video techniques.     

Basolateral plasma membrane localization of ouabain-sensitive sodium transport sites in the secretory epithelium of the avian salt gland

The distribution of Na+ pump sites (Na+-K+-ATPase) in the secretory epithelium of the avian salt gland was demonstrated by freeze-dry autoradiographic analysis of [(3)H] ouabain binding sites. Kinetic studies indicated that near saturation of tissue binding sites occurred when slices of salt glands from salt-stressed ducks were exposed to 2.2 μM ouabain (containing 5 μCi/ml [(3)H]ouabain) for 90 min. Washing with label-free Ringer's solution for 90 min extracted only 10% of the inhibitor, an amount which corresponded to ouabain present in the tissue spaces labeled by [(14)C]insulin. Increasing the KCl concentration of the incubation medium reduced the rate of ouabain binding but not the maximal amount bound. In contrast to the low level of ouabain binding to salt glands of ducks maintained on a freshwater regimen, exposure to a salt water diet led to a more than threefold increase in binding within 9-11 days. This increase paralleled the similar increment in Na+-K+-ATPase activity described previously. [(3)H]ouabain binding sites were localized autoradiographically to the folded basolateral plasma membrane of the principal secretory cells. The luminal surfaces of these cells were unlabeled. Mitotically active peripheral cells were also unlabeled. The cell-specific pattern of [(3)H]ouabain binding to principal secretory cells and the membrane-specific localization of binding sites to the nonluminal surfaces of these cells were identical to the distribution of Na+-K+-ATPase as reflected by the cytochemical localization of ouabain-sensitive and K+-dependent nitrophenyl phosphatase activity. The relationship between the nonluminal localization of Na+-K+-ATPase and the possible role of the enzyme n NaCl secretion is considered in the light of physiological data on electrolyte transport in salt glands and other secretory epithelia.     

Metabolism of adenine nucleotides by ectoenzymes of vascular endothelial and smooth-muscle cells in culture.

1. Pig aortic endothelial and smooth-muscle cells in culture rapidly catabolize exogenous ATP, ADP or AMP. 2. In both cell types catabolism is due to Mg2+-stimulated ectoenzymes. 3. Inhibition and substrate-specificity studies suggest that both cell types possess three distinct ectonucleotidases, namely nucleoside triphosphatase (EC 3.6.1.15), nucleoside diphosphatase (EC 3.6.1.6) and 5'-nucleotidase (EC 3.1.3.5), as well as nucleoside diphosphate kinase (EC 2.7.4.6). 4. These ectonucleotidase systems could be of importance in the regulation of neurotransmission, blood platelet function and vasodilation.     

Determination of plasmid copy number by fluorescence densitometry

A simple and reliable method for the determination of plasmid copy numbers by direct fluorescence densitometry of ethidium bromide-stained electrophoretic gels was developed. In developing the method, the following parameters were evaluated and controlled: plasmid DNA trapping in the linear chromosomal DNA, staining-destaining kinetics for ethidium bromide, linearity of the fluorescence response, and the effect of the molecular topology of DNA on ethidium bromide binding to DNA in agarose.