Attachment of Neisseria gonorrhoeae to the cellular pilus receptor CD46: identification of domains important for bacterial adherence

Pili of Neisseria gonorrhoeae mediate binding of the bacteria to human host cells. Membrane cofactor protein (MCP or CD46), a human cell-surface protein involved in regulation of complement activation, acts as a cellular pilus receptor. In this work, we examined which domains of CD46 mediate bacterial adherence. The CD46 expression was quantified and characterized in human epithelial cell lines. N. gonorrhoeae showed the highest adherence to ME180 cells, which have BC1 as the dominant phenotype. The BC isoforms of CD46 were expressed in all cell lines tested. The adherence was not enhanced by high expression of other isoforms, showing that the BC domain of CD46 is important in adherence of N. gonorrhoeae to human cells. To characterize the pilus-binding site within the CD46 molecule, a set of CD46–BC1 deletion constructs were transfected into COS-7 cells. Piliated N. gonorrhoeae attached well to CD46–BC1-expressing COS-7 cells. We show that the complement control protein repeat 3 (CCP-3) and the serine–threonine–proline (STP)-rich domain of CD46 are important for efficient adherence to host cells. Further, partial deletion of the cytoplasmic tail of CD46 results in low bacterial binding, indicating that the cytoplasmic tail takes part in the process of establishing a stable interaction between N. gonorrhoeae and host cells.     

Complement deficiency ameliorates collagen-induced arthritis in mice

Collagen-induced arthritis (CIA) is an experimental animal model of human rheumatoid arthritis being characterized by synovitis and progressive destruction of cartilage and bone. CIA is induced by injection of heterologous or homologous collagen type II in a susceptible murine strain. DBA/1J mice deficient of complement factors C3 (C3(-/-)) and factor B (FB(-/-)) were generated to elucidate the role of the complement system in CIA. When immunized with bovine collagen type II emulsified in CFA, control mice developed severe arthritis and high CII-specific IgG Ab titers. In contrast, the C3(-/-) and FB(-/-) were highly resistant to CIA and displayed decreased CII-specific IgG Ab response. A repeated bovine collagen type II exposure 3 wk after the initial immunization led to an increase in the Ab response in all mice and triggered arthritis also in the complement-deficient mice. Although the arthritic score of the C3(-/-) mice was low, the arthritis in FB(-/-) mice ranked intermediate with regard to C3(-/-) and control mice. We conclude that complement activation by both the classical and the alternative pathway plays a deleterious role in CIA.     

Gold-coated capillary based 2,4-dichlorophenoxyacetic acid chemi-luminescent assays: possibilities towards multianalysis

The application of gold-coated glass capillaries for the design of a sensitive chemiluminescent immunoassay for 2,4-dichlorophenoxyacetic acid (2,4-D) is reported. The gold coating on the glass capillaries has been partially characterized and its effect on enhancing the signal intensity has been measured. A simple photo-multiplier tube-based photon detector is used for this purpose. At least three times improvement in the signal intensity is observed compared to uncoated glass capillaries, with a consequent improvement in the sensitivity of detection. Using such gold-coated glass capillaries, 2,4-D in the range 10(-9) to 10(-13) mol/l is detectable at a precision of +/-15% (CV%) and a limit of detection of 10(-15) mol/l is achievable. The possibility of using such gold-coated capillaries with a portable multianalytical set-up for field studies is also demonstrated.     

Temporal and spatial patterns in recruitment and succession of a temperate marine fouling assemblage: a comparison of static panels and boat hulls during the boating season

Protection against biofouling is essential for efficient operation of boats and ships. Restrictions on the use of traditional, toxic antifouling coatings call for new less toxic methods. Future antifouling strategies will likely be based on more specific action against dominant foulers and will require more detailed information about spatial and temporal differences in fouling communities on artificial substrates. In this study, the recruitment and succession of fouling organisms was examined on artificial (PMMA) panels exposed to natural flow speeds on the Skagerrak coast (Sweden). The recruitment of foulers on static panels was then compared to fouling allowed to develop on boat hulls in surveys of new nontoxic coatings. The temporal and spatial variation in recruitment was examined on a monthly interval within the boating season, from May-September. Furthermore, the succession of the fouling community was examined during the same interval. A total of 12 sessile invertebrate species was recorded on the static panels with dominant foulers being the barnacle Balanus improvisus (July-September) and the mussel Mytilus edulis (June-August). The mean abundance during peak settlement on panels after 1 month's deployment was 370 +/- 134 individuals dm-2 for B. improvisus and 340 +/- 415 individuals dm-2 for M. edulis. The succession of foulers on the panels led to a dominance of M. edulis (maximum of 7470 +/- 2830 individuals dm-2) over B. improvisus (maximum of 2295 +/- 680 individuals dm-2). This was in contrast to the fouling development on boat hulls, examined after 4 months exposure in 3 successive years, where B. improvisus was the dominant species (mean abundance 222 +/- 104 and maximum 713 +/- 527 individuals dm-2). Some boats were covered to an extent of almost 100% by B. improvisus with only a few M. edulis (mean abundance 20 +/- 16). The biomass of fouling on boat hulls varied from average dry weights of 1.3 +/- 1 to 13 +/- 5 g dm-2. These results show that extrapolation from static panels (common in evaluation of antifouling performance) to fouling on boat hulls may be problematic.     

Long-term study of the ecology of wild Atlantic salmon smolts in a small Norwegian river

Backcalculated lengths at the end of the first growth season in wild Atlantic salmon Salmo salar differed significantly between parr smolting at age 1, 2 and 3 years over a period of 11 years (i.e. 1983–1993). Mean body lengths of the respective age groups at the end of the first growth period were 11·1, 6·2 and 4·7 cm, respectively. The mean percentage distribution of fish smolting at age 1, 2 and 3 was 14, 78 and 7%, and the mean smolt age was 1·95 years. Mean lengths at smolting of age groups 1, 2 and 3 were 13·6, 15·8 and 17·5 cm, respectively. Females outnumbered males among the downstream migrating smolts with a mean sex ratio (females/ males) estimated at 1·61, with a significant female surplus in 7 of the 11 years sampled. Of the smolts sampled, 14% exhibited enlarged gonads indicative of parr maturation, and all were males (37% of the parr males sampled). Mean annual smolt density from 1975 to 1996 was 13·4 individuals 100 m⁻² ranging between 0·3–31 smolts 100 m⁻². Mean densities (100 m⁻²) of the smolts aged 1, 2 and 3 years were 1·5, 9·3 and 0·9 fish, respectively. Mean annual biomass for the 22-year period (1975–1996) was estimated at 437 g 100 m⁻², with a range of variation from 136 to 683 g 100 m⁻². Smolt age 2 made up 81% of the mean annual biomass (355 g 100 m⁻²) and smolt age 1 and 3, 8% (35 g 100 m⁻²) and 11% (47 g 100 m⁻²), respectively.     

Screening for species potentially sensitive to habitat fragmentation

FORUM is a lighter channel of communication between readers and contributors: it aims to stimulate discussion and debate, particularly by presenting new ideas and by suggesting alternative interpretations to the more formal research papers published in ECOGRAPHY and elsewhere. A lighter prose is encouraged and no summary is required. Contributions should be concise and to the point, with a relatively short bibliography. Formal research papers, however short, will not be considered.     

LCA of concrete and steel building frames

The effects on the external environment of seven concrete and steel building frames representative of present-day building technology in Sweden were analysed using LCA methodology. Objects of the study included frame construction and supplementary materials. Several-storey offices and dwellings were studied. The functional unit was defined as one average m² of floor area during the lifetime of the building. Inventory data were elaborated for concrete and steel production, the building site, service life, demolition and final disposal. Parameters included were raw material use, energy use, emissions to air, emissions to water and waste generation. The inventory results were presented and evaluated as such, in addition to an interpretation by using three quantitative impact assessment methods. Parameters that weighed heavily were use of fossil fuels, CO₂, electricity, SO_x ² NO_x ² alloy materials and waste, depending on what assessment method was used. Over the life cycle, building production from cradle to gate accounted for about the same contribution to total environmental loads as maintenance and replacement of heat losses through external walls during service life, whereas demolition and final disposal accounted for a considerably lower contribution.     

Patterns in species associations in plant communities: the importance of scale

Abstract. Present discussions on competitive interactions and the occurrence of predictable patterns in species composition – including assembly rules – are likely to benefit from appropriate analyses of the spatial structure in plant communities. We suggest such an analysis when we specifically want to detect scale regions where fine-scale local processes may affect the spatial pattern of species composition. We combine indirect ordination in the form of Detrended Correspondence Analysis (DCA) and geostatistics in the form of variography. The species abundance data in the sampled quadrats are summarized as positions on the axes in the ordination. Each axis is used as a regionalized variable in the variography to obtain the spatial dependence of the quadrats. The spatial pattern found will suggest the relevant scale region in which to perform an analysis of species associations. A significant spatial dependence (the ‘range’ in geostatistical jargon) will define the size of a sampling plot that will minimize both the problem of being too small and thus having the risk of oversampling of e.g. clonal individuals and of being too large which will risk including individuals that do not interact. We also suggest that plots are spaced at least a ‘range’ apart to insure spatial and statistical independence. Comparisons of species compositions in such plots will reveal any positive or negative associations between species on a scale where these should reflect species-species interactions. To illustrate the method it is applied to three different data sets from two different plant communities.     

Effects of fine‐scale genetic structure on male mating success in gynodioecious Beta vulgaris ssp. maritima

Plant mating systems are known to influence population genetic structure because pollen and seed dispersal are often spatially restricted. However, the reciprocal outcomes of population structure on the dynamics of polymorphic mating systems have received little attention. In gynodioecious sea beet (Beta vulgaris ssp. maritima), three sexual types co‐occur: females carrying a cytoplasmic male sterility (CMS) gene, hermaphrodites carrying a non‐CMS cytoplasm and restored hermaphrodites that carry CMS genes and nuclear restorer alleles. This study investigated the effects of fine‐scale genetic structure on male reproductive success of the two hermaphroditic forms. Our study population was strongly structured and characterized by contrasting local sex‐ratios. Pollen flow was constrained over short distances and depended on local plant density. Interestingly, restored hermaphrodites sired significantly more seedlings than non‐CMS hermaphrodites, despite the previous observation that the former produce pollen of lower quality than the latter. This result was explained by the higher frequency of females in the local vicinity of restored (CMS) hermaphrodites as compared to non‐CMS hermaphrodites. Population structure thus strongly influences individual fitness and may locally counteract the expected effects of selection, suggesting that understanding fine scale population processes is central to predicting the evolution of gender polymorphism in angiosperms.