Biological invasions and the conservation of biodiversity.

Consideration of definitions of 'biological invasion' and 'biodiversity' shows why invasions have recently generated great interest among conservationists. Many studies show that invasion numbers have increased drastically over the last five centuries, that this exponential increase is not levelling off, and that human activities are the only reason for the phenomenon. Many mechanisms are portrayed in an evolutionary framework and their consequences for biodiversity are described at three levels of life--gene, species and ecosystem. Examples from islands show that insular ecosystems are especially prone to damage from invasions; they also serve as 'laboratories' to elucidate the nature of invasion impacts. An important management approach--eradication--is discussed. Eradicating invaders not only aids understanding of their impacts on native species but also in understanding how ecosystems function. In fact, biological invasions can be seen as 'experiments', providing a rare opportunity to help answer certain fundamental scientific questions.

[The neuronal zootype]. / Le zootype neuronal.

We present an hypothesis, derived from the zootype concept of Slack, Holland and Graham. The main point of this hypothesis is to postulate that the primordial function of the zootype genes is to design an appropriate neuronal network in bilaterian animals, by controlling the genes involved in the specificity of the axon pathways. This would be the primary function of the zootype genes in development and their primitive function in evolution. The hypothesis is discussed in view of the current knowledge on the Hox genes, their evolution, their genomic organisation, their expression and their targets.

Early-branching or fast-evolving eukaryotes? An answer based on slowly evolving positions.

The current paradigm of eukaryotic evolution is based primarily on comparative analysis of ribosomal RNA sequences. It shows several early-emerging lineages, mostly amitochondriate, which might be living relics of a progressive assembly of the eukaryotic cell. However, the analysis of slow-evolving positions, carried out with the newly developed slow-fast method, reveals that these lineages are, in terms of nucleotide substitution, fast-evolving ones, misplaced at the base of the tree by a long branch attraction artefact. Since the fast-evolving groups are not always the same, depending on which macromolecule is used as a marker, this explains most of the observed incongruent phylogenies. The current paradigm of eukaryotic evolution thus has to be seriously re-examined as the eukaryotic phylogeny is presently best summarized by a multifurcation. This is consistent with the Big Bang hypothesis that all extant eukaryotic lineages are the result of multiple cladogeneses within a relatively brief period, although insufficiency of data is also a possible explanation for the lack of resolution. For further resolution, rare evolutionary events such as shared insertions and/or deletions or gene fusions might be helpful.

The origin of red algae and the evolution of chloroplasts.

Chloroplast structure and genome analyses support the hypothesis that three groups of organisms originated from the primary photosynthetic endosymbiosis between a cyanobacterium and a eukaryotic host: green plants (green algae + land plants), red algae and glaucophytes (for example, Cyanophora). Although phylogenies based on several mitochondrial genes support a specific green plants/red algae relationship, the phylogenetic analysis of nucleus-encoded genes yields inconclusive, sometimes contradictory results. To address this problem, we have analysed an alternative nuclear marker, elongation factor 2, and included new red algae and protist sequences. Here we provide significant support for a sisterhood of green plants and red algae. This sisterhood is also significantly supported by a multi-gene analysis of a fusion of 13 nuclear markers (5,171 amino acids). In addition, the analysis of an alternative fusion of 6 nuclear markers (1,938 amino acids) indicates that glaucophytes may be the closest relatives to the green plants/red algae group. Thus, our study provides evidence from nuclear markers for a single primary endosymbiosis at the origin of these groups, and supports a kingdom Plantae comprising green plants, red algae and glaucophytes.

Biochemical analysis of the interaction between elongation factor 1alpha and alpha/beta-tubulins from a ciliate, Tetrahymena pyriformis.

The interaction between elongation factor 1alpha (EF-1alpha) and alpha/beta-tubulins has been analyzed in vivo and in vitro. An association of both alpha- and beta-tubulins with EF-1alpha in the lysate of Tetrahymena pyriformis was detected by co-immunoprecipitation analysis. In contrast, in vitro biomolecular interaction analysis with glutathione S-transferase (GST) fusion proteins revealed that GST-beta-tubulin, but not GST-alpha-tubulin, can bind to GST-EF-1alpha. Two beta-tubulin binding sites have been identified to reside in the domains I and III of EF-1alpha. In addition, beta-tubulin itself seems to have two distinct interaction sites for each of the domains. Since domain II of EF-1alpha did not interact with beta-tubulin, we have re-evaluated the phylogenetic status of ciliates using EF-1alpha sequences devoid of domain II. The phylogenetic tree thus obtained was significantly different from that inferred from the whole sequence of EF-1alpha, suggesting the presence of functional constraints on the molecular evolution of EF-1alpha.

Unusually high evolutionary rate of the elongation factor 1 alpha genes from the Ciliophora and its impact on the phylogeny of eukaryotes.

The elongation factor 1 alpha (EF-1 alpha) has become widely employed as a phylogenetic marker for studying eukaryotic evolution. However, a disturbing problem, the artifactual polyphyly of ciliates, is always observed. It has been suggested that the addition of new sequences will help to circumvent this problem. Thus, we have determined 15 new ciliate EF-1 alpha sequences, providing for a more comprehensive taxonomic sampling of this phylum. These sequences have been analyzed together with a representation of eukaryotic sequences using distance-, parsimony-, and likelihood-based phylogenetic methods. Such analyses again failed to recover the monophyly of Ciliophora. A study of the substitution rate showed that ciliate EF-1 alpha genes exhibit a high evolutionary rate, produced in part by an increased number of variable positions. This acceleration could be related to alterations of the accessory functions acquired by this protein, likely to those involving interactions with the cytoskeleton, which is very modified in the Ciliophora. The high evolutionary rate of these sequences leads to an artificial basal emergence of some ciliates in the eukaryotic tree by effecting a long-branch attraction artifact that produces an asymmetric topology for the basal region of the tree. The use of a maximum-likelihood phylogenetic method (which is less sensitive to long-branch attraction) and the addition of sequences to break long branches allow retrieval of more symmetric topologies, which suggests that the asymmetric part of the tree is most likely artifactual. Therefore, the sole reliable part of the tree appears to correspond to the apical symmetric region. These kinds of observations suggest that the general eukaryotic evolution might have consisted of a massive radiation followed by an increase in the evolutionary rates of certain groups that emerge artificially as early branches in the asymmetric base of the tree. Ciliates in the case of the EF-1 alpha genes would offer clear evidence for this hypothesis.

The neuronal zootype. An hypothesis.

We present an hypothesis, derived from the zootype concept of Slack, Holland and Graham. The main point of this hypothesis is to postulate that the primordial function of the zootype genes is to design an appropriate neuronal network in bilaterian animals, by controlling the genes involved in the specificity of the axon pathways. This would be the primary function of the zootype genes in development and their primitive function in evolution. The hypothesis is discussed in view of the current knowledge on the Hox genes, their evolution, their genomic organization, their expression and their targets.

Evidence for loss of mitochondria in Microsporidia from a mitochondrial-type HSP70 in Nosema locustae.

In molecular phylogenies based on ribosomal RNA, three amitochondriate protist lineages, Microsporidia, Metamonada (including diplomonads) and Parabasala (including trichomonads), are the earliest offshoots of the eukaryotic tree. As an explantation for the lack of mitochondria in these organisms, the hypothesis that they have diverged before the mitochondrial endosymbiosis is preferred to the less parsimonious hypothesis of several independent losses of the organelle. Nevertheless, if they had descended from mitochondrion-containing ancestors, it may be possible to find in their nuclear DNA genes that derive from the endosymbiont which gave rise to mitochondria. Based on similar evidence, secondary losses of mitochondria have recently been suggested for Entamoeba histolytica and for Trichomonas vaginalis. In this study, we have isolated a gene encoding a chaperone protein (HSP70, 70 kDa heat shock protein) from the microspordian Nosema locustae. In phylogenetic trees, this HSP70 was located within a group of sequences that in other lineages is targetted to the mitochondrial compartment, itself included in the proteobacterial clade. In addition, the N. locustae protein contained the GDAW(V) motif shared by mitochondrial and proteobacterial sequences, with only one conservative substitution. Moreover, microsporidia, a phylum which was assumed to emerge close to the base of the eukaryotic tree, appears as the sister-group of fungi in the HSP70 phylogeny, in agreement with some ultrastructural characters and phylogenies based on alpha- and beta-tubulins. Loss of mitochondria, now demonstrated for several amitochondriate groups, indicates that the common ancestor of all the extant eukaryotic species could have been a mitochondriate eukaryote.

Presence of a mitochondrial-type 70-kDa heat shock protein in Trichomonas vaginalis suggests a very early mitochondrial endosymbiosis in eukaryotes.

Molecular phylogenetic analyses, based mainly on ribosomal RNA, show that three amitochondriate protist lineages, diplomonads, microsporidia, and trichomonads, emerge consistently at the base of the eukaryotic tree before groups having mitochondria. This suggests that these groups could have diverged before the mitochondrial endosymbiosis. Nevertheless, since all these organisms live in anaerobic environments, the absence of mitochondria might be due to secondary loss, as demonstrated for the later emerging eukaryote Entamoeba histolytica. We have now isolated from Trichomonas vaginalis a gene encoding a chaperone protein (HSP70) that in other lineages is addressed to the mitochondrial compartment. The phylogenetic reconstruction unambiguously located this HSP70 within a large set of mitochondrial sequences, itself a sister-group of alpha-purple bacteria. In addition, the T. vaginalis protein exhibits the GDAWV sequence signature, so far exclusively found in mitochondrial HSP70 and in proteobacterial dnaK. Thus mitochondrial endosymbiosis could have occurred earlier than previously assumed. The trichomonad double membrane-bounded organelles, the hydrogenosomes, could have evolved from mitochondria.

Cortical memory in Paramecium: a theoretical approach to the structural heredity.

Structural heredity is frequently encountered in ciliates. In particular, during division of Paramecium the cortical characteristics of the 2 daughter cells are inherited from those of the mother cell. Genesis of new structures is therefore only possible if there are pre-existing older structures. We attempted to simulate this phenomenon with a model already used to describe the structure and morphogenesis of the cortex of Paramecium. We show that to initiate spatial organization it is enough to choose special starting conditions which comprise the primer of the structure, even if highly localized and at a low concentration of morphogens. This property does not question the basic concepts of genomic heredity.

How many nucleotides are required to resolve a phylogenetic problem? The use of a new statistical method applicable to available sequences.

The evolution of bootstrap proportions (BP) with sequence length was studied using a 28S ribosomal RNA data set. For different sequence lengths, informative sites were jackknifed several times. Bootstrapping was subsequently performed on each of these subsamples. For each node, BPs so obtained were plotted against sequence length, showing the evolution of the robustness with increasing number of informative sites. For robust nodes (BP of 100%), the pattern of BPs is unvarying and is described by a simple function BP = 100 (1-e-b(x-x')), where x is the number of informative sites and b and x' are two parameters estimated using a nonlinear regression procedure. When a node has a BP < 100% and the pattern of BPs fits this function, it is possible to estimate the number of informative sites required to obtain a given average BP. The method also identifies nonrobust nodes (nonascending clusters of BP dots), for which it seems to be more cost effective and fruitful to turn to other species and/or genes rather than to continue sequencing longer gene lengths from the same species to reach a BP of 95%.

Species sampling has a major impact on phylogenetic inference.

Representative properties of gnathostome species of a rich 28S rRNA data base were studied through the analysis of the fluctuations they provoked in bootstrap proportions (BPs) of nodes of parsimonious trees. Using original programs which permit BP comparison between different trees, it is empirically demonstrated that 4- to 24-species-trees are highly sensitive to species sampling: the inferences obtained from subsets of 4, 8, 16, or 24 species are not congruent with the whole set of 31 species. Study of trees obtained from exhaustively sampling all combinations of single species taken from each presumed monophyletic group shows precisely the impact of each species on the BP of each node. This procedure also shows that the impact of species changes within a given group on tree BPs is localized to its two or three neighboring nodes. The observation of differing impacts of species emphasizes the importance of sampling several species per presumed monophyletic group. It is also concluded that it is necessary to sample several successive outgroups and that the impact of a species on BPs depends mainly on the sampling context. Before undertaking extensive sequencing, the impact of species should be more often considered, since its effect on BPs is stronger than previously thought.

A scaffold for basal body patterning revealed by a monoclonal antibody in the hypotrich ciliate Paraurostyla weissei.

In the hypotrich ciliate Paraurostyla weissei, the infraciliature consists of basal bodies on which cilia are anchored together with associated dense material and microtubular rootlets. This is renewed at each morphogenesis, which occurs both in the fully differentiated cell and in a transient dedifferentiated stage, the zygocyst. In both situations, the cell must reconstitute its typical ciliature by properly patterning its basal bodies. Insights into these morphogenetic processes were obtained through an immunocytochemical study using an antipericentriolar material antibody displaying remarkable properties. In Paraurostyla, this antibody decorates the electron-dense material associated with the basal bodies in the interphase cell (as it does with centrosomes on metazoan cells) but, during morphogenesis, the antibody decorates a transient system of tracks which appear prior to basal body patterning and along which basal bodies will later be distributed. In all cases, tracks are initiated close to parental organelles and then elongate to form a system linking the anlagen together during their migration. During zygocyst morphogenesis, they extend along three main cellular meridians. As this antibody decorates an antigen associated with early steps of morphogenesis, it visualizes a proteinaceous system upstream to basal body patterning and provides a structural continuity between parental and newly assembled basal body systems.

[Hip arthroplasty and thromboembolic complications]. / Arthroplastie de hanche et complications thrombo-emboliques.

Orthopedic départment of Centre hospitalo-universitaire de Brest use one upon another two procedures: from 1974 to 1984, for 1287 cases the prevention of thromboembolic complications is done with the help of subcutaneous heparin at standard dose during 12 days; the clinical diagnosis in confirmed by an isotopic phlebography and isotopic lung scan. In 1986 and 1987, for 391 cases this prevention is done with the help of subcutaneous heparin in adapted doses during 21 days; the clinical diagnosis is confirmed by X ray phlebography and isotopic lung scan. The frequency of thromboembolic complications has been 4.6% in the first period and 1.2% in the second period. This results confirm the role of heparin in preventing post operative thromboembolic complications after total hip replacement. The results after knee arthroplasty are not so good.

MPF and cyclin: modelling of the cell cycle minimum oscillator.

The cell cycle appears to be controlled by the interplay between two protein complexes, MPF and cyclin. Their interactions play an essential role in the structure of the oscillator governing the cell cycle. There seems to be no general agreement on this latter crucial question. Two different mechanisms are proposed: (i) cyclin and p34 kinase combine to form an oligomer with MPF activity; (ii) cyclin enzymatically activates the passage from inactive pre-MPF to active MPF, with the postulate that MPF initiates cyclin degradation. We have modelled these two hypotheses to see whether both actually lead to oscillatory behaviour. The p34-cyclin oligomerization does so without any difficulty. With the second mechanism, however, the strict hypothesis that cyclin degradation is activated by MPF must be re-examined: the system only oscillates if, in disappearing, the MPF and the cyclin react with each other stoichiometrically. The model also demonstrates that it is useless to seek cyclic control of cyclin proteolysis.