Competition and dispersal in the pea aphid: clonal variation and correlations across traits

Abstract.  1. The presence of an across-species trade-off between dispersal ability and competitive ability has been proposed as a mechanism that facilitates coexistence. It is not clear if a similar trade-off exists within species. Such a trade-off would constrain the evolution of either trait and, given appropriate selection pressures, promote local adaptation in these traits.
2. This study found substantial levels of heritable variation in competitive ability of the pea aphid, Acyrthosiphon pisum Harris (Homoptera: Aphididae), measured in terms of relative survival when reared with a single clone of the vetch aphid, Megoura viciae Buckton (Homoptera: Aphididae).
3. Pea aphids can move to new patches by either flying (longer distance dispersal) or walking (local dispersal) from plant to plant. There was considerable clonal variation in dispersal ability, measured in terms of the proportion of winged offspring produced, and ability to survive away from their host plant.
4. Winged individuals showed longer off-plant survival times than wingless forms of the same pea aphid clone.
5. There was no evidence of a relationship between clonal competitive ability and either measure of dispersal ability, although the power of the test is limited by the number of pea aphid clones used in the trial.
6. However, there was a positive correlation between clonal fecundity and the proportion of winged offspring produced. Although speculative, it is suggested that clones that are more likely to either overwhelm their host plant or attract higher numbers of natural enemies as a result of having higher fecundity are more likely to produce winged morphs.     

Model selection and information theory in geographical ecology

Aim   Although parameter estimates are not as affected by spatial autocorrelation as Type I errors, the change from classical null hypothesis significance testing to model selection under an information theoretic approach does not completely avoid problems caused by spatial autocorrelation. Here we briefly review the model selection approach based on the Akaike information criterion (AIC) and present a new routine for Spatial Analysis in Macroecology (SAM) software that helps establishing minimum adequate models in the presence of spatial autocorrelation.
Innovation    We illustrate how a model selection approach based on the AIC can be used in geographical data by modelling patterns of mammal species in South America represented in a grid system ( n  = 383) with 2° of resolution, as a function of five environmental explanatory variables, performing an exhaustive search of minimum adequate models considering three regression methods: non-spatial ordinary least squares (OLS), spatial eigenvector mapping and the autoregressive (lagged-response) model. The models selected by spatial methods included a smaller number of explanatory variables than the one selected by OLS, and minimum adequate models contain different explanatory variables, although model averaging revealed a similar rank of explanatory variables.
Main conclusions    We stress that the AIC is sensitive to the presence of spatial autocorrelation, generating unstable and overfitted minimum adequate models to describe macroecological data based on non-spatial OLS regression. Alternative regression techniques provided different minimum adequate models and have different uncertainty levels. Despite this, the averaged model based on Akaike weights generates consistent and robust results across different methods and may be the best approach for understanding of macroecological patterns.     

In vitro three-dimensional modelling of human ovarian surface epithelial cells

Objectives:  Ninety percent of malignant ovarian cancers are epithelial and thought to arise from the ovarian surface epithelium (OSE). We hypothesized that biological characteristics of primary OSE cells would more closely resemble OSE in vivo if established as three-dimensional (3D) cultures.
Materials and methods:  OSE cells were cultured as multicellular spheroids (MCS) (i) in a rotary cell culture system (RCCS) and (ii) on polyHEMA-coated plastics. The MCSs were examined by electron microscopy and compared to OSE from primary tissues and cells grown in 2D. Annexin V FACS analysis was used to evaluate apoptosis and expression of extracellular matrix (ECM) proteins was analysed by immunohistochemical staining.
Results:  On polyHEMA-coated plates, OSE spheroids had defined internal architecture. RCCS MCSs had disorganized structure and higher proportion of apoptotic cells than polyHEMA MCSs and the same cells grown in 2D culture. In 2D, widespread expression of AE1/AE3, laminin and vimentin were undetectable by immunohistochemistry, whereas strong expression of these proteins was observed in the same cells grown in 3D culture and in OSE on primary tissues.
Conclusions:  Physiological and biological features of OSE cells grown in 3D culture more closely resemble characteristics of OSE cells in vivo than when grown by classical 2D approaches. It is likely that establishing in vitro 3D OSE models will lead to greater understanding of the mechanisms of neoplastic transformation in epithelial ovarian cancers.     

Impact of oxygen environment on mesenchymal stem cell expansion and chondrogenic differentiation

Introduction:  In vitro expansion and differentiation of mesenchymal stem cells (MSC) rely on specific environmental conditions, and investigations have demonstrated that one crucial factor is oxygen environment.
Objectives:  In order to understand the impact of oxygen tension on MSC culture and chondrogenic differentiation in vitro , we developed a mathematical model of these processes and applied it in predicting optimal assays.
Methods and results:  We compared ovine MSCs under physiologically low and atmospheric oxygen tension. Low oxygen tension improved their in vitro population growth as demonstrated by monoclonal expansion and colony forming assays. Moreover, it accelerated induction of the chondrogenic phenotype in subsequent three-dimensional differentiation cultures. We introduced a hybrid stochastic multiscale model of MSC organization in vitro . The model assumes that cell adaptation to non-physiological high oxygen tension reversibly changes the structure of MSC populations with respect to differentiation. In simulation series, we demonstrated that these changes profoundly affect chondrogenic potential of the populations. Our mathematical model provides a consistent explanation of our experimental findings.
Conclusions:  Our approach provides new insights into organization of MSC populations in vitro. The results suggest that MSC differentiation is largely reversible and that lineage plasticity is restricted to stem cells and early progenitors. The model predicts a significant impact of short-term low oxygen treatment on MSC differentiation and optimal chondrogenic differentiation at 10–11% pO₂.     

Colonic stem cell data are consistent with the immortal model of stem cell division under non-random strand segregation

Objectives:  Colonic stem cells are thought to reside towards the base of crypts of the colon, but their numbers and proliferation mechanisms are not well characterized. A defining property of stem cells is that they are able to divide asymmetrically, but it is not known whether they always divide asymmetrically (immortal model) or whether there are occasional symmetrical divisions (stochastic model). By measuring diversity of methylation patterns in colon crypt samples, a recent study found evidence in favour of the stochastic model, assuming random segregation of stem cell DNA strands during cell division. Here, the effect of preferential segregation of the template strand is considered to be consistent with the 'immortal strand hypothesis', and explore the effect on conclusions of previously published results.
Materials and methods:  For a sample of crypts, it is shown how, under the immortal model, to calculate mean and variance of the number of unique methylation patterns allowing for non-random strand segregation and compare them with those observed.
Results:  The calculated mean and variance are consistent with an immortal model that incorporates non-random strand segregation for a range of stem cell numbers and levels of preferential strand segregation.
Conclusions:  Allowing for preferential strand segregation considerably alters previously published conclusions relating to stem cell numbers and turnover mechanisms. Evidence in favour of the stochastic model may not be as strong as previously thought.     

Biased competition between Lgr5 intestinal stem cells driven by oncogenic mutation induces clonal expansion

The concept of ‘field cancerization’ describes the clonal expansion of genetically altered, but morphologically normal cells that predisposes a tissue to cancer development. Here, we demonstrate that biased stem cell competition in the mouse small intestine can initiate the expansion of such clones. We quantitatively analyze how the activation of oncogenic K-ras in individual Lgr5⁺ stem cells accelerates their cell division rate and creates a biased drift towards crypt clonality. K-ras mutant crypts then clonally expand within the epithelium through enhanced crypt fission, which distributes the existing Paneth cell niche over the two new crypts. Thus, an unequal competition between wild-type and mutant intestinal stem cells initiates a biased drift that leads to the clonal expansion of crypts carrying oncogenic mutations.     

Functional expression of human HMG-CoA reductase in Saccharomyces cerevisiae: a system to analyse normal and mutated versions of the enzyme in the context of statin treatment

Aims:  Statins – inhibitors of the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase – are known to reduce blood cholesterol levels. In this paper, we present a Saccharomyces cerevisiae expression system, which enables quick evaluation of the sensitivity of the wild-type and/or mutant forms of human HMG-CoA reductase towards statins or other drugs.
Methods and results:  We analysed the sequence of the HMG-CoA reductase gene in DNA extracted from blood samples of 16 patients with cardiovascular disorders. We applied the yeast system to examine the sensitivity of the wild-type and mutated versions of the hHMG-CoA reductase to different types of statins.
Conclusion:  The yeast and mammalian HMG-CoA reductases demonstrate structural and functional conservation, and expression of human HMG-CoA reductase in yeast complements the lethal phenotype of strains lacking the HMG1 and HMG2 genes.
Significance and Impact of the Study:  These data indicate that a yeast expression system can serve to study the influence of selected mutations in human HMG-CoA reductase on the sensitivity of the enzyme to commonly prescribed statins. Our results suggest that this model system is suitable for the development and selection of lipid-lowering drugs as well as for the examination of DNA sequence variations in the context of statin therapy.     

High potential of adhesion to abiotic and biotic materials in fish aquaculture facility by Vibrio alginolyticus strains

Aims:  The ability of Vibrio alginolyticus strains isolated from Sparus aurata and Dicentrarchus labrax nursery to adhere to epithelial cell lines (Hep-2 and Caco-2), fish mucus and their ability to form a biofilm on different surfaces (glass, polystyrene, polyethylene and polyvinyl-chloride) was investigated in this study.
Methods and Results:  The extracellular products were rich in enzymes and the strains were haemolytic on Wagatsuma agar and possessed several hydrolytic exoenzymes such as proteases, DNase and lipases. Most strains tested were multiresistant to the 17 antibiotics tested including those used in the farm to treat vibriosis.
Conclusions:  These bacteria were able to form a biofilm on all the surfaces tested and the cell density was the highest on the PVC surface followed by that on the glass slides, polystyrene and the polyethylene surface. More than 50% of the tested strains were adhesive to the epithelial cell lines (Hep-2 and Caco-2).
Significance and Impact of the Study:  These properties allow these bacteria to survive, proliferate and persist in all stages of fish rearing nursery even after seawater treatment with UV light.     

Obstructor-A is required for epithelial extracellular matrix dynamics, exoskeleton function, and tubulogenesis

The epidermis and internal tubular organs, such as gut and lungs, are exposed to a hostile environment. They form an extracellular matrix to provide epithelial integrity and to prevent contact with pathogens and toxins. In arthropods, the cuticle protects, shapes, and enables the functioning of organs. During development, cuticle matrix is shielded from premature degradation; however, underlying molecular mechanisms are poorly understood. Previously, we identified the conserved obstructor multigene-family, which encodes chitin-binding proteins. Here we show that Obstructor-A is required for extracellular matrix dynamics in cuticle forming organs. Loss of obstructor-A causes severe defects during cuticle molting, wound protection, tube expansion and larval growth control. We found that Obstructor-A interacts and forms a core complex with the polysaccharide chitin, the cuticle modifier Knickkopf and the chitin deacetylase Serpentine. Knickkopf protects chitin from chitinase-dependent degradation and deacetylase enzymes ensure extracellular matrix maturation. We provide evidence that Obstructor-A is required to control the presence of Knickkopf and Serpentine in the extracellular matrix. We propose a model suggesting that Obstructor-A coordinates the core complex for extracellular matrix protection from premature degradation. This mechanism enables exoskeletal molting, tube expansion, and epithelial integrity. The evolutionary conservation suggests a common role of Obstructor-A and homologs in coordinating extracellular matrix protection in epithelial tissues of chitinous invertebrates.     

Hedgehog signaling controls homeostasis of adult intestinal smooth muscle

The Hedgehog (Hh) pathway plays multiple patterning roles during development of the mammalian gastrointestinal tract, but its role in adult gut function has not been extensively examined. Here we show that chronic reduction in the combined epithelial Indian (Ihh) and Sonic (Shh) hedgehog signal leads to mislocalization of intestinal subepithelial myofibroblasts, loss of smooth muscle in villus cores and muscularis mucosa as well as crypt hyperplasia. In contrast, chronic over-expression of Ihh in the intestinal epithelium leads to progressive expansion of villus smooth muscle, but does not result in reduced epithelial proliferation. Together, these mouse models show that smooth muscle populations in the adult intestinal lamina propria are highly sensitive to the level of Hh ligand. We demonstrate further that Hh ligand drives smooth muscle differentiation in primary intestinal mesenchyme cultures and that cell-autonomous Hh signal transduction in C3H10T1/2 cells activates the smooth muscle master regulator Myocardin (Myocd) and induces smooth muscle differentiation. The rapid kinetics of Myocd activation by Hh ligands as well as the presence of an unusual concentration of Gli sties in this gene suggest that regulation of Myocd by Hh might be direct. Thus, these data indicate that Hh is a critical regulator of adult intestinal smooth muscle homeostasis and suggest an important link between Hh signaling and Myocd activation. Moreover, the data support the idea that lowered Hh signals promote crypt expansion and increased epithelial cell proliferation, but indicate that chronically increased Hh ligand levels do not dampen crypt proliferation as previously proposed.     

Comparative demographic impacts of `info-chemicals' and exploitative competition: an empirical test using Daphnia magna

1. Crowding `info-chemicals', metabolites released into the environment that act as signals to conspecifics as well as other organisms, have often been shown to cause phenotypic plasticity in cladoceran life-history traits including morphology, reproductive strategy and sex induction. Effects on population demography and diversity, however, have not been examined directly.
2. Replicate laboratory populations of the zooplankter, Daphnia magna, were started with 250 juveniles and allowed to develop for 8 weeks in 10 L flow-through aquaria with food levels of 1 mg CL^–1. Two crowded water treatments from different clonal sources were compared with untreated water as a control.
3. The presence of crowding chemicals affected initial population structure, through reduction in parthenogenetic female body lengths, brood sizes and juvenile densities. However, the timing of population density maxima and subsequent population structure, clonal composition and diversity were similar among all treatments.
4. Clonal richness remained slightly higher in untreated populations because of increased prevalence of rare clones.
5. Exploitation competition had greater impact on population structure than the relative concentration of crowding chemicals, indicating that density dependent effects were mediated primarily by competition for common resources.
6. Crowding info-chemicals may play a greater role in community structure than in single species population structure because of taxonomic diversity in species' response that may be unrelated to the source species density.     

The equilibrium assumption in estimating the parameters of metapopulation models

1.  The construction of a predictive metapopulation model includes three steps: the choice of factors affecting metapopulation dynamics, the choice of model structure, and finally parameter estimation and model testing.
2.  Unless the assumption is made that the metapopulation is at stochastic quasi-equilibrium and unless the method of parameter estimation of model parameters uses that assumption, estimates from a limited amount of data will usually predict a trend in metapopulation size.
3.  This implicit estimation of a trend occurs because extinction-colonization stochasticity, possibly amplified by regional stochasticity, leads to unequal numbers of observed extinction and colonization events during a short study period.
4.  Metapopulation models, such as those based on the logistic regression model, that rely on observed population turnover events in parameter estimation are sensitive to the implicit estimation of a trend.
5.  A new parameter estimation method, based on Monte Carlo inference for statistically implicit models, allows an explicit decision about whether metapopulation quasi-stability is assumed or not.
6. Our confidence in metapopulation model parameter estimates that have been produced from only a few years of data is decreased by the need to know before parameter estimation whether the metapopulation is in quasi-stable state or not.
7. The choice of whether metapopulation stability is assumed or not in parameter estimation should be done consciously. Typical data sets cover only a few years and rarely allow a statistical test of a possible trend. While making the decision about stability one should consider any information about the landscape history and species and metapopulation characteristics.     

Mechanisms involved in the photosensitized inactivation of Acanthamoeba palestinensis trophozoites

Aims:  To advance our understanding of the mechanisms involved in the RLP068 phthalocyanine-photosensitized inactivation of Acanthamoeba palestinensis trophozoites through a precise identification of the targets of the photoprocess in both the cytosolic and mitochondrial compartments.
Methods and Results:  We followed the activities of selected marker enzymes as well as we performed fluorescence and transmission electron microscopy investigations of the alterations induced by the photoprocess in the fine structure of subcellular compartments. RLP068 is preferentially located in the contractile vacuole: the fluorescence in that site is particularly evident in the unirradiated cells and becomes more diffused after irradiation. Electron microscopic analysis of photosensitized A. palestinensis cells clearly shows that the swelling of trophozoites and the appearance of vacuoles spread throughout the cytoplasm after phototreatment. The activity of a typical cytoplasmic enzyme, such as lactate dehydrogenase, underwent a 35% decrease as a consequence of the photoprocess, reflecting the photodamage induced by migrating phthalocyanine molecules in their micro-environment.
Conclusions:  The presence of multiple targets for the phthalocyanine-photosensitized process is of utmost importance because this pattern of cell damage makes it unlikely that photoresistant A. palestinensis strains are gradually selected or mutagenic phenomena are developed as a consequence of the photoinduced damage.
Significance and Impact of the Study:  Photosensitization via phthalocyanines appears to represent an efficient and safe approach for achieving a close control of the population of a potentially pathogenic protozoan such as A. palestinensis , opening new perspectives for the disinfection of microbiologically polluted waters.     

Three clonal types of urothelium with different capacities for replication

Objectives:  Similar to other epithelia, urothelium in vivo has a hierarchal organization of cells each with specific gradients of differentiation. While distinct cell types have been described as important in bladder cancer in vitro , clonal and proliferative capacities of normal urothelial cells have not been characterized.
Materials and methods:  Three cell types and colony types were identified from primary porcine urothelial culture. Proliferative activity, patterns of apoptosis and differentiation, colony forming efficiency and ability to change phenotype with passage were determined and compared.
Results:  Small, T-I colonies with large flattened (type-1) cells had low levels of proliferation and high levels of apoptosis. Large T-III colonies had a central area of small (type-3) cells surrounded by type-1 and type-2 cells. Proliferation and apoptosis were asymmetrically distributed in the periphery of T-II and T-III colonies. T-III colonies proved to be significantly more clonogenic and proliferative. With appropriate induction, type-1 cells were able to proliferate upon passage and form type-3 cells, yet long-term culture demonstrated that progeny of type-1 cells appeared to have inherited a clonogenic handicap.
Conclusions:  Type-3 cells in the centre of T-III colonies appear to harbour stem-like qualities with a relatively low proliferative and apoptotic index at homeostasis and the ability to become highly proliferative upon passage. This study demonstrates that distinct urothelial cell types with differing clonal capacities can be isolated from the bladder and these cells may have implications for tissue engineering and carcinogenesis.     

Continuous growth of the giant grass Zizaniopsis bonariensis in subtropical wetlands

1.  Zizaniopsis bonariensis (giant grass) is an emergent macrophyte species endemic to subtropical wetlands in Brazil, Argentina, Paraguay and Uruguay. In this study, we show the effects of its continuous clonal reproduction and its 'phalanx' growth strategy in the Taim Wetland (southern Brazil).
2. The continuous clonal growth of this 3-m high grass gave rise to the emergence of 11.7 new shoots m⁻² per month and a high total above ground production (2870 g dry weight m⁻² year⁻¹). The biomass of the new shoots emerging every month formed a wave of growth, moderated by only weak seasonal variation.
3. We show its phalanx growth strategy by analysing the variations in population density and shoot height within a transect through the stand canopy. The inverse relation between density and height from the border to the interior indicates self-regulation of biomass.
4. The plants modified their environment, enhancing resistance to drought within the stand and thus facilitating their dominance. This positive feedback suggests that the dominance of the plant might constitute an alternative state in subtropical wetlands.