Conditional Maximum Likelihood Estimation Following a Group Sequential Test

We consider estimation after a group sequential test. An estimator that is unbiased or has small bias may have substantial conditional bias (Troendle and Yu, 1999, Coburger and Wassmer, 2001). In this paper we derive the conditional maximum likelihood estimators of both the primary parameter and a secondary parameter, and investigate their properties within a conditional inference framework. The method applies to both the usual and adaptive group sequential test designs. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Integrated palaeoecological and historical data in the service of fine-resolution land use and ecological change assessment during the last 1000 years in Rõuge, southern Estonia

Aims Our aim is to reconstruct decadal scale development of historical landscapes during the last 1000 years by means of fossil pollen analysis of annually laminated lake sediments, and detailed historical maps and documents. Location Lake Rõuge Tõugjärv (Estonia), a small lake with annually laminated lake sediments situated in a dense prehistoric setting. Methods The chronology of the palaeodata is based on the annual laminations supported by AMS ¹⁴C and ²¹⁰Pb dating and ¹³⁷Cs, ²⁴¹Am, and spheroidal carbonaceous particle marker horizons. The time‐scale and resolution allows fine sampling (the pollen samples generally comprise 3.5 years) and vegetation change reconstruction. Relevant source area of pollen (RSAP) of the lake was estimated, and the statistical zonation, rate of change, palynological richness, and DCA and PCA ordinations were generated on the basis of the pollen data. The historical calibration data set (maps, numerical information on population, domestic stock, farmland division, etc.) is based on archival material preserved in the Estonian Historical Archives. Results The topmost part (0–180 cm) of the sediment column of Lake Rõuge Tõugjärv, covering the last 1000 years, is visibly laminated carbonaceous gyttja. The varve chronology extends from ad 2000 to ad 1339, with a cumulative ± 9‐year error estimate. Beyond this the chronology is extrapolated using the ¹⁴C date and varve age–depth estimations. The simulation of the RSAP of Lake Tõugjärv shows that the major portion of the pollen loading originating from local vegetation is derived from plants growing within 2000 m of the sampling site. The pollen record divides into five statistically significant subgroups, which fall on the PCA plot into three clusters reflecting the general openness–closedness of the landscape. During the period between ad 1000 and 1200 (RT 1) the Rõuge area was generally wooded with birch, spruce and pine forests. The advancement of extensive farming gradually opened up the landscape between ad 1200 and 1650 (RT 2 and RT 3). The maximum openness of the landscape was reached between ad 1650 and 1875 (RT 4), with the most open period in the late eighteenth century. Historical maps from 1684 and 1870–99 and available quantitative data on population, domestic stock, farmland division, etc. show the same trend. The pollen data covering the last 125 years, and maps from 1935 and 1995, show the reduction of arable land in RSAP of the lake under investigation and the reduction of open land to an extent comparable with the end of the seventeenth century. Main conclusions The formation and development of the cultural landscape at Rõuge over the last 1000 years is characterized by rapid changes in floristic richness and rates of vegetation change attributed to certain historic processes in the RSAP. Five phases of landscape and social development are clearly distinguished during the last 1000 years. The decadal scale vegetation response to human‐induced forcing agrees with historical maps and documents and could be used for past landscapes prior to the period with solid historical data.     

Phyletic coevolution between subterranean rodents of the genus Ctenomys (Rodentia: Hystricognathi) and nematodes of the genus Paraspidodera (Heterakoidea: Aspidoderidae) in the Neotropics: temporal and evolutionary implications

Coevolution was studied in six species of rodents of the genus Ctenomys and their parasitic nematodes of the genus Paraspidodera , collected in Bolivia. Representatives of the families Octodontidae and Caviidae were used as outgroups for the mammals, and nematodes from caviids were used as outgroups of the nematodes from ctenomyids. For the nematodes, quantitative and qualitative morphological characteristics of both males and females and electrophoretic characters of both sexes were used to generate phylogenetic hypotheses of evolutionary relationships of the OTUs occurring in hosts of different species. Concordance estimates of cladograms generated from biochemical—genetic and morphological data of the nematodes show a percentage incongruence (Mickevich-Farris Incongruence Statistic or I _MF) of 8.23% in the character sets. Parsimony mapping, testing concordance of topologies between the trees derived from both analysis of both morphological and biochemical—genetic data indicates an overall agreement of 82.3°. Comparisons of topologies of the host and parasite cladograms, as measured with parsimony mapping, showed 70.8% concordance, indicating substantially more cospeciation than host-switching in the Ctenomys-Paraspidodera host-parasite system. Nematodes of the genus Paraspidodera appear to have invaded the Ctenomys lineage from an origin in caviids sometime before the ctenomyids began to diversify in early Pleistocene time.     

Phylogeny and biogeography of the Vitrinidae (Gastropoda: Stylommatophora)

The phylogeny of the Vitrinidae is reconstructed in a cladistic analysis based on characters of the genitalia, the copulation behaviour and the radula. The genera with an atrial stimulator turned out to be the earliest branches of the Vitrinidae, whereas the genera with a glandula amatoria form a monophyletic, taxonomically apomorphic group. The differences between the proposed phylogeny and previous hypotheses are discussed. The ancestral areas of the Vitrinidae and its sister group, the limacoid slugs Boettgerillidae–Limacidae–Agriolimacidae, are estimated using weighted ancestral area analysis. The Vitrinidae and the limacoid slugs might have originated by a vicariance event between Central Europe and the Near East. © 2002 The Linnean Society of London. Zoological Journal of the Linnean Society , 2002, 134 , 347–358.     

Introducing the Multivariate Dale Model in Population‐Based Genetic Association Studies

Until recently, the most common parametric approaches to study the combined effects of several genetic polymorphisms located within a gene or in a small genomic region are, at the genotype level, logistic regressions and at the haplotype level, haplotype analyses. An alternative modeling approach, based on the case/control principle, is to regard exposures (e.g., genetic data such as derived from Single Nucleotide Polymorphisms – SNPs) as random and disease status as fixed and to use a marginal multivariate model that accounts for inter‐relationships between exposures. One such model is the multivariate Dale model. This model is based on multiple logistic regressions. That is why the model, applied in a case/control setting, leads to straightforward interpretations that are similar to those drawn in a classical logistic modeling framework. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Ecology and biogeography of plant communities associated with the post Plio-Pleistocene relict Rhododendron ponticum subsp. baeticum in southern Spain

Aims Rhododendron ponticum L. is reputed to be a post Plio‐Pleistocene relict plant species with a disjunct distribution that comprises the Iberian Peninsula to the west and the Euxinian region plus some restricted Mediterranean areas to the east. We analysed the ecological range (of subsp. baeticum) in the western area (Aljibe Mountains, north of the Strait of Gibraltar) to understand the factors determining the present area limitation. Location Sierra del Aljibe, north of the Strait of Gibraltar (Iberian Peninsula). Methods We selected 20 riparian sites where R. ponticum is common, and compiled data on the ecological diversity of associated woody species and ferns. We established a 500‐m main transect in each site, along the stream or river course, in which we placed five 20‐m‐long plots at regular intervals. We recorded physiographic habitat features, woody plants and fern abundance, and the number of R. ponticum individuals. Results Rhododendron ponticum in southern Spain is restricted to riparian forests in acidic soils (pH 4.0–6.4), and is mainly found on the banks of inclined and enclosed streams. In our inventory we recorded 59 woody taxa and 12 ferns, with R. ponticum being the dominant species of the understorey (mean abundance 78.6%). The communities are characterized by a high incidence of the humid warm temperate element, both in number of species (18.8 ± 3.7 per site) and abundance; meanwhile, the presence of the modern Mediterranean element (mean number of species 3.4 ± 3.8 per site) appears to be favoured by disturbance. These ecological–historical groups of taxa also show distinct patterns of typological habit, frequency of endemism, infrageneric diversity and geographical range. Populations of R. ponticum are characterized by a very variable density of seedlings in many sites, and the virtual lack of juveniles. Main conclusions Riparian forests of the Aljibe Mountains constitute a refuge for R. ponticum where the species persists, but populations appear to be in decline. The narrow ecological range of R. ponticum in the area strongly contrasts with its wide amplitude in the eastern natural area, mainly the Euxinian region, where R. ponticum probably finds better conditions due to the environmental heterogeneity of the region, and the lack of a hot dry season.     

Gene assignment by means of somatic cell hybrids: a new method for the analysis of data

Summary A statistical approach to the interpretation of data from gene assignment with somatic cell hybrids is presented. The observed data are analysed under a variety of hypotheses. The fit to the hypotheses is compared by means of the likelihood obtained under a given hypothesis. Two of these hypotheses are related to fundamental questions: is a gene responsible for the enzyme observation and if so, is that gene located on a specific chromosome or could it change its position and be sometimes on chromosome j and, in another hybrid line, on chromosome k? The other hypotheses concern the assignment of the gene to just one of the chromosomes.To improve the traditional data analysis approach we considered additional information: the uncertainties and possible errors of laboratory methods in all our calculations and the length of the donor chromosomes in connection with one specific hypothesis.This method allows us to account for the reliability of the investigation methods and the nature of the hybrid lines involved. Data can be evaluated at different error probabilities within a realistic range in order to compare and discuss results.     

Effects of phorbol esters on endothelial cell microfilaments: Laser scanning confocal microscopy and quantitative morphometry of dose dependent changes

Phorbol esters are known to alter microfilaments but it is not clear if the changes correspond to modulation of the phosphoinositide turnover/protein kinase C system. The novel technique of laser scanning confocal epifluorescence was used to study fiber orientation in phorbol ester treated cells. We treated endothelial cells with control agents and agents known to stimulate protein kinase C: 4 alpha-phorbol, phorbol 12-myristate 13-acetate (PMA), phorbol dibutyrate (PDB), or lipopolysaccharide. After incubation with the test agents, the endothelial cell microfilaments were stained with rhodamine pholloidin and viewed by conventional epifluorescence and by laser scanning confocal epifluorescence microscopy. The images obtained by the confocal microscopy corresponded to a thin optical section through the cells, 300 nm or more in thickness. The microfilaments extended predominantly in the plane of focus. After exposure of the cells to phorbol esters, the stress fibers became more nearly parallel in arrangement or were shortened, but remained in the plane of focus. The modification of microfilaments in response to phorbol esters was quantitated by a single blind analysis. In order to compare the morphological changes with a biochemical action of the phorbol esters, we measured phosphoinositide turnover. The dose-dependence of morphological changes was compared and contrasted to the dose-dependent effect of phorbol esters on bradykinin-stimulated phosphoinositide turnover. PMA had about the same EC50 (1-5 nM) for both biochemical and morphological processes. PDB was less potent in inducing the disruption of microfilament structure than in inhibiting phosphoinositide turnover. Lipopolysaccharide was ineffective in inducing a morphological change under these conditions. A simple activation of protein kinase C is insufficient to explain the dose-dependent effects of phorbol esters. Thus a morphometric analysis can help distinguish the potency of cytoskeleton modulators.     

Molecular evolution among someDrosophila species groups as indicated by two-dimensional electrophoresis

Summary The evolutionary and phylogenetic relationships of sevenDrosophila species groups (represented byD. melanogaster, D. mulleri, D. mercatorum, D. robusta, D. virilis, D. immigrans, D. funebris, andD. melanica) were investigated by the use of two-dimensional electrophoresis. The resulting phylogeny is congruent with the current views of evolution among these groups based on morphological characters and immunological distances. Previous studies indicated that the ability of one-dimensional electrophoresis to resolve relationships between distantly related taxa extended to about the Miocene [25 million years (Myr) ago], but the present study demonstrates that two-dimensional electrophoresis is a useful indicator of phylogeny even back to the Paleocene (65 Myr ago). In addition, two-dimensional electrophoresis is shown to be a useful technique for detecting slowly evolving structural proteins such as actins and tropomyosins.     

Sexual dimorphism in the early Miocene species ofProconsul from the Kisingiri Formation of east Africa: A morphometric examination using multivariate statistics

This paper examines the pattern(s) of sexual dimorphism within the upper dentition ofProconsul specimens from the early Miocene of east Africa. These fossils are compared against the corresponding dentition ofPan troglodytes andGorilla gorilla using principal components and cluster analyses. This paper demonstrates that both sexes ofPan andGorilla are characterized by their own distinctive shape patterns. It is also demonstrated that someProconsul specimens examined here display a pattern that is dissimilar from otherProconsul specimens also examined. This suggests that at least two species ofProconsul may have to be recognized as having lived in this region during the early Miocene. The identification of distinct patterns withinProconsul also suggests that their overall shape and size range are more similar toPan than toGorilla.