@article{4173, abstract = {Background: Zebrafish (D. rerio) has become a powerful and widely used model system for the analysis of vertebrate embryogenesis and organ development. While genetic methods are readily available in zebrafish, protocols for two dimensional (2D) gel electrophoresis and proteomics have yet to be developed. Results: As a prerequisite to carry out proteomic experiments with early zebrafish embryos, we developed a method to efficiently remove the yolk from large batches of embryos. This method enabled high resolution 2D gel electrophoresis and improved Western blotting considerably. Here, we provide detailed protocols for proteomics in zebrafish from sample preparation to mass spectrometry (MS), including a comparison of databases for MS identification of zebrafish proteins. Conclusion: The provided protocols for proteomic analysis of early embryos enable research to be taken in novel directions in embryogenesis.}, author = {Link, Vinzenz and Shevchenko, Andrej and Heisenberg, Carl-Philipp J}, journal = {BMC Developmental Biology}, pages = {1 -- 9}, publisher = {BioMed Central}, title = {{Proteomics of early zebrafish embryos}}, doi = {10.1186/1471-213X-6-1}, volume = {6}, year = {2006}, } @article{4176, abstract = {During vertebrate gastrulation, a well-orchestrated series of morphogenetic changes leads to the formation of the three germ layers: the ectoderm, mesoderm and endoderm. The analysis of gene expression patterns during gastrulation has been central to the identification of genes involved in germ layer formation. However, many proteins are regulated on a translational or post-translational level and are thus undetectable by gene expression analysis. Therefore, we developed a 2D-gel-based comparative proteomic approach to target proteins involved in germ layer morphogenesis during zebrafish gastrulation. Proteomes of ectodermal and mesendodermal progenitor cells were compared and 35 significantly regulated proteins were identified by mass spectrometry, including several proteins with predicted functions in cytoskeletal organization. A comparison of our proteomic results with data obtained in an accompanying microarray-based gene expression analysis revealed no significant overlap, confirming the complementary nature of proteomics and transcriptomics. The regulation of ezrin2, which was identified based on a reduction in spot intensity in mesendodermal cells, was independently validated. Furthermore, we show that ezrin2 is activated by phosphorylation in mesendodermal cells and is required for proper germ layer morphogenesis. We demonstrate the feasibility of proteomics in zebrafish, concluding that proteomics is a valuable tool for analysis of early development.}, author = {Link, Vinzenz and Carvalho, Lara and Castanon, Irinka and Stockinger, Petra and Shevchenko, Andrej and Heisenberg, Carl-Philipp J}, journal = {Journal of Cell Science}, number = {10}, pages = {2073 -- 2083}, publisher = {Company of Biologists}, title = {{Identification of regulators of germ layer morphogenesis using proteomics in zebrafish}}, doi = {10.1242/jcs.02928}, volume = {119}, year = {2006}, } @article{4178, abstract = {Detailed reconstruction of the spatiotemporal history of embryonic cells is key to understanding tissue formation processes but is often complicated by the large number of cells involved, particularly so in vertebrates. Through a combination of high-resolution time-lapse lineage tracing and antibody staining, we have analyzed the movement of mesencephalic and metencephalic cell populations in the early zebrafish embryo. To facilitate the analysis of our cell tracking data, we have created TracePilot, a software tool that allows interactive manipulation and visualization of tracking data. We demonstrate its utility by showing novel visualizations of cell movement in the developing zebrafish brain. TracePilot (http://www.mpi-cbg.de/tracepilot) is Java-based, available free of charge, and has a program structure that allows the incorporation of additional analysis tools.}, author = {Langenberg, Tobias and Dracz, Tadeusz and Oates, Andrew and Heisenberg, Carl-Philipp J and Brand, Michael}, journal = {Developmental Dynamics}, number = {4}, pages = {928 -- 933}, publisher = {Wiley-Blackwell}, title = {{Analysis and visualization of cell movement in the developing zebrafish brain}}, doi = {10.1002/dvdy.20692}, volume = {235}, year = {2006}, } @article{4184, abstract = {Epithelial morphogenesis depends on coordinated changes in cell shape, a process that is still poorly understood. During zebrafish epiboly and Drosophila dorsal closure, cell-shape changes at the epithelial margin are of critical importance. Here evidence is provided for a conserved mechanism of local actin and myosin 2 recruitment during theses events. It was found that during epiboly of the zebrafish embryo, the movement of the outer epithelium (enveloping layer) over the yolk cell surface involves the constriction of marginal cells. This process depends on the recruitment of actin and myosin 2 within the yolk cytoplasm along the margin of the enveloping layer. Actin and myosin 2 recruitment within the yolk cytoplasm requires the Ste20-like kinase Msn1, an orthologue of Drosophila Misshapen. Similarly, in Drosophila, actin and myosin 2 localization and cell constriction at the margin of the epidermis mediate dorsal closure and are controlled by Misshapen. Thus, this study has characterized a conserved mechanism underlying coordinated cell-shape changes during epithelial morphogenesis.}, author = {Köppen, Mathias and Fernández, Beatriz and Carvalho, Lara and Jacinto, António and Heisenberg, Carl-Philipp J}, journal = {Development}, number = {14}, pages = {2671 -- 2681}, publisher = {Company of Biologists}, title = {{Coordinated cell-shape changes control epithelial movement in zebrafish and Drosophila}}, doi = {doi: 10.1242/dev.02439}, volume = {133}, year = {2006}, } @article{4218, abstract = {The molecular and cellular mechanisms governing cell motility and directed migration in response to the chemokine SDF-1 are largely unknown. Here, we demonstrate that zebrafish primordial germ cells whose migration is guided by SDF-1 generate bleb-like protrusions that are powered by cytoplasmic flow. Protrusions are formed at sites of higher levels of free calcium where activation of myosin contraction occurs. Separation of the acto-myosin cortex from the plasma membrane at these sites is followed by a flow of cytoplasm into the forming bleb. We propose that polarized activation of the receptor CXCR4 leads to a rise in free calcium that in turn activates myosin contraction in the part of the cell responding to higher levels of the ligand SDF-1. The biased formation of new protrusions in a particular region of the cell in response to SDF-1 defines the leading edge and the direction of cell migration.}, author = {Blaser, Heiko and Reichman Fried, Michal and Castanon, Irinka and Dumstrei, Karin and Marlow, Florence and Kawakami, Koichi and Solnica Krezel, Lilianna and Heisenberg, Carl-Philipp J and Raz, Erez}, journal = {Developmental Cell}, number = {5}, pages = {613 -- 627}, publisher = {Cell Press}, title = {{Migration of zebrafish primordial germ cells: A role for myosin contraction and cytoplasmic flow}}, doi = {10.1016/j.devcel.2006.09.023}, volume = {11}, year = {2006}, } @article{4235, abstract = {The accompanying letter by H. Ortega addresses some points with respect to our previous article (de Vladar and González, 2004). He begins addressing some arguments about our interpretations of the parameters that are hand-waving rather than scientific. However, he is mostly concerned with the asymptotic behaviour of the tumour-immunity system when therapy is present. Dr. Ortega repeats calculations of our system where he either (i) reaches the same conclusions as we do or (ii) reaches different but mistaken conclusions. In any case, he intends to pose destructive critiques to our paper without providing a real contribution to the field, much less an enrichment to our model.}, author = {Vladar, Harold and González, J.}, journal = {Journal of Theoretical Biology}, number = {1}, pages = {162--163}, publisher = {Elsevier}, title = {{Dynamic response of cancer under the influence of immunological activity and therapy}}, doi = {10.1016/j.jtbi.2005.11.016}, volume = {240}, year = {2006}, } @article{4237, abstract = {The growth function of populations is central in biomathematics. The main dogma is the existence of density-dependence mechanisms, which can be modelled with distinct functional forms that depend on the size of the Population. One important class of regulatory functions is the theta-logistic, which generalizes the logistic equation. Using this model as a motivation, this paper introduces a simple dynamical reformulation that generalizes many growth functions. The reformulation consists of two equations, one for population size, and one for the growth rate. Furthermore, the model shows that although population is density-dependent, the dynamics of the growth rate does not depend either on population size, nor on the carrying capacity. Actually, the growth equation is uncoupled from the population size equation, and the model has only two parameters, a Malthusian parameter rho and a competition coefficient theta. Distinct sign combinations of these parameters reproduce not only the family of theta-logistics, but also the van Bertalanffy, Gompertz and Potential Growth equations, among other possibilities. It is also shown that, except for two critical points, there is a general size-scaling relation that includes those appearing in the most important allometric theories, including the recently proposed Metabolic Theory of Ecology. With this model, several issues of general interest are discussed such as the growth of animal population, extinctions, cell growth and allometry, and the effect of environment over a population. (c) 2005 Elsevier Ltd. All rights reserved.}, author = {de Vladar, Harold}, journal = {Journal of Theoretical Biology}, number = {2}, pages = {245 -- 256}, publisher = {Elsevier}, title = {{Density-dependence as a size-independent regulatory mechanism}}, doi = {10.1016/j.jtbi.2005.05.014}, volume = {238}, year = {2006}, } @article{4248, abstract = {In finite populations, genetic drift generates interference between selected loci, causing advantageous alleles to be found more often on different chromosomes than on the same chromosome, which reduces the rate of adaptation. This “Hill–Robertson effect” generates indirect selection to increase recombination rates. We present a new method to quantify the strength of this selection. Our model represents a new beneficial allele (A) entering a population as a single copy, while another beneficial allele (B) is sweeping at another locus. A third locus affects the recombination rate between selected loci. Using a branching process model, we calculate the probability distribution of the number of copies of A on the different genetic backgrounds, after it is established but while it is still rare. Then, we use a deterministic model to express the change in frequency of the recombination modifier, due to hitchhiking, as A goes to fixation. We show that this method can give good estimates of selection for recombination. Moreover, it shows that recombination is selected through two different effects: it increases the fixation probability of new alleles, and it accelerates selective sweeps. The relative importance of these two effects depends on the relative times of occurrence of the beneficial alleles.}, author = {Roze, Denis and Nicholas Barton}, journal = {Genetics}, number = {3}, pages = {1793 -- 1811}, publisher = {Genetics Society of America}, title = {{The Hill-Robertson effect and the evolution of recombination}}, doi = {10.1534/genetics.106.058586 }, volume = {173}, year = {2006}, } @misc{4250, abstract = {A recent analysis has shown that divergence between human and chimpanzee varies greatly across the genome. Although this is consistent with ‘hybridisation’ between the diverging human and chimp lineages, such observations can be explained more simply by the null model of allopatric speciation.}, author = {Nicholas Barton}, booktitle = {Current Biology}, number = {16}, pages = {647 -- 650}, publisher = {Cell Press}, title = {{Evolutionary Biology: How did the human species form?}}, doi = {10.1016/j.cub.2006.07.032}, volume = {16}, year = {2006}, } @article{4345, abstract = {Der Artikel beschäftigt sich mit dem Konzept der Bibliothek 2.0 (bzw. Library 2.0). Er skizziert anhand einiger Beispiele die Entwicklung zum Web 2.0 und beschreibt, wie Web 2.0-Technologien und -Anwendungen in Bibliotheken eingesetzt werden. Im Mittelpunkt stehen Social-Tagging-Systeme, benutzerorientierte Erweiterungen von Bibliothekskatalogen und Dokumentenservern sowie der Einsatz von Weblogs an Bibliotheken. Ferner werden neue Anforderungen an Bibliothekare diskutiert.}, author = {Danowski, Patrick and Heller, Lambert}, journal = {Bibliotheksdienst}, number = {11}, pages = {1250 -- 1271}, publisher = {De Gruyter}, title = {{Bibliothek 2.0 - Die Bibliothek der Zukunft?}}, doi = {10.1515/bd.2006.40.11.1259}, volume = {40}, year = {2006}, } @article{4351, abstract = {BACKGROUND: Character mapping on phylogenies has played an important, if not critical role, in our understanding of molecular, morphological, and behavioral evolution. Until very recently we have relied on parsimony to infer character changes. Parsimony has a number of serious limitations that are drawbacks to our understanding. Recent statistical methods have been developed that free us from these limitations enabling us to overcome the problems of parsimony by accommodating uncertainty in evolutionary time, ancestral states, and the phylogeny. RESULTS: SIMMAP has been developed to implement stochastic character mapping that is useful to both molecular evolutionists, systematists, and bioinformaticians. Researchers can address questions about positive selection, patterns of amino acid substitution, character association, and patterns of morphological evolution. CONCLUSION: Stochastic character mapping, as implemented in the SIMMAP software, enables users to address questions that require mapping characters onto phylogenies using a probabilistic approach that does not rely on parsimony. Analyses can be performed using a fully Bayesian approach that is not reliant on considering a single topology, set of substitution model parameters, or reconstruction of ancestral states. Uncertainty in these quantities is accommodated by using MCMC samples from their respective posterior distributions.}, author = {Jonathan Bollback}, journal = {BMC Bioinformatics}, publisher = {BioMed Central}, title = {{SIMMAP: stochastic character mapping of discrete traits on phylogenies}}, doi = {10.1186/1471-2105-7-88}, volume = {7}, year = {2006}, } @article{4352, abstract = {Anopheles darlingi is the primary malaria vector in Latin America, and is especially important in Amazonian Brazil. Historically, control efforts have been focused on indoor house spraying using a variety of insecticides, but since the mid-1990s there has been a shift to patient treatment and focal insecticide fogging. Anopheles darlingi was believed to have been significantly reduced in a gold-mining community, Peixoto de Azevedo (in Mato Grosso State), in the early 1990s by insecticide use during a severe malaria epidemic. In contrast, although An. darlingi was eradicated from some districts of the city of Belem (the capital of Para State) in 1968 to reduce malaria, populations around the water protection area in the eastern district were treated only briefly. To investigate the population structure of An. darlingi including evidence for a population bottleneck in Peixoto, we analyzed eight microsatellite loci of 256 individuals from seven locations in Brazil: three in Amapa State, three in Para State, and one in Mato Grosso State. Allelic diversity and mean expected heterozygosity were high for all populations (mean number alleles/locus and H(E) were 13.5 and 0.834, respectively) and did not differ significantly between locations. Significant heterozygote deficits were associated with linkage disequilibrium, most likely due to either the Wahlund effect or selection. We found no evidence for a population bottleneck in Peixoto, possibly because the reduction was not extreme enough to be detected. Overall estimates of long-term N(e) varied from 92.4 individuals under the linkage disequilibrium model to infinity under the heterozygote excess model. Fixation indices and analysis of molecular variance demonstrated significant differentiation between locations north and south of the Amazon River, suggesting a degree of genetic isolation between them, attributed to isolation by distance.}, author = {Conn, Jan E and Vineis, Joseph H and Jonathan Bollback and Onyabe, David Y and Wilkerson, Richard C and Povoa, Marinete M}, journal = {The American Journal of Tropical Medicine and Hygiene}, number = {5}, pages = {798 -- 806}, publisher = {American Society of Tropical Medicine and Hygiene}, title = {{Population structure of the malaria vector Anopheles darlingi in a malaria-endemic region of eastern Amazonian Brazil}}, volume = {74}, year = {2006}, } @inproceedings{4359, abstract = {We introduce field constraint analysis, a new technique for verifying data structure invariants. A field constraint for a field is a formula specifying a set of objects to which the field can point. Field constraints enable the application of decidable logics to data structures which were originally beyond the scope of these logics, by verifying the backbone of the data structure and then verifying constraints on fields that cross-cut the backbone in arbitrary ways. Previously, such cross-cutting fields could only be verified when they were uniquely determined by the backbone, which significantly limits the range of analyzable data structures. Field constraint analysis permits non-deterministic field constraints on cross-cutting fields, which allows the verificiation of invariants for data structures such as skip lists. Non-deterministic field constraints also enable the verification of invariants between data structures, yielding an expressive generalization of static type declarations. The generality of our field constraints requires new techniques. We present one such technique and prove its soundness. We have implemented this technique as part of a symbolic shape analysis deployed in the context of the Hob system for verifying data structure consistency. Using this implementation we were able to verify data structures that were previously beyond the reach of similar techniques.}, author = {Wies, Thomas and Kuncak, Viktor and Lam, Patrick and Podelski, Andreas and Rinard, Martin}, booktitle = {7th International Conference on Verification, Model Checking, and Abstract Interpretation}, location = {Charleston, SC, United States}, pages = {157 -- 173}, publisher = {Springer}, title = {{Field constraint analysis}}, doi = {10.1007/11609773_11}, volume = {3855}, year = {2006}, } @inproceedings{4373, abstract = {This paper attempts to improve our understanding of timed languages and their relation to timed automata. We start by giving a constructive proof of the folk theorem stating that timed languages specified by the past fragment of mitl, can be accepted by deterministic timed automata. On the other hand we provide a proof that certain languages expressed in the future fragment of mitl are not deterministic, and analyze the reason for this asymmetry.}, author = {Maler, Oded and Nickovic, Dejan and Pnueli, Amir}, booktitle = {Third International Conference on Formal Modeling and Analysis of Timed Systems}, location = {Uppsala, Sweden}, pages = {2 -- 16}, publisher = {Springer}, title = {{Real time temporal logic: Past, present, future}}, doi = {10.1007/11603009_2}, volume = {3829}, year = {2006}, } @inproceedings{4374, abstract = {We show how to transform formulae written in the real-time temporal logic MITL into timed automata that recognize their satisfying models. This compositional construction is much simpler than previously known and can be easily implemented.}, author = {Maler, Oded and Nickovic, Dejan and Pnueli, Amir}, booktitle = {4th International Conference on Formal Modeling and Analysis of Timed Systems}, location = {Paris, France}, pages = {274 -- 289}, publisher = {Springer}, title = {{From MITL to timed automata}}, doi = {10.1007/11867340_20}, volume = {4202}, year = {2006}, } @inproceedings{4401, abstract = {We propose a general framework of secrecy and preservation of secrecy for labeled transition systems. Our definition of secrecy is parameterized by the distinguishing power of the observer, the properties to be kept secret, and the executions of interest, and captures a multitude of definitions in the literature. We define a notion of secrecy preserving refinement between systems by strengthening the classical trace-based refinement so that the implementation leaks a secret only when the specification also leaks it. We show that secrecy is in general not definable in μ-calculus, and thus not expressible in specification logics supported by standard model-checkers. However, we develop a simulation-based proof technique for establishing secrecy preserving refinement. This result shows how existing refinement checkers can be used to show correctness of an implementation with respect to a specification.}, author = {Alur, Rajeev and Cerny, Pavol and Zdancewic, Steve}, booktitle = {33rd International Colloquium on Automata, Languages and Programming}, pages = {107 -- 118}, publisher = {Springer}, title = {{Preserving secrecy under refinement}}, doi = {10.1007/11787006_10}, volume = {4052}, year = {2006}, } @inproceedings{4406, abstract = {We propose and evaluate a new algorithm for checking the universality of nondeterministic finite automata. In contrast to the standard algorithm, which uses the subset construction to explicitly determinize the automaton, we keep the determinization step implicit. Our algorithm computes the least fixed point of a monotone function on the lattice of antichains of state sets. We evaluate the performance of our algorithm experimentally using the random automaton model recently proposed by Tabakov and Vardi. We show that on the difficult instances of this probabilistic model, the antichain algorithm outperforms the standard one by several orders of magnitude. We also show how variations of the antichain method can be used for solving the language-inclusion problem for nondeterministic finite automata, and the emptiness problem for alternating finite automata.}, author = {De Wulf, Martin and Doyen, Laurent and Thomas Henzinger and Raskin, Jean-François}, pages = {17 -- 30}, publisher = {Springer}, title = {{Antichains: A new algorithm for checking universality of finite automata}}, doi = {10.1007/11817963_5}, volume = {4144}, year = {2006}, } @inproceedings{4431, abstract = {We summarize some current trends in embedded systems design and point out some of their characteristics, such as the chasm between analytical and computational models, and the gap between safety-critical and best-effort engineering practices. We call for a coherent scientific foundation for embedded systems design, and we discuss a few key demands on such a foundation: the need for encompassing several manifestations of heterogeneity, and the need for constructivity in design. We believe that the development of a satisfactory Embedded Systems Design Science provides a timely challenge and opportunity for reinvigorating computer science.}, author = {Thomas Henzinger and Sifakis, Joseph}, pages = {1 -- 15}, publisher = {Springer}, title = {{The embedded systems design challenge}}, doi = {10.1007/11813040_1}, volume = {4085}, year = {2006}, } @inproceedings{4432, abstract = {We add freeze quantifiers to the game logic ATL in order to specify real-time objectives for games played on timed structures. We define the semantics of the resulting logic TATL by restricting the players to physically meaningful strategies, which do not prevent time from diverging. We show that TATL can be model checked over timed automaton games. We also specify timed optimization problems for physically meaningful strategies, and we show that for timed automaton games, the optimal answers can be approximated to within any degree of precision.}, author = {Thomas Henzinger and Prabhu, Vinayak S}, pages = {1 -- 17}, publisher = {Springer}, title = {{Timed alternating-time temporal logic}}, doi = {10.1007/11867340_1}, volume = {4202}, year = {2006}, } @inproceedings{4436, abstract = {We present an assume-guarantee interface algebra for real-time components. In our formalism a component implements a set of task sequences that share a resource. A component interface consists of an arrival rate function and a latency for each task sequence, and a capacity function for the shared resource. The interface specifies that the component guarantees certain task latencies depending on assumptions about task arrival rates and allocated resource capacities. Our algebra defines compatibility and refinement relations on interfaces. Interface compatibility can be checked on partial designs, even when some component interfaces are yet unknown. In this case interface composition computes as new assumptions the weakest constraints on the unknown components that are necessary to satisfy the specified guarantees. Interface refinement is defined in a way that ensures that compatible interfaces can be refined and implemented independently. Our algebra thus formalizes an interface-based design methodology that supports both the incremental addition of new components and the independent stepwise refinement of existing components. We demonstrate the flexibility and efficiency of the framework through simulation experiments.}, author = {Thomas Henzinger and Matic, Slobodan}, pages = {253 -- 266}, publisher = {IEEE}, title = {{An interface algebra for real-time components}}, doi = {10.1109/RTAS.2006.11}, year = {2006}, }