Validation of membrane protein topology models by oxidative labeling and mass spectrometry

Computer-assisted topology predictions are widely used to build low-resolution structural models of integral membrane proteins (IMPs). Experimental validation of these models by traditional methods is labor intensive and requires modifications that might alter the IMP native conformation. This work employs oxidative labeling coupled with mass spectrometry (MS) as a validation tool for computer-generated topology models. ·OH exposure introduces oxidative modifications in solvent-accessible regions, whereas buried segments (e.g., transmembrane helices) are non-oxidizable. The Escherichia coli protein WaaL (O-antigen ligase) is predicted to have 12 transmembrane helices and a large extramembrane domain (Pérez et al., Mol. Microbiol. 2008, 70, 1424). Tryptic digestion and LC-MS/MS were used to map the oxidative labeling behavior of WaaL. Met and Cys exhibit high intrinsic reactivities with ·OH, making them sensitive probes for solvent accessibility assays. Overall, the oxidation pattern of these residues is consistent with the originally proposed WaaL topology. One residue (M151), however, undergoes partial oxidation despite being predicted to reside within a transmembrane helix. Using an improved computer algorithm, a slightly modified topology model was generated that places M151 closer to the membrane interface. On the basis of the labeling data, it is concluded that the refined model more accurately reflects the actual topology of WaaL. We propose that the combination of oxidative labeling and MS represents a useful strategy for assessing the accuracy of IMP topology predictions, supplementing data obtained in traditional biochemical assays. In the future, it might be possible to incorporate oxidative labeling data directly as constraints in topology prediction algorithms.

BRITE: Universal Topology Generation from a User's Perspective

Effective engineering of the Internet is predicated upon a detailed understanding of issues such as the large-scale structure of its underlying physical topology, the manner in which it evolves over time, and the way in which its constituent components contribute to its overall function. Unfortunately, developing a deep understanding of these issues has proven to be a challenging task, since it in turn involves solving difficult problems such as mapping the actual topology, characterizing it, and developing models that capture its emergent behavior. Consequently, even though there are a number of topology models, it is an open question as to how representative the topologies they generate are of the actual Internet. Our goal is to produce a topology generation framework which improves the state of the art and is based on design principles which include representativeness, inclusiveness, and interoperability. Representativeness leads to synthetic topologies that accurately reflect many aspects of the actual Internet topology (e.g. hierarchical structure, degree distribution, etc.). Inclusiveness combines the strengths of as many generation models as possible in a single generation tool. Interoperability provides interfaces to widely-used simulation and visualization applications such as ns and SSF. We call such a tool a universal topology generator. In this paper we discuss the design, implementation and usage of the BRITE universal topology generation tool that we have built. We also describe the BRITE Analysis Engine, BRIANA, which is an independent piece of software designed and built upon BRITE design goals of flexibility and extensibility. The purpose of BRIANA is to act as a repository of analysis routines along with a user–friendly interface that allows its use on different topology formats.

A Framework for Realistic and Systematic Multicast Performance Evaluation

Previous multicast research often makes commonly accepted but unverifed assumptions on network topologies and group member distribution in simulation studies. In this paper, we propose a framework to systematically evaluate multicast performance for different protocols. We identify a series of metrics, and carry out extensive simulation studies on these metrics with different topological models and group member distributions for three case studies. Our simulation results indicate that realistic topology and group membership models are crucial to accurate multicast performance evaluation. These results can provide guidance for multicast researchers to perform realistic simulations, and facilitate the design and development of multicast protocols.

TOPSIG : Topology Preserving Document Signatures

Performance comparisons between File Signatures and Inverted Files for text retrieval have previously shown several significant shortcomings of file signatures relative to inverted files. The inverted file approach underpins most state-of-the-art search engine algorithms, such as Language and Probabilistic models. It has been widely accepted that traditional file signatures are inferior alternatives to inverted files. This paper describes TopSig, a new approach to the construction of file signatures. Many advances in semantic hashing and dimensionality reduction have been made in recent times, but these were not so far linked to general purpose, signature file based, search engines. This paper introduces a different signature file approach that builds upon and extends these recent advances. We are able to demonstrate significant improvements in the performance of signature file based indexing and retrieval, performance that is comparable to that of state of the art inverted file based systems, including Language models and BM25. These findings suggest that file signatures offer a viable alternative to inverted files in suitable settings and positions the file signatures model in the class of Vector Space retrieval models.

Oblique-Shock-Wave/Boundary-Layer Interaction Using Near-Wall Reynolds-Stress Models

A synthesis is presented of the predictive capability of a family of near-wall wall-normal free Reynolds stress models (which are completely independent of wall topology, i.e., of the distance fromthe wall and the normal-to-thewall orientation) for oblique-shock-wave/turbulent-boundary-layer interactions. For the purpose of comparison, results are also presented using a standard low turbulence Reynolds number k–ε closure and a Reynolds stress model that uses geometric wall normals and wall distances. Studied shock-wave Mach numbers are in the range MSW = 2.85–2.9 and incoming boundary-layer-thickness Reynolds numbers are in the range Reδ0 = 1–2×106. Computations were carefully checked for grid convergence. Comparison with measurements shows satisfactory agreement, improving on results obtained using a k–ε model, and highlights the relative importance of redistribution and diffusion closures, indicating directions for future modeling work.

Maximal structuring of acyclic process models

This article addresses the transformation of a process model with an arbitrary topology into an equivalent structured process model. In particular, this article studies the subclass of process models that have no equivalent well-structured representation but which, nevertheless, can be partially structured into their maximally-structured representation. The transformations are performed under a behavioral equivalence notion that preserves the observed concurrency of tasks in equivalent process models. The article gives a full characterization of the subclass of acyclic process models that have no equivalent well-structured representation, but do have an equivalent maximally-structured one, as well as proposes a complete structuring method. Together with our previous results, this article completes the solution of the process model structuring problem for the class of acyclic process models.

Structuring acyclic process models

This article studies the problem of transforming a process model with an arbitrary topology into an equivalent well-structured process model. While this problem has received significant attention, there is still no full characterization of the class of unstructured process models that can be transformed into well-structured ones, nor an automated method for structuring any process model that belongs to this class. This article fills this gap in the context of acyclic process models. The article defines a necessary and sufficient condition for an unstructured acyclic process model to have an equivalent well-structured process model under fully concurrent bisimulation, as well as a complete structuring method. The method has been implemented as a tool that takes process models captured in the BPMN and EPC notations as input. The article also reports on an empirical evaluation of the structuring method using a repository of process models from commercial practice.

Structuring acyclic process models

This paper addresses the problem of transforming a process model with an arbitrary topology into an equivalent well-structured process model. While this problem has received significant attention, there is still no full characterization of the class of unstructured process models that can be transformed into well-structured ones, nor an automated method to structure any process model that belongs to this class. This paper fills this gap in the context of acyclic process models. The paper defines a necessary and sufficient condition for an unstructured process model to have an equivalent structured model under fully concurrent bisimulation, as well as a complete structuring method.

Impact of mobility and topology on information diffusion in MANETs

In some delay-tolerant communication systems such as vehicular ad-hoc networks, information flow can be represented as an infectious process, where each entity having already received the information will try to share it with its neighbours. The random walk and random waypoint models are popular analysis tools for these epidemic broadcasts, and represent two types of random mobility. In this paper, we introduce a simulation framework investigating the impact of a gradual increase of bias in path selection (i.e. reduction of randomness), when moving from the former to the latter. Randomness in path selection can significantly alter the system performances, in both regular and irregular network structures. The implications of these results for real systems are discussed in details.

Optimization models for expanding a railway’s theoretical capacity

Changing the topology of a railway network can greatly affect its capacity. Railway networks however can be altered in a multitude of different ways. As each way has significant immediate and long term financial ramifications, it is a difficult task to decide how and where to expand the network. In response some railway capacity expansion models (RCEM) have been developed to help capacity planning activities, and to remove physical bottlenecks in the current railway system. The exact purpose of these models is to decide given a fixed budget, where track duplications and track sub divisions should be made, in order to increase theoretical capacity most. These models are high level and strategic, and this is why increases to the theoretical capacity is concentrated upon. The optimization models have been applied to a case study to demonstrate their application and their worth. The case study evidently shows how automated approaches of this nature could be a formidable alternative to current manual planning techniques and simulation. If the exact effect of track duplications and sub-divisions can be sufficiently approximated, this approach will be very applicable.

NUVIEW:software for display and interactive manipulation of nucleic acid models

The NUVIEW software package allows skeletal models of any double helical nucleic acid molecule to be displayed out a graphics monitor and to apply various rotations, translations and scaling transformations interactively, through the keyboard. The skeletal model is generated by connecting any pair of representative points, one from each of the bases in the basepair. In addition to the above mentioned manipulations, the base residues can be identified by using a locator and the distance between any pair of residues can be obtained. A sequence based color coded display allows easy identification of sequence repeats, such as runs of Adenines. The real time interactive manipulation of such skeletal models for large DNA/RNA double helices, can be used to trace the path of the nucleic acid chain in three dimensions and hence get a better idea of its topology, location of linear or curved regions, distances between far off regions in the sequence etc. A physical picture of these features will assist in understanding the relationship between base sequence, structure and biological function in nucleic acids.

Heritability of brain network topology in 853 twins and siblings

Anatomical brain networks change throughout life and with diseases. Genetic analysis of these networks may help identify processes giving rise to heritable brain disorders, but we do not yet know which network measures are promising for genetic analyses. Many factors affect the downstream results, such as the tractography algorithm used to define structural connectivity. We tested nine different tractography algorithms and four normalization methods to compute brain networks for 853 young healthy adults (twins and their siblings). We fitted genetic structural equation models to all nine network measures, after a normalization step to increase network consistency across tractography algorithms. Probabilistic tractography algorithms with global optimization (such as Probtrackx and Hough) yielded higher heritability statistics than 'greedy' algorithms (such as FACT) which process small neighborhoods at each step. Some global network measures (probtrackx-derived GLOB and ST) showed significant genetic effects, making them attractive targets for genome-wide association studies.

Playing Games on Sets and Models

The most prominent objective of the thesis is the development of the generalized descriptive set theory, as we call it. There, we study the space of all functions from a fixed uncountable cardinal to itself, or to a finite set of size two. These correspond to generalized notions of the universal Baire space (functions from natural numbers to themselves with the product topology) and the Cantor space (functions from natural numbers to the {0,1}-set) respectively. We generalize the notion of Borel sets in three different ways and study the corresponding Borel structures with the aims of generalizing classical theorems of descriptive set theory or providing counter examples. In particular we are interested in equivalence relations on these spaces and their Borel reducibility to each other. The last chapter shows, using game-theoretic techniques, that the order of Borel equivalence relations under Borel reduciblity has very high complexity. The techniques in the above described set theoretical side of the thesis include forcing, general topological notions such as meager sets and combinatorial games of infinite length. By coding uncountable models to functions, we are able to apply the understanding of the generalized descriptive set theory to the model theory of uncountable models. The links between the theorems of model theory (including Shelah's classification theory) and the theorems in pure set theory are provided using game theoretic techniques from Ehrenfeucht-Fraïssé games in model theory to cub-games in set theory. The bottom line of the research declairs that the descriptive (set theoretic) complexity of an isomorphism relation of a first-order definable model class goes in synch with the stability theoretical complexity of the corresponding first-order theory. The first chapter of the thesis has slightly different focus and is purely concerned with a certain modification of the well known Ehrenfeucht-Fraïssé games. There we (me and my supervisor Tapani Hyttinen) answer some natural questions about that game mainly concerning determinacy and its relation to the standard EF-game

A Deterministic Optimization Approach to Protein Sequence Design Using Continuous Models

Determining the sequence of amino acid residues in a heteropolymer chain of a protein with a given conformation is a discrete combinatorial problem that is not generally amenable for gradient-based continuous optimization algorithms. In this paper we present a new approach to this problem using continuous models. In this modeling, continuous "state functions" are proposed to designate the type of each residue in the chain. Such a continuous model helps define a continuous sequence space in which a chosen criterion is optimized to find the most appropriate sequence. Searching a continuous sequence space using a deterministic optimization algorithm makes it possible to find the optimal sequences with much less computation than many other approaches. The computational efficiency of this method is further improved by combining it with a graph spectral method, which explicitly takes into account the topology of the desired conformation and also helps make the combined method more robust. The continuous modeling used here appears to have additional advantages in mimicking the folding pathways and in creating the energy landscapes that help find sequences with high stability and kinetic accessibility. To illustrate the new approach, a widely used simplifying assumption is made by considering only two types of residues: hydrophobic (H) and polar (P). Self-avoiding compact lattice models are used to validate the method with known results in the literature and data that can be practically obtained by exhaustive enumeration on a desktop computer. We also present examples of sequence design for the HP models of some real proteins, which are solved in less than five minutes on a single-processor desktop computer Some open issues and future extensions are noted.