Structural and functional analysis of p84N5

Normal development and tissue homeostasis requires the carefully orchestrated balance between cell proliferation and cell death. Cell cycle checkpoints control the extent of cell proliferation. Cell death is coordinated through the activation of a cell suicide pathway that results in the morphologically recognizable form of death, apoptosis. Tumorigenesis requires that the balance between these two pathways be disrupted. The tumor suppressor protein Rb has not only been shown to be involved in the enforcement of cell cycle checkpoints, but has also been implicated in playing a role in the regulation of apoptosis. The manner in which Rb enforces cell cycle checkpoints has been well studied; however, its involvement in the regulation of apoptosis is still very unclear. p84N5 is a novel nuclear death domain containing protein that has been shown to interact with the N-terminus of Rb. The fact that it contains a death domain and the fact that it is nuclear localized possibly provides the first known mechanism for apoptotic signaling from the nucleus. The following study tested the hypothesis that the novel exclusively nuclear death domain containing protein p84N5 is an important mediator of programmed cell death and that its apoptotic function is reliant upon its nuclear localization and is regulated by unique functional domains within the p84N5 protein. We identified the p84N5 nuclear localization signal (NLS), eliminated it, and tested the functional significance of nuclear localization by using wild type and mutant sequences fused to EGFP-C1 (Clontech) to create wild type GFPN5 and subsequent mutants. The results of these assays demonstrated exclusive nuclear localization of GFPN5 is required for normal p84N5 induced apoptosis. We further conducted large-scale mutagenesis of the GFPN5 construct to identify a minimal region within p84N5 capable of interacting with Rb. We were able to identify a minimal sequence containing p84N5 amino acids 318 to 464 that was capable of interacting with Rb in co-immunoprecipitation assays. We continued by conducting a structural and functional analysis to identify the region or regions within p84N5 responsible for inducing apoptosis. Point mutations and small-scale deletions within the death domain of p84N5 lessened the effect but did not eliminate p84N5-induced cytotoxicity. Further analysis revealed that the minimal sequence of 318 to 464 of p84N5 was capable of inducing apoptosis to a similar degree as wild-type GFPN5 protein. Since amino acids 318 to 464 of p84N5 are capable of inducing apoptosis and interacting with Rb, we propose possible mechanisms whereby p84N5 may function in a Rb regulated manner. These results demonstrate that p84N5 induced apoptosis is reliant upon its nuclear localization and is regulated by unique functional domains within the p84N5 protein. ^

Heregulin -beta 1 directly associates with RhoA via the immunoglobulin -like domain thereby modulating breast cancer metastasis-related properties

Heregulins constitute a family of growth factors belonging to the epidermal growth factor (EGF) family. Breast cancers that overexpress specific members of the EGF receptor family (EGFR, ErbB2, ErbB3, ErbB4) have increased metastatic potential, and Heregulin-β1 (HRGβ1), a ligand for ErbB3 and ErbB4, has also been shown to induce metastasis-related properties in breast cancer cells in vitro. The secreted form of the HRGβ1 is composed of five distinct structural domains, including the N-terminal domain, an immunoglobulin-like domain (IgG-like), a glycosylation domain, an EGF-like domain, and a β1-specific domain. Of these, the EGF-like domain is well characterized for its function in metastasis-related properties as well as its structure. However, the contributions of the other HRGβ1 domains in breast cancer metastasis remains unclear. ^ To investigate this, HRGβ1 proteins with targeted domain deletions were purified and subjected to assays for metastasis-related properties, including aggregation, invasion, activation of EGFR family members, and motility of breast cancer cells. These assays showed that retaining the EGF-like domain of HRGβ1 is important for activation of EGFRs. Interestingly, the HRGβ1 protein lacking the IgG-like domain (NGEB) led to a decrease in breast cancer cell motility, indicating the IgG-like domain modulates cell motility, an important step in cancer metastasis. ^ To understand the underlying mechanisms, I performed protein sequence and structural analysis of HRGβ1 and identified that the IgG-like domain of HRGβ1 shares sequence homology and three-dimensional structural similarity with the IgG-like domain of TRIO. TRIO is a cytoplasmic protein that directly associates with RhoA, a GTPase involved in cell reorganization and cell motility. Therefore, I hypothesized that HRGβ1 may translocate inside the breast cancer cells through receptor mediated endocytosis and bind to RhoA via its IgG-like domain. I show wild type HRGβ1 but not NGEB binds RhoA in vitro and in vivo, leading to RhoA activation. Inhibition of HRG-β1 internalization via endocytosis disrupted HRGβ1 binding to RhoA. Additionally, breast cancer cell motility induced by HRG-β1 is reduced after treatment with inhibitors to both endocytosis and RhoA function, similar to levels seen with NGEB treatment. ^ Thus, in addition to the well-known role of HRGβ1 as an extracellular stimulator of the EGFR family members, HRGβ1 also functions within the cell as a binding partner and activator of RhoA to modulate cancer cell motility. ^

Paleomagnetic analysis of ODP Site 133-820

Paleomagnetic analysis of sediment samples from Ocean Drilling Program (ODP) Leg 133, Site 820, 10 km from the outer edge of the Great Barrier Reef, is undertaken to investigate the mineral magnetic response to environmental (sea level) changes. Viscous remanent magnetization (VRM) of both multidomain and near-superparamagnetic origin is prevalent and largely obscures the primary remanence, except in isolated high-magnetization zones. The Brunhes/Matuyama boundary cannot be identified, but is expected to be below 120 mbsf. The only evidence that exists for a geomagnetic excursion occurs at about 33 mbsf (-135 k.y.). Only one-half the cores were oriented, and many suffered from internal rotation about the core axis, caused by coring and/or slicing. The decay of magnetic remanence below the surface layer (0-2 mbsf) is attributed to sulfate reduction processes. The magnetic susceptibility (K) record is central for describing and understanding the magnetic properties of the sediments, and their relationship to glacio-eustatic fluctuations in sea level. Three prominent magnetic susceptibility peaks, at about 7, 32, and 64 mbsf, are superimposed on a background of smaller susceptibility oscillations. Fluctuations in susceptibility and remanence in the ôbackgroundö zone are controlled predominantly by variations in the concentration, rather than the composition of ferrimagnetics, with carbonate dilution playing an important role (type-A properties). The sharp susceptibility maxima occur at the start of the marine transgressions following low stands in sea level (high d18O, glacial maxima), and are characterized by a stable single-domain remanence, with a significant contribution from ultra-fine, superparamagnetic grains (type-C properties). During the later marine transgression, the susceptibility gradually returns to low values and the remanence is carried by stable single-domain magnetite (type-B properties). The A, B, and C types of sediment have distinctive ARM/K ratios. Throughout most of the sequence a strong inverse correlation exists between magnetic susceptibility and both CaCO3 and d18O variations. However, in the sharp susceptibility peaks (early transgression), more complex phase relationships are apparent among these parameters. In particular, the K-d18O correlation switches to positive, then reverts to negative during the course of the late transgression, indicating that two distinct mechanisms are responsible for the K-d18O correlation. Lower in the sequence, where sea-level-controlled cycles of upward-coarsening sediments, we find that the initial, mud phase of each cycle has been enriched in high-coercivity magnetic material, which is indicative of more oxic conditions. The main magnetic characteristics of the sediments are thought to reflect sea-level-controlled variations in the sediment source regions and related run-off conditions. Some preliminary evidence is seen that biogenic magnetite may play a significant role in the magnetization of these sediments.

Analysis of the iron oxide assemblages in the ANDRILL AND-1B drill core, Ross Sea, Antarctica

The AND-1B drill core recovered a 13.57 million year Miocene through Pleistocene record from beneath the McMurdo Ice Shelf in Antarctica (77.9°S, 167.1°E). Varying sedimentary facies in the 1285 m core indicate glacial-interglacial cyclicity with the proximity of ice at the site ranging from grounding of ice in 917 m of water to ice free marine conditions. Broader interpretation of climatic conditions of the wider Ross Sea Embayment is deduced from provenance studies. Here we present an analysis of the iron oxide assemblages in the AND-1B core and interpret their variability with respect to wider paleoclimatic conditions. The core is naturally divided into an upper and lower succession by an expanded 170 m thick volcanic interval between 590 and 760 m. Above 590 m the Plio-Pleistocene glacial cycles are diatom rich and below 760 m late Miocene glacial cycles are terrigenous. Electron microscopy and rock magnetic parameters confirm the subdivision with biogenic silica diluting the terrigenous input (fine pseudo-single domain and stable single domain titanomagnetite from the McMurdo Volcanic Group with a variety of textures and compositions) above 590 m. Below 760 m, the Miocene section consists of coarse-grained ilmenite and multidomain magnetite derived from Transantarctic Mountain lithologies. This may reflect ice flow patterns and the absence of McMurdo Volcanic Group volcanic centers or indicate that volcanic centers had not yet grown to a significant size. The combined rock magnetic and electron microscopy signatures of magnetic minerals serve as provenance tracers in both ice proximal and distal sedimentary units, aiding in the study of ice sheet extent and dynamics, and the identification of ice rafted debris sources and dispersal patterns in the Ross Sea sector of Antarctica.

Fossil pollen record of composite sediment core TMD from Tso Moriri, analysis of modern surface pollen samples, mean annual precipitation reconstruction, and digitisation and recalibration of different discussed palaeoclimate proxy records

This paper presents a new fossil pollen record from Tso Moriri (32°54'N, 78°19'E, 4512 m a.s.l.) and seeks to reconstruct changes in mean annual precipitation (MAP) during the last 12,000 years. This high-alpine lake occupies an area of 140 km**2 in a glacial-tectonic valley in the northwestern Himalaya. The region has a cold climate, with a MAP <300 mm, and open vegetation. The hydrology is controlled by the Indian Summer Monsoon (ISM), but winter westerly-associated precipitation also affects the regional water balance. Results indicate that precipitation levels varied significantly during the Holocene. After a rapid increase in MAP, a phase of maximum humidity was reached between ca. 11 to 9.6 cal ka BP, followed by a gradual decline in MAP. This trend parallels the reduction in the Northern Hemisphere summer insolation. Comparison of different palaeoclimate proxy records reveal evidence for a stronger Holocene decrease in precipitation in the northern versus the southern parts of the ISM domain. The long-term trend of ISM weakening is overlaid with several short periods of greater dryness, which are broadly synchronous with the North Atlantic cold spells, suggesting reduced amounts of westerly-associated winter precipitation. Compared to the mid and late Holocene, it appears that westerlies had a greater influence on the western parts of the ISM domain during the early Holocene. During this period, the westerly-associated summer precipitation belt was positioned at Mediterranean latitudes and amplified the ISM-derived precipitation. The Tso Moriri pollen record and moisture reconstructions also suggest that changes in climatic conditions affected the ancient Harappan Civilisation, which flourished in the greater Indus Valley from approximately 5.2 to 3 cal ka BP. The prolonged Holocene trend towards aridity, punctuated by an interval of increased dryness (between ca. 4.5 to 4.3 cal ka BP), may have pushed the Mature Harappan urban settlements (between ca. 4.5 to 3.9 cal ka BP) to develop more efficient agricultural practices to deal with the increasingly acute water shortages. The amplified aridity associated with North Atlantic cooling between ca. 4 to 3.6 and around 3.2 cal ka BP further hindered local agriculture, possibly causing the deurbanisation that occurred from ca. 3.9 cal ka BP and eventual collapse of the Harappan Civilisation between ca. 3.5 to 3 cal ka BP.

Analysis of jitter influence in fast frequency measurements

This paper presents a theoretical analysis of possible jitter impact in application of numeric criterion for fastmeasurement of frequency by coincidence principle. The primary goal is the generation of a signal containing a known amount of each jitter components. This signal was used for testing signals with regular pulse trains. Initially, jitter components are analyzed and modeled individually. Next, sequences for combining different kinds of jitter are modeled, simulated and evaluated. Jitter model simulation in Matlab is utilized to show the independence of frequencymeasurement results on the total jitter present in the reference and desired pulse trains independently. A good agreement between previously introduced theory of fastmeasurement of frequency and simulation in jitter presence is verified; these results allows to engineers use the numeric criterion for fastmeasurement of frequency in spite to interactions among jitter components in various applications for frequency domain sensors.

Introducing non-linear analysis into sustained speech characterization to improve sleep apnea detection

We present a novel approach for detecting severe obstructive sleep apnea (OSA) cases by introducing non-linear analysis into sustained speech characterization. The proposed scheme was designed for providing additional information into our baseline system, built on top of state-of-the-art cepstral domain modeling techniques, aiming to improve accuracy rates. This new information is lightly correlated with our previous MFCC modeling of sustained speech and uncorrelated with the information in our continuous speech modeling scheme. Tests have been performed to evaluate the improvement for our detection task, based on sustained speech as well as combined with a continuous speech classifier, resulting in a 10% relative reduction in classification for the first and a 33% relative reduction for the fused scheme. Results encourage us to consider the existence of non-linear effects on OSA patients' voices, and to think about tools which could be used to improve short-time analysis.

Dynamic Analysis of high Speed Railway Traffic Loads on Ballasted Track

This paper reports the studies carried out to develop and calibrate the optimal models for the objectives of this work. In particular, quarter bogie model for vehicle, rail-wheel contact with Lagrangian multiplier method, 2D spatial discretization were selected as the optimal decisions. Furthermore, the 3D model of coupled vehicle-track also has been developed to contrast the results obtained in the 2D model. The calculations were carried out in the time domain and envelopes of relevant results were obtained for several track profiles and speed ranges. Distributed elevation irregularities were generated based on power spectral density (PSD) distributions. The results obtained include the wheel-rail contact forces, forces transmitted to the bogie by primary suspension. The latter loads are relevant for the purpose of evaluating the performance of the infrastructure

Broadband Channel Long Delay Cluster Measurements and Analysis at 2.4GHz in Subway Tunnels

The delay caused by the reflected ray in broadband communication has a great influence on the communications in subway tunnel. This paper presents measurements taken in subway tunnels at 2.4 GHz, with 5 MHz bandwidth. According to propagation characteristics of tunnel, the measurements were carried out with a frequency domain channel sounding technique, in three typical scenarios: line of sight (LOS), Non-line-of-sight (NLOS) and far line of sight (FLOS), which lead to different delay distributions. Firstly IFFT was chosen to get channel impulse response (CIR) h(t) from measured three-dimensional transfer functions. Power delay profile (PDP) was investigated to give an overview of broadband channel model. Thereafter, a long delay caused by the obturation of tunnel is observed and investigated in all the scenarios. The measurements show that the reflection can be greatly remained by the tunnel, which leads to long delay cluster where the reflection, but direct ray, makes the main contribution for radio wave propagation. Four important parameters: distribution of whole PDP power, first peak arriving time, reflection cluster duration and PDP power distribution of reflection cluster were studied to give a detailed description of long delay characteristic in tunnel. This can be used to ensure high capacity communication in tunnels

Global analysis of constraint logic programs

This article presents and illustrates a practical approach to the dataow analysis of constraint logic programming languages using abstract interpretation. It is rst argued that from the framework point of view it suces to propose relatively simple extensions of traditional analysis methods which have already been proved useful and practical and for exist. This is shown by proposing a simple extension of Bruynooghes traditional framework which allows it to analyze constraint logic programs. Then and using this generalized framework two abstract domains and their required abstract functions are presented the rst abstract domain approximates deniteness information and the second one freeness. Finally an approach for cobining those domains is proposed The two domains and their combination have been implemented and used in the analysis of CLP and Prolog III applications. Results from this implementation showing its performance and accuracy are also presented

Automated attribute inference in complex service workflows based on sharing analysis

The properties of data and activities in business processes can be used to greatly facilítate several relevant tasks performed at design- and run-time, such as fragmentation, compliance checking, or top-down design. Business processes are often described using workflows. We present an approach for mechanically inferring business domain-specific attributes of workflow components (including data Ítems, activities, and elements of sub-workflows), taking as starting point known attributes of workflow inputs and the structure of the workflow. We achieve this by modeling these components as concepts and applying sharing analysis to a Horn clause-based representation of the workflow. The analysis is applicable to workflows featuring complex control and data dependencies, embedded control constructs, such as loops and branches, and embedded component services.

Exploiting goal independence in the analysis of logic programs

This paper illustrates the use of a top-down framework to obtain goal independent analyses of logic programs, a task which is usually associated with the bottom-up approach. While it is well known that the bottomup approach can be used, through the magic set transformation, for goal dependent analysis, it is less known that the top-down approach can be used for goal independent analysis. The paper describes two ways of doing the latter. We show how the results of a goal independent analysis can be used to speed up subsequent goal dependent analyses. However this speed-up may result in a loss of precisión. The influence of domain characteristics on this precisión is discussed and an experimental evaluation using a generic top-down analyzer is described.

Goal dependent vs goal independent analysis of logic programs

Goal independent analysis of logic programs is commonly discussed in the context of the bottom-up approach. However, while the literature is rich in descriptions of top-down analysers and their application, practical experience with bottom-up analysis is still in a preliminary stage. Moreover, the practical use of existing top-down frameworks for goal independent analysis has not been addressed in a practical system. We illustrate the efficient use of existing goal dependent, top-down frameworks for abstract interpretation in performing goal independent analyses of logic programs much the same as those usually derived from bottom-up frameworks. We present several optimizations for this flavour of top-down analysis. The approach is fully implemented within an existing top-down framework. Several implementation tradeoffs are discussed as well as the influence of domain characteristics. An experimental evaluation including a comparison with a bottom-up analysis for the domain Prop is presented. We conclude that the technique can offer advantages with respect to standard goal dependent analyses.

Program analysis, debugging and optimization using the ciao system preprocessor

We present a tutorial overview of Ciaopp, the Ciao system preprocessor. Ciao is a public-domain, next-generation logic programming system, which subsumes ISO-Prolog and is specifically designed to a) be highly extensible via librarles and b) support modular program analysis, debugging, and optimization. The latter tasks are performed in an integrated fashion by Ciaopp. Ciaopp uses modular, incremental abstract interpretation to infer properties of program predicates and literals, including types, variable instantiation properties (including modes), non-failure, determinacy, bounds on computational cost, bounds on sizes of terms in the program, etc. Using such analysis information, Ciaopp can find errors at compile-time in programs and/or perform partial verification. Ciaopp checks how programs cali system librarles and also any assertions present in the program or in other modules used by the program. These assertions are also used to genérate documentation automatically. Ciaopp also uses analysis information to perform program transformations and optimizations such as múltiple abstract specialization, parallelization (including granularity control), and optimization of run-time tests for properties which cannot be checked completely at compile-time. We illustrate "hands-on" the use of Ciaopp in all these tasks. By design, Ciaopp is a generic tool, which can be easily tailored to perform these and other tasks for different LP and CLP dialects.

A generic framework for context-sensitive analysis of modular programs

Context-sensitive analysis provides information which is potentially more accurate than that provided by context-free analysis. Such information can then be applied in order to validate/debug the program and/or to specialize the program obtaining important improvements. Unfortunately, context-sensitive analysis of modular programs poses important theoretical and practical problems. One solution, used in several proposals, is to resort to context-free analysis. Other proposals do address context-sensitive analysis, but are only applicable when the description domain used satisfies rather restrictive properties. In this paper, we argüe that a general framework for context-sensitive analysis of modular programs, Le., one that allows using all the domains which have proved useful in practice in the non-modular setting, is indeed feasible and very useful. Driven by our experience in the design and implementation of analysis and specialization techniques in the context of CiaoPP, the Ciao system preprocessor, in this paper we discuss a number of design goals for context-sensitive analysis of modular programs as well as the problems which arise in trying to meet these goals. We also provide a high-level description of a framework for analysis of modular programs which does substantially meet these objectives. This framework is generic in that it can be instantiated in different ways in order to adapt to different contexts. Finally, the behavior of the different instantiations w.r.t. the design goals that motivate our work is also discussed.