Relaxin stimulates leukocyte adhesion and migration through a relaxin receptor LGR7-dependent mechanism

Leukocytes are critical effectors of inflammation and tumor biology. Chemokine-like factors produced by such inflammatory sites are key mediators of tumor growth that activate leukocytic recruitment and tumor infiltration and suppress immune surveillance. Here we report that the endocrine peptide hormone, relaxin, is a regulator of leukocyte biology with properties important in recruitment to sites of inflammation. This study uses the human monocytic cell line THP-1 and normal human peripheral blood mononuclear cells to define a novel role for relaxin in regulation of leukocyte adhesion and migration. Our studies indicate that relaxin promotes adenylate cyclase activation, substrate adhesion, and migratory capacity of mononuclear leukocytes through a relaxin receptor LGR7-dependent mechanism. Relaxin-stimulated cAMP accumulation was observed to occur primarily in non-adherent cells. Relaxin stimulation results in increased substrate adhesion and increased migratory activity of leukocytes. In addition, relaxin-stimulated substrate adhesion resulted in enhanced chemotaxis to monocyte chemoattractant protein-1. These responses in THP-1 and peripheral blood mononuclear cells are relaxin dose-dependent and proportional to cAMP accumulation. We further demonstrate that LGR7 is critical for mediating these biological responses by use of RNA interference lentiviral short hairpin constructs. In summary, we provide evidence that relaxin is a novel leukocyte stimulatory agent with properties affecting adhesion and chemomigration

The ontological deficiencies of process modeling in practice

Business process modeling is widely regarded as one of the most popular forms of conceptual modeling. However, little is known about the capabilities and deficiencies of process modeling grammars and how existing deficiencies impact actual process modeling practice. This paper is a first contribution towards a theory-driven, exploratory empirical investigation of the ontological deficiencies of process modeling with the industry standard Business Process Modeling Notation (BPMN). We perform an analysis of BPMN using a theory of ontological expressiveness. Through a series of semi-structured interviews with BPMN adopters we explore empirically the actual use of this grammar. Nine ontological deficiencies related to the practice of modeling with BPMN are identified, for example, the capture of business rules and the specification of process decompositions. We also uncover five contextual factors that impact on the use of process modeling grammars, such as tool support and modeling conventions. We discuss implications for research and practice, highlighting the need for consideration of representational issues and contextual factors in decisions relating to BPMN adoption in organizations.

Modeling, stability analysis and control of microgrid

With the increase in the level of global warming, renewable energy based distributed generators (DGs) will increasingly play a dominant role in electricity production. Distributed generation based on solar energy (photovoltaic and solar thermal), wind, biomass, mini-hydro along with use of fuel cells and micro turbines will gain considerable momentum in the near future. A microgrid consists of clusters of load and distributed generators that operate as a single controllable system. The interconnection of the DG to the utility/grid through power electronic converters has raised concern about safe operation and protection of the equipments. Many innovative control techniques have been used for enhancing the stability of microgrid as for proper load sharing. The most common method is the use of droop characteristics for decentralized load sharing. Parallel converters have been controlled to deliver desired real power (and reactive power) to the system. Local signals are used as feedback to control converters, since in a real system, the distance between the converters may make the inter-communication impractical. The real and reactive power sharing can be achieved by controlling two independent quantities, frequency and fundamental voltage magnitude. In this thesis, an angle droop controller is proposed to share power amongst converter interfaced DGs in a microgrid. As the angle of the output voltage can be changed instantaneously in a voltage source converter (VSC), controlling the angle to control the real power is always beneficial for quick attainment of steady state. Thus in converter based DGs, load sharing can be performed by drooping the converter output voltage magnitude and its angle instead of frequency. The angle control results in much lesser frequency variation compared to that with frequency droop. An enhanced frequency droop controller is proposed for better dynamic response and smooth transition between grid connected and islanded modes of operation. A modular controller structure with modified control loop is proposed for better load sharing between the parallel connected converters in a distributed generation system. Moreover, a method for smooth transition between grid connected and islanded modes is proposed. Power quality enhanced operation of a microgrid in presence of unbalanced and non-linear loads is also addressed in which the DGs act as compensators. The compensator can perform load balancing, harmonic compensation and reactive power control while supplying real power to the grid A frequency and voltage isolation technique between microgrid and utility is proposed by using a back-to-back converter. As utility and microgrid are totally isolated, the voltage or frequency fluctuations in the utility side do not affect the microgrid loads and vice versa. Another advantage of this scheme is that a bidirectional regulated power flow can be achieved by the back-to-back converter structure. For accurate load sharing, the droop gains have to be high, which has the potential of making the system unstable. Therefore the choice of droop gains is often a tradeoff between power sharing and stability. To improve this situation, a supplementary droop controller is proposed. A small signal model of the system is developed, based on which the parameters of the supplementary controller are designed. Two methods are proposed for load sharing in an autonomous microgrid in rural network with high R/X ratio lines. The first method proposes power sharing without any communication between the DGs. The feedback quantities and the gain matrixes are transformed with a transformation matrix based on the line R/X ratio. The second method involves minimal communication among the DGs. The converter output voltage angle reference is modified based on the active and reactive power flow in the line connected at point of common coupling (PCC). It is shown that a more economical and proper power sharing solution is possible with the web based communication of the power flow quantities. All the proposed methods are verified through PSCAD simulations. The converters are modeled with IGBT switches and anti parallel diodes with associated snubber circuits. All the rotating machines are modeled in detail including their dynamics.

Statistical modeling to determine resonant frequency information present in combustion chamber pressure signals

A statistical modeling method to accurately determine combustion chamber resonance is proposed and demonstrated. This method utilises Markov-chain Monte Carlo (MCMC) through the use of the Metropolis-Hastings (MH) algorithm to yield a probability density function for the combustion chamber frequency and find the best estimate of the resonant frequency, along with uncertainty. The accurate determination of combustion chamber resonance is then used to investigate various engine phenomena, with appropriate uncertainty, for a range of engine cycles. It is shown that, when operating on various ethanol/diesel fuel combinations, a 20% substitution yields the least amount of inter-cycle variability, in relation to combustion chamber resonance.

Flow modeling in a novel non-perfusion conical bioreactor

We have developed a bioreactor vessel design which has the advantages of simplicity and ease of assembly and disassembly, and with the appropriately determined flow rate, even allows for a scaffold to be suspended freely regardless of its weight. This article reports our experimental and numerical investigations to evaluate the performance of a newly developed non-perfusion conical bioreactor by visualizing the flow through scaffolds with 45° and 90° fiber lay down patterns. The experiments were conducted at the Reynolds numbers (Re) 121, 170, and 218 based on the local velocity and width of scaffolds. The flow fields were captured using short-time exposures of 60 µm particles suspended in the bioreactor and illuminated using a thin laser sheet. The effects of scaffold fiber lay down pattern and Reynolds number were obtained and correspondingly compared to results obtained from a computational fluid dynamics (CFD) software package. The objectives of this article are twofold: to investigate the hypothesis that there may be an insufficient exchange of medium within the interior of the scaffold when using our non-perfusion bioreactor, and second, to compare the flows within and around scaffolds of 45° and 90° fiber lay down patterns. Scaffold porosity was also found to influence flow patterns. It was therefore shown that fluidic transport could be achieved within scaffolds with our bioreactor design, being a non-perfusion vessel. Fluid velocities were generally same of the same or one order lower in magnitude as compared to the inlet flow velocity. Additionally, the 90° fiber lay down pattern scaffold was found to allow for slightly higher fluid velocities within, as compared to the 45° fiber lay down pattern scaffold. This was due to the architecture and pore arrangement of the 90° fiber lay down pattern scaffold, which allows for fluid to flow directly through (channel-like flow).

Modeling crash outcome probabilities at rural intersections: Application of hierarchical binomial logistic models

It is important to examine the nature of the relationships between roadway, environmental, and traffic factors and motor vehicle crashes, with the aim to improve the collective understanding of causal mechanisms involved in crashes and to better predict their occurrence. Statistical models of motor vehicle crashes are one path of inquiry often used to gain these initial insights. Recent efforts have focused on the estimation of negative binomial and Poisson regression models (and related deviants) due to their relatively good fit to crash data. Of course analysts constantly seek methods that offer greater consistency with the data generating mechanism (motor vehicle crashes in this case), provide better statistical fit, and provide insight into data structure that was previously unavailable. One such opportunity exists with some types of crash data, in particular crash-level data that are collected across roadway segments, intersections, etc. It is argued in this paper that some crash data possess hierarchical structure that has not routinely been exploited. This paper describes the application of binomial multilevel models of crash types using 548 motor vehicle crashes collected from 91 two-lane rural intersections in the state of Georgia. Crash prediction models are estimated for angle, rear-end, and sideswipe (both same direction and opposite direction) crashes. The contributions of the paper are the realization of hierarchical data structure and the application of a theoretically appealing and suitable analysis approach for multilevel data, yielding insights into intersection-related crashes by crash type.

Governors highway safety associations and transportation planning: Exploratory factor analysis and structural equation modeling

The Intermodal Surface Transportation Efficiency Act (ISTEA) of 1991 mandated the consideration of safety in the regional transportation planning process. As part of National Cooperative Highway Research Program Project 8-44, "Incorporating Safety into the Transportation Planning Process," we conducted a telephone survey to assess safety-related activities and expertise at Governors Highway Safety Associations (GHSAs), and GHSA relationships with metropolitan planning organizations (MPOs) and state departments of transportation (DOTs). The survey results were combined with statewide crash data to enable exploratory modeling of the relationship between GHSA policies and programs and statewide safety. The modeling objective was to illuminate current hurdles to ISTEA implementation, so that appropriate institutional, analytical, and personnel improvements can be made. The study revealed that coordination of transportation safety across DOTs, MPOs, GHSAs, and departments of public safety is generally beneficial to the implementation of safety. In addition, better coordination is characterized by more positive and constructive attitudes toward incorporating safety into planning.

Modeling crash types: New insights into the effects of covariates on crashes at rural intersections

Many studies focused on the development of crash prediction models have resulted in aggregate crash prediction models to quantify the safety effects of geometric, traffic, and environmental factors on the expected number of total, fatal, injury, and/or property damage crashes at specific locations. Crash prediction models focused on predicting different crash types, however, have rarely been developed. Crash type models are useful for at least three reasons. The first is motivated by the need to identify sites that are high risk with respect to specific crash types but that may not be revealed through crash totals. Second, countermeasures are likely to affect only a subset of all crashes—usually called target crashes—and so examination of crash types will lead to improved ability to identify effective countermeasures. Finally, there is a priori reason to believe that different crash types (e.g., rear-end, angle, etc.) are associated with road geometry, the environment, and traffic variables in different ways and as a result justify the estimation of individual predictive models. The objectives of this paper are to (1) demonstrate that different crash types are associated to predictor variables in different ways (as theorized) and (2) show that estimation of crash type models may lead to greater insights regarding crash occurrence and countermeasure effectiveness. This paper first describes the estimation results of crash prediction models for angle, head-on, rear-end, sideswipe (same direction and opposite direction), and pedestrian-involved crash types. Serving as a basis for comparison, a crash prediction model is estimated for total crashes. Based on 837 motor vehicle crashes collected on two-lane rural intersections in the state of Georgia, six prediction models are estimated resulting in two Poisson (P) models and four NB (NB) models. The analysis reveals that factors such as the annual average daily traffic, the presence of turning lanes, and the number of driveways have a positive association with each type of crash, whereas median widths and the presence of lighting are negatively associated. For the best fitting models covariates are related to crash types in different ways, suggesting that crash types are associated with different precrash conditions and that modeling total crash frequency may not be helpful for identifying specific countermeasures.

Computer simulation and modeling in railway applications

An electrified railway system includes complex interconnections and interactions of several subsystems. Computer simulation is the only viable means for system evaluation and analysis. This paper discusses the difficulties and requirements of effective simulation models for this specialized industrial application; and the development of a general-purpose multi-train simulator.

Petri net modeling of spatial and temporal constraints in track allocation problem at railway stations

The track allocation problem (TAP) at a multi-track, multi-platform mainline railway station is defined by the station track layout and service timetable, which implies combinations of spatial and temporal conflicts. Feasible solutions are available from either traditional planning or advanced intelligent searching methods and their evaluations with respect to operational requirements are essential for the operators. To facilitate thorough analysis, a timed Coloured Petri Nets (CPN) model is presented here to encapsulate the inter-relationships of the spatial and temporal constraints in the TAP.

Modeling the stepped potential discharge of primary alkaline battery cathodes

A novel model for the potentiostatic discharge of primary alkaline battery cathodes is presented. The model is used to simulate discharges resulting from the stepped potential electrochemical spectroscopy (SPECS) of primary alkaline battery cathodes cathodes, and the results are validated with experimental data. We show that a model based on a single (or mean) reaction framework can be used to simulate multi-reaction discharge behaviour and we develop a consistent functional modification to the kinetic equation of the model that allows for this to occur. The model is used to investigate the effects that the initial exchange current density, i00, and the diffusion coefficient for protons in electrolytic manganese dioxide (EMD), DH+, have on SPECS discharge. The behaviour observed is consistent with the idea that individual reduction reactions, within the multi-reaction, reduction behaviour of EMD, have distinct i00 and DH+ values.

Multi-criteria ranking and receptor modelling of airborne fine particles at three sites in the Pearl River Delta region of China

The multi-criteria decision making methods, Preference METHods for Enrichment Evaluation (PROMETHEE) and Graphical Analysis for Interactive Assistance (GAIA), and the two-way Positive Matrix Factorization (PMF) receptor model were applied to airborne fine particle compositional data collected at three sites in Hong Kong during two monitoring campaigns held from November 2000 to October 2001 and November 2004 to October 2005. PROMETHEE/GAIA indicated that the three sites were worse during the later monitoring campaign, and that the order of the air quality at the sites during each campaign was: rural site > urban site > roadside site. The PMF analysis on the other hand, identified 6 common sources at all of the sites (diesel vehicle, fresh sea salt, secondary sulphate, soil, aged sea salt and oil combustion) which accounted for approximately 68.8 ± 8.7% of the fine particle mass at the sites. In addition, road dust, gasoline vehicle, biomass burning, secondary nitrate, and metal processing were identified at some of the sites. Secondary sulphate was found to be the highest contributor to the fine particle mass at the rural and urban sites with vehicle emission as a high contributor to the roadside site. The PMF results are broadly similar to those obtained in a previous analysis by PCA/APCS. However, the PMF analysis resolved more factors at each site than the PCA/APCS. In addition, the study demonstrated that combined results from multi-criteria decision making analysis and receptor modelling can provide more detailed information that can be used to formulate the scientific basis for mitigating air pollution in the region.

The ghrelin receptor isoforms (GHS-R1a and GHS-R1b) and GPR39 : an investigation into receptor dimerisation

Prostate cancer is the second most common cause of cancer-related deaths in Western males. Current diagnostic, prognostic and treatment approaches are not ideal and advanced metastatic prostate cancer is incurable. There is an urgent need for improved adjunctive therapies and markers for this disease. GPCRs are likely to play a significant role in the initiation and progression of prostate cancer. Over the last decade, it has emerged that G protein coupled receptors (GPCRs) are likely to function as homodimers and heterodimers. Heterodimerisation between GPCRs can result in the formation of novel pharmacological receptors with altered functional outcomes, and a number of GPCR heterodimers have been implicated in the pathogenesis of human disease. Importantly, novel GPCR heterodimers represent potential new targets for the development of more specific therapeutic drugs. Ghrelin is a 28 amino acid peptide hormone which has a unique n-octanoic acid post-translational modification. Ghrelin has a number of important physiological roles, including roles in appetite regulation and the stimulation of growth hormone release. The ghrelin receptor is the growth hormone secretagogue receptor type 1a, GHS-R1a, a seven transmembrane domain GPCR, and GHS-R1b is a C-terminally truncated isoform of the ghrelin receptor, consisting of five transmembrane domains. Growing evidence suggests that ghrelin and the ghrelin receptor isoforms, GHS-R1a and GHS-R1b, may have a role in the progression of a number of cancers, including prostate cancer. Previous studies by our research group have shown that the truncated ghrelin receptor isoform, GHS-R1b, is not expressed in normal prostate, however, it is expressed in prostate cancer. The altered expression of this truncated isoform may reflect a difference between a normal and cancerous state. A number of mutant GPCRs have been shown to regulate the function of their corresponding wild-type receptors. Therefore, we investigated the potential role of interactions between GHS-R1a and GHS-R1b, which are co-expressed in prostate cancer and aimed to investigate the function of this potentially new pharmacological receptor. In 2005, obestatin, a 23 amino acid C-terminally amidated peptide derived from preproghrelin was identified and was described as opposing the stimulating effects of ghrelin on appetite and food intake. GPR39, an orphan GPCR which is closely related to the ghrelin receptor, was identified as the endogenous receptor for obestatin. Recently, however, the ability of obestatin to oppose the effects of ghrelin on appetite and food intake has been questioned, and furthermore, it appears that GPR39 may in fact not be the obestatin receptor. The role of GPR39 in the prostate is of interest, however, as it is a zinc receptor. Zinc has a unique role in the biology of the prostate, where it is normally accumulated at high levels, and zinc accumulation is altered in the development of prostate malignancy. Ghrelin and zinc have important roles in prostate cancer and dimerisation of their receptors may have novel roles in malignant prostate cells. The aim of the current study, therefore, was to demonstrate the formation of GHS-R1a/GHS-R1b and GHS-R1a/GPR39 heterodimers and to investigate potential functions of these heterodimers in prostate cancer cell lines. To demonstrate dimerisation we first employed a classical co-immunoprecipitation technique. Using cells co-overexpressing FLAG- and Myc- tagged GHS-R1a, GHS-R1b and GPR39, we were able to co-immunoprecipitate these receptors. Significantly, however, the receptors formed high molecular weight aggregates. A number of questions have been raised over the propensity of GPCRs to aggregate during co-immunoprecipitation as a result of their hydrophobic nature and this may be misinterpreted as receptor dimerisation. As we observed significant receptor aggregation in this study, we used additional methods to confirm the specificity of these putative GPCR interactions. We used two different resonance energy transfer (RET) methods; bioluminescence resonance energy transfer (BRET) and fluorescence resonance energy transfer (FRET), to investigate interactions between the ghrelin receptor isoforms and GPR39. RET is the transfer of energy from a donor fluorophore to an acceptor fluorophore when they are in close proximity, and RET methods are, therefore, applicable to the observation of specific protein-protein interactions. Extensive studies using the second generation bioluminescence resonance energy transfer (BRET2) technology were performed, however, a number of technical limitations were observed. The substrate used during BRET2 studies, coelenterazine 400a, has a low quantum yield and rapid signal decay. This study highlighted the requirement for the expression of donor and acceptor tagged receptors at high levels so that a BRET ratio can be determined. After performing a number of BRET2 experimental controls, our BRET2 data did not fit the predicted results for a specific interaction between these receptors. The interactions that we observed may in fact represent ‘bystander BRET’ resulting from high levels of expression, forcing the donor and acceptor into close proximity. Our FRET studies employed two different FRET techniques, acceptor photobleaching FRET and sensitised emission FRET measured by flow cytometry. We were unable to observe any significant FRET, or FRET values that were likely to result from specific receptor dimerisation between GHS-R1a, GHS-R1b and GPR39. While we were unable to conclusively demonstrate direct dimerisation between GHS-R1a, GHS-R1b and GPR39 using several methods, our findings do not exclude the possibility that these receptors interact. We aimed to investigate if co-expression of combinations of these receptors had functional effects in prostate cancers cells. It has previously been demonstrated that ghrelin stimulates cell proliferation in prostate cancer cell lines, through ERK1/2 activation, and GPR39 can stimulate ERK1/2 signalling in response to zinc treatments. Additionally, both GHS-R1a and GPR39 display a high level of constitutive signalling and these constitutively active receptors can attenuate apoptosis when overexpressed individually in some cell types. We, therefore, investigated ERK1/2 and AKT signalling and cell survival in prostate cancer the potential modulation of these functions by dimerisation between GHS-R1a, GHS-R1b and GPR39. Expression of these receptors in the PC-3 prostate cancer cell line, either alone or in combination, did not alter constitutive ERK1/2 or AKT signalling, basal apoptosis or tunicamycin-stimulated apoptosis, compared to controls. In summary, the potential interactions between the ghrelin receptor isoforms, GHS-R1a and GHS-R1b, and the related zinc receptor, GPR39, and the potential for functional outcomes in prostate cancer were investigated using a number of independent methods. We did not definitively demonstrate the formation of these dimers using a number of state of the art methods to directly demonstrate receptor-receptor interactions. We investigated a number of potential functions of GPR39 and GHS-R1a in the prostate and did not observe altered function in response to co-expression of these receptors. The technical questions raised by this study highlight the requirement for the application of extensive controls when using current methods for the demonstration of GPCR dimerisation. Similar findings in this field reflect the current controversy surrounding the investigation of GPCR dimerisation. Although GHS-R1a/GHS-R1b or GHS-R1a/GPR39 heterodimerisation was not clearly demonstrated, this study provides a basis for future investigations of these receptors in prostate cancer. Additionally, the results presented in this study and growing evidence in the literature highlight the requirement for an extensive understanding of the experimental method and the performance of a range of controls to avoid the spurious interpretation of data gained from artificial expression systems. The future development of more robust techniques for investigating GPCR dimerisation is clearly required and will enable us to elucidate whether GHS-R1a, GHS-R1b and GPR39 form physiologically relevant dimers.

Bitrate modeling of scalable videos using quantization parameter, frame rate and spatial resolution

The quality and bitrate modeling is essential to effectively adapt the bitrate and quality of videos when delivered to multiplatform devices over resource constraint heterogeneous networks. The recent model proposed by Wang et al. estimates the bitrate and quality of videos in terms of the frame rate and quantization parameter. However, to build an effective video adaptation framework, it is crucial to incorporate the spatial resolution in the analytical model for bitrate and perceptual quality adaptation. Hence, this paper proposes an analytical model to estimate the bitrate of videos in terms of quantization parameter, frame rate, and spatial resolution. The model can fit the measured data accurately which is evident from the high Pearson correlation. The proposed model is based on the observation that the relative reduction in bitrate due to decreasing spatial resolution is independent of the quantization parameter and frame rate. This modeling can be used for rate-constrained bit-stream adaptation scheme which selects the scalability parameters to optimize the perceptual quality for a given bandwidth constraint.