Similarity between class A and class B G-protein-coupled receptors exemplified through calcitonin gene-related peptide receptor modelling and mutagenesis studies

Modelling class B G-protein-coupled receptors (GPCRs) using class A GPCR structural templates is difficult due to lack of homology. The plant GPCR, GCR1, has homology to both class A and class B GPCRs. We have used this to generate a class A-class B alignment, and by incorporating maximum lagged correlation of entropy and hydrophobicity into a consensus score, we have been able to align receptor transmembrane regions. We have applied this analysis to generate active and inactive homology models of the class B calcitonin gene-related peptide (CGRP) receptor, and have supported it with site-directed mutagenesis data using 122 CGRP receptor residues and 144 published mutagenesis results on other class B GPCRs. The variation of sequence variability with structure, the analysis of polarity violations, the alignment of group-conserved residues and the mutagenesis results at 27 key positions were particularly informative in distinguishing between the proposed and plausible alternative alignments. Furthermore, we have been able to associate the key molecular features of the class B GPCR signalling machinery with their class A counterparts for the first time. These include the [K/R]KLH motif in intracellular loop 1, [I/L]xxxL and KxxK at the intracellular end of TM5 and TM6, the NPXXY/VAVLY motif on TM7 and small group-conserved residues in TM1, TM2, TM3 and TM7. The equivalent of the class A DRY motif is proposed to involve Arg(2.39), His(2.43) and Glu(3.46), which makes a polar lock with T(6.37). These alignments and models provide useful tools for understanding class B GPCR function.

Self-training from labeled features for sentiment analysis

Sentiment analysis concerns about automatically identifying sentiment or opinion expressed in a given piece of text. Most prior work either use prior lexical knowledge defined as sentiment polarity of words or view the task as a text classification problem and rely on labeled corpora to train a sentiment classifier. While lexicon-based approaches do not adapt well to different domains, corpus-based approaches require expensive manual annotation effort. In this paper, we propose a novel framework where an initial classifier is learned by incorporating prior information extracted from an existing sentiment lexicon with preferences on expectations of sentiment labels of those lexicon words being expressed using generalized expectation criteria. Documents classified with high confidence are then used as pseudo-labeled examples for automatical domain-specific feature acquisition. The word-class distributions of such self-learned features are estimated from the pseudo-labeled examples and are used to train another classifier by constraining the model's predictions on unlabeled instances. Experiments on both the movie-review data and the multi-domain sentiment dataset show that our approach attains comparable or better performance than existing weakly-supervised sentiment classification methods despite using no labeled documents.

Development of areolae and growth of the peripheral prothallus in the crustose lichen Rhizocarpon geographicum:an image analysis study

Areolae of the crustose lichen Rhizocarpon geographicum (L.) DC., are present on the peripheral prothallus (marginal areolae) and also aggregate to form confluent masses in the centre of the thallus (central areolae). To determine the relationships between these areolae and whether growth of the peripheral prothallus is dependent on the marginal areolae, the density, morphology, and size frequency distributions of marginal areolae were measured in 23 thalli of R. geographicum in north Wales, UK using image analysis (Image J). Size and morphology of central areolae were also studied across the thallus. Marginal areolae were small, punctate, and occurred in clusters scattered over the peripheral prothallus while central areolae were larger and had a lobed structure. The size-class frequency distributions of the marginal and central areolae were fitted by power-law and log-normal models respectively. In 16 out of 23 thalli, central areolae close to the outer edge were larger and had a more complex lobed morphology than those towards the thallus centre. Neither mean width nor radial growth rate (RaGR) of the peripheral prothallus were correlated with density, diameter, or area fraction of marginal areolae. The data suggest central areolae may develop from marginal areolae as follows: (1) marginal areolae develop in clusters at the periphery and fuse to form central areolae, (2) central areolae grow exponentially, and (3) crowding of central areolae results in constriction and fragmentation. In addition, growth of the peripheral prothallus may be unrelated to the marginal areolae. © 2013 Springer Science+Business Media Dordrecht.

Toward the prediction of class I and II mouse major histocompatibility complex-peptide-binding affinity:in silico bioinformatic step-by-step guide using quantitative structure-activity relationships

Quantitative structure-activity relationship (QSAR) analysis is a cornerstone of modern informatics. Predictive computational models of peptide-major histocompatibility complex (MHC)-binding affinity based on QSAR technology have now become important components of modern computational immunovaccinology. Historically, such approaches have been built around semiqualitative, classification methods, but these are now giving way to quantitative regression methods. We review three methods--a 2D-QSAR additive-partial least squares (PLS) and a 3D-QSAR comparative molecular similarity index analysis (CoMSIA) method--which can identify the sequence dependence of peptide-binding specificity for various class I MHC alleles from the reported binding affinities (IC50) of peptide sets. The third method is an iterative self-consistent (ISC) PLS-based additive method, which is a recently developed extension to the additive method for the affinity prediction of class II peptides. The QSAR methods presented here have established themselves as immunoinformatic techniques complementary to existing methodology, useful in the quantitative prediction of binding affinity: current methods for the in silico identification of T-cell epitopes (which form the basis of many vaccines, diagnostics, and reagents) rely on the accurate computational prediction of peptide-MHC affinity. We have reviewed various human and mouse class I and class II allele models. Studied alleles comprise HLA-A*0101, HLA-A*0201, HLA-A*0202, HLA-A*0203, HLA-A*0206, HLA-A*0301, HLA-A*1101, HLA-A*3101, HLA-A*6801, HLA-A*6802, HLA-B*3501, H2-K(k), H2-K(b), H2-D(b) HLA-DRB1*0101, HLA-DRB1*0401, HLA-DRB1*0701, I-A(b), I-A(d), I-A(k), I-A(S), I-E(d), and I-E(k). In this chapter we show a step-by-step guide into predicting the reliability and the resulting models to represent an advance on existing methods. The peptides used in this study are available from the AntiJen database (http://www.jenner.ac.uk/AntiJen). The PLS method is available commercially in the SYBYL molecular modeling software package. The resulting models, which can be used for accurate T-cell epitope prediction, will be made are freely available online at the URL http://www.jenner.ac.uk/MHCPred.

A predictor of membrane class:discriminating α-helical and β-barrel membrane proteins from non-membranous proteins

Accurate protein structure prediction remains an active objective of research in bioinformatics. Membrane proteins comprise approximately 20% of most genomes. They are, however, poorly tractable targets of experimental structure determination. Their analysis using bioinformatics thus makes an important contribution to their on-going study. Using a method based on Bayesian Networks, which provides a flexible and powerful framework for statistical inference, we have addressed the alignment-free discrimination of membrane from non-membrane proteins. The method successfully identifies prokaryotic and eukaryotic α-helical membrane proteins at 94.4% accuracy, β-barrel proteins at 72.4% accuracy, and distinguishes assorted non-membranous proteins with 85.9% accuracy. The method here is an important potential advance in the computational analysis of membrane protein structure. It represents a useful tool for the characterisation of membrane proteins with a wide variety of potential applications.

Quantitative prediction of mouse class I MHC peptide binding affinity using support vector machine regression (SVR) models

Background - The binding between peptide epitopes and major histocompatibility complex proteins (MHCs) is an important event in the cellular immune response. Accurate prediction of the binding between short peptides and the MHC molecules has long been a principal challenge for immunoinformatics. Recently, the modeling of MHC-peptide binding has come to emphasize quantitative predictions: instead of categorizing peptides as "binders" or "non-binders" or as "strong binders" and "weak binders", recent methods seek to make predictions about precise binding affinities. Results - We developed a quantitative support vector machine regression (SVR) approach, called SVRMHC, to model peptide-MHC binding affinities. As a non-linear method, SVRMHC was able to generate models that out-performed existing linear models, such as the "additive method". By adopting a new "11-factor encoding" scheme, SVRMHC takes into account similarities in the physicochemical properties of the amino acids constituting the input peptides. When applied to MHC-peptide binding data for three mouse class I MHC alleles, the SVRMHC models produced more accurate predictions than those produced previously. Furthermore, comparisons based on Receiver Operating Characteristic (ROC) analysis indicated that SVRMHC was able to out-perform several prominent methods in identifying strongly binding peptides. Conclusion - As a method with demonstrated performance in the quantitative modeling of MHC-peptide binding and in identifying strong binders, SVRMHC is a promising immunoinformatics tool with not inconsiderable future potential.

New horizons in mouse immunoinformatics:reliable in silico prediction of mouse class I histocompatibility major complex peptide binding affinity

Quantitative structure–activity relationship (QSAR) analysis is a main cornerstone of modern informatic disciplines. Predictive computational models, based on QSAR technology, of peptide-major histocompatibility complex (MHC) binding affinity have now become a vital component of modern day computational immunovaccinology. Historically, such approaches have been built around semi-qualitative, classification methods, but these are now giving way to quantitative regression methods. The additive method, an established immunoinformatics technique for the quantitative prediction of peptide–protein affinity, was used here to identify the sequence dependence of peptide binding specificity for three mouse class I MHC alleles: H2–Db, H2–Kb and H2–Kk. As we show, in terms of reliability the resulting models represent a significant advance on existing methods. They can be used for the accurate prediction of T-cell epitopes and are freely available online (http://www.jenner.ac.uk/MHCPred).

Coupling in silico and in vitro analysis of peptide-MHC binding:a bioinformatic approach enabling prediction of superbinding peptides and anchorless epitopes

The ability to define and manipulate the interaction of peptides with MHC molecules has immense immunological utility, with applications in epitope identification, vaccine design, and immunomodulation. However, the methods currently available for prediction of peptide-MHC binding are far from ideal. We recently described the application of a bioinformatic prediction method based on quantitative structure-affinity relationship methods to peptide-MHC binding. In this study we demonstrate the predictivity and utility of this approach. We determined the binding affinities of a set of 90 nonamer peptides for the MHC class I allele HLA-A*0201 using an in-house, FACS-based, MHC stabilization assay, and from these data we derived an additive quantitative structure-affinity relationship model for peptide interaction with the HLA-A*0201 molecule. Using this model we then designed a series of high affinity HLA-A2-binding peptides. Experimental analysis revealed that all these peptides showed high binding affinities to the HLA-A*0201 molecule, significantly higher than the highest previously recorded. In addition, by the use of systematic substitution at principal anchor positions 2 and 9, we showed that high binding peptides are tolerant to a wide range of nonpreferred amino acids. Our results support a model in which the affinity of peptide binding to MHC is determined by the interactions of amino acids at multiple positions with the MHC molecule and may be enhanced by enthalpic cooperativity between these component interactions.

G-protein-coupled receptor solubilization and purification for biophysical analysis and functional studies, in the total absence of detergent

G-protein coupled receptors (GPCRs) constitute the largest class of membrane proteins and are a major drug target. A serious obstacle to studying GPCR structure/function characteristics is the requirement to extract the receptors from their native environment in the plasma membrane, coupled with the inherent instability of GPCRs in the detergents required for their solubilization. In the present study, we report the first solubilization and purification of a functional GPCR [human adenosine A2A receptor (A2AR)], in the total absence of detergent at any stage, by exploiting spontaneous encapsulation by styrene maleic acid (SMA) co-polymer direct from the membrane into a nanoscale SMA lipid particle (SMALP). Furthermore, the A2AR-SMALP, generated from yeast (Pichia pastoris) or mammalian cells, exhibited increased thermostability (∼5°C) compared with detergent [DDM (n-dodecyl-β-D-maltopyranoside)]-solubilized A2AR controls. The A2AR-SMALP was also stable when stored for prolonged periods at 4°C and was resistant to multiple freeze-thaw cycles, in marked contrast with the detergent-solubilized receptor. These properties establish the potential for using GPCR-SMALP in receptor-based drug discovery assays. Moreover, in contrast with nanodiscs stabilized by scaffold proteins, the non-proteinaceous nature of the SMA polymer allowed unobscured biophysical characterization of the embedded receptor. Consequently, CD spectroscopy was used to relate changes in secondary structure to loss of ligand binding ([³H]ZM241385) capability. SMALP-solubilization of GPCRs, retaining the annular lipid environment, will enable a wide range of therapeutic targets to be prepared in native-like state to aid drug discovery and understanding of GPCR molecular mechanisms.

A Sensitive Metric of Class Cohesion

Metrics estimate the quality of different aspects of software. In particular, cohesion indicates how well the parts of a system hold together. A metric to evaluate class cohesion is important in object-oriented programming because it gives an indication of a good design of classes. There are several proposals of metrics for class cohesion but they have several problems (for instance, low discrimination). In this paper, a new metric to evaluate class cohesion is proposed, called SCOM, which has several relevant features. It has an intuitive and analytical formulation, what is necessary to apply it to large-size software systems. It is normalized to produce values in the range [0..1], thus yielding meaningful values. It is also more sensitive than those previously reported in the literature. The attributes and methods used to evaluate SCOM are unambiguously stated. SCOM has an analytical threshold, which is a very useful but rare feature in software metrics. We assess the metric with several sample cases, showing that it gives more sensitive values than other well know cohesion metrics.

Computer-aided System of Semantic Text Analysis of a Technical Specification

The given work is devoted to development of the computer-aided system of semantic text analysis of a technical specification. The purpose of this work is to increase efficiency of software engineering based on automation of semantic text analysis of a technical specification. In work it is offered and investigated the model of the analysis of the text of the technical project is submitted, the attribute grammar of a technical specification, intended for formalization of limited Russian is constructed with the purpose of analysis of offers of text of a technical specification, style features of the technical project as class of documents are considered, recommendations on preparation of text of a technical specification for the automated processing are formulated. The computer-aided system of semantic text analysis of a technical specification is considered. This system consists of the following subsystems: preliminary text processing, the syntactic and semantic analysis and construction of software models, storage of documents and interface.

Using the e-Class Open Platform for e-Learning Support at the TEI of Larissa, Greece

In the current paper we firstly give a short introduction on e-learning platforms and review the case of the e-class open e-learning platform being used by the Greek tertiary education sector. Our analysis includes strategic selection issues and outcomes in general and operational and adoption issues in the case of the Technological Educational Institute (TEI) of Larissa, Greece. The methodology is being based on qualitative analysis of interviews with key actors using the platform, and statistical analysis of quantitative data related to adoption and usage in the relevant populations. The author has been a key actor in all stages and describes his insights as an early adopter, diffuser and innovative user. We try to explain the issues under consideration using existing past research outcomes and we also arrive to some conclusions and points for further research.

Computer Support of Semantic Text Analysis of a Technical Specification on Designing Software

The given work is devoted to development of the computer-aided system of semantic text analysis of a technical specification. The purpose of this work is to increase efficiency of software engineering based on automation of semantic text analysis of a technical specification. In work it is offered and investigated a technique of the text analysis of a technical specification is submitted, the expanded fuzzy attribute grammar of a technical specification, intended for formalization of limited Russian language is constructed with the purpose of analysis of offers of text of a technical specification, style features of the technical specification as class of documents are considered, recommendations on preparation of text of a technical specification for the automated processing are formulated. The computer-aided system of semantic text analysis of a technical specification is considered. This system consist of the following subsystems: preliminary text processing, the syntactic and semantic analysis and construction of software models, storage of documents and interface.

On Two Saigo’s Fractional Integral Operators in the Class of Univalent Functions

2000 Mathematics Subject Classification: Primary 26A33, 30C45; Secondary 33A35

Applications of the Owa-Srivastava Operator to the Class of K-Uniformly Convex Functions

2000 Mathematics Subject Classification: Primary 30C45, 26A33; Secondary 33C15