A novel grass pollen allergen mimotope identified by phage display peptide library inhibits allergen-human IgE antibody interaction

The aim of this study was to investigate the molecular basis of human IgE-allergen interaction by screening a phage-displayed peptide library with an allergen-specific human IgE-mimicking monoclonal antibody (mAb). A mAb that reacted with major grass pollen allergens was successfully identified and shown to inhibit human IgE-allergen interaction. Biopanning of a phage-displayed random peptide library with this mAb yielded a 12 amino acid long mimotope. A synthetic peptide based on this 12-mer mimotope inhibited mAb and human IgE binding to grass pollen extracts. Our results indicate that such synthetic peptide mimotopes of allergens have potential as novel therapeutic agents. © 2001 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.

Monoclonal antibody screening of a phage-displayed random peptide library reveals mimotopes of chemokine receptor CCR5: Implications for the tertiary structure of the receptor and for an n-terminal binding site for HIV-1 gp120

The chemokine receptor CCR5 contains seven transmembrane-spanning domains. It binds chemokines and acts as co-receptor for macrophage (m)-tropic (or R5) strains of HIV-1. Monoclonal antibodies (mAb) to CCR5, 3A9 and 5C7, were used for biopanning a nonapeptide cysteine (C)-constrained phage-displayed random peptide library to ascertain contact residues and define tertiary structures of possible epitopes on CCR5. Reactivity of antibodies with phagotopes was established by enzyme-linked immunosorbent assay (ELISA). mAb 3A9 identified a phagotope C-HASIYDFGS-C (3A9/1), and 5C7 most frequently identified C-PHWLRDLRV-C (5C7/1). Corresponding peptides were synthesized. Phagotopes and synthetic peptides reacted in ELISA with corresponding antibodies and synthetic peptides inhibited antibody binding to the phagotopes. Reactivity by immunofluorescence of 3A9 with CCR5 was strongly inhibited by the corresponding peptide. Both mAb 3A9 and 5C7 reacted similarly with phagotopes and the corresponding peptide selected by the alternative mAb. The sequences of peptide inserts of phagotopes could be aligned as mimotopes of the sequence of CCR5. For phage 3A9/1, the motif SIYD aligned to residues at the N terminus and FG to residues on the first extracellular loop; for 5C7/1, residues at the N terminus, first extracellular loop, and possibly the third extracellular loop could be aligned and so would contribute to the mimotope. The synthetic peptides corresponding to the isolated phagotopes showed a CD4-dependent reactivity with gp120 of a primary, m-tropic HIV-1 isolate. Thus reactivity of antibodies raised to CCR5 against phage-displayed peptides defined mimotopes that reflect binding sites for these antibodies and reveal a part of the gp120 binding sites on CCR5.

A polymerase chain reaction-based method for isolating clones from a complimentary DNA library in sheep

The sheep (Ovis aries) is favored by many musculoskeletal tissue engineering groups as a large animal model because of its docile temperament and ease of husbandry. The size and weight of sheep are comparable to humans, which allows for the use of implants and fixation devices used in human clinical practice. The construction of a complimentary DNA (cDNA) library can capture the expression of genes in both a tissue- and time-specific manner. cDNA libraries have been a consistent source of gene discovery ever since the technology became commonplace more than three decades ago. Here, we describe the construction of a cDNA library using cells derived from sheep bones based on the pBluescript cDNA kit. Thirty clones were picked at random and sequenced. This led to the identification of a novel gene, C12orf29, which our initial experiments indicate is involved in skeletal biology. We also describe a polymerase chain reaction-based cDNA clone isolation method that allows the isolation of genes of interest from a cDNA library pool. The techniques outlined here can be applied in-house by smaller tissue engineering groups to generate tools for biomolecular research for large preclinical animal studies and highlights the power of standard cDNA library protocols to uncover novel genes.

Creating a global law graduate: The need, benefits and practical approaches to internationalise the curriculum

The increasingly integrated world has facilitated important international and trans-border trends, such as a progressively connected global economy, a significant growth in transnational business transactions and an increase in global regulation of global issues. Such globalisation has had a transformational impact on the legal profession in a number of ways. These include the need to provide advice on issues or transactions that have a transnational or international element; the increasing globalisation of large law firms; and the delivery of offshore services by legal service providers. This means that not only do law graduates need to be prepared to practice in an increasingly globalised economy and legal profession, there will also be new career opportunities available to them which require understanding of international law, for example in emerging international institutions and non-government organisations. Accordingly there is a need to ensure that law students develop the knowledge and skills they will require to succeed in a globalised legal profession. That is, there is a need to internationalise the law curriculum. This paper provides an insight into the recent progression of law schools in internationalising the law curriculum and provides practical avenues and strategies for the increased integration of international law, foreign law and a comparative perspective into core subjects which will develop the graduates’ knowledge and skills in international and foreign law, in order to enhance their ability to succeed as legal professionals in a globalised world.

Indonesian library and information science professionals’ understanding of Library 2.0: A pilot study

The literature around Library 2.0 remains largely theoretical with few empirical studies and is particularly limited in developing countries such as Indonesia. This study addresses this gap and aims to provide information about the current state of knowledge on Indonesian LIS professionals’ understanding of Library 2.0. The researchers used qualitative and quantitative approaches for this study, asking thirteen closed- and open-ended questions in an online survey. The researchers used descriptive and in vivo coding to analyze the responses. Through their analysis, they identified three themes: technology, interactivity, and awareness of Library 2.0. Respondents demonstrated awareness of Library 2.0 and a basic understanding of the roles of interactivity and technology in libraries. However, overreliance on technology used in libraries to conceptualize Library 2.0 without an emphasis on its core characteristics and principles could lead to the misalignment of limited resources. The study results will potentially strengthen the research base for Library 2.0 practice as well as inform LIS curriculum in Indonesia so as to develop practitioners who are able to adapt to users’ changing needs and expectations. It is expected that the preliminary data from this study could be used to design a much larger and more complex future research project in this area.

LibVM: An architecture for shared library sandboxing

Many software applications extend their functionality by dynamically loading libraries into their allocated address space. However, shared libraries are also often of unknown provenance and quality and may contain accidental bugs or, in some cases, deliberately malicious code. Most sandboxing techniques which address these issues require recompilation of the libraries using custom tool chains, require significant modifications to the libraries, do not retain the benefits of single address-space programming, do not completely isolate guest code, or incur substantial performance overheads. In this paper we present LibVM, a sandboxing architecture for isolating libraries within a host application without requiring any modifications to the shared libraries themselves, while still retaining the benefits of a single address space and also introducing a system call inter-positioning layer that allows complete arbitration over a shared library’s functionality. We show how to utilize contemporary hardware virtualization support towards this end with reasonable performance overheads and, in the absence of such hardware support, our model can also be implemented using a software-based mechanism. We ensure that our implementation conforms as closely as possible to existing shared library manipulation functions, minimizing the amount of effort needed to apply such isolation to existing programs. Our experimental results show that it is easy to gain immediate benefits in scenarios where the goal is to guard the host application against unintentional programming errors when using shared libraries, as well as in more complex scenarios, where a shared library is suspected of being actively hostile. In both cases, no changes are required to the shared libraries themselves.

Analysis of a Genomic DNA Expression Library of Mycobacterium tuberculosis Using Tuberculosis Patient Sera: Evidence for Modulation of Host Immune Response

DNA obtained from a human sputum isolate of Mycobacterium tuberculosis, NTI-64719, which showed extensive dissemination in the guinea pig model resulting in a high score for virulence was used to construct an expression library in the lambda ZAP vector. The size of DNA inserts in the library ranged from 1 to 3 kb, and recombinants represented 60% of the total plaques obtained. When probed with pooled serum from chronically infected tuberculosis patients, the library yielded 176 recombinants with a range of signal intensities. Among these, 93 recombinants were classified into 12 groups on the basis of DNA hybridization experiments, The polypeptides synthesized by the recombinants were predominantly LacZ fusion proteins, Serum obtained from patients who were clinically diagnosed to be in the early phase of M. tuberculosis infection was used to probe the 176 recombinants obtained. interestingly, some recombinants that gave very strong signals in the original screen did not react with early-phase serum; conversely, others whose signals were extremely weak in the original screen gave very intense signals with serum from recently infected patients, This indicates the differential nature of either the expression of these antigens or the immune response elicited by them as a function of disease progression.

State Library Queensland

Sheona Thomson writes about the transformation of a public institution in relation to a study on post-occupancy. "Brisbanites like me, with memories of long hours of study in the former buildings of the State Library of Queensland, can only marvel at the living institution we have in our city today. For most of the 80s, our bookish pursuits were hosted in the fustily intimate reading rooms of Centennial Hall, the late 1950s extension to the nineteenth-century building (formerly housing the state museum) by F. D. G. Stanley in William Street on the north bank of the Brisbane River. At the time, the wheels of an expansive cultural ambition were turning, and piece by piece on the south bank of the river the rambling Queensland Cultural Centre was realized. The fourth stage of the complex opened in 1988 as the new home for the State Library and for many years after, countless studious, transient folk whiled away time in the deep interiors of the straight-faced concrete and glass edifice by Robin Gibson and Partners..."

Pedagogical tools designed to promote engagement and active learning in the biomedical sciences and allied health disciplines

Perceiving students, science students especially, as mere consumers of facts and information belies the importance of a need to engage them with the principles underlying those facts and is counter-intuitive to the facilitation of knowledge and understanding. Traditional didactic lecture approaches need a re-think if student classroom engagement and active learning are to be valued over fact memorisation and fact recall. In our undergraduate biomedical science programs across Years 1, 2 and 3 in the Faculty of Health at QUT, we have developed an authentic learning model with an embedded suite of pedagogical strategies that foster classroom engagement and allow for active learning in the sub-discipline area of medical bacteriology. The suite of pedagogical tools we have developed have been designed to enable their translation, with appropriate fine-tuning, to most biomedical and allied health discipline teaching and learning contexts. Indeed, aspects of the pedagogy have been successfully translated to the nursing microbiology study stream at QUT. The aims underpinning the pedagogy are for our students to: (1) Connect scientific theory with scientific practice in a more direct and authentic way, (2) Construct factual knowledge and facilitate a deeper understanding, and (3) Develop and refine their higher order flexible thinking and problem solving skills, both semi-independently and independently. The mindset and role of the teaching staff is critical to this approach since for the strategy to be successful tertiary teachers need to abandon traditional instructional modalities based on one-way information delivery. Face-to-face classroom interactions between students and lecturer enable realisation of pedagogical aims (1), (2) and (3). The strategy we have adopted encourages teachers to view themselves more as expert guides in what is very much a student-focused process of scientific exploration and learning. Specific pedagogical strategies embedded in the authentic learning model we have developed include: (i) interactive lecture-tutorial hybrids or lectorials featuring teacher role-plays as well as class-level question-and-answer sessions, (ii) inclusion of “dry” laboratory activities during lectorials to prepare students for the wet laboratory to follow, (iii) real-world problem-solving exercises conducted during both lectorials and wet laboratory sessions, and (iv) designing class activities and formative assessments that probe a student’s higher order flexible thinking skills. Flexible thinking in this context encompasses analytical, critical, deductive, scientific and professional thinking modes. The strategic approach outlined above is designed to provide multiple opportunities for students to apply principles flexibly according to a given situation or context, to adapt methods of inquiry strategically, to go beyond mechanical application of formulaic approaches, and to as much as possible self-appraise their own thinking and problem solving. The pedagogical tools have been developed within both workplace (real world) and theoretical frameworks. The philosophical core of the pedagogy is a coherent pathway of teaching and learning which we, and many of our students, believe is more conducive to student engagement and active learning in the classroom. Qualitative and quantitative data derived from online and hardcopy evaluations, solicited and unsolicited student and graduate feedback, anecdotal evidence as well as peer review indicate that: (i) our students are engaging with the pedagogy, (ii) a constructivist, authentic-learning approach promotes active learning, and (iii) students are better prepared for workplace transition.

When film, fashion & interiors collide: Designing the After Darkly Graduate Fashion Show

In 2013 QUT Interior Design and Fashion Disciplines partnered to design the Catwalk for the QUT After Darkly Graduate Fashion Show. The ephemeral work (catwalk canopy and cinematic affects) was developed through collaboration between the authors based upon an undergraduate interior design unit ‘Filmic Interiors’ in which students were tasked with designing a fashion show. Filmic Interiors exploited the potential of film to influence, understand, and develop novel interior spaces through consideration of mise-en-scene, cinematic effects and atmospheric design strategies engaged by key film directors Jean Pierre Jeunet and Darren Aronofsky. The design outcome represents a hybridisation of student design proposals, contemplating both film and emerging collections from graduate fashion students. The work explored a number of iterations each testing material qualities and immaterial cinematic affects, as a means to develop new space. The process was led by experimentation undertaken by the designers through previous studio explorations surrounding the theme of ‘Strange Space’ and design practice ‘Making Strange’(Lindquist & Pytel, 2012). In doing so, the work paralleled the material formations of ‘obsessive collections’ and ‘making do’ evident in Jeunet’s scenography, rendering uncanny hybrid space (Ezra, 2008). Evocation of the immaterial found in much of director Aronofsky’s work, also became critical in the atmospheric experience intended for the show. This paper explores the process of collaboration and material experimentation in design, approached through a filmic lens. It provides insight into what happens when one enters into what can be termed an ‘ecology of production’, whereby the experimental making becomes the collaborative agent between designers, disciplines, and between stage and spectators. Finally it underlines the importance of ‘finding the work’ through material making and testing rather than through more controlled formalistic responses.

Rapid and efficient screening of a Representational Difference Analysis library using reverse Southern hybridisation: Identification of genetic differences between Haemophilus parasuis isolates

Representational Difference Analysis (RDA) is an established technique used for isolation of specific genetic differences between or within bacterial species. This method was used to investigate the genetic basis of serovar-specificity and the relationship between serovar and virulence in Haemophilus parasuis. An RDA clone library of 96 isolates was constructed using H. parasuis strains H425(P) (serovar 12) and HS1967 (serovar 4). To screen such a large clone library to determine which clones are strain-specific would typically involved separately labelling each clone for use in Southern hybridisation against genomic DNA from each of the strains. In this study, a novel application of reverse Southern hybridisation was used to screen the RDA library: genomic DNA from each strain was labelled and used to probe the library to identify strain-specific clones. This novel approach represents a significant improvement in methodology that is rapid and efficient.

Identification and immunogenicity of an immunodominant mimotope of Avibacterium paragallinarum from a phage display peptide library

Avibacterium paragallinarum is the causative agent of infectious coryza. The protective antigens of this important pathogen have not yet been clearly identified. In this paper, we applied phage display technique to screen the immunodominant mimotopes of a serovar A strain of A. paragallinarum by using a random 12-peptide library, and evaluated the immunogenicity in chickens of the selected mimotope. Polyclonal antibody directed against A. paragallinarum strain 0083 (serovar A) was used as the target antibody and phage clones binding to this target were screened from the 12-mer random peptide library. More than 50% of the phage clones selected in the third round carried the consensus peptide motif sequence A-DP(M)L. The phage clones containing the peptide motif reacted with the target antibody and this interaction could be blocked, in a dose-dependent manner, by A. paragallinarum. One of the peptide sequences, YGLLAVDPLFKP, was selected and the corresponding oligonucleotide sequence was synthesized and then inserted into the expression vector pFliTrx. The recombinant plasmid was transferred into an expression host Escherichia coli GI826 by electroporation, resulting in a recombinant E. coli expressing the peptide on the bacterial surface. Intramuscular injection of the epitope-expressing recombinant bacteria into chickens induced a specific serological response to serovar A. A. paragallinarum. The chickens given the recombinant E. coli showed significant protection against challenge with A. paragallinarum 0083. These results indicated a potential for the use of the mimotope in the development of molecular vaccines for infectious coryza.

Product-library-derived specification parameters in building information models augmentation

This paper conceptualizes a framework for bridging the BIM (building information modelling)-specifications divide through augmenting objects within BIM with specification parameters derived from a product library. We demonstrate how model information, enriched with data at various LODs (levels of development), can evolve simultaneously with design and construction using different representation of a window object embedded in a wall as lifecycle phase exemplars at different levels of granularity. The conceptual standpoint is informed by the need for exploring a methodological approach which extends beyond current limitations of current modelling platforms in enhancing the information content of BIM models. Therefore, this work demonstrates that BIM objects can be augmented with construction specification parameters leveraging product libraries.