Plasmid DNA purification via the use of a dual affinity protein

Methods are presented for the production, affinity purification and analysis of plasmid DNA (pDNA). Batch fermentation is used for the production of the pDNA, and expanded bed chromatography, via the use of a dual affinity glutathione S-transferase (GST) fusion protein, is used for the capture and purification of the pDNA. The protein is composed of GST, which displays affinity for glutathione immobilized to a solid-phase adsorbent, fused to a zinc finger transcription factor, which displays affinity for a target 9-base pair sequence contained within the target pDNA. A Picogreen™ fluorescence assay and/or anx ethidium bromide agarose gel electrophoresis assay can be used to analyze the eluted pDNA.

Using DNA as a drug : challenges and opportunities for process engineers

The maturing of the biotechnology industry and a focus on productivity has seen a shift from discovery science to small-scale bench-top research to higher productivity, large scale production. Health companies are aggressively expanding their biopharmaceutical interests, an expansion which is facilitated by biochemical and bioprocess engineering. An area of continuous growth is vaccines. Vaccination will be a key intervention in the case of an influenza pandemic. The global manufacturing capacity for fast turn around vaccines is currently woefully inadequate at around 300 million shots. As the prevention of epidemics requires > 80 % vaccination, in theory the world should currently be aiming for the ability to produce around 5.3 billion vaccines. Presented is a production method for the creation of a fast turn around DNA vaccine. A DNA vaccine could have a production time scale of as little as two weeks. This process has been harnessed into a pilot scale production system for the creation of a pre-clinical grade malaria vaccine in a collaborative project with the Coppel Lab, Department of Microbiology, Monash University. In particular, improvements to the fermentation, chromatography and delivery stages will be discussed. Consideration will then be given as to how the fermentation stage affects the mid and downstream processing stages.

Computational micromodel for epigenetic mechanisms

Characterization of the epigenetic profile of humans since the initial breakthrough on the human genome project has strongly established the key role of histone modifications and DNA methylation. These dynamic elements interact to determine the normal level of expression or methylation status of the constituent genes in the genome. Recently, considerable evidence has been put forward to demonstrate that environmental stress implicitly alters epigenetic patterns causing imbalance that can lead to cancer initiation. This chain of consequences has motivated attempts to computationally model the influence of histone modification and DNA methylation in gene expression and investigate their intrinsic interdependency. In this paper, we explore the relation between DNA methylation and transcription and characterize in detail the histone modifications for specific DNA methylation levels using a stochastic approach.

Cell type-dependent, infection-induced, aberrant DNA methylation in gastric cancer

Epigenetic changes correspond to heritable modifications of the chromatin structure, which do not involve any alteration of the DNA sequence but nonetheless affect gene expression. These mechanisms play an important role in cell differentiation, but aberrant occurrences are also associated with a number of diseases, including cancer and neural development disorders. In particular, aberrant DNA methylation induced by H. Pylori has been found to be a significant risk factor in gastric cancer. To investigate the sensitivity of different genes and cell types to this infection, a computational model of methylation in gastric crypts is developed. In this article, we review existing results from physical experiments and outline their limitations, before presenting the computational model and investigating the influence of its parameters.

Computational analysis of epigenetic information in human DNA sequences

Over the last few years, investigations of human epigenetic profiles have identified key elements of change to be Histone Modifications, stable and heritable DNA methylation and Chromatin remodeling. These factors determine gene expression levels and characterise conditions leading to disease. In order to extract information embedded in long DNA sequences, data mining and pattern recognition tools are widely used, but efforts have been limited to date with respect to analyzing epigenetic changes, and their role as catalysts in disease onset. Useful insight, however, can be gained by investigation of associated dinucleotide distributions. The focus of this paper is to explore specific dinucleotides frequencies across defined regions within the human genome, and to identify new patterns between epigenetic mechanisms and DNA content. Signal processing methods, including Fourier and Wavelet Transformations, are employed and principal results are reported.

DNA molecules and future blockbuster therapeutics

Deoxyribonucleic acid molecules are heralding a new generation of reverse - engineered biopharmaceuticals. In terms of potential application in gene medicine, plasmid DNA (pDNA) vectors have exceptional therapeutic and immunological profiles as they are free from safety concerns associated with viral vectors, display non-toxicity and are simpler to develop. This presentation will discuss the potential applications of pDNA molecules in vaccine development and gene therapy, pilot-scale production of pDNA-based biopharmaceuticals and the controlled delivery of therapeutic sequences in biodegradable polymers to different target cells via the nasal route.

Design and synthesis of a screening library using the natural product scaffold 3-chloro-4-hydroxyphenylacetic acid

The fungal metabolite 3-chloro-4-hydroxyphenylacetic acid (1) was utilized in the generation of a unique drug-like screening library using parallel solution-phase synthesis. A 20-membered amide library (3–22) was generated by first converting 1 to methyl (3-chloro-4-hydroxyphenyl)acetate (2), then reacting this scaffold with a diverse series of primary amines via a solvent-free aminolysis procedure. The structures of the synthetic analogues (3–22) were elucidated by spectroscopic data analysis. The structures of compounds 8, 12, and 22 were confirmed by single X-ray crystallographic analysis. All compounds were evaluated for cytotoxicity against a human prostate cancer cell line (LNCaP) and for antiparasitic activity toward Trypanosoma brucei brucei and Plasmodium falciparum and showed no significant activity at 10 μM. The library was also tested for effects on the lipid content of LNCaP and PC-3 prostate cancer cells, and it was demonstrated that the fluorobenzyl analogues (12–14) significantly reduced cellular phospholipid and neutral lipid levels.

Knowledge unlatched: A global library consortium model for funding open access scholarly books: Full report on the Proof of Concept Pilot 2014

This special issue of Cultural Science Journal is devoted to the report of a groundbreaking experiment in re-coordinating global markets for specialist scholarly books and enabling the knowledge commons: the Knowledge Unlatched proof-of-concept pilot. The pilot took place between January 2012 and September 2014. It involved libraries, publishers, authors, readers and research funders in the process of developing and testing a global library consortium model for supporting Open Access books. The experiment established that authors, librarians, publishers and research funding agencies can work together in powerful new ways to enable open access; that doing so is cost effective; and that a global library consortium model has the potential dramatically to widen access to the knowledge and ideas contained in book-length scholarly works.

Establishing a participatory library model: A Grounded Theory study

Purpose The purpose of this research is to explore the idea of the participatory library in higher education settings. This research aims to address the question, what is a participatory university library? Design/methodology/approach Grounded theory approach was adopted. In-depth individual interviews were conducted with two diverse groups of participants including ten library staff members and six library users. Data collection and analysis were carried out simultaneously and complied with Straussian grounded theory principles and techniques. Findings Three core categories representing the participatory library were found including “community”, “empowerment”, and “experience”. Each category was thoroughly delineated via sub-categories, properties, and dimensions that all together create a foundation for the participatory library. A participatory library model was also developed together with an explanation of model building blocks that provide a deeper understanding of the participatory library phenomenon. Research limitations The research focuses on a specific library system, i.e., academic libraries. Therefore, the research results may not be very applicable to public, special, and school library contexts. Originality/value This is the first empirical study developing a participatory library model. It provides librarians, library managers, researchers, library students, and the library community with a holistic picture of the contemporary library.

(Digital library) education or (digital) library education? An Australian perspective

Education for Library and Information professionals in the digital environment has been an important discussion point the world over. However, before designing and implementing a programme for digital library education, it is prudent that the skills and knowledge required to work in this environment are identified to enable informed decisions to be made. Hitherto, there has been very little research which has sought the opinion of both educators and practitioners on this topic, and none with a wide geographical coverage of Australia. This paper presents the key findings of research undertaken at Tallinn University in the first half of 2009.

Using a free online questionnaire to determine the skills, competencies and knowledge required to work in a digital library environment in Australia

This paper discusses the data collection technique used to determine the skills and knowledge required of academic librarians working in a digital library environment in Australia. The research was undertaken as part of the researcher’s master’s thesis conducted at Tallinn University. The data collection instrument used was a freely available online survey tool, and its advantages and disadvantages are discussed in terms of the desired outcomes and circumstances surrounding the thesis project. Decisions regarding the design of the questionnaire are also discussed.

The Library beyond (just) the book: Collaboration and co-working spaces for new technology and new technological practices

Libraries have often been first adopters of many new technological innovations, such as, punch cards, computers, barcodes, and e-book readers. It is thus not surprising that many libraries have embraced the advent of the internet as an opportunity to move away from just being repositories of books, towards becoming ideas stores and local network hubs for entrepreneurial thinking and new creative practices. This presentation will look at the case of “The Edge” – an initiative of the State Library of Queensland in Brisbane, Australia, to establish a digital culture centre and learning environment deliberately designed for the co-creation and co-construction of knowledge. This initiative illustrates the potential role of libraries as testing grounds for new technologies and technological practices, which is particularly relevant in the context of the NBN rollout across Australia. It also provides an example of new engagement strategies for innovative co-working spaces that are a vital element in a trend that sees professionals, creatives and designers leave their traditional places of work and embrace the city as their office.