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.

Convergence-guaranteed time-varying RRT path planning for profiling floats in 4-Dimensional flow

This paper presents an extension to the Rapidly-exploring Random Tree (RRT) algorithm applied to autonomous, drifting underwater vehicles. The proposed algorithm is able to plan paths that guarantee convergence in the presence of time-varying ocean dynamics. The method utilizes 4-Dimensional, ocean model prediction data as an evolving basis for expanding the tree from the start location to the goal. The performance of the proposed method is validated through Monte-Carlo simulations. Results illustrate the importance of the temporal variance in path execution, and demonstrate the convergence guarantee of the proposed methods.

Soluble NSF attachment protein receptor molecular mimicry by a Legionella pneumophila Dot/Icm effector

Upon infection, Legionella pneumophila uses the Dot/Icm type IV secretion system to translocate effector proteins from the Legionella-containing vacuole (LCV) into the host cell cytoplasm. The effectors target a wide array of host cellular processes that aid LCV biogenesis, including the manipulation of membrane trafficking. In this study, we used a hidden Markov model screen to identify two novel, non-eukaryotic soluble NSF attachment protein receptor (SNARE) homologs: the bacterial Legionella SNARE effector A (LseA) and viral SNARE homolog A proteins. We characterized LseA as a Dot/Icm effector of L. pneumophila, which has close homology to the Qc-SNARE subfamily. The lseA gene was present in multiple sequenced L. pneumophila strains including Corby and was well distributed among L. pneumophila clinical and environmental isolates. Employing a variety of biochemical, cell biological and microbiological techniques, we found that farnesylated LseA localized to membranes associated with the Golgi complex in mammalian cells and LseA interacted with a subset of Qa-, Qb- and R-SNAREs in host cells. Our results suggested that LseA acts as a SNARE protein and has the potential to regulate or mediate membrane fusion events in Golgi-associated pathways.

Tensor reduction for user profiling in personalized recommender systems

User profiling is the process of constructing user models which represent personal characteristics and preferences of customers. User profiles play a central role in many recommender systems. Recommender systems recommend items to users based on user profiles, in which the items can be any objects which the users are interested in, such as documents, web pages, books, movies, etc. In recent years, multidimensional data are getting more and more attention for creating better recommender systems from both academia and industry. Additional metadata provides algorithms with more details for better understanding the interactions between users and items. However, most of the existing user/item profiling techniques for multidimensional data analyze data through splitting the multidimensional relations, which causes information loss of the multidimensionality. In this paper, we propose a user profiling approach using a tensor reduction algorithm, which we will show is based on a Tucker2 model. The proposed profiling approach incorporates latent interactions between all dimensions into user profiles, which significantly benefits the quality of neighborhood formation. We further propose to integrate the profiling approach into neighborhoodbased collaborative filtering recommender algorithms. Experimental results show significant improvements in terms of recommendation accuracy.

Criminal profiling

One of the more widely recognized and practiced subspecialities within forensic criminology is that of criminal profiling. It has a long history, as detailed in Turvey (2008a). It also boasts a small library of distinct literature, with different methods and subspecialities all its own. Criminal profiling is a practice that has seen increasing popular and media attention over the past several decades. It has been depicted in popular fiction such as films like Silence of the Lambs (1991) and television programs like Criminal Minds (2005– present). It has also been applied in a number of high profile cases, including the “Washington Snipers” (see Turvey and McGrath, 2005, for an extended discussion of profiling and the media in the D.C. Sniper case). As a result, students of criminology commonly express an interest in studying criminal profiling with a view to becoming profilers themselves.

Criminal profiling : behavioural consistency, the homology assumption and case linkage

Criminal profiling is an investigative tool used around the world to infer the personality and behavioural characteristics of an offender based on their crime. Case linkage, the process of determining discreet connections between crimes of the same offender, is a practice that falls under the general banner of criminal profiling and has been widely criticized. Two theories, behavioural consistency and the homology assumption, are examined and their impact on profiling in general and case linkage specifically is discussed...

Green-fluorescent protein facilitates rapid in vivo detection of genetically transformed plant cells

Early detection of plant transformation events is necessary for the rapid establishment and optimization of plant transformation protocols. We have assessed modified versions of the green fluorescent protein (GFP) from Aequorea victoria as early reporters of plant transformation using a dissecting fluorescence microscope with appropriate filters. Gfp-expressing cells from four different plant species (sugarcane, maize, lettuce, and tobacco) were readily distinguished, following either Agrobacterium-mediated or particle bombardment-mediated transformation. The identification of gfp-expressing sugarcane cells allowed for the elimination of a high proportion of non-expressing explants and also enabled visual selection of dividing transgenic cells, an early step in the generation of transgenic organisms. The recovery of transgenic cell clusters was streamlined by the ability to visualize gfp-expressing tissues in vitro.

Profiling unlicensed drivers and riders in Queensland. Report to Queensland Department of Transport and Main Roads

Unlicensed driving remains a serious problem for road safety, despite ongoing improvements in traffic law enforcement practices and technology. This report examines de-identified traffic infringement and sanction histories for drivers in Queensland who had lost their licence between 1st January 2003 and 31st December 2008. A total of 546,117 Queensland drivers were identified. Key areas discussed include the prevalence of unlicensed driving and the extent to which particular offences were detected amongst drivers with a licence sanction or disqualified licence.

Lipid-modified azurin of Neisseria meningitidis is a copper protein localized on the outer membrane surface and not regulated by FNR

The laz gene of Neisseria meningitidis is predicted to encode a lipid-modified azurin (Laz). Laz is very similar to azurin, a periplasmic protein, which belongs to the copper-containing proteins in the cupredoxin superfamily. In other bacteria, azurin is an electron donor to nitrite reductase, an important enzyme in the denitrifying process. It is not known whether Laz could function as an electron transfer protein in this important pathogen. Laz protein was heterologously expressed in Escherichia coli and purified. Electrospray mass spectrometry indicated that the Laz protein contains one copper ion. Laz was shown to be redox-active in the presence of its redox center copper ion. When oxidized, Laz exhibits an intense blue colour and absorbs visible light around 626 nm. The absorption is lost when exposed to diethyldithiocarbamate, a copper chelating agent. Polyclonal antibodies were raised against purified Laz for detecting expression of Laz under different growth conditions and to determine the orientation of Laz on the outer membrane. The expression of Laz under microaerobic and microaerobic denitrifying conditions was slightly higher than that under aerobic conditions. However, the expression of Laz was similar between the wild type strain and an fnr mutant, suggesting that Fumarate/Nitrate reduction regulator (FNR) does not regulate the expression of Laz despite the presence of a partial FNR box upstream of the laz gene. We propose that some Laz protein is exposed on the outer membrane surface of N. meningitidis as the αLaz antibodies can increase killing by complement in a capsule deficient N. meningitidis strain, in a dose-dependent fashion.

The structural basis of DNA binding by the single-stranded DNA-binding protein from Sulfolobus solfataricus

Canonical single-stranded DNA-binding proteins (SSBs) from the oligosaccharide/oligonucleotide-binding (OB) domain family are present in all known organisms and are critical for DNA replication, recombination and repair. The SSB from the hyperthermophilic crenarchaeote Sulfolobus solfataricus (SsoSSB) has a ‘simple’ domain organization consisting of a single DNA-binding OB fold coupled to a flexible C-terminal tail, in contrast with other SSBs in this family that incorporate up to four OB domains. Despite the large differences in the domain organization within the SSB family, the structure of the OB domain is remarkably similar all cellular life forms. However, there are significant differences in the molecular mechanism of ssDNA binding. We have determined the structure of the SsoSSB OB domain bound to ssDNA by NMR spectroscopy. We reveal that ssDNA recognition is modulated by base-stacking of three key aromatic residues, in contrast with the OB domains of human RPA and the recently discovered human homologue of SsoSSB, hSSB1. We also demonstrate that SsoSSB binds ssDNA with a footprint of five bases and with a defined binding polarity. These data elucidate the structural basis of DNA binding and shed light on the molecular mechanism by which these ‘simple’ SSBs interact with ssDNA.

Who are our repeating students? Profiling students persisting after failure

Research on attrition has focused on the economic significance of low graduation rates in terms of costs to students (fees that do not culminate in a credential) and impact on future income. For a student who fails a unit and repeats the unit multiple times, the financial impact is significant and lasting (Bexley, Daroesman, Arkoudis & James 2013). There are obvious advantages for the timely completion of a degree, both for the student and the institution. Advantages to students include fee minimisation, enhanced engagement opportunities, effectual pathway to employment and a sense of worth, morale and cohort-identity benefits. Work undertaken by the QUT Analytics Project in 2013 and 2014 explored student engagement patterns capturing a variety of data sources and specifically, the use of LMS amongst students in 804 undergraduate units in one semester. Units with high failure rates were given further attention and it was found that students who were repeating a unit were less likely to pass the unit than students attempting it for the first time. In this repeating cohort, academic and behavioural variables were consistently more significant in the modelling than were any demographic variables, indicating that a student’s performance at university is far more impacted by what they do once they arrive than it is by where they come from. The aim of this poster session is to examine the findings and commonalities of a number of case studies that articulated the engagement activities of repeating students (which included collating data from Individual Unit Reports, academic and peer advising programs and engagement with virtual learning resources). Understanding the profile of the repeating student cohort is therefore as important as considering the characteristics of successful students so that the institution might be better placed to target the repeating students and make proactive interventions as early as possible.

Importance of literature profiling: An archival analysis with illustrative examples for IS researchers

The importance of a thorough and systematic literature review has long been recognised across academic domains as critical to the foundation of new knowledge and theory evolution. Driven by an exponentially growing body of knowledge in the IS discipline, there has been a recent influx of guidance on how to conduct a literature review. As literature reviews are emerging as a standalone research method in itself, increasingly these method focused guidelines are of great interest, receiving acceptance at top tier IS publication outlets. Nevertheless, the finer details which offer justification for the selected content, and the effective presentation of supporting data has not been widely discussed in these method papers to date. This paper addresses this gap by exploring the concept of ‘literature profiling’ while arguing that it is a key aspect of a comprehensive literature review. The study establishes the importance of profiling for managing aspects such as quality assurance, transparency and the mitigation of selection bias. And then discusses how profiling can provide a valid basis for data analysis based on the attributes of selected literature. In essence, this study has conducted an archival analysis of literature (predominately from the IS domain) to present its main argument; the value for literature profiling, with supporting exemplary illustrations.

Architecture for service profiling

Service oriented architecture is gaining momentum. However, in order to be successful, the proper and up-to-date description of services is required. Such a description may be provided by service profiling mechanisms, such as one presented in this article. Service profile can be defined as an up-to-date description of a subset of non-functional properties of a service. It allows for service comparison on the basis of non-functional parameters, and choosing the service which is most suited to the needs of a user. In this article the notion of a service profile along with service profiling mechanism is presented as well as the architecture of a profiling system. © 2006 IEEE.

Evaluation of protein bait laced with various insecticides on the Queensland fruit fly (Diptera: Tephritidae): Attraction, feeding, mortality and bait persistence

The use of malathion in fruit fly protein bait sprays has raised serious concerns due to its adverse effects on non-target organisms. This has necessitated the evaluation of novel reduced-risk compounds. This study evaluated the effects of spinosad, fipronil, malathion and chlorpyrifos mixed with fruit fly protein bait (Mauri Pinnacle protein®) on attraction, feeding and mortality of the Queensland fruit fly, Bactrocera tryoni (Froggatt). The effects of outdoor weathering of these mixtures on fly mortality were also determined. In field-cage experiment, protein-starved flies showed the same level of attraction to baits containing spinosad, fipronil, malathion, chlorpyrifos and protein alone used as control. Female protein-starved flies were deterred from feeding on baits containing malathion and chlorpyrifos compared to baits containing spinosad, fipronil and protein alone. Baits containing malathion and chlorpyrifos caused higher fly mortality and rapid fly knock down than spinosad and fipronil. However, spinosad acted slowly and caused an increase in fly mortality over time, causing up to 90% fly mortality after 72-h. Baits containing malathion and chlorpyrifos, applied on citrus leaves and weathered outdoors, had longer residual effectiveness in killing flies than spinosad and fipronil. Residual effectiveness of the spinosad bait mixture waned significantly after 3 days of outdoor weathering. Results suggest that spinosad and fipronil can be potential alternatives for malathion in protein bait sprays.

Autosomal dominant familial calcium pyrophosphate dihydrate deposition disease is caused by mutation in the transmembrane protein ANKH

Familial autosomal dominant calcium pyrophosphate dihydrate (CPPD) chondrocalcinosis has previously been mapped to chromosome 5pl5. We have identified a mutation in the ANKH gene that segregates with the disease in a family with this condition. ANKH encodes a putative transmembrane inorganic pyrophosphate (PPi) transport channel. We postulate that loss of function of ANKH causes elevated extracellular PPi levels, predisposing to CPPD crystal deposition.