Redesigning gfp Reporter System for Utilization in Clostridium Difficile

Clostridium difficile (C. difficile) is a gram-positive bacterium that comprises part of the healthy human gut microbiome. When it gains sufficient access to peptides, C. difficile flourishes and releases tissue-damaging toxins, which cause inflammation of the colon that can develop into a Clostridium difficile Infection (CDI).10 The Ivey Laboratory believes that the best tactic in preventing CDIs is stopping peptide ingestion, which theoretically could be accomplished by manipulating the oligopeptide permease (App) system.7 In order to verify that altering the App system would successfully impede peptide uptake, first the expression of the app Promoter Region (appProR) of C. difficile’s DNA needs to be better understood. This characterization can be accomplished by fusing appProR to the gfp-reporter gene, which codes for Green Fluorescent Protein (GFP). GFP emits green fluorescent light when exposed to blue or ultraviolet light, and the degree of fluorescence can be used to quantify the gene expression of whatever DNA sequence to which the gfp-reporter gene is fused.9 The specific aim of this project was to incorporate the appProR-gfp-reporter gene complex first into Eschericheria coli (E. coli), and then into Bacillus subtilis (B. subtilis). Those two bacterial species were chosen as hosts for the transformations, for E. coli and B. subtilis are known for being more receptive to recombinant DNA techniques than C. difficile.22 By ligating the appProR-gfp-reporter gene sequence of pUA321 to pG+host4, the resulting plasmid, pUA625, contained a broad enough host range to transform both gram-negative E. coli and gram-positive B. subtilis. Those successful transformations indicate that pUA625 could be integrated into C. difficile in the future, an achievement which would lead to a better understanding of the expression of C. difficile’s App system

Beyond Marbury : Jurisdictional Self-Dealing In Seminole Tribe

In Seminole Tribe v. Florida, the Supreme Court held that the Constitution\u27s Article III embodies a principle of state sovereign immunity which so constrains the federal judicial power that it prohibits Congress from granting federal courts subject matter jurisdiction over private lawsuits to enforce Article I legislation against states. At the same time, however, and again in Idaho v. Coeur d\u27Alene Tribe, the Court reaffirmed its own Ex parte Young doctrine, under which the Court itself unilaterally granted federal courts subject matter jurisdiction over private lawsuits to coerce states to comply with federal law despite state sovereign immunity. Neither in Seminole Tribe nor in Coeur d\u27Alene Tribe did the Court explain how Article HI could so limit Congress\u27s power to grant subject matter jurisdiction in such lawsuits, while leaving intact the Court\u27s own power to do so under Ex parte Young. Arguing that Ex parte Young and the Young doctrine rest on no affirmative principle of law, Professor Fitzgerald suggests that Young\u27s survival of Seminole Tribe reflects the Court\u27s claim for itself of a freestanding judicial power to regulate access to federal courts for private lawsuits challenging state interests--a judicial power that now outstrips both ordinary judicial review and also Congress\u27s constitutional authority over the federal courts

Suspecting the States: Supreme Court Review of State-Court State-Law Judgments

At the Supreme Court these days, it is unfashionable to second-guess states\u27 fealty to federal law without real proof that they are ignoring it. As the Court declared in Alden v. Maine: We are unwilling to assume the States will refuse to honor the Constitution or obey the binding laws of the United States. The good faith of the States thus provides an important assurance that \u27this Constitution, and the Laws of the United States which shall be made in Pursuance thereof ... shall be the supreme Law of the Land.\u27 Accordingly, without proof that a state has systematic[ally] shirked its supremacy clause duty to honor Article I legislation, the Court appears unwilling to enforce compliance in a particular case. Likewise, the Court makes Congress prove a broad pattern of federal-law transgressions by many states before it can hold any state accountable to individuals for violating their constitutional rights. Indeed, a handful of even egregious anecdotes simply cannot overcome the Court\u27s presumption that all states can be trusted to meet their federal-law duties

Structure and Behaviour of Virtual Organisation Breeding Environments

This paper provides an outline of a formal approach that we are developing for modelling Virtual Organisations (VOs) and their Breeding Environments (VBEs). We propose different levels of representation for the functional structures and processes that VBEs and VOs involve, which are independent of the specificities of the infrastructures (organisational and technical) that support the functioning of VBEs. This allows us to reason about properties of tasks performed within VBEs and services provided through VOs without committing to the way in which they are implemented

The application of microfiltration as a partial sterilisation technique for the reduction of psychrotrophic spore forming bacteria from viscous dairy feeds

The use of microfiltration as an alternative to pasteurisation to reduce the microbial load of raw skimmed milk is a well established technology. However, its application in reducing bacteria from highly viscous dairy based solutions has not due to issues of low flux and high fouling tendency. This study involves the application of microfiltration to remove spores from high solids content Milk Protein Isolate (MPI) solutions. MPI feeds were inoculated with Bacillus mycoides spores a safer alternative to Bacillus cereus, a psychrotrophic spore forming bacteria found in dairy feeds. Suitable protocols for MPI resolubilisation, Bacillus mycoides cell and spore preparations were established and the membranes, MPI and spores were fully characterised by scanning electron microscopy (SEM), particle size distribution, rheology and pure water flux (PWF) measurements. Feed and permeate samples collected during experiments were analysed for solids content by oven drying, protein content using the Bradford assay and spore content using PetrifilmTM Aerobic count plates. To try and determine an optimum protocol for MPI filtration, a variety of filtration rig set-ups, modules and membranes were tested. Experiments were carried out at different MPI concentrations (4 – 16 wt%), cross flow velocities (CFV’s) (0.7 – 2.0 m s-1) and transmembrane pressures (TMP’s ) (1 and 2 bar). The filtration of 15 wt% MPI proved challenging. The best set of results were obtained using the 12.0 μm membrane at 1.4 m s-1, producing a 27 LMH flux, 96.5% protein transmission and a 2.1 log spore reduction. These results indicate that large pore ceramic microfiltration may be a suitable technology to replace or augment pasteurisation for high solids content dairy feeds. The effect of backwashing using different durations and frequencies was investigated. Backwashing parameters of 10 seconds every 5 minutes at 1 bar were found to be the most effective. The optimum cleaning regime found for MPI fouled ceramic membranes involved a long rinsing backflush at 1 bar, acid and alkali steps without backwashing, which produced a 99.6% flux recovery.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Blended Stakeholder Participation for Responsible Information Systems Research

Researchers often conduct information systems (IS) research under the assumption that technology use leads to positive outcomes for different stakeholders. However, many IS studies demonstrate limited evidence of having engaged with the stakeholders that they claim benefit and speak on behalf of. Therefore, we can unsurprisingly find many examples in which technology use failed to make the world a better place or, worse still, contributed towards unintended negative outcomes. Given these concerns, calls have recently emerged for responsible research and innovation (RRI) studies in IS to understand how different stakeholder groups can have a voice in complex socio-technical issues. In this paper, we take steps towards addressing this call by presenting case study findings from a responsible IS research project that combined “blended” face-to-face and online participatory techniques. The case study relates to a large-scale consultation in a 24-month European project that involved 30 countries. The project engaged over 1,500 stakeholders in co-creating future research agendas for the European Union. We discuss case study findings using Stilgoe, Owen, and Macnaghten’s (2013) framework and reflect on lessons learned for responsible IS research going forward

Student Ensemble: Chamber Music Cor-tet

Kemp Recital HallNovember 5, 2015Thursday Evening8:00 p.m

Cognitive Processes and Memory Differences in Recall and Recognition in Adults

Eyewitness testimony is critical in both criminal court and civil court, so determining the most reliable method to gain information from witnesses is imperative. Past research in this area has focused on false memory, assisted recall, stress, and event perception. A gap exists in the current literature regarding the best method to gain the most accuracy in recall. The purpose of this study was to evaluate free recall, cued recall, and recognition, in an attempt to examine the accuracy of eyewitness memory. The study utilized a quantitative design to assess the accuracy of eyewitness memory as measured by results on free-recall, cued recall, and recognition tests. The theoretical foundation for this study was the theory of information processing, which contends that information is processed in stages and combines visual cognition, memory, and memory recall; therefore, this theory applies to the study by helping determine the most accurate way for individuals to recall events. Introduction to Psychology students were shown a video, then asked to recall what they saw using either free recall, cued recall, or recognition. A one-way between-subjects analysis of variance was utilized to determine whether there were significant differences in the number of items recalled as a function of recall format. Results suggested that participants were more accurate with the utilization of recognition techniques for recall, as opposed to the free or cued-recall. The importance of evaluating effective methods to promote accurate eyewitness testimony is to advance forensic science. The implications for social change include the ability to have more effective methods to gain accurate eye-witness testimony, thereby assisting with proper outcomes during trials

Fire Effects on Three Trophic Levels in a Central Arkansas Grassland

We studied the effect of a late growing-season fire on the plant and foliar arthropod communities in a naturally occurring grassland. In central Arkansas, these grasslands are common on south-facing slopes where shallow soils and hot/dry weather conditions during the summer cannot support the growth of a forest community. Patches of grassland were burned in the autumn (4 November, late growing season), often the time of natural fires in Arkansas, and compared to unburned areas. Fire increased the biomass of forbs and decreased the biomass of grasses, although overall biomass was not different between treatments. Among the foliar arthropods, herbivores were significantly reduced by burning, especially the Homoptera. Carnivorous arthropods as a whole were not affected by burning, although spiders showed a small but significant reduction. The response of arthropods to fire occurred almost one year after the burn, showing that fire effects can be delayed for a substantial period of time. This experiment shows that fire occurring during the natural burning period in Arkansas can have substantial effects on grasslands communities. The response of plants in Arkansas is similar to that of plants in nearby grasslands on the Great Plains and southeastern United States which also show a great increase in forbs under late growing season burning regimes. The changes seen in this experiment demonstrate that the suppression of fire by humans has probably modified the structure of Arkansas grasslands. With the increasing use of fire as a management tool in Arkansas, changes to grassland systems are likely to be profound