Feasibility of Inducing Overlap Immunologic Competence in Gallinaceous Birds with Ascardia dissimilis and A. galli

Chickens and turkeys are routinely infected with the roundworms Ascaridia galli and A. dissimilis, respectively. The current study was conducted to gather basic information on these worms and to determine whether heterologous infections (chicken worms in turkeys and turkey worms in chickens) would be successful. Chickens and turkeys were obtained at day of hatch, brooded to 7 days of age, and placed in pens (25/pen) according to infection as received at 7 days of age: homologous, heterologous and control (no infection). Bird weights, mortalities, and feed efficiencies were monitored for 3 weeks postinfection, at which time all birds were killed for parasite collection and counting. Feed efficiency, a parameter more adequately measured in large-scale studies, did not vary between experimental groups. Bird weights at necropsy varied significantly (P \u3c 0.05) between groups only for the turkeys, with homologous infection (A. dissimilis) birds weighing less than controls. All induced, homologous, and heterologous infections were successful. Rates of establishment, however, were significantly (P \u3c 0.05) depressed for each heterologous model. Total A. dissimilis numbers were only 55% as great as those for A. galli in chickens [geometric means (GMs) of 13.2 versus 24.2], and total A. galli numbers were only 56% as great as A. dissimilis numbers in turkeys (GMs of 5.6 versus 10.0). Given the fact that heterologous infections were successful, albeit reduced, in both types of birds (infections that included tissue-phase forms), additional studies are planned to determine whether these infections might induce interspecies (overlapping) immune competence in the host and aid in reducing natural parasitisms to levels with no economic impact

Malaria intervention scale-up in Africa : effectiveness predictions for health programme planning tools, based on dynamic transmission modelling

Scale-up of malaria prevention and treatment needs to continue to further important gains made in the past decade, but national strategies and budget allocations are not always evidence-based. Statistical models were developed summarizing dynamically simulated relations between increases in coverage and intervention impact, to inform a malaria module in the Spectrum health programme planning tool.; The dynamic Plasmodium falciparum transmission model OpenMalaria was used to simulate health effects of scale-up of insecticide-treated net (ITN) usage, indoor residual spraying (IRS), management of uncomplicated malaria cases (CM) and seasonal malaria chemoprophylaxis (SMC) over a 10-year horizon, over a range of settings with stable endemic malaria. Generalized linear regression models (GLMs) were used to summarize determinants of impact across a range of sub-Sahara African settings.; Selected (best) GLMs explained 94-97 % of variation in simulated post-intervention parasite infection prevalence, 86-97 % of variation in case incidence (three age groups, three 3-year horizons), and 74-95 % of variation in malaria mortality. For any given effective population coverage, CM and ITNs were predicted to avert most prevalent infections, cases and deaths, with lower impacts for IRS, and impacts of SMC limited to young children reached. Proportional impacts were larger at lower endemicity, and (except for SMC) largest in low-endemic settings with little seasonality. Incremental health impacts for a given coverage increase started to diminish noticeably at above ~40 % coverage, while in high-endemic settings, CM and ITNs acted in synergy by lowering endemicity. Vector control and CM, by reducing endemicity and acquired immunity, entail a partial rebound in malaria mortality among people above 5 years of age from around 5-7 years following scale-up. SMC does not reduce endemicity, but slightly shifts malaria to older ages by reducing immunity in child cohorts reached.; Health improvements following malaria intervention scale-up vary with endemicity, seasonality, age and time. Statistical models can emulate epidemiological dynamics and inform strategic planning and target setting for malaria control

Functionalizing conjugative systems to deliver CRISPR nucleases for targeted bacterial killing

The interactions between humans and microbes are intimately important to human health, with both commensal and pathogenic bacteria affecting homeostasis and disease. Increasing concern over antibiotic resistance in bacterial pathogens represents a significant threat to human health, and use of traditional antibiotics to treat infections can be detrimental to commensal bacteria as well as pathogens, demonstrating a need for more specific antibacterial reagents. RNA-guided CRISPR nucleases, which can target and cleave genomes of interest, are a potential tool for specific bacterial targeting. A key limitation to the use of CRISPR antimicrobials is effective and robust delivery to the target bacteria. My thesis addresses this key issue by functionalizing conjugative systems to deliver a CRISPR nuclease for bacterial killing. First, a plasmid containing an arabinose-inducible TevCas9 nuclease that is mobilizable in-trans by an RK2-based conjugative system was constructed. Inclusion of the RK2-based conjugative system in-cis on the same plasmid was shown to greatly increase the conjugation frequency over time. Furthermore, when conjugating in liquid we observed that providing glass beads to increase surface area for biofilm development and cell-cell contact significantly improved conjugation frequency. Crucially, conjugated TevCas9 was able to kill Salmonella enterica with up to 99% efficiency, depending on the sgRNA provided. Next, to explore the importance of conjugative systems for delivery, a database containing thousands of conjugative systems identified from gut metagenomic data was constructed. From this database, a conjugative system of 54 kb native to the Citrobacter genus was constructed de novo. This conjugative plasmid, p20298-15a, showed 30-fold increased conjugation frequency to Citrobacter rodentium than to Escherichia coli, and was capable of conjugation to several additional Citrobacter strains. The p20298-15a plasmid was then functionalized to clone the arabinose-inducible CRISPR-TevCas9 system, which was able to target and kill C. rodentium. Importantly, the construction and engineering of p20298 shows that large genetic systems found in metagenomic data sets can be synthesized and functionalized. Overall, this thesis demonstrates the effective use of conjugative systems as a delivery mechanism for CRISPR-based antimicrobials for the targeted killing of bacteria

Finding the trigger to Iapetus' odd global albedo pattern: Dynamics of dust from Saturn's irregular satellites

The leading face of Saturn's moon Iapetus, Cassini Regio, has an albedo only one tenth that on its trailing side. The origin of this enigmatic dichotomy has been debated for over forty years, but with new data, a clearer picture is emerging. Motivated by Cassini radar and imaging observations, we investigate Soter's model of dark exogenous dust striking an originally brighter Iapetus by modeling the dynamics of the dark dust from the ring of the exterior retrograde satellite Phoebe under the relevant perturbations. In particular, we study the particles' probabilities of striking Iapetus, as well as their expected spatial distribution on the Iapetian surface. We find that, of the long-lived particles (greater than about 5 microns), most particle sizes (greater than about 10 microns) are virtually certain to strike Iapetus, and their calculated distribution on the surface matches up well with Cassini Regio's extent in its longitudinal span. The satellite's polar regions are observed to be bright, presumably because ice is deposited there. Thus, in the latitudinal direction we estimate polar dust deposition rates to help constrain models of thermal migration invoked to explain the bright poles (Spencer & Denk 2010). We also analyze dust originating from other irregular outer moons, determining that a significant fraction of that material will eventually coat Iapetus--perhaps explaining why the spectrum of Iapetus' dark material differs somewhat from that of Phoebe. Finally we track the dust particles that do not strike Iapetus, and find that most land on Titan, with a smaller fraction hitting Hyperion. As has been previously conjectured, such exogenous dust, coupled with Hyperion's chaotic rotation, could produce Hyperion's roughly isotropic, moderate-albedo surface.Comment: Accepted for publication in Icaru

A Simple Method for Synchronising Multiple IMUs Using the Magnetometer

This paper presents a novel method to synchronise multiple IMU (inertial measurement units) devices using their onboard magnetometers. The method described uses an external electromagnetic pulse to create a known event measured by the magnetometer of multiple IMUs and in turn used to synchronise these devices. The method is applied to 4 IMU devices decreasing their de-synchronisation from 270ms when using only the RTC (real time clock) to 40ms over a 1 hour recording. It is proposed that this can be further improved to approximately 3ms by increasing the magnetometer’s sample frequency from 25Hz to 300Hz

Neuron-Specific HuR-Deficient Mice Spontaneously Develop Motor Neuron Disease

Human Ag R (HuR) is an RNA binding protein in the ELAVL protein family. To study the neuron-specific function of HuR, we generated inducible, neuron-specific HuR-deficient mice of both sexes. After tamoxifen-induced deletion of HuR, these mice developed a phenotype consisting of poor balance, decreased movement, and decreased strength. They performed significantly worse on the rotarod test compared with littermate control mice, indicating coordination deficiency. Using the grip-strength test, it was also determined that the forelimbs of neuron-specific HuR-deficient mice were much weaker than littermate control mice. Immunostaining of the brain and cervical spinal cord showed that HuR-deficient neurons had increased levels of cleaved caspase-3, a hallmark of cell apoptosis. Caspase-3 cleavage was especially strong in pyramidal neurons and α motor neurons of HuR-deficient mice. Genome-wide microarray and real-time PCR analysis further indicated that HuR deficiency in neurons resulted in altered expression of genes in the brain involved in cell growth, including trichoplein keratin filament-binding protein, Cdkn2c, G-protein signaling modulator 2, immediate early response 2, superoxide dismutase 1, and Bcl2. The additional enriched Gene Ontology terms in the brain tissues of neuron-specific HuR-deficient mice were largely related to inflammation, including IFN-induced genes and complement components. Importantly, some of these HuR-regulated genes were also significantly altered in the brain and spinal cord of patients with amyotrophic lateral sclerosis. Additionally, neuronal HuR deficiency resulted in the redistribution of TDP43 to cytosolic granules, which has been linked to motor neuron disease. Taken together, we propose that this neuron-specific HuR-deficient mouse strain can potentially be used as a motor neuron disease model

Neurodegenerative brain changes are associated with area deprivation in the United Kingdom: findings from the Brains for Dementia Research study

Socioeconomic disadvantage is associated with greater risk of dementia. This has been theorised to reflect inequalities in cognitive reserve, healthcare access, lifestyle, and other health factors which may contribute to the clinical manifestation of dementia. We aimed to assess whether area deprivation in the United Kingdom was associated with greater risk or severity of the specific neurodegenerative diseases which lead to dementia in a multi-centre cohort with autopsy assessment. Participants underwent clinical assessment prior to brain tissue donation post-mortem. Each then underwent detailed, standardised neuropathological assessment. National area deprivation statistics were derived for each participant’s neighbourhood, for use as a predictor in binary and ordinal logistic models assessing the respective presence and severity of staging of key neuropathological changes, adjusting for theorised confounders. Individuals from among the 20% most deprived neighbourhoods in the United Kingdom had significantly higher neurofibrillary tangle and neuritic plaque staging, and increased risk of cerebral amyloid angiopathy. These findings were not explained by a greater risk of diabetes or hypertension, APOE genotype, alcohol misuse or tobacco smoking, sex, or age differences. A sensitivity analysis conditioning on baseline cognitive impairment did not meaningfully change the observed association. Socioeconomic disadvantage may contribute to dementia incidence through a greater severity of specific neuropathological changes (neurofibrillary tangles, neuritic plaques, and cerebral amyloid angiopathy), independent of other indirect influences. Mechanisms through which deprivation is associated with these require further exploration

Testing for Vertical Inequity in Property Tax Systems

Models for testing assessor performance have been widely discussed in the literature. Many have been used in practice. The purpose of this study are to evaluate the performance of existing models and to propose two new models. We find that existing models can be used correctly to test for inequity when their functional form is consistent with the pattern of the assessment-sales ratio data. Results from the application of different models show inconsistencies since the appropriate functional form may vary for different data sets. The new models have the ability to emulate the forms of the existing models as well as handle more complex relationships.