Acute phase proteins and biomarkers for health in chickens

Acute phase proteins (APPs) are proteins synthesised predominantly in the liver, whose plasma concentrations increase (positive APP) or decrease (negative APP) as a result of infection, inflammation, trauma and tissue injury. They also change as a result of the introduction of immunogens such as bacterial lipopolysaccharide (LPS), turpentine and vaccination. While publications on APPs in chickens are numerous, the limited availability of anti-sera and commercial ELISAs has resulted in a lot of information on only a few APPs. Disease is a threat to the poultry industry, as pathogens have the potential to evolve, spread and cause rapid onset of disease that is detrimental to the welfare of birds. Low level, sub-acute disease with non-specific, often undiagnosed causes can greatly affect bird health and growth and impact greatly on productivity and profitability. Developing and validating methods to measure and characterise APPs in chickens will allow these proteins to be used diagnostically for monitoring flock health. Using immune parameters such as APPs that correlate with disease resistance or improvements in production and welfare will allow the use of APPs as selection parameters for breeding to be evaluated. For APPs to be useful parameters on which to evaluate chicken health, information on normal APP concentrations is required. Ceruloplasmin (Cp) and PIT54 concentrations were found to be much lower in healthy birds form commercial production farms than the reported normal values obtained from the literature. These APPs were found to be significantly higher in culled birds from a commercial farm and Cp, PIT54 and ovotransferrin (Ovt) were significantly higher in birds classified as having obvious gait defects. Using quantitative shotgun proteomics to identify the differentially abundant proteins between three pools: highly acute phase (HAP), acute phase (AP) and non-acute phase (NAP), generated data from which a selection of proteins, based on the fold difference between the three pools was made. These proteins were targeted on a individual samples alongside proteins known to be APPs in chickens or other species: serum amyloid A (SAA), C-reactive protein (CRP), Ovt, apolipoprotein A-I (apo-AI), transthyretin (Ttn), haemopexin (Hpx) and PIT54. Together with immunoassay data for SAA, Ovt, alpha-1-acid glycoprotein (AGP) and Cp the results of this research reveal that SAA is the only major APP in chickens. Ovotransferrin and AGP behave as moderate APPs while PIT54 and Cp are minor APPs. Haemopexin was not significantly different between the three acute phase groups. Apolipoprotein AI and Ttn were significantly lower in the HAP and AP groups and as such can be classed as negative APPs. In an effort to identify CRP, multiple anti-sera cross reacting with CRP from other species were used and a phosphorylcholine column known to affinity purify CRP were used. Enriched fractions containing low molecular weight proteins, elutions from the affinity column together with HAP, AP and NAP pooled samples were applied to a Q-Exactive Hybrid Quadrupole–Orbitrap mass spectrometer (Thermo Scientific) for Shotgun analysis and CRP was not identified. It would appear that CRP is not present as a plasma protein constitutively or during an APR in chickens and as such is not an APP in this species. Of the proteins targeted as possible novel biomarkers of the APR in chickens mannan binding lectin associated serine protease-2, α-2-HS-glycoprotein (fetuin) and major facilitator superfamily domain-containing protein 10 were reduced in abundance in the HAP group, behaving as negative biomarkers. Myeloid protein and putative ISG(12)2 were positively associated with the acute phase being significantly higher in the HAP and AP groups. The protein cathepsin D was significantly higher in both HAP and AP compared to the NAP indicating that of all the proteins targeted, this appears to have the most potential as a biomarker of the acute phase, as it was significantly increased in the AP as well as the HAP group. To evaluate APPs and investigate biomarkers of intestinal health, a study using re-used poultry litter was undertaken. The introduction of litter at 12 days of age did not significantly increase any APPs measured using immunoassays and quantitative proteomics at 3, 6 and 10 days post introduction. While no APP was found to be significantly different between the challenged and control groups at anytime point, the APPs AGP, SAA and Hpx did increase over time in all birds. The protein apolipoprotein AIV (apo-AIV) was targeted as a possible APP and because of its reported role in controlling satiety. An ELISA was developed, successfully validated and used to measure apo-AIV in this study. While no significant differences in apo-AIV plasma concentrations between challenged and control groups were identified apo-AIV plasma concentrations did change significantly between certain time points in challenged and control groups. Apoliporotein AIV does not appear to behave as an APP in chickens, as it was not significantly different between acute phase groups. The actin associated proteins villin and gelsolin were investigated as possible biomarkers of intestinal health. Villin was found not to be present in the plasma of chickens and as such not a biomarker target. Gelsolin was found not to be differentially expressed during the acute phase or as a result of intestinal challenge. Finally a proteomic approach was undertaken to investigate gastrocnemius tendon (GT) rupture in broiler chickens with a view of elucidating to and identify proteins associated with risk of rupture. A number of proteins were found to be differentially expressed between tendon pools and further work would enable further detailing of these findings. In conclusion this work has made a number of novel findings and addressed a number of data poor areas. The area of chicken APPs research has stagnated over the last 15 years with publications becoming repetitive and reliant on a small number of immunoassays. This work has sought to characterise the classic APPs in chickens, and use a quantitative proteomic approach to measure and categorise them. This method was also used to take a fresh approach to biomarker identification for both the APR and intestinal health. The development and validation of assays for Ovt and apo-AIV and the shotgun data mean that these proteins can be further characterised in chickens with a view of applying their measurement to diagnostics and selective breeding programs.

Acute Transient Stress Induced Adrenal Hypertrophy and Adrenal Medullary Hyperactivity

Objectives: Adrenal gland hypertrophy can be related to acute stress with abnormal adrenal function tests. It may not always need treatment. Material and methods: An acute presentation of adrenal gland hypertrophy following an abdominal emergency, with subsequent hypoadrenalism was investigated. Results: Adrenal medullary and cortical function fully recovered without treatment. Conclusions: We postulate that the adrenal glands became enlarged and hypertrophied during an acute stress event, possibly caused by acute adrenal medullary hypersecretion and subsequent cortical hyposecretion. A wait and watch policy should be followed if no other clinical symptoms and signs of adrenal disease are present. CT scan remains an important diagnostic tool.

Making a home: the difficulty of dwelling in Genesis and its intertexts

Using an inter-disciplinary range of research on the home-space, home-making practices and the concept of ‘dwelling’, I achieve a new understanding of a central thematic concern in Genesis: its characters’ struggle to build stable, lasting homes upon the earth. Genesis starts with a lost home-space named Eden, before progressing towards other temporary dwellings such as the ark Noah builds, and Abraham’s tents. The biblical ‘home’ is constructed from a mix of materials: the birth of children, divine instructions and journeys, dreams, homemaking acts and so on. Alongside social scientific criticism, this thesis uses literary and midrashic intertexts as a way into re-imagining the ‘unhomely’ experiences of certain characters, or drawing out tensions in acts such as home-unmaking or homecomings. The investigation of the concept of ‘home’ in Genesis contributes to the study of this space more widely, as well as reinterpreting familiar biblical themes such as identity, family and community.

Hypoglyceamia in a Patient with a Solitary Fibrous Tumour

Objective: To illustrate an unusual mechanism causing hypoglycaemia. Material and methods: A 76-year-old man presented with episodes of agitation and confusion and was resuscitated with oral glucose gel when found to be hypoglycaemic. Results: A CT scan for an abdominal mass confirmed a solitary fibrous tumour (SFT). The sarcoma multidisciplinary team suggested conservative management. The patient's episodic hypoglycaemia was managed with diet modification including corn-based starch, scheduled snacks and dexamethasone. Glucose levels were within normal range at discharge from hospital. The patient was referred to the palliative care team for follow-up. Conclusion: SFTs causing non-islet cell tumour hypoglycaemia are difficult to treat. KEYWORDS

Accumulation de dose à partir de champs de déformation 4D appliqués aux traitements au CyberKnife et à l'IMRT

Le cancer pulmonaire est la principale cause de décès parmi tous les cancers au Canada. Le pronostic est généralement faible, de l'ordre de 15% de taux de survie après 5 ans. Les déplacements internes des structures anatomiques apportent une incertitude sur la précision des traitements en radio-oncologie, ce qui diminue leur efficacité. Dans cette optique, certaines techniques comme la radio-chirurgie et la radiothérapie par modulation de l'intensité (IMRT) visent à améliorer les résultats cliniques en ciblant davantage la tumeur. Ceci permet d'augmenter la dose reçue par les tissus cancéreux et de réduire celle administrée aux tissus sains avoisinants. Ce projet vise à mieux évaluer la dose réelle reçue pendant un traitement considérant une anatomie en mouvement. Pour ce faire, des plans de CyberKnife et d'IMRT sont recalculés en utilisant un algorithme Monte Carlo 4D de transport de particules qui permet d'effectuer de l'accumulation de dose dans une géométrie déformable. Un environnement de simulation a été développé afin de modéliser ces deux modalités pour comparer les distributions de doses standard et 4D. Les déformations dans le patient sont obtenues en utilisant un algorithme de recalage déformable d'image (DIR) entre les différentes phases respiratoire générées par le scan CT 4D. Ceci permet de conserver une correspondance de voxels à voxels entre la géométrie de référence et celles déformées. La DIR est calculée en utilisant la suite ANTs («Advanced Normalization Tools») et est basée sur des difféomorphismes. Une version modifiée de DOSXYZnrc de la suite EGSnrc, defDOSXYZnrc, est utilisée pour le transport de particule en 4D. Les résultats sont comparés à une planification standard afin de valider le modèle actuel qui constitue une approximation par rapport à une vraie accumulation de dose en 4D.

Happy and Other Stories

The characters in “Happy,” “The Price of Independence,” “Itch,” and “Guillotine” all struggle with their loss of power and ability to successfully navigate their own lives. Though different genders, ages, and worlds are rendered each character must choose to either face their conflict head on or submit to the external pressure present. “The Price of Independence” and “Itch” highlight the precariousness of relationships and how one relationship, whether it be romantic or platonic, can change everything. “Happy” and “Guillotine” feature characters struggling from within, they are separated from the world around them and their failures force a spotlight on the misconceptions of mental health care.

Dysautonomia: an Explanation for the Medically Unexplained?

Objective: To identify dysautonomia as a collection of conditions with variable presentation that may be mistaken for medically unexplained symptoms. Methods: Case series. Results: Tilt table testing and 24 h electrocardiographic monitoring provided useful diagnostic tools to confirm the diagnosis. Conclusion: A greater awareness and recognition of the disorders that result from dysautonomia and recognition of the disability that results from these disorders will improve patients’ quality of life.

Investigating novel approaches to the detection of virus neutralising antibodies to rabies and Rift Valley fever virus

Serosurveillance is a powerful tool fundamental to understanding infectious disease dynamics. The presence of virus neutralising antibody (VNAb) in sera is considered the best evidence of infection, or indeed vaccination, and the gold standard serological assay for their detection is the virus neutralisation test (VNT). However, VNTs are labour intensive, costly and time consuming. In addition, VNTs for the detection of antibodies to highly pathogenic viruses require the use of high containment facilities, restricting the application of these assays to the few laboratories with adequate facilities. As a result, robust serological data on such viruses are limited. In this thesis I develop novel VNTs for the detection of VNAb to two important, highly pathogenic, zoonotic viruses; rabies and Rift Valley fever virus (RVFV). The pseudotype-based neutralisation test developed in this study allows for the detection of rabies VNAb without the requirement for high containment facilities. This assay was utilised to investigate the presence of rabies VNAb in animals from a variety of ecological settings. In this thesis I present evidence of natural rabies infection in both domestic dogs and lions from rabies endemic settings. The assay was further used to investigate the kinetics of VNAb response to rabies vaccination in a cohort of free-roaming dogs. The RVFV neutralisation assay developed herein utilises a recombinant luciferase expressing RVFV, which allows for rapid, high-throughput serosurveillance of this important neglected pathogen. In this thesis I present evidence of RVFV infection in a variety of domestic and wildlife species from Northern Tanzania, in addition to the detection of low-level transmission of RVFV during interepidemic periods. Additionally, the investigation of a longitudinal cohort of domestic livestock also provided evidence of rapid waning of RVF VNAb following natural infection. Collectively, the serological data presented in this thesis are consistent with existing data in the literature generated using the gold standard VNTs. Increasing the availability of serological assays will allow the generation of robust serological data, which are imperative to enhancing our understanding of the complex, multi-host ecology of these two viruses.

Test Anxiety, Coping, Gender and Academic Performance in English Exams: A Study of Finnish Upper Secondary School Students

In this MA thesis, test anxiety related to English exams among Finnish upper secondary school students was studied. In addition, the ways students try to cope with test anxiety were investigated. The purpose of the study was to investigate gender differences in test anxiety, the effects of test anxiety on academic performance and relationships between test anxiety, academic performance and coping strategies. Test anxiety and coping strategies were analysed as scores of questionnaire responses. Coping strategies comprised of three categories – task-orientation and preparation, seeking social support and avoidance. Academic performance was analysed as teacher ratings of general performance in English exams. In total 67 subjects were studied. The subjects were Finnish general upper secondary school students. The data were collected by using online questionnaires. This data were mainly quantitative, but also qualitative elements were included. The quantitative data were analysed by using statistical methods. The results showed that females experienced statistically significantly more test anxiety than males. In addition, a statistically significant correlation was found between test anxiety levels and academic performance ratings of the subjects: the higher the test anxiety score, the lower the academic performance rating. A meaningful correlation was found between test anxiety and seeking social support as a coping strategy: a higher test anxiety score was related to using social support as a coping strategy. However, no relationships were found between academic performance and the three coping strategies when quantitative and qualitative data were analysed. Therefore, different coping strategies per se did not seem to be related to academic performance, but instead it was assumed that the effectiveness of coping strategies is dependent on individual differences. In order to obtain more generalisable results and to gain more understanding of test anxiety and coping with it, a larger number of subjects form different areas of Finland and of different ages could be examined in future studies. Moreover, cross-national and cross-cultural studies could provide valuable information. As a practical recommendation for educational purposes, the results of this study indicated that a more individualised approach is needed.

Translation – The fifth language skill? A comparison of the role of translation in Finnish lukio and the International Baccalaureate Diploma Programme

This study attempts to answer the question “Should translation be considered a fifth language skill?” by examining and comparing the use of translation as a language learning and assessment method in the national Finnish lukio curriculum and the curriculum of the International Baccalaureate Diploma Programme (IBDP). Furthermore, the students’ ability to translate and their opinions on the usefulness of translation in language learning will be examined. The students’ opinions were gathered through a questionnaire that was given to 156 students studying in either lukio or the IBDP in Turku and Rovaniemi. I present and compare the role of translation in selected language teaching and learning methods and approaches, and discuss the effectiveness of translation as a language learning method and an assessment method. The theoretical discussion provides the basis for examining the role of translation as a language learning method and an assessment method in the curricula and final examinations of both education programs. The analysis of the two curricula indicated that there is a significant difference in the use of translation, as translation is used as a language learning method and as an assessment method in lukio, but is not used in either form in the IB. The data obtained through the questionnaire indicated that there is a difference in the level of language competence between the lukio and IB students and suggested that the curriculum in which the student studies has some effect on his/her cognitive use of translation, ability to translate and opinions concerning the usefulness of translation in language learning. The results indicated that both groups of students used translation, along with their mother tongue, as a cognitive language learning method, and, contrary to the expectations set by the analysis of the two curricula, the IB students performed better in the translation exercises than lukio students. Both groups of students agreed that translation is a useful language learning method, and indicated that the most common dictionaries they use are bilingual Internet dictionaries. The results suggest that translation is a specific skill that requires teaching and practice, and that perhaps the translation exercises used in lukio should be developed from translating individual words and phrases to translating cultural elements. In addition, the results suggest that perhaps the IB curriculum should include the use of translation exercises (e.g., communicative translation exercises) in order to help students learn to mediate between languages and cultures rather than learn languages in isolation from each other.

A Worldwide History Spanning Genocide for the Sake of Corporate Profit

A Worldwide History Spanning Genocide for the Sake of Corporate Profit is the title of a thesis consisting of four stories. Crosky’s Knife is presented first for the sake of allowing potential readers to embrace a sense of amiable audience engagement and is based on a true story of a veteran of one of the many global conflicts currently raged on behalf of freedom. The three pieces following this feint are reworked versions of stories written from the heart and delivered to machines. As it presents numerous aspects of reality that the average person may not wish to consider, doing so with shockingly casual acceptance of such horror and/or banality, the conscious reception of the duty of engagement and possible appreciation of the text is not advised. Knives, rabid dogs, severed tongues, and a downpour of malnourished Iraqi babies are components intrinsic to the direction of this thesis.

Empirical evidence of bias in the design of experimental stroke studies: a metaepidemiologic approach

Background and Purpose: At least part of the failure in the transition from experimental to clinical studies in stroke has been attributed to the imprecision introduced by problems in the design of experimental stroke studies. Using a metaepidemiologic approach, we addressed the effect of randomization, blinding, and use of comorbid animals on the estimate of how effectively therapeutic interventions reduce infarct size. Methods: Electronic and manual searches were performed to identify meta-analyses that described interventions in experimental stroke. For each meta-analysis thus identified, a reanalysis was conducted to estimate the impact of various quality items on the estimate of efficacy, and these estimates were combined in a meta meta-analysis to obtain a summary measure of the impact of the various design characteristics. Results: Thirteen meta-analyses that described outcomes in 15 635 animals were included. Studies that included unblinded induction of ischemia reported effect sizes 13.1% (95% CI, 26.4% to 0.2%) greater than studies that included blinding, and studies that included healthy animals instead of animals with comorbidities overstated the effect size by 11.5% (95% CI, 21.2% to 1.8%). No significant effect was found for randomization, blinded outcome assessment, or high aggregate CAMARADES quality score. Conclusions: We provide empirical evidence of bias in the design of studies, with studies that included unblinded induction of ischemia or healthy animals overestimating the effectiveness of the intervention. This bias could account for the failure in the transition from bench to bedside of stroke therapies.

Contrasts: Quartets and Art Songs of the Nineteenth Century

The nineteenth-century Romantic era saw the development and expansion of many vocal and instrumental forms that had originated in the Classical era. In particular, the German lied and French mélodie matured as art forms, and they found a kind of equilibrium between piano and vocal lines. Similarly, the nineteenth-century piano quartet came into its own as a form of true chamber music in which all instruments participated equally in the texture. Composers such as Robert Schumann, Johannes Brahms, and Gabriel Fauré offer particularly successful examples of both art song and piano quartets that represent these genres at their highest level of artistic complexity. Their works have become the cornerstones of the modern collaborative pianist’s repertoire. My dissertation explored both the art songs and the piano quartets of these three composers and studied the different skills needed by a pianist performing both types of works. This project included the following art song cycles: Robert Schumann’s Dichterliebe, Gabriel Fauré’s Poème d’un Jour, and Johannes Brahms’ Zigeunerlieder. I also performed Schumann’s Piano Quartet in E-flat Major, Op. 47, Fauré’s Piano Quartet in C minor, Op. 15, and Brahms’ Piano Quartet in G minor, Op. 25. My collaborators included: Zachariah Matteson, violin and viola; Kristin Bakkegard, violin; Molly Jones, cello; Geoffrey Manyin, cello; Karl Mitze, viola; Emily Riggs, soprano, and Matthew Hill, tenor. This repertoire was presented over the course of three recitals on February 13, 2015, December 11, 2015, March 25, 2016 at the University of Maryland’s Gildenhorn Recital Hall. These recitals can be found in the Digital Repository at the University of Maryland (DRUM).

Good laboratory practice: preventing introduction of bias at the bench

Background and Purpose—As a research community, we have failed to demonstrate that drugs which show substantial efficacy in animal models of cerebral ischemia can also improve outcome in human stroke. Summary of Review—Accumulating evidence suggests this may be due, at least in part, to problems in the design, conduct and reporting of animal experiments which create a systematic bias resulting in the overstatement of neuroprotective efficacy. Conclusions—Here, we set out a series of measures to reduce bias in the design, conduct and reporting of animal experiments modeling human stroke.