Bioluminescence imaging of chronic Trypanosoma cruzi infections reveals tissue-specific parasite dynamics and heart disease in the absence of locally persistent infection.

Chronic Trypanosoma cruzi infections lead to cardiomyopathy in 20-30% of cases. A causal link between cardiac infection and pathology has been difficult to establish because of a lack of robust methods to detect scarce, focally distributed parasites within tissues. We developed a highly sensitive bioluminescence imaging system based on T. cruzi expressing a novel luciferase that emits tissue-penetrating orange-red light. This enabled long-term serial evaluation of parasite burdens in individual mice with an in vivo limit of detection of significantly less than 1000 parasites. Parasite distributions during chronic infections were highly focal and spatiotemporally dynamic, but did not localize to the heart. End-point ex vivo bioluminescence imaging allowed tissue-specific quantification of parasite loads with minimal sampling bias. During chronic infections, the gastro-intestinal tract, specifically the colon and stomach, was the only site where T. cruzi infection was consistently observed. Quantitative PCR-inferred parasite loads correlated with ex vivo bioluminescence and confirmed the gut as the parasite reservoir. Chronically infected mice developed myocarditis and cardiac fibrosis, despite the absence of locally persistent parasites. These data identify the gut as a permissive niche for long-term T. cruzi infection and show that canonical features of Chagas disease can occur without continual myocardium-specific infection

Doctor of Philosophy

dissertationIn an extension of research demonstrating the need for systematic behavioral change through intervention in public schools, the present study explored effects of positive behavior interventions and supports for improved student performance and reduced reactionary management for school staff. Districts participating in a statewide training initiative in Utah provided 10 target schools for study. These target schools collected data and participated in a series of training activities prior to implementation. Implementation included the establishment of a multidisciplinary team. Outcome data included office disciplinary referrals, fidelity of implementation scales, reported satisfaction, positive reinforcement, administrative time and cost, and proficiency in high-stakes testing. The participant schools demonstrated statistically significant positive outcomes with reduced rates of reported negative student behavior, including tardies. No statistically significant differences between baseline and treatment were observed in high-stakes academic assessment measures. Recommendations for future research include the selection of more sensitive measures and disaggregated analysis

Confirmation that “Brachyspira hampsonii” clade I (Canadian strain 30599) causes mucohemorrhagic diarrhea and colitis in experimentally infected pigs

BACKGROUND: “Brachyspira hampsonii”, discovered in North America in 2010 associated with dysentery-like illness, is an economically relevant swine pathogen resulting in decreased feed efficiency and increased morbidity, mortality and medication usage. “B. hampsonii” clade II strain 30446 has been shown to be causally associated with mucohemorrhagic diarrhea and colitis. Our objectives were to determine if “Brachyspira hampsonii” clade I strain 30599 is pathogenic to pigs, and to evaluate the relative diagnostic performance of three ante mortem sampling methodologies (direct PCR on feces, PCR on rectal GenoTube Livestock swabs, Brachyspira culture from rectal swabs). Five-week old pigs were intragastrically inoculated thrice with 10(8) genomic equivalents "B. hampsonii" (n = 12), or served as sham controls (n = 6). Feces were sampled and consistency assessed daily. Necropsies were performed 24 h after peak clinical signs. RESULTS: One pig died due to unrelated illness. Nine of 11 inoculated pigs, but no controls, developed mucoid or mucohemorrhagic diarrhea (MHD). Characteristic lesions of swine dysentery were observed in large intestine. “B. hampsonii” strain 30599 DNA was detected by qPCR in feces of all inoculated pigs for up to 6 days prior to the onset of MHD. The organism was isolated from the feces and colons of pigs demonstrating MHD, but not from controls. B. intermedia was isolated from inoculated pigs without MHD, and from 5 of 6 controls. CONCLUSIONS: We conclude that “Brachyspira hampsonii” clade I strain 30599 is pathogenic and causes mucohemorrhagic diarrhea and colitis in susceptible pigs. Moreover, the three sampling methodologies performed similarly. GenoTube Livestock, a forensic swab designed to preserve DNA during shipping is a useful tool especially in settings where timely transport of diagnostic samples is challenging

16-O-methylcafestol is present in ground roast Arabica coffees: Implications for authenticity testing

High-field and low-field proton NMR spectroscopy were used to analyse lipophilic extracts from ground roast coffees. Using a sample preparation method that produced concentrated extracts, a small marker peak at 3.16 ppm was observed in 30 Arabica coffees of assured origin. This signal has previously been believed absent from Arabicas, and has been used as a marker for detecting adulteration with robusta. Via 2D 600 MHz NMR and LC-MS, 16-O-methylcafestol and 16-O-methylkahweol were detected for the first time in Arabica roast coffee and shown to be responsible for the marker peak. Using low-field NMR, robusta in Arabica could be detected at levels of the order of 1-2% w/w. A surveillance study of retail purchased "100% Arabica" coffees found that 6 out of 60 samples displayed the 3.16 ppm marker signal to a degree commensurate with adulteration at levels of 3-30% w/w

Total neoadjuvant therapy in oesophageal and gastro-oesophageal junctional adenocarcinoma

Adenocarcinoma of the oesophagus and gastro-oesophageal junction represent a large burden of cancer death in the Western World with an increasing incidence. In the past two decades, the overall survival of patients on a potentially curative treatment pathway has more than doubled due to the addition of perioperative oncological therapies to surgery. However, patients often fail to respond to oncological treatment or struggle to complete their treatment after surgery. In this review, we discuss the current evidence for total neoadjuvant therapy and options for assessment of treatment response.</p

Exploring the processes involved in seeking help from a general practitioner for young people who have been at risk of suicid

General practitioners (GPs) play a key role in the early identification and management of suicide risk in young people. However, little is known about the processes involved in how, when and why a young person decides to seek help from their GP. Eight young people, aged 17–23, took part in semi-structured interviews exploring their experiences of help seeking when feeling suicidal. Data were analysed using framework analysis. The analysis identified three main themes and seven subthemes. The main themes explored were: understanding when to seek help from a GP, barriers and facilitating factors at the GP consultation, and help seeking as a non-linear and dynamic process. The processes involved in how, when and why young people seek help from a GP when feeling suicidal were found to be dynamic and to fluctuate over time. Help seeking was initially related to how young people were able to understand and articulate their distress, the availability of informal support networks, and their perception of the GP as a source of help. During a GP consultation, help seeking was influenced by how safe and supported the young people felt. Perceived GP training, communication and validation of young people’s concerns were important factors to help facilitate this process. Subsequent help seeking was influenced by prior experience of GP consultations and the availability of alternative support

The Unequal Treatment of Animals by Species and Practice in the United States : a Moral and Legal Dilemma

Todos los estados de los Estados Unidos de América disponen de una compleja red de leyes encaminadas a la protección de los algunos animales frente a la crueldad y al abandono. Dependiendo de la gravedad del delito y otros factores, la ley estatal puede considerar la crueldad hacia los animales como una infracción, un delito menor o un delito mayor. Sin embargo, no todos los animales reciben el mismo tratamiento. Sólo el tiempo dirá si los animales de granja (o animales de laboratorio) lleguen algún día a recibir el mismo nivel de protección legal, por las mismas razones filosóficas, que nuestros animales de compañía.Every state in the United States of America has a complex web of laws protecting at least some animals from cruelty and neglect. Depending on the severity of the crime and other factors, state law may consider animal cruelty an infraction, a misdemeanor, or a felony. However, not all the animals receive the same treatment. Only time will tell if farm animals (or laboratory animals) will ever receive the same level of legal protection, for the same philosophical reasons, as our companion animals

A mitigation hierarchy approach for managing sea turtle captures in small-scale fisheries

The mitigation hierarchy has been proposed as an overarching framework for managing fisheries and reducing marine megafauna bycatch, but requires empirical application to show its practical utility. Focusing on a small-scale fishing community in Peru as a case study system, we test how the mitigation hierarchy can support efforts to reduce captures of sea turtles in gillnets and link these actions to broader goals for biodiversity. We evaluate three management scenarios by drawing on ecological risk assessment (ERA) and qualitative management strategy evaluation to assess trade-offs between biological, economic, and social considerations. The turtle species of management focus include leatherback turtle Dermochelys coriacea, green turtle Chelonia mydas, and olive ridley turtle Lepidochelys olivacea. Adopting a mixed-methods iterative approach to data collection, we undertook a literature review to collate secondary data on the fishery and the species of turtles captured. We then collected primary data to fill the knowledge gaps identified, including establishing the spatial extent of the fishery and calculating turtle capture rates for the fishery. We identified and evaluated the potential risk that the fishery poses to each turtle species within Pacific East regional management units using a qualitative ERA. Finally, we evaluated potential management strategies to reduce turtle captures, incorporating stakeholder preference from questionnaire-based surveys and considering preliminary estimates of trends across a range of performance indicators. We illustrate how the proposed framework can integrate existing knowledge on an issue of marine megafauna captures, and incorporate established decision-making processes to help identify data gaps. This supports a holistic assessment of management strategies toward biodiversity goals standardized across fisheries and scales

Physiological and Psychological Effects of Deception on Pacing Strategy and Performance: A Review

The aim of an optimal pacing strategy during exercise is to enhance performance whilst ensuring physiological limits are not surpassed, which has been shown to result in a metabolic reserve at the end of the exercise. There has been debate surrounding the theoretical models that have been proposed to explain how pace is regulated, with more recent research investigating a central control of exercise regulation. Deception has recently emerged as a common, practical approach to manipulate key variables during exercise. There are a number of ways in which deception interventions have been designed, each intending to gain particular insights into pacing behaviour and performance. Deception methodologies can be conceptualised according to a number of dimensions such as deception timing (prior to or during exercise), presentation frequency (blind, discontinuous or continuous) and type of deception (performance, biofeedback or environmental feedback). However, research evidence on the effects of deception has been perplexing and the use of complex designs and varied methodologies makes it difficult to draw any definitive conclusions about how pacing strategy and performance are affected by deception. This review examines existing research in the area of deception and pacing strategies, and provides a critical appraisal of the different methodological approaches used to date. It is hoped that this analysis will inform the direction and methodology of future investigations in this area by addressing the mechanisms through which deception impacts upon performance and by elucidating the potential application of deception techniques in training and competitive settings