Evaluation of the diagnostic potential of a novel field immunomagnetic separation-immunochromatographic lateral flow assay to detect Mycobacterium bovis cells in European badger (Meles meles) faeces

The European badger (Meles meles) is a natural reservoir for Mycobacterium bovis, the causative agent of Bovine Tuberculosis, and has consequently been implicated in transmission of the disease to cattle. This study describes application of a novel M. bovis-specific immunochromatographic (lateral flow) assay in combination with immunomagnetic separation (IMS-LFD), to test badger faeces samples. In total, 441 faeces samples from badgers of unknown disease status collected from latrines at 110 badger setts throughout Northern Ireland (NI) and 100 faeces samples from badgers of known infection status from Great Britain (GB) were tested. Faeces (approx. 1g) was homogenised in 9 ml phosphate buffered saline, filtered (70 µm), and then 6-8 ml subjected to the IMS-LFD test. Residual clarified faecal homogenates were subjected to automated IMS followed by MGIT™ liquid culture (AIMS-MGIT™ culture) and qPCR (AIMS-qPCR). Evidence for the presence of M. bovis was obtained for 78 (18%), 61 (14%) and 140 (32%) of 441 NI badger faeces samples, and 10 (10%), 41 (41%) and 56 (56%) of 100 GB badger faeces samples, by IMS-LFD, AIMS-MGIT culture and AIMS-qPCR tests, respectively. The IMS-LFD test was less sensitive than AIMS-qPCR for detection of M. bovis and was, therefore, detecting badgers shedding high numbers of M. bovis in their faeces only. However, these ‘super shedders’ may be primarily responsible for the spread of Bovine Tuberculosis so are, therefore, an important target. This non-invasive test could form the basis of a field surveillance tool to indicate infected badger groups which are actively spreading M. bovis.

THE SAAF STUDY: evaluation of the Safeguarding Children Assessment and Analysis Framework (SAAF), compared with management as usual, for improving outcomes for children and young people who have experienced, or are at risk of, maltreatment: study protocol for a randomised controlled trial

Background: Serious case reviews and research studies have indicated weaknesses in risk assessments conducted by child protection social workers. Social workers are adept at gathering information but struggle with analysis and assessment of risk. The Department for Education wants to know if the use of a structured decision-making tool can improve child protection assessments of risk.

Methods/design: This multi-site, cluster-randomised trial will assess the effectiveness of the Safeguarding Children Assessment and Analysis Framework (SAAF). This structured decision-making tool aims to improve social workers' assessments of harm, of future risk and parents' capacity to change. The comparison is management as usual.

Inclusion criteria: Children's Services Departments (CSDs) in England willing to make relevant teams available to be randomised, and willing to meet the trial's training and data collection requirements.

Exclusion criteria: CSDs where there were concerns about performance; where a major organisational restructuring was planned or under way; or where other risk assessment tools were in use.

Six CSDs are participating in this study. Social workers in the experimental arm will receive 2 days training in SAAF together with a range of support materials, and access to limited telephone consultation post-training. The primary outcome is child maltreatment. This will be assessed using data collected nationally on two key performance indicators: the first is the number of children in a year who have been subject to a second Child Protection Plan (CPP); the second is the number of re-referrals of children because of related concerns about maltreatment. Secondary outcomes are: i) the quality of assessments judged against a schedule of quality criteria and ii) the relationship between the three assessments required by the structured decision-making tool (level of harm, risk of (re) abuse and prospects for successful intervention).

Discussion: This is the first study to examine the effectiveness of SAAF. It will contribute to a very limited literature on the contribution that structured decision-making tools can make to improving risk assessment and case planning in child protection and on what is involved in their effective implementation.

Evaluation of the Retinal Vasculature in Hypertension and Chronic Kidney Disease in an elderly population of Irish nuns

Background: Chronic kidney disease (CKD) and hypertension are global public health problems associated with considerable morbidity, premature mortality and attendant healthcare costs. Previous studies have highlighted that non-invasive examination of the retinal microcirculation can detect microvascular pathology that is associated with systemic disorders of the circulatory system such as hypertension. We examined the associations between retinal vessel caliber (RVC) and fractal dimension (DF), with both hypertension and CKD in elderly Irish nuns.

Methods: Data from 1233 participants in the cross-sectional observational Irish Nun Eye Study (INES) were assessed from digital photographs with a standardized protocol using computer-assisted software. Multivariate regression analyses were used to assess associations with hypertension and CKD, with adjustment for age, body mass index (BMI), refraction, fellow eye RVC, smoking, alcohol consumption, ischemic heart disease (IHD), cerebrovascular accident (CVA), diabetes and medication use.

Results: In total, 1122 (91%) participants (mean age: 76.3 [range: 56-100] years) had gradable retinal images of sufficient quality for blood vessel assessment. Hypertension was significantly associated with a narrower central retinal arteriolar equivalent (CRAE) in a fully adjusted analysis (P = 0.002; effect size= -2.16 μm; 95% confidence intervals [CI]: -3.51, -0.81 μm). No significant associations between other retinal vascular parameters and hypertension or between any retinal vascular parameters and CKD were found.

Conclusions: Individuals with hypertension have significantly narrower retinal arterioles which may afford an earlier opportunity for tailored prevention and treatment options to optimize the structure and function of the microvasculature, providing additional clinical utility. No significant associations between retinal vascular parameters and CKD were detected.

Psychometric evaluation of the Student Authorship Questionnaire: a confirmatory factor analysis approach

This research provides new insights into the measurement of students’ authorial identity and its potential for minimising the incidence of unintentional plagiarism by providing evidence about the psychometric properties of the Student Authorship Questionnaire (SAQ). Exploratory and confirmatory factor analyses (EFA and CFA) are employed to investigate the measurement properties of the scales which comprise the SAQ using data collected from accounting students. The results provide limited psychometric support in favour of the factorial structure of the SAQ and raise a number of questions regarding the instrument’s robustness and generalisability across disciplines. An alternative model derived from the EFA outperforms the SAQ model with regard to its psychometric properties. Explanations for these findings are proffered and avenues for future research suggested.

Process Evaluation of the Roots of Empathy Program

This paper presents the findings of a qualitative process evaluation of the Roots of Empathy (ROE) programme. ROE is a universal, classroom-based intervention, which aims to enhance social and emotional learning of primary (elementary) school children. Effective delivery of such complex social interventions in real-world settings requires in-depth knowledge and understanding of factors that interact to influence implementation and fidelity. A case study methodology was employed with six schools, to explore the views of key actors and stakeholders involved in the delivery and receipt of the programme. Overall, ROE was delivered with high fidelity and the programme was viewed positively across the schools. However, one issue was the varied level of interest and awareness of the programme from parents.

Volatile terpenoids and C13 norisoprenoids in wines: development of rapid methods of analysis and evaluation of the sesquiterpenoids biological potential

Vitis vinifera L., the most widely cultivated fruit crop in the world, was the starting point for the development of this PhD thesis. This subject was exploited following on two actual trends: i) the development of rapid, simple, and high sensitive methodologies with minimal sample handling; and ii) the valuation of natural products as a source of compounds with potential health benefits. The target group of compounds under study were the volatile terpenoids (mono and sesquiterpenoids) and C13 norisoprenoids, since they may present biological impact, either from the sensorial point of view, as regards to the wine aroma, or by the beneficial properties for the human health. Two novel methodologies for quantification of C13 norisoprenoids in wines were developed. The first methodology, a rapid method, was based on the headspace solid-phase microextraction combined with gas chromatography-quadrupole mass spectrometry operating at selected ion monitoring mode (HS-SPME/GC-qMS-SIM), using GC conditions that allowed obtaining a C13 norisoprenoid volatile signature. It does not require any pre-treatment of the sample, and the C13 norisoprenoid composition of the wine was evaluated based on the chromatographic profile and specific m/z fragments, without complete chromatographic separation of its components. The second methodology, used as reference method, was based on the HS-SPME/GC-qMS-SIM, allowing the GC conditions for an adequate chromatographic resolution of wine components. For quantification purposes, external calibration curves were constructed with β-ionone, with regression coefficient (r2) of 0.9968 (RSD 12.51 %) and 0.9940 (RSD of 1.08 %) for the rapid method and for the reference method, respectively. Low detection limits (1.57 and 1.10 μg L-1) were observed. These methodologies were applied to seventeen white and red table wines. Two vitispirane isomers (158-1529 L-1) and 1,1,6-trimethyl-1,2-dihydronaphthalene (TDN) (6.42-39.45 μg L-1) were quantified. The data obtained for vitispirane isomers and TDN using the two methods were highly correlated (r2 of 0.9756 and 0.9630, respectively). A rapid methodology for the establishment of the varietal volatile profile of Vitis vinifera L. cv. 'Fernão-Pires' (FP) white wines by headspace solid-phase microextraction combined with comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (HS-SPME/GCxGC-TOFMS) was developed. Monovarietal wines from different harvests, Appellations, and producers were analysed. The study was focused on the volatiles that seem to be significant to the varietal character, such as mono and sesquiterpenic compounds, and C13 norisoprenoids. Two-dimensional chromatographic spaces containing the varietal compounds using the m/z fragments 93, 121, 161, 175 and 204 were established as follows: 1tR = 255-575 s, 2tR = 0,424-1,840 s, for monoterpenoids, 1tR = 555-685 s, 2tR = 0,528-0,856 s, for C13 norisoprenoids, and 1tR = 695-950 s, 2tR = 0,520-0,960 s, for sesquiterpenic compounds. For the three chemical groups under study, from a total of 170 compounds, 45 were determined in all wines, allowing defining the "varietal volatile profile" of FP wine. Among these compounds, 15 were detected for the first time in FP wines. This study proposes a HS-SPME/GCxGC-TOFMS based methodology combined with classification-reference sample to be used for rapid assessment of varietal volatile profile of wines. This approach is very useful to eliminate the majority of the non-terpenic and non-C13 norisoprenic compounds, allowing the definition of a two-dimensional chromatographic space containing these compounds, simplifying the data compared to the original data, and reducing the time of analysis. The presence of sesquiterpenic compounds in Vitis vinifera L. related products, to which are assigned several biological properties, prompted us to investigate the antioxidant, antiproliferative and hepatoprotective activities of some sesquiterpenic compounds. Firstly, the antiradical capacity of trans,trans-farnesol, cis-nerolidol, α-humulene and guaiazulene was evaluated using chemical (DPPH• and hydroxyl radicals) and biological (Caco-2 cells) models. Guaiazulene (IC50= 0.73 mM) was the sesquiterpene with higher scavenger capacity against DPPH•, while trans,trans-farnesol (IC50= 1.81 mM) and cis-nerolidol (IC50= 1.48 mM) were more active towards hydroxyl radicals. All compounds, with the exception of α-humulene, at non-cytotoxic levels (≤ 1 mM), were able to protect Caco-2 cells from oxidative stress induced by tert-butyl hydroperoxide. The activity of the compounds under study was also evaluated as antiproliferative agents. Guaiazulene and cis-nerolidol were able to more effectively arrest the cell cycle in the S-phase than trans,trans-farnesol and α-humulene, being the last almost inactive. The relative hepatoprotection effect of fifteen sesquiterpenic compounds, presenting different chemical structures and commonly found in plants and plant-derived foods and beverages, was assessed. Endogenous lipid peroxidation and induced lipid peroxidation with tert-butyl hydroperoxide were evaluated in liver homogenates from Wistar rats. With the exception of α-humulene, all the sesquiterpenic compounds under study (1 mM) were effective in reducing the malonaldehyde levels in both endogenous and induced lipid peroxidation up to 35% and 70%, respectively. The developed 3D-QSAR models, relating the hepatoprotection activity with molecular properties, showed good fit (R2LOO > 0.819) with good prediction power (Q2 > 0.950 and SDEP < 2%) for both models. A network of effects associated with structural and chemical features of sesquiterpenic compounds such as shape, branching, symmetry, and presence of electronegative fragments, can modulate the hepatoprotective activity observed for these compounds. In conclusion, this study allowed the development of rapid and in-depth methods for the assessment of varietal volatile compounds that might have a positive impact on sensorial and health attributes related to Vitis vinifera L. These approaches can be extended to the analysis of other related food matrices, including grapes and musts, among others. In addition, the results of in vitro assays open a perspective for the promising use of the sesquiterpenic compounds, with similar chemical structures such as those studied in the present work, as antioxidants, hepatoprotective and antiproliferative agents, which meets the current challenges related to diseases of modern civilization.

Promoting self-regulated learning in technology enhanced learning environments : keeping a digital track of the learning process

Tese de doutoramento (co-tutela), Psicologia (Psicologia da Educação), Faculdade de Psicologia da Universidade de Lisboa, Faculdade de Psicologia e de Ciências da Educação da Universidade de Coimbra, Technial University of Darmstadt, 2014

Evaluation of the anti-arenaviral activity of the subtilisin kexin isozyme-1/site-1 protease inhibitor PF-429242.

The cellular protease subtilisin kexin isozyme-1 (SKI-1)/site-1 protease (S1P) is implicated in the proteolytic processing of the viral envelope glycoprotein precursor (GPC) of arenaviruses, a step strictly required for production of infectious progeny. The small molecule SKI-1/S1P inhibitor PF-429242 was shown to have anti-viral activity against Old World arenaviruses. Here we extended these studies and show that PF-429242 also inhibits GPC processing and productive infection of New World arenaviruses, making PF-429242 a broadly active anti-arenaviral drug. In combination therapy, PF-429242 potentiated the anti-viral activity of ribavirin, indicating a synergism between the two drugs. A hallmark of arenaviruses is their ability to establish persistent infection in vitro and in vivo. Notably, PF-429242 was able to efficiently and rapidly clear persistent infection by arenaviruses. Interruption of drug treatment did not result in re-emergence of infection, indicating that PF-429242 treatment leads to virus extinction.

Evaluation of Machine Learning Classifiers for Mobile Network Intrusion Detection Systems

Mobile malwares are increasing with the growing number of Mobile users. Mobile malwares can perform several operations which lead to cybersecurity threats such as, stealing financial or personal information, installing malicious applications, sending premium SMS, creating backdoors, keylogging and crypto-ransomware attacks. Knowing the fact that there are many illegitimate Applications available on the App stores, most of the mobile users remain careless about the security of their Mobile devices and become the potential victim of these threats. Previous studies have shown that not every antivirus is capable of detecting all the threats; due to the fact that Mobile malwares use advance techniques to avoid detection. A Network-based IDS at the operator side will bring an extra layer of security to the subscribers and can detect many advanced threats by analyzing their traffic patterns. Machine Learning(ML) will provide the ability to these systems to detect unknown threats for which signatures are not yet known. This research is focused on the evaluation of Machine Learning classifiers in Network-based Intrusion detection systems for Mobile Networks. In this study, different techniques of Network-based intrusion detection with their advantages, disadvantages and state of the art in Hybrid solutions are discussed. Finally, a ML based NIDS is proposed which will work as a subsystem, to Network-based IDS deployed by Mobile Operators, that can help in detecting unknown threats and reducing false positives. In this research, several ML classifiers were implemented and evaluated. This study is focused on Android-based malwares, as Android is the most popular OS among users, hence most targeted by cyber criminals. Supervised ML algorithms based classifiers were built using the dataset which contained the labeled instances of relevant features. These features were extracted from the traffic generated by samples of several malware families and benign applications. These classifiers were able to detect malicious traffic patterns with the TPR upto 99.6% during Cross-validation test. Also, several experiments were conducted to detect unknown malware traffic and to detect false positives. These classifiers were able to detect unknown threats with the Accuracy of 97.5%. These classifiers could be integrated with current NIDS', which use signatures, statistical or knowledge-based techniques to detect malicious traffic. Technique to integrate the output from ML classifier with traditional NIDS is discussed and proposed for future work.

An impact evaluation of the Ontario ban on smoking in schools and on school property : does the ban on smoking influence high school students' intentions to smoke cigarettes?

The Ontario Tobacco Control Act of 1994 imposed a total ban on smoking in schools, and on school property for every school in the province. The imposition of this policy created problems for school administrators. For instance, students who were smoking on walkways and properties adjacent to school boundaries, clashed with neighbouring property owners who were angry about the resulting damage and disruption. The enforcement of this policy consumes valuable resources at each school; therefore, knowledge about the impact of the policy is important. If effective, this policy has the potential to improve the health of students over their lifetime, by preventing or delaying smoking behaviour. Alternatively, an ineffective policy will continue to create administrative problems for the school and serve no legitimate purpose. Therefore, knowledge about the impact of the smoking ban policy on students' smoking intentions assists policy makers and school administrators in their understanding of the policy's impact within the schools. This research provided an impact evaluation of the ban on smoking in schools and on school property in Ontario. A total of 2069 students, from five high schools, in the Niagara Region, provided complete responses to a survey, designed to test whether smoking intentions were affected by the imposition of the policy. The study used Ajzen's theory of planned behaviour (Ajzen, 1991), specifically, the perceived behavioural control measure, to gain some understanding of students' perceptions of control over smoking imposed by the ban. The findings indicate the policy has the potential to influence students' overall smoking intentions. The ban on smoking policy was found to be a significant predictor of the smoking intentions of high school students. As well, attitude, social norms, and perceptions of control were significant predictors of smoking intentions. Exploratory findings also indicated differences between the control beliefs of students from different high schools, indicating potential differences in the enforcement of the smoking ban between schools. The findings also support the utility of the theory of planned behaviour as a methodology for evaluating the influence of punitive policies. This research study should be continued by utilizing the full theory of planned behaviour, including two phases of data collection and the measurement of actual smoking behaviour.

The development, assessment, and implementation of an evaluation of the EMPOWER Program at shelters for abused women /

The development, assessment, and implementation of a program evaluation instrument was carried out to evaluate the impact and efficacy of the EMPOWER Program. This intervention was created to educate residents at a shelter for abused women with an anticipated outcome of prevention. Participants included the staff and residents at 2 shelters in Southern Ontario. Client pre, post and follow-up measures were obtained and analyzed statistically and using keyword content analysis. A single staff measure was obtained and summarized using keyword content analysis. Qualitative results were suggestive of important change in participants. All women in the post and follow-up measures believed their participation in the EMPOWER Program provided them with the knowledge, skills, and confidence to avoid abusive relationships in the fliture. This transformational impact was repeatedly expressed in both resident and staff feedback. Limitations of this research, as well as suggestions for future study were discussed.