Exploring investigative question generation

A failure to conduct effective investigative interviews can have drastic consequences such as wrongful convictions or the inability to prevent terrorist attacks. One method of judging the efficacy of an interview is the ability to distinguish between honest and deceptive interviewees. Many techniques claim to improve the ability to detect deception, such as the CCE technique. However, little research has focused on the conditions that might enhance the ability to generate investigatively useful questions. In series of experiments we sought to identify the underlying dimensions of question quality. Initially, we investigated unexpectedness, finding that it was a useful dimension but dependent on the content of the questions (Chapters 2 and 3). In Chapter 5 we used a bottom-up approach to develop a 3-dimensional model of question quality. These dimensions were investigative relevance, unpredictability and type of knowledge probed. The model proved to be effective in predicting the outcome of real-world investigate interviews (Chapter 6). We also aimed to investigate the factors which might affect question generation ability. The scope of episodic information inherently available to the interviewer was shown to be a context-dependent factor affecting the ability to generate useful questions (Chapters 4 and 5). Training, via a short instructional video, was also shown to improve question generating ability (Chapter 4). Additionally, the veracity of the interviewee and the expertise of the question generator affected ability, though this was only detected by novice judges (Chapter 5). The findings presented in this thesis have implications for the investigative community, suggesting that deliberate attention should be paid to the phrasing of key interview questions in order to ensure that they are relevant, unpredictable and probing episodic knowledge. Additionally, the findings may inform current research that is focused on developing technology designed to assist investigative interviewers

Dysfunctional Crohn's Disease-Associated NOD2 Polymorphisms Cannot be Reliably Predicted on the Basis of RIPK2 Binding or Membrane Association.

Polymorphisms in NOD2 represent the single greatest genetic risk factor for the development of Crohn's disease. Three different non-synonomous NOD2 polymorphisms - R702W, G908R, and L1007fsincC - account for roughly 80% of all NOD2-associated cases of Crohn's disease and are reported to result in a loss of receptor function in response to muramyl dipeptide (MDP) stimulation. Loss of NOD2 signaling can result from a failure to detect ligand; alterations in cellular localization; and changes in protein interactions, such as an inability to interact with the downstream adaptor protein RIPK2. Using an overexpression system, we analyzed ~50 NOD2 polymorphisms reportedly connected to Crohn's disease to determine if they also displayed loss of function and if this could be related to alterations in protein localization and/or association with RIPK2. Just under half the polymorphisms displayed a significant reduction in signaling capacity following ligand stimulation, with nine of them showing near complete ablation. Only two polymorphisms, R38M and R138Q, lost the ability to interact with RIPK2. However, both these polymorphisms still associated with cellular membranes. In contrast, L248R, W355stop, L550V, N825K, L1007fsinC, L1007P, and R1019stop still bound RIPK2, but showed impaired membrane association and were unable to signal in response to MDP. This highlights the complex contributions of NOD2 polymorphisms to Crohn's disease and reiterates the importance of both RIPK2 binding and membrane association in NOD2 signaling. Simply ascertaining whether or not NOD2 polymorphisms bind RIPK2 or associate with cellular membranes is not sufficient for determining their signaling competency.The authors would like to thank Joe Boyle for helpful discussion. This work was funded by a Wellcome Trust CDF (WT085090MA) to TPM and the Medical Research Council (U105960399). RP was a BBSRC doctoral training student.This is the final version of the article. It first appeared from Frontiers via http://dx.doi.org/10.3389/fimmu.2015.0052

Blau syndrome polymorphisms in NOD2 identify nucleotide hydrolysis and helical domain 1 as signalling regulators.

Understanding how single nucleotide polymorphisms (SNPs) lead to disease at a molecular level provides a starting point for improved therapeutic intervention. SNPs in the innate immune receptor nucleotide oligomerisation domain 2 (NOD2) can cause the inflammatory disorders Blau Syndrome (BS) and early onset sarcoidosis (EOS) through receptor hyperactivation. Here, we show that these polymorphisms cluster into two primary locations: the ATP/Mg(2+)-binding site and helical domain 1. Polymorphisms in these two locations may consequently dysregulate ATP hydrolysis and NOD2 autoinhibition, respectively. Complementary mutations in NOD1 did not mirror the NOD2 phenotype, which indicates that NOD1 and NOD2 are activated and regulated by distinct methods.This work was funded by a Wellcome Trust CDF (WT085090MA).This is the published version. It was originally published by Elsevier on behalf of FEBS Letters, at http://www.sciencedirect.com/science/article/pii/S001457931400578X

Insights into the molecular basis of the NOD2 signalling pathway.

The cytosolic pattern recognition receptor NOD2 is activated by the peptidoglycan fragment muramyl dipeptide to generate a proinflammatory immune response. Downstream effects include the secretion of cytokines such as interleukin 8, the upregulation of pro-interleukin 1β, the induction of autophagy, the production of antimicrobial peptides and defensins, and contributions to the maintenance of the composition of the intestinal microbiota. Polymorphisms in NOD2 are the cause of the inflammatory disorder Blau syndrome and act as susceptibility factors for the inflammatory bowel condition Crohn's disease. The complexity of NOD2 signalling is highlighted by the observation that over 30 cellular proteins interact with NOD2 directly and influence or regulate its functional activity. Previously, the majority of reviews on NOD2 function have focused upon the role of NOD2 in inflammatory disease or in its interaction with and response to microbes. However, the functionality of NOD2 is underpinned by its biochemical interactions. Consequently, in this review, we have taken the opportunity to address the more 'basic' elements of NOD2 signalling. In particular, we have focused upon the core interactions of NOD2 with protein factors that influence and modulate the signal transduction pathways involved in NOD2 signalling. Further, where information exists, such as in relation to the role of RIP2, we have drawn comparison with the closely related, but functionally discrete, pattern recognition receptor NOD1. Overall, we provide a comprehensive resource targeted at understanding the complexities of NOD2 signalling.T.P.M. was supported by a Wellcome Trust Career Development Fellowship (WT085090MA). J.P.B. and R.P. were supported by BBSRC Doctoral Training Grants.This is the final published version. It first appeared at: http://rsob.royalsocietypublishing.org/content/4/12/140178

Interaction between NOD2 and CARD9 involves the NOD2 NACHT and the linker region between the NOD2 CARDs and NACHT domain.

NOD2 activation by muramyl dipeptide causes a proinflammatory immune response in which the adaptor protein CARD9 works synergistically with NOD2 to drive p38 and c-Jun N-terminal kinase (JNK) signalling. To date the nature of the interaction between NOD2 and CARD9 remains undetermined. Here we show that this interaction is not mediated by the CARDs of NOD2 and CARD9 as previously suggested, but that NOD2 possesses two interaction sites for CARD9; one in the CARD-NACHT linker and one in the NACHT itself.This work was funded by a Wellcome Trust Career Development Fellowship (WT085090MA) to TPM and a Medical Research Council grant (U117565398) to KR. RP and JPB were supported by BBSRC Doctoral Training Grants.This is the final published version of the article, which can also be found on the publisher's website at: http://www.sciencedirect.com/science/article/pii/S0014579314004979

Research and Innovation for and with Adolescent Young Carers to Influence Policy and Practice—The European Union Funded “ME-WE” Project

Young carers are children and adolescents who provide care to other family members or friends, taking over responsibilities that are usually associated with adulthood. There is emerging but still scarce knowledge worldwide about the phenomenon of young carers and the impact of a caring role on their health, social and personal development spheres. This paper provides an overview of the main results from the ME-WE project, which is the first European research and innovation project dedicated to adolescent young carers (AYCs) (15–17 years). The project methods relied on three main activities: (1) a systematization of knowledge (by means of a survey to AYCs, country case studies, Delphi study, literature review); (2) the co-design, implementation and evaluation of a primary prevention intervention addressing AYCs’ mental health (by means of Blended Learning Networks and a clinical trial in six European countries); (3) the implementation of knowledge translation actions for dissemination, awareness, advocacy and lobbying (by means of national and international stakeholder networks, as well as traditional and new media). Project results substantially contributed to a better understanding of AYCs’ conditions, needs and preferences, defined tailored support intervention (resilient to COVID-19 related restrictions), and significant improvements in national and European policies for AYCs

The coiled-coil of ATG16L1 is highly conserved across vertebrates.

<p>(A) Cross-species alignment of the coiled-coil (human residues M126 – A207) of ATG16L1 reveals levels of sequence identity with the human sequence of between 73% and 100%. The percentage identity listed is in comparison with the human sequence. The common names and database identifies for each sequence are listed in Materials and Methods. (B) The syntenic position of <i>Atg16L1</i> is well conserved across species. The three adjacent upstream and downstream genese to <i>Atg16L1</i> are displayed. Genes are denoted by individual blocks; yellow indicates a position on the forward strand and green a position on the reverse strand. Gene identities are as follows: ATG16L1 – autophagy related protein 16 isoform 1; SAG – S-antigen, retina and pineal gland; DGKD – diacylglycerol kinase delta; USP40 – ubiquitin specific peptidase 40; INPP5D – inositol polyphosphate-5-phosphatase; NEU2 – sialidase 2; NGEF - neuronal guanine exchange factor; TRPV2 – transient receptor potential cation channel, subfamily V, member 2; UBB – ubiquitin B; AC106876.2 – uncharacterised; 00570 (ENSMODG00000000570) – uncharacterised; 23152 (ENSMODG00000023152) – uncharacterised, BLASTp indicates a possible ortholog of glyceraldehydes 3-phosphate dehydrogenase.</p

The coiled-coil of human ATG16L1 is alpha helical.

<p>(A) PSIPRED prediction of the secondary structure of the minimal coiled-coil domain (residues M126 – A207; CCD3) of human ATG16L1. Sequencing numbering corresponds to the human protein, alpha helices are marked as pink cylinders and the confidence level of the prediction for each residue is provided by the blue bars. (B) Circular dichroism analysis of CCD3. Mean residual ellipicity (θ) plotted against wavelength (nm) indicates a helical protein.</p

Characterisation of the oligomeric status of the human ATG16L1 coiled-coil domain.

<p>(A) Size exclusion chromatography of CCD3. CCD3 – red trace; Blue Dextran – blue trace; Molecular size markers – black trace (mass labelled in kDa). (B) Native-PAGE analysis of CCD3. 1 µg of CCD3 was analysed by Native-PAGE both before (lane 1) and after (lane 2) freeze/thawing at −80°C. M – NativeMark™ protein standards (Life Tehnologies). (C) Nanospray Electrospray Ionisation Mass Spectroscopy (ESI-MS) analysis of CCD3 under conditions that preserve non-covalent interactions. CCD3 dimers are detected in the low m/z range corresponding to charge states of +11 to +7. There is no evidence of any additional species of CCD3. (D) Analytical ultracentrifugation sedimentation velocity data. Interference optical signal distributions of CCD3FH at time intervals of 320 s at a rotor speed of 50,000 rpm and a temperature of 20°C, with systematic noise subtracted. The residuals are from the fit with the hybrid discrete/continuous model. Component sedimentation coefficient distributions showed a dominant population of the dimeric species, with a uniform frictional ratio of Fk,w = 1.73. The final r.m.s.d. was 0.006.</p