Errors in the identification of question types in investigative interviews of children

This study examined the incidence and nature of the errors made by trainee coders during their coding of question types in interviews in which children disclosed abuse. Three groups of trainees (online, postgraduate and police) studied the coding manual before practising their question coding. After this practice, participants were given two-page field transcripts to code in which children disclosed abuse. Their coding was assessed for accuracy; any errors were analysed thematically. The overall error rate was low, and police participants made the fewest errors. Analysis of the errors revealed four common misunderstandings: (1) the use of a ‘wh’ question always denotes a specific cued-recall question; (2) ‘Tell me’ always constitutes an open-ended question; (3) open-ended questions cannot include specific detail; and (4) specific questions cannot elicit elaborate responses. An analysis of coding accuracy in the one group who were able to practise question coding over time revealed that practice was essential for trainees to maintain their accuracy. Those who did not practise decreased in coding accuracy. This research shows that trainees need more than a coding manual; they must demonstrate their understanding of question codes through practice training tasks. Misunderstandings about questions need to be elicited and corrected so that accurate codes are used in future tasks

Motif affinity and mass spectrometry proteomic approach for the discovery of cellular AMPK targets: identification of mitochondrial fission factor as a new AMPK substrate

AMP-activated protein kinase (AMPK) is a key cellular energy sensor and regulator of metabolic homeostasis. Although it is best known for its effects on carbohydrate and lipid metabolism, AMPK is implicated in diverse cellular processes, including mitochondrial biogenesis, autophagy, and cell growth and proliferation. To further our understanding of energy homeostasis through AMPK-dependent processes, the design and application of approaches to identify and characterise novel AMPK substrates are invaluable. Here, we report an affinity proteomicstrategy for the discovery and validation of AMPK targets using an antibody to isolate proteins containing the phospho-AMPK substrate recognition motif from hepatocytes that had been treated with pharmacological AMPK activators. We identified 57 proteins that were uniquely enriched in the activator-treated hepatocytes, but were absent in hepatocytes lacking AMPK. We focused on two candidates, cingulin and mitochondrial fission factor (MFF), and further characterised/validated them as AMPK-dependent targets by immunoblotting with phosphorylation site-specific antibodies. A small-molecule AMPK activator caused transient phosphorylation of endogenous cingulin at S137 in intestinal Caco2 cells. Multiple splice-variants of MFF appear to express in hepatocytes and we identified a common AMPK-dependent phospho-site (S129) in all the 3 predominant variants spanning the mass range and a short variant-specific site (S146). Collectively, our proteomic-based approach using a phospho-AMPK substrate antibody in combination with genetic models and selective AMPK activators will provide a powerful and reliable platform for identifying novel AMPK-dependent cellular targets

Skeletal muscle AMPK is essential for the maintenance of FNDC5 expression

Fibronectin type III domain‐containing protein 5 (FNDC5) expression is controlled by the transcriptional co‐activator, peroxisome proliferator‐activated receptor gamma, coactivator 1 alpha (PGC1α). FNDC5 expression has been shown to be increased in muscle in response to endurance exercise in some but not all studies, therefore a greater understanding of the mechanisms controlling this process are needed. The AMP‐activated protein kinase (AMPK) is activated by exercise in an intensity dependent manner and is an important regulator of PGC1α activity; therefore, we explored the role of AMPK in the regulation of FNDC5 using AMPK β1β2 double muscle‐null mice (AMPK DMKO), which lack skeletal muscle AMPK activity. We found that FNDC5 expression is dramatically reduced in resting muscles of AMPK DMKO mice compared to wild‐type littermates. In wild‐type mice, activating phosphorylation of AMPK was elevated immediately post contraction and was abolished in muscle from AMPK DMKO mice. In contrast, PGC1α was increased in both wild‐type and AMPK DMKO mice 3 h post contraction but FNDC5 protein expression was not altered. Lastly, acute or chronic activation of AMPK with the pharmacological AMPK activator AICAR did not increase PGC1α or FNDC5 expression in muscle. These data indicate that skeletal muscle AMPK is required for the maintenance of basal FNDC5 expression

Genomes and Virulence Factors of Novel Bacterial Pathogens Causing Bleaching Disease in the Marine Red Alga Delisea pulchra

Nautella sp. R11, a member of the marine Roseobacter clade, causes a bleaching disease in the temperate-marine red macroalga, Delisea pulchra. To begin to elucidate the molecular mechanisms underpinning the ability of Nautella sp. R11 to colonize, invade and induce bleaching of D. pulchra, we sequenced and analyzed its genome. The genome encodes several factors such as adhesion mechanisms, systems for the transport of algal metabolites, enzymes that confer resistance to oxidative stress, cytolysins, and global regulatory mechanisms that may allow for the switch of Nautella sp. R11 to a pathogenic lifestyle. Many virulence effectors common in phytopathogenic bacteria are also found in the R11 genome, such as the plant hormone indole acetic acid, cellulose fibrils, succinoglycan and nodulation protein L. Comparative genomics with non-pathogenic Roseobacter strains and a newly identified pathogen, Phaeobacter sp. LSS9, revealed a patchy distribution of putative virulence factors in all genomes, but also led to the identification of a quorum sensing (QS) dependent transcriptional regulator that was unique to pathogenic Roseobacter strains. This observation supports the model that a combination of virulence factors and QS-dependent regulatory mechanisms enables indigenous members of the host alga's epiphytic microbial community to switch to a pathogenic lifestyle, especially under environmental conditions when innate host defence mechanisms are compromised

The responses of brown macroalgae to environmental change from local to global scales: direct versus ecologically mediated effects

In many temperate regions, brown macroalgae fulfil essential ecosystem services such as the provision of structure, the fixation of nutrients and carbon, and the production of biomass and oxygen. Their populations in many regions around the globe have declined and/or spatially shifted in recent decades. In this review we highlight the potential global and regional drives of these changes, describe the status of regionally particularly important brown macroalgal species, and describe the capacity of interactions among abiotic and biotic factors to amplify or buffer environmental pressure on brown macroalgae. We conclude with a consideration of possible management and restoration measures

Invasive Evaluation for Coronary Vasospasm

Vasospastic angina (VSA) occurs at rest and on exertion, with transient electrocardiographic ischemic changes. VSA presents with spontaneous coronary artery spasm (CAS); it has been associated with stable angina, acute coronary syndromes, and sudden cardiac death. CAS can be identified in normal arteries or non-obstructive coronary atherosclerosis, but is also prevalent in patients with coronary artery disease. The diagnosis is made with invasive coronary reactivity testing with provocation using acetylcholine (Ach). Epicardial spasms can be visualized through coronary angiography as a reversible epicardial vessel narrowing, while the diagnosis of microvascular spasm can be made when angina symptoms and ECG changes happen following intracoronary Ach without epicardial spasm. Identification of CAS allows for risk stratification and specific therapies targeting endothelial dysfunction and paradoxical vascular smooth muscle cell constriction. Therapies include calcium channel blockers as monotherapy or in a combination of a dihydropyridine and non-dihydropyridine. Short-acting nitrates offer acute symptomatic relief but long-acting nitrates should be used sparingly. This current update on invasive evaluation of VSA discusses unified Ach protocols

Toward a New Technology and Policy Program (TPP) Curriculum

The mission of the MIT Technology and Policy Program (TPP) is: “Provide an integrative education to scientists and engineers who wish to lead in the development and implementation of responsible strategies and policies for exploitation of technology for the benefit of their communities” (Hastings, 2000). Embedded in the TPP mission statement are several educational requirements: (1) a comprehensive and diverse set of solid analytical skills needed to develop and assess strategies and policies, (2) the flexibility to manage the conflicting interests and values that are present at all stages of the policy process, and (3) the ability to provide leadership at each stage in the policy process. With these concepts in mind, the TPP Curriculum Development Committee will work to place TPP at the forefront of educating the “leaders (researchers and practitioners) of the fields of technology and policy studies” (Hastings, 2000)