Screening and classifying small-molecule inhibitors of amyloid formation using ion mobility spectrometry-mass spectrometry

The search for therapeutic agents that bind specifically to precursor protein conformations and inhibit amyloid assembly is an important challenge. Identifying such inhibitors is difficult because many protein precursors of aggregation are partially folded or intrinsically disordered, which rules out structure-based design. Furthermore, inhibitors can act by a variety of mechanisms, including specific or nonspecific binding, as well as colloidal inhibition. Here we report a high-throughput method based on ion mobility spectrometry–mass spectrometry (IMS–MS) that is capable of rapidly detecting small molecules that bind to amyloid precursors, identifying the interacting protein species and defining the mode of inhibition. Using this method we have classified a variety of small molecules that are potential inhibitors of human ​islet amyloid polypeptide (​hIAPP) aggregation or ​amyloid-beta 1-40 aggregation as specific, nonspecific, colloidal or non-interacting. We also demonstrate the ability of IMS–MS to screen for inhibitory small molecules in a 96-well plate format and use this to discover a new inhibitor of ​hIAPP amyloid assembly

An in vivo platform to select and evolve aggregation-resistant proteins

Protein biopharmaceuticals are highly successful, but their utility is compromised by their propensity to aggregate during manufacture and storage. As aggregation can be triggered by non-native states, whose population is not necessarily related to thermodynamic stability, prediction of poorly-behaving biologics is difficult, and searching for sequences with desired properties is labour-intensive and time-consuming. Here we show that an assay in the periplasm of E. coli linking aggregation directly to antibiotic resistance acts as a sensor for the innate (un-accelerated) aggregation of antibody fragments. Using this assay as a directed evolution screen, we demonstrate the generation of aggregation resistant scFv sequences when reformatted as IgGs. This powerful tool can thus screen and evolve ‘manufacturable’ biopharmaceuticals early in industrial development. By comparing the mutational profiles of three different immunoglobulin scaffolds, we show the applicability of this method to investigate protein aggregation mechanisms important to both industrial manufacture and amyloid disease

High sensitivity and specificity of a 5-analyte protein and microRNA biosignature for identification of active tuberculosis.

Objectives: Non-sputum-based tests to accurately identify active tuberculosis (TB) disease and monitor response to therapy are urgently needed. This study examined the biomarker capacity of a panel of plasma proteins alone, and in conjunction with a previously identified miRNA signature, to identify active TB disease. Methods: The expression of nine proteins (IP-10, MCP-1, sTNFR1, RANTES, VEGF, IL-6, IL-10, TNF and Eotaxin) was measured in the plasma of 100 control subjects and 100 TB patients, at diagnosis (treatment naïve) and over the course of treatment (1-, 2- and 6-month intervals). The diagnostic performance of the nine proteins alone, and with the miRNA, was assessed. Results: Six proteins were significantly up-regulated in the plasma of TB patients at diagnosis compared to controls. Receiver operator characteristic curve analysis demonstrated that IP-10 with an AUC = 0.874, sensitivity of 75% and specificity of 87% was the best single biomarker candidate to distinguish TB patients from controls. IP-10 and IL-6 levels fell significantly within one month of commencing treatment and may have potential as indicators of a positive response to therapy. The combined protein and miRNA panel gave an AUC of 1.00. A smaller panel of only five analytes (IP-10, miR-29a, miR-146a, miR-99b and miR-221) showed an AUC = 0.995, sensitivity of 96% and specificity of 97%. Conclusions: A novel combination of miRNA and proteins significantly improves the sensitivity and specificity as a biosignature over single biomarker candidates and may be useful for the development of a non-sputum test to aid the diagnosis of active TB disease

Towards simulated morality systems: Role-playing games as artificial societies

Computer role-playing games (RPGs) often include a simulated morality system as a core design element. Games' morality systems can include both god's eye view aspects, in which certain actions are inherently judged by the simulated world to be good or evil, as well as social simulations, in which non-player characters (NPCs) react to judgments of the player's and each others' activities. Games with a larger amount of social simulation have clear affinities to multi-agent systems (MAS) research on artificial societies. They differ in a number of key respects, however, due to a mixture of pragmatic game-design considerations and their typically strong embeddedness in narrative arcs, resulting in many important aspects of moral systems being represented using explicitly scripted scenarios rather than through agent-based simulations. In this position paper, we argue that these similarities and differences make RPGs a promising challenge domain for MAS research, highlighting features such as moral dilemmas situated in more organic settings than seen in game-theoretic models of social dilemmas, and heterogeneous representations of morality that use both moral calculus systems and social simulation. We illustrate some possible approaches using a case study of the morality systems in the game The Elder Scrolls IV: Oblivion

Small Tympanic Membrane Perforations in the Inferior Quadrants Do Not Impact the Manubrium Vibration in Guinea Pigs

BACKGROUND: It has been believed that location of the perforation has a significant impact on hearing loss. However, recent studies have demonstrated that the perforation sites had no impact on hearing loss. We measured the velocity and pattern of the manubrium vibration in guinea pigs with intact and perforated eardrum using a laser Doppler vibrometer in order to determine the effects of different location perforations on the middle ear transfer functions. METHODS: Two bullas from 2 guinea pigs were used to determine stability of the umbo velocities, and 12 bullas from six guinea pigs to determine the effects of different location perforations on sound transmission. The manubrium velocity was measured at three points on the manubrium in the frequencies of 0.5-8 kHz before and after a perforation was made. The sites of perforations were in anterior-inferior (AI) quadrants of left ears and posterior-inferior (PI) quadrants of right ears. RESULTS: The manubrium vibration velocity losses were noticed in the perforated ears only below 1.5 kHz. The maximum velocity loss was about 7 dB at 500 Hz with the PI perforation. No significant difference in the velocity loss was found between AI and PI perforations. The average ratio of short process velocity to the umbo velocity was approximately 0.5 at all frequencies. No significant differences were found before and after perforation at all frequencies (p>0.05) except 7 kHz (p = 0.004) for both AI and PI perforations. CONCLUSIONS: The manubrium vibration velocity losses from eardrum perforation were frequency-dependent and the largest losses occur at low frequencies. Manubrium velocity losses caused by small acute inferior perforations in guinea pigs have no significant impact on middle ear sound transmission at any frequency tested. The manubrium vibration axis may be perpendicular to the manubrium below 8 kHz in guinea pigs

Improving detection of familial hypercholesterolaemia in primary care using electronic audit and nurse-led clinics

RATIONALE, AIMS AND OBJECTIVES: In the UK fewer than 15% of familial hypercholesterolemia (FH) cases are diagnosed, representing a major gap in coronary heart disease prevention. We wished to support primary care doctors within the Medway Clinical Commissioning Group (CCG) to implement NICE guidance (CG71) and consider the possibility of FH in adults who have raised total cholesterol concentrations, thereby improving the detection of people with FH. METHODS: Utilizing clinical decision support software (Audit+) we developed an FH Audit Tool and implemented a systematic audit of electronic medical records within GP practices, first identifying all patients diagnosed with FH or possible FH and next electronically flagging patients with a recorded total cholesterol of >7.5 mmol L(-1) or LDL-C > 4.9 mmol L(-1) (in adults), for further assessment. After a 2-year period, a nurse-led clinic was introduced to screen more intensely for new FH index cases. We evaluated if these interventions increased the prevalence of FH closer to the expected prevalence from epidemiological studies. RESULTS: The baseline prevalence of FH within Medway CCG was 0.13% (1 in 750 persons). After 2 years, the recorded prevalence of diagnosed FH increased by 0.09% to 0.22% (1 in 450 persons). The nurse advisor programme ran for 9 months (October 2013-July 2014) and during this time, the recorded prevalence of patients diagnosed with FH increased to 0.28% (1 in 357 persons) and the prevalence of patients 'at risk and unscreened' reduced from 0.58% to 0.14%. CONCLUSIONS: Our study shows that two simple interventions increased the detection of FH. This systematic yet simple electronic case-finding programme with nurse-led review allowed the identification of new index cases, more than doubling the recorded prevalence of detected disease to 1 in 357 (0.28%). This study shows that primary care has an important role in identifying patients with this condition

Exploring Novel Game Spaces with Fluidic Games

AISB: Games AI & VR, 4 pages, 4 figures, game design, tools, creativityWith the growing integration of smartphones into our daily lives, and their increased ease of use, mobile games have become highly popular across all demographics. People listen to music, play games or read the news while in transit or bridging gap times. While mobile gaming is gaining popularity, mobile expression of creativity is still in its early stages. We present here a new type of mobile app -- fluidic games -- and illustrate our iterative approach to their design. This new type of app seamlessly integrates exploration of the design space into the actual user experience of playing the game, and aims to enrich the user experience. To better illustrate the game domain and our approach, we discuss one specific fluidic game, which is available as a commercial product. We also briefly discuss open challenges such as player support and how generative techniques can aid the exploration of the game space further

An eHealth-Care Driven Perspective on 5G Networks and Infrastructure

This work describes the advancements that next generation mobile networks can bring to emergency services on the basis of a fully 5G enabled medical emergency response scenario. An ambulance service combining autonomous driving, advanced on-board patient monitoring, remote diagnosis and remote control from the hospital is introduced, allowing increased levels of care during patient transport and improved early diagnosis, thus enhancing patient survival rates. Furthermore, it is shown that such an ambulance service requires a variety of different traffic types that can only be supported concurrently and with guaranteed quality of service by a high-performance network fulfilling all 5G key performance indicators. The scenario described combines a multitude of aspects and applications enabled by 5G mobile communications, including autonomous driving, ultra-high definition video streaming, tactile remote interaction and continuous sensing, into a compelling showcase for a 5G enabled future. A centralized radio access 5G network with space division multiplexed optical fronthaul using analog radio-over-fiber and optical beamforming is analyzed, fully supporting SDN and NFV for advanced network slicing and quality of service guarantee.</p

Combining cardiovascular and pupil features using k-nearest neighbor classifiers to assess task demand, social context and sentence accuracy during listening

In daily life, both acoustic factors and social context can affect listening effort investment. In laboratory settings, information about listening effort has been deduced from pupil and cardiovascular responses independently. The extent to which these measures can jointly predict listening-related factors is unknown. Here we combined pupil and cardiovascular features to predict acoustic and contextual aspects of speech perception. Data were collected from 29 adults (mean  =  64.6 years, SD  =  9.2) with hearing loss. Participants performed a speech perception task at two individualized signal-to-noise ratios (corresponding to 50% and 80% of sentences correct) and in two social contexts (the presence and absence of two observers). Seven features were extracted per trial: baseline pupil size, peak pupil dilation, mean pupil dilation, interbeat interval, blood volume pulse amplitude, pre-ejection period and pulse arrival time. These features were used to train k-nearest neighbor classifiers to predict task demand, social context and sentence accuracy. The k-fold cross validation on the group-level data revealed above-chance classification accuracies: task demand, 64.4%; social context, 78.3%; and sentence accuracy, 55.1%. However, classification accuracies diminished when the classifiers were trained and tested on data from different participants. Individually trained classifiers (one per participant) performed better than group-level classifiers: 71.7% (SD  =  10.2) for task demand, 88.0% (SD  =  7.5) for social context, and 60.0% (SD  =  13.1) for sentence accuracy. We demonstrated that classifiers trained on group-level physiological data to predict aspects of speech perception generalized poorly to novel participants. Individually calibrated classifiers hold more promise for future applications