Adaption of the ex vivo mycobacterial growth inhibition assay for use with murine lung cells.

In the absence of a correlate(s) of protection against human tuberculosis and a validated animal model of the disease, tools to facilitate vaccine development must be identified. We present an optimised ex vivo mycobacterial growth inhibition assay (MGIA) to assess the ability of host cells within the lung to inhibit mycobacterial growth, including Bacille Calmette-Guérin (BCG) and Mycobacterium tuberculosis (MTB) Erdman. Growth of BCG was reduced by 0.39, 0.96 and 0.73 log10 CFU following subcutaneous (s.c.) BCG, intranasal (i.n.) BCG, or BCG s.c. + mucosal boost, respectively, versus naïve mice. Comparatively, a 0.49 (s.c.), 0.60 (i.n.) and 0.81 (s.c. + mucosal boost) log10 reduction in MTB CFU was found. A BCG growth inhibitor, 2-thiophenecarboxylic acid hydrazide (TCH), was used to prevent quantification of residual BCG from i.n. immunisation and allow accurate MTB quantification. Using TCH, a further 0.58 log10 reduction in MTB CFU was revealed in the i.n. group. In combination with existing methods, the ex vivo lung MGIA may represent an important tool for analysis of vaccine efficacy and the immune mechanisms associated with vaccination in the organ primarily affected by MTB disease

Exploring the use of strategic frameworks in dental practice

This paper explores the use of strategic frameworks in NHS and private dental practice. It reviews the policy context of dentistry and suggests the challenges in this context will require dental practices to prioritise understanding and engagement with a strategic approach. A strategic approach will be required in order to enhance and improve performance. Two specific strategic frameworks will be explored in terms of their relevance to NHS and private dental practic

Intracellular replication of the well-armed pathogen Burkholderia pseudomallei.

The Burkholderia genus contains a group of soil-dwelling Gram-negative organisms that are prevalent in warm and humid climates. Two species in particular are able to cause disease in animals, B. mallei primarily infects Equus spp. and B. pseudomallei (BPS), that is able to cause potentially life-threatening disease in humans. BPS is naturally resistant to many antibiotics and there is no vaccine available. Although not a specialised human pathogen, BPS possesses a large genome and many virulence traits that allow it to adapt and survive very successfully in the human host. Key to this survival is the ability of BPS to replicate intracellularly. In this review we highlight recent advances in our understanding of the intracellular survival of BPS, including how it overcomes host immune defenses and other challenges to establish its niche and then spread the infection. Knowledge of these mechanisms increases our capacity for therapeutic interventions against a well-armed foe

Thermal refraction: implications for subglacial heat flux

First published online: 12 May 2021In this study, we explore small-scale (~1 to 20 km) thermal-refractive effects on basal geothermal heat flux (BGHF) at subglacial boundaries resulting from lateral thermal conductivity contrasts associated with subglacial topography and geologic contacts. We construct a series of two-dimensional, conductive, steady-state models that exclude many of the complexities of ice sheets in order to demonstrate the effect of thermal refraction. We show that heat can preferentially flow into or around a subglacial valley depending on the thermal conductivity contrast with underlying bedrock, with anomalies of local BGHF at the ice–bedrock interface between 80 and 120% of regional BGHF and temperature anomalies on the order of ±15% for the typical range of bedrock conductivities. In the absence of bed topography, subglacial contacts can produce significant heat flux and temperature anomalies that are locally extensive (>10 km). Thermal refraction can result in either an increase or decrease in the likelihood of melting and ice-sheet stability depending on the conductivity contrast and bed topography. While our models exclude many of the physical complexities of ice behavior, they illustrate the need to include refractive effects created by realistic geology into future glacial models to improve the prediction of subglacial melting and ice viscosity.Simon Willcocks, Derrick Hasterok, Samuel Jenning

A Method to Improve the Early Stages of the Robotic Process Automation Lifecycle

The robotic automation of processes is of much interest to organizations. A common use case is to automate the repetitive manual tasks (or processes) that are currently done by back-office staff through some information system (IS). The lifecycle of any Robotic Process Automation (RPA) project starts with the analysis of the process to automate. This is a very time-consuming phase, which in practical settings often relies on the study of process documentation. Such documentation is typically incomplete or inaccurate, e.g., some documented cases never occur, occurring cases are not documented, or documented cases differ from reality. To deploy robots in a production environment that are designed on such a shaky basis entails a high risk. This paper describes and evaluates a new proposal for the early stages of an RPA project: the analysis of a process and its subsequent design. The idea is to leverage the knowledge of back-office staff, which starts by monitoring them in a non-invasive manner. This is done through a screen-mousekey- logger, i.e., a sequence of images, mouse actions, and key actions are stored along with their timestamps. The log which is obtained in this way is transformed into a UI log through image-analysis techniques (e.g., fingerprinting or OCR) and then transformed into a process model by the use of process discovery algorithms. We evaluated this method for two real-life, industrial cases. The evaluation shows clear and substantial benefits in terms of accuracy and speed. This paper presents the method, along with a number of limitations that need to be addressed such that it can be applied in wider contexts.Ministerio de Economía y Competitividad TIN2016-76956-C3-2-

High-throughput analysis of Yersinia pseudotuberculosis gene essentiality in optimised in vitro conditions, and implications for the speciation of Yersinia pestis.

BACKGROUND: Yersinia pseudotuberculosis is a zoonotic pathogen, causing mild gastrointestinal infection in humans. From this comparatively benign pathogenic species emerged the highly virulent plague bacillus, Yersinia pestis, which has experienced significant genetic divergence in a relatively short time span. Much of our knowledge of Yersinia spp. evolution stems from genomic comparison and gene expression studies. Here we apply transposon-directed insertion site sequencing (TraDIS) to describe the essential gene set of Y. pseudotuberculosis IP32953 in optimised in vitro growth conditions, and contrast these with the published essential genes of Y. pestis. RESULTS: The essential genes of an organism are the core genetic elements required for basic survival processes in a given growth condition, and are therefore attractive targets for antimicrobials. One such gene we identified is yptb3665, which encodes a peptide deformylase, and here we report for the first time, the sensitivity of Y. pseudotuberculosis to actinonin, a deformylase inhibitor. Comparison of the essential genes of Y. pseudotuberculosis with those of Y. pestis revealed the genes whose importance are shared by both species, as well as genes that were differentially required for growth. In particular, we find that the two species uniquely rely upon different iron acquisition and respiratory metabolic pathways under similar in vitro conditions. CONCLUSIONS: The discovery of uniquely essential genes between the closely related Yersinia spp. represent some of the fundamental, species-defining points of divergence that arose during the evolution of Y. pestis from its ancestor. Furthermore, the shared essential genes represent ideal candidates for the development of novel antimicrobials against both species

Amperometric and spectrophotometric determination of carbaryl in natural waters and commercial formulations

The work presented describes the development and evaluation of two flow-injection analysis (FIA) systems for the automated determination of carbaryl in spiked natural waters and commercial formulations. Samples are injected directly into the system where they are subjected to alkaline hydrolysis thus forming 1-naphthol. This product is readily oxidised at a glassy carbon electrode. The electrochemical behaviour of 1-naphthol allows the development of an FIA system with an amperometric detector in which 1-naphthol determination, and thus measurement of carbaryl concentration, can be performed. Linear response over the range 1.0×10–7 to 1.0×10–5 mol L–1, with a sampling rate of 80 samples h–1, was recorded. The detection limit was 1.0×10–8 mol L–1. Another FIA manifold was constructed but this used a colorimetric detector. The methodology was based on the coupling of 1-naphthol with phenylhydrazine hydrochloride to produce a red complex which has maximum absorbance at 495 nm. The response was linear from 1.0×10–5 to 1.5×10–3 mol L–1 with a detection limit of 1.0×10–6 mol L–1. Sample-throughput was about 60 samples h–1. Validation of the results provided by the two FIA methodologies was performed by comparing them with results from a standard HPLC–UV technique. The relative deviation was <5%. Recovery trials were also carried out and the values obtained ranged from 97.0 to 102.0% for both methods. The repeatability (RSD, %) of 12 consecutive injections of one sample was 0.8% and 1.6% for the amperometric and colorimetric systems, respectively

Towards an OpenSource Logger for the Analysis of RPA Projects

Process automation typically begins with the observation of humans conducting the tasks that will be eventually automated. Sim ilarly, successful RPA projects require a prior analysis of the undergo ing processes which are being executed by humans. The process of col lecting this type of information is known as user interface (UI) logging since it records the interaction against a UI. Main RPA platforms (e.g., Blueprism and UIPath) incorporate functionalities that allow the record ing of these UI interactions. However, the records that these platforms generate lack some functionalities that large-scale RPA projects require. Besides, they are only understandable by the proper RPA platforms. This paper presents an extensible and multi-platform OpenSource UI logger that generate UI logs in a standard format. This system collects information from all the computers it is running on and sends it to a central server for its processing. Treatment of the collected information will allow the creation of an enriched UI log which can be used, among others purposes, for smart process analysis, machine learning training, the creation of RPA robots, or, being more general, for task mining .Ministerio de Economía y Competitividad TIN2016-76956-C3-2-R (POLOLAS)Junta de Andalucía CEI-12-TIC021Centro para el Desarrollo Tecnol´ogico Industrial (CDTI) P011-19/E0

UK Housing Market: Time Series Processes with Independent and Identically Distributed Residuals

The paper examines whether a univariate data generating process can be identified which explains the data by having residuals that are independent and identically distributed, as verified by the BDS test. The stationary first differenced natural log quarterly house price index is regressed, initially with a constant variance and then with a conditional variance. The only regression function that produces independent and identically distributed standardised residuals is a mean process based on a pure random walk format with Exponential GARCH in mean for the conditional variance. There is an indication of an asymmetric volatility feedback effect but higher frequency data is required to confirm this. There could be scope for forecasting the index but this is tempered by the reduction in the power of the BDS test if there is a non-linear conditional variance process