CHO genome mining for synthetic promoter design

Synthetic promoters are an attractive alternative for use in mammalian hosts such as CHO cells as they can be designed de novo with user-defined functionalities. In this study, we describe and validate a method for bioprocess-directed design of synthetic promoters utilizing CHO genomic sequence information. We designed promoters with two objective features, (i) constitutive high-level recombinant gene transcription, and (ii) upregulated transcription under mild hypothermia or late-stage culture. CHO genes varying in transcriptional activity were selected based on a comparative analysis of RNA-Seq transcript levels in normal and biphasic cultures in combination with estimates of mRNA half-life from published genome scale datasets. Discrete transcription factor regulatory elements (TFREs) upstream of these genes were informatically identified and functionally screened in vitro to identify a subset of TFREs with the potential to support high activity recombinant gene transcription during biphasic cell culture processes. Two libraries of heterotypic synthetic promoters with varying TFRE combinations were then designed in silico that exhibited a maximal 2.5-fold increase in transcriptional strength over the CMV-IE promoter after transient transfection into host CHO-K1 cells. A subset of synthetic promoters was then used to create stable transfectant pools using CHO-K1 cells under glutamine synthetase selection. Whilst not achieving the maximal 2.5-fold increase in productivity over stable pools harboring the CMV promoter, all stably transfected cells utilizing synthetic promoters exhibited increased reporter production - up to 1.6-fold that of cells employing CMV, both in the presence or absence of intron A immediately downstream of the promoter. The increased productivity of stably transfected cells harboring synthetic promoters was maintained during fed-batch culture, with or without a transition to mild hypothermia at the onset of stationary phase. Our data exemplify that it is important to consider both host cell and intended bioprocess contexts as design criteria in the de novo construction of synthetic genetic parts for mammalian cell engineering

Urban groundwater quality in sub-Saharan Africa: current status and implications for water security and public health

Groundwater resources are important sources of drinking water in Africa, and they are hugely important in sustaining urban livelihoods and supporting a diverse range of commercial and agricultural activities. Groundwater has an important role in improving health in sub-Saharan Africa (SSA). An estimated 250 million people (40% of the total) live in urban centres across SSA. SSA has experienced a rapid expansion in urban populations since the 1950s, with increased population densities as well as expanding geographical coverage. Estimates suggest that the urban population in SSA will double between 2000 and 2030. The quality status of shallow urban groundwater resources is often very poor due to inadequate waste management and source protection, and poses a significant health risk to users, while deeper borehole sources often provide an important source of good quality drinking water. Given the growth in future demand from this finite resource, as well as potential changes in future climate in this region, a detailed understanding of both water quantity and quality is required to use this resource sustainably. This paper provides a comprehensive assessment of the water quality status, both microbial and chemical, of urban groundwater in SSA across a range of hydrogeological terrains and different groundwater point types. Lower storage basement terrains, which underlie a significant proportion of urban centres in SSA, are particularly vulnerable to contamination. The relationship between mean nitrate concentration and intrinsic aquifer pollution risk is assessed for urban centres across SSA. Current knowledge gaps are identified and future research needs highlighted

Subjective cues to deception/honesty in a high stakes situation: An exploratory approach

This is an Accepted Manuscript of an article published by Taylor & Francis in The Journal of Psychology: Interdisciplinary and Applied on 7/5/2014 available online: http://wwww.tandfonline.com/10.1080/00223980.2014.911140The low ecological validity of much of the research on deception detection is a limitation recognised by researchers in the field. Consequently, the present studies investigated subjective cues to deception using the real life, high stakes situation of people making public appeals for help with missing or murdered relatives. It was expected that cues related to affect would be particularly salient in this context. Study 1 was a qualitative investigation identifying cues to deception reportedly used by people accurate at detecting deception. Studies 2 and 3 were then empirical investigations which mainly employed the cues reported in Study 1. A number of subjective cues were found to discriminate between honest and deceptive appeals, including some previously unidentified cues, and cues likely to be context-specific. Most could be categorised under the themes of authenticity of emotion, and negative and positive affective reactions to the appealer. It is concluded that some cues to deception may emerge only in real life, high stakes situations; however, it is argued that some of these may be influenced by observers’ perceptions of the characteristics of offenders, rather than acts of deception per se.ESRC grant number ES/I90316X/

Constraint rule-based programming of norms for electronic institutions

Peer reviewedPostprin

Monitoring changes in human activity during the COVID-19 shutdown in Las Vegas using infrasound microbarometers

While studies of urban acoustics are typically restricted to the audio range, anthropogenic activity also generates infrasound (<20 Hz, roughly at the lower end of the range of human hearing). Shutdowns related to the COVID-19 pandemic unintentionally created ideal conditions for the study of urban infrasound and low frequency audio (20-500 Hz), as closures reduced human-generated ambient noise, while natural signals remained relatively unaffected. An array of infrasound sensors deployed in Las Vegas, NV, provides data for a case study in monitoring human activity during the pandemic through urban acoustics. The array records a sharp decline in acoustic power following the temporary shutdown of businesses deemed nonessential by the state of Nevada. This decline varies spatially across the array, with stations close to McCarran International Airport generally recording the greatest declines in acoustic power. Further, declines in acoustic power fluctuate with the time of day. As only signals associated with anthropogenic activity are expected to decline, this gives a rough indication of periodicities in urban acoustics throughout Las Vegas. The results of this study reflect the city's response to the pandemic and suggest spatiotemporal trends in acoustics outside of shutdowns

Synthesis, biological profiling and mechanistic studies of 4-aminoquinoline-based heterodimeric compounds with dual trypanocidal–antiplasmodial activity.

YesDual submicromolar trypanocidal–antiplasmodial compounds have been identified by screening and chemical synthesis of 4-aminoquinoline-based heterodimeric compounds of three different structural classes. In Trypanosoma brucei, inhibition of the enzyme trypanothione reductase seems to be involved in the potent trypanocidal activity of these heterodimers, although it is probably not the main biological target. Regarding antiplasmodial activity, the heterodimers seem to share the mode of action of the antimalarial drug chloroquine, which involves inhibition of the haem detoxification process. Interestingly, all of these heterodimers display good brain permeabilities, thereby being potentially useful for late stage human African trypanosomiasis. Future optimization of these compounds should focus mainly on decreasing cytotoxicity and acetylcholinesterase inhibitory activity

Multi-layered Ruthenium-modified Bond Coats for Thermal Barrier Coatings

Diffusional approaches for fabrication of multi-layered Ru-modified bond coats for thermal barrier coatings have been developed via low activity chemical vapor deposition and high activity pack aluminization. Both processes yield bond coats comprising two distinct B2 layers, based on NiAl and RuAl, however, the position of these layers relative to the bond coat surface is reversed when switching processes. The structural evolution of each coating at various stages of the fabrication process has been and subsequent cyclic oxidation is presented, and the relevant interdiffusion and phase equilibria issues in are discussed. Evaluation of the oxidation behavior of these Ru-modified bond coat structures reveals that each B2 interlayer arrangement leads to the formation of α-Al 2 O 3 TGO at 1100°C, but the durability of the TGO is somewhat different and in need of further improvement in both cases

Human annoyance, acceptability and concern as responses to vibration from the construction of light rapid transit lines in residential environments

The aim of this paper is to investigate the use of different self-reported measures for assessing the human response to environmental vibration from the construction of an urban LRT (Light Rapid Transit) system. The human response to environmental stressors such as vibration and noise is often expressed in terms of exposure–response relationships that describe annoyance as a function of the magnitude of the vibration. These relationships are often the basis of noise and vibration policy and the setting of limit values. This paper examines measures other than annoyance by expressing exposure–response relationships for vibration in terms of self-reported concern about property damage and acceptability. The exposure–response relationships for concern about property damage and for acceptability are then compared with those for annoyance. It is shown that concern about property damage occurs at vibration levels well below those where there is any risk of damage. Earlier research indicated that concern for damage is an important moderator of the annoyance induced. Acceptability, on the other hand, might be influenced by both annoyance and concern, as well as by other considerations. It is concluded that exposure–response relationships expressing acceptability as a function of vibration exposure could usefully complement existing relationships for annoyance in future policy decisions regarding environmental vibration. The results presented in this paper are derived from data collected through a socio-vibration survey (N = 321) conducted for the construction of an urban LRT in the United Kingdom

Microstructure, Processing, Performance Relationships for High Temperature Coatings

HVOF coating have shown high resistance to corrosion in fossil energy applications and it is generally accepted that mechanical failure, e.g. cracking or spalling, ultimately will determine coating lifetime. The high velocity oxygen-fuel method (HVOF) of applying coatings is one of the most commercially viable and allows the control of various parameters including powder particle velocity and temperature which influence coating properties, such as residual stress, bond coat strength and microstructure. Methods of assessing the mechanical durability of coatings are being developed in order to explore the relationship between HVOF spraying parameters and the mechanical properties of the coating and coating bond strength. The room temperature mechanical strength, as well as the resistance of the coating to cracking/spalling during thermal transients, is of considerable importance. Eddy current, acoustic emission and thermal imaging methods are being developed to detect coating failure during thermal cycling tests and room temperature tensile tests. Preliminary results on coating failure of HVOF FeAl coatings on carbon steel, as detected by eddy current measurements during thermal cycling, are presented. The influence of HVOF coating parameters of iron aluminides - applied to more relevant structural steels, like 316 SS and Grade 91 steel, - on coating durability will be explored once reliable methods for identification of coating failure have been developed