Methods for detecting PER2::LUCIFERASE bioluminescence rhythms in freely moving mice [preprint]

Circadian rhythms are driven by daily oscillations of gene expression. An important tool for studying cellular and tissue rhythms is the use of a gene reporter, such as bioluminescence from the reporter gene luciferase controlled by a rhythmically expressed gene of interest. Here we describe methods that allow measurement of bioluminescence from a freely-moving mouse housed in a standard cage. Using a LumiCycle In Vivo (Actimetrics), we determined conditions that allow detection of circadian rhythms of bioluminescence from the PER2 reporter, PER2::LUC, in freely behaving mice. We tested delivery of D-luciferin via a subcutaneous minipump and in the drinking water. Further, we demonstrate that a synthetic luciferase substrate, CycLuc1, can support circadian rhythms of bioluminescence, even when delivered at a lower concentration than D-luciferin. We share our analysis scripts and suggestions for further improvements in this method. This approach will be straightforward to apply to mice with tissue-specific reporters, allowing insights into responses of specific peripheral clocks to perturbations such as environmental or pharmacological manipulations

Lake drying and livelihood dynamics in Lake Chad: unravelling the mechanisms, contexts and responses

This article examines lake drying and livelihood dynamics in the context of multiple stressors through a case study of the ‘‘Small Lake Chad’’ in the Republic of Chad. Livelihoods research in regions experiencing persistent lake water fluctuations has largely focused on the wellbeing and security of lakeshore dwellers. Little is known about the mechanisms through which lake drying shapes livelihood drawbacks and opportunities, and whether locally evolved responses are enhancing livelihoods. Here we address these gaps using empirical, mixed-methods field research couched within the framework of livelihoods and human well-being contexts. The analysis demonstrates that limited opportunities outside agriculture, the influx of mixed ethnic migrants and the increasing spate of violence all enhance livelihood challenges. Livelihood opportunities centre on the renewal effects of seasonal flood pulses on lake waters and the learning opportunities triggered by past droughts. Although drying has spurred new adaptive behaviours predicated on seasonality, traditional predictive factors and the availability of assets, responses have remained largely reactive. The article points to where lake drying fits amongst changes in the wider socioeconomic landscape in which people live, and suggests that awareness of the particularities of the mechanisms that connect lake drying to livelihoods can offer insights into the ways local people might be assisted by governments and development actors

Arms Racing, Military Build-Ups and Dispute Intensity: Evidence from the Greek-Turkish Rivalry, 1985-2020

Arms races are linked in the public conscience to potential violence. Following gas discoveries in eastern Mediterranean, Greece and Turkey nearly came to blows in August 2020 and both states have enacted military expansion plans, further risking escalation. We present a novel approach to study the effect of military build-ups on dispute intensity, using monthly data on Turkish incursions into Greek-claimed airspace. Because airspace claims feature strongly in the dispute, these contestations represent an appropriate measure of the intensity with which Turkey pursues the conflict. Theoretically, we suggest that bilateral factors drive this intensity. We argue that increased Greek military capabilities deter incursions whereas increased Turkish military capabilities fuel them. Results from time-series models support the second expectation. Consequently, the study provides a novel methodological approach to studying interstate conflict intensity and shines new light on escalation dynamics in the Greek-Turkish dispute

Effects of Cage Position and Light Transmission on Home Cage Activity and Circadian Entrainment in Mice

Light is known to exert powerful effects on behavior and physiology, including upon the amount and distribution of activity across the day/night cycle. Here we use home cage activity monitoring to measure the effect of differences in home cage light spectrum and intensity on key circadian activity parameters in mice. Due to the relative positioning of any individually ventilated cage (IVC) with regard to the animal facility lighting, notable differences in light intensity occur across the IVC rack. Although all mice were found to be entrained, significant differences in the timing of activity onset and differences in activity levels were found between mice housed in standard versus red filtering cages. Furthermore, by calculating the effective irradiance based upon the known mouse photopigments, a significant relationship between light intensity and key circadian parameters are shown. Perhaps unsurprisingly given the important role of the circadian photopigment melanopsin in circadian entrainment, melanopic illuminance is shown to correlate more strongly with key circadian activity parameters than photopic lux. Collectively, our results suggest that differences in light intensity may reflect an uncharacterized source of variation in laboratory rodent research, with potential consequences for reproducibility. Room design and layout vary within and between facilities, and caging design and lighting location relative to cage position can be highly variable. We suggest that cage position should be factored into experimental design, and wherever possible, experimental lighting conditions should be characterized as a way of accounting for this source of variation

Nonlinear optimal control for Synchronous Reluctance Machines

A nonlinear H-infinity (optimal) control approach is proposed for the problem of control of Synchronous Reluctance Machines (SRMs). Approximate linearization is applied to the dynamic model of the Synchronous Reluctance Machine, round a local operating point. To accomplish this linearization Taylor series expansion and the computation of the associated Jacobian matrices are performed. The robustness of the control scheme assures that the modelling error due to truncation of higher order terms from the Taylor expansion will be compensated. Next, an H-infinity feedback controller is designed. After solving an algebraic Riccati equation at each iteration of the control algorithm, the feedback gain is computed. Lyapunov stability analysis proves that the control loop satisfies an H-infinity tracking performance criterion. This in turn signifies elevated robustness to model uncertainty and external perturbations. Moreover, under moderate conditions it is proven that the control loop is globally asymptotically stable