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

Methods for Detecting PER2:LUCIFERASE Bioluminescence Rhythms in Freely Moving Mice

Circadian rhythms are driven by daily oscillations of gene expression. An important tool for studying cellular and tissue circadian 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 circadian 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. The LumiCycle In Vivo applies a background subtraction that corrects for effects of room temperature on photomultiplier tube (PMT) output. We tested delivery of d-luciferin via a subcutaneous minipump and in the drinking water. We demonstrate spikes in bioluminescence associated with drinking bouts. 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, and can support longer-term studies. A small difference in phase of the PER2::LUC bioluminescence rhythms, with females phase leading males, can be detected with this technique. 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

Replication data for: Does the Diversionary Use of Force Threaten the Democratic Peace? Assessing the Effect of Economic Growth on Interstate Conflict, 1921- 2001

A democratic leader, anticipating a ‘‘rally ‘round the flag effect,’’ may have an incentive to divert attention from domestic economic problems by becoming involved in military conflict abroad, undermining Immanuel Kant’s prescription for ‘‘perpetual peace.’’ We assess the risk to the democratic peace by evaluating this diversionary incentive within a general dyadic model of interstate conflict, 1921–2001, using both directed and nondirected analyses. Our results indicate that economic conditions do affect the likelihood that a democracy, but not an autocracy, will initiate a fatal militarized dispute, even against another democracy. Economic growth rates sufficiently low to negate the democratic peace are, however, rare; and the behavior of five powerful democracies raises further doubts about the importance of diversions. We find no significant evidence that a bad economy makes a democratic state less likely to be targeted by others, nor does the timing of legislative elections influence the decision of democratic leaders to use force. Alth ough economic conditions affect the likelihood of a fatal dispute for democracies, the influence is sufficiently small that Kant’s hope for a more peaceful world does not seem misplaced. A copy of the article is available here http://www.bama.ua.edu/~joneal/ISQ2006_data

ASTER and Landsat ETM+ images applied to sugarcane yield forecast

This paper proposes a method to support sugarcane yield forecast using vegetation spectral indices, principal component analysis and historic yield data. The study area is located in the State of Sao Paulo, Brazil, and is divided into 11 production plots (108.75 ha), where sugarcane of the RB85 5536 variety is cultivated on red latossol (oxissol-type) soil and flat topography. The data employed in the study include radiometrically and geometrically corrected enhanced thermatic mapper Plus (ETM+)/Landsat-7 and ASTER/Terra images, acquired in June and April 2001, respectively, and historic harvest data measured in 2000 and 2001. The method comprises several steps: (a) enhancement of specific spectral responses of vegetation constituents; (b) reduction of spectral dimensions with prioritization of information and weighing of parameters related to foliar area; the data processed through these steps are reduced to a single image (the synthesis image), from which the mean DN (digital number) per cultivated area is calculated; (c) the image DNs are subsequently transformed into ton of stalk per hectare (t ha(-1)) through normalization, which requires knowledge of the previous year's yield for the cultivated production plots under analysis. Yield estimates using the method showed greater precision in comparison to the ubiquitous visual methods employed by the sugarcane agro-industry in Brazil. Using factual productivity data of the year 2000 harvest only, the method achieved estimate errors varying between 2.57% and 5.65%, compared with 9.06% expected by the sugar factory; whereas using data from the year 2001 harvest, error margins were remarkably lower, around 1%.27194057406