Recombinant canine single chain insulin analogues: Insulin receptor binding capacity and ability to stimulate glucose uptake

Virtually all diabetic dogs require exogenous insulin therapy to control their hyperglycaemia. In the UK, the only licensed insulin product currently available is a purified porcine insulin preparation. Recombinant insulin is somewhat problematic in terms of its manufacture, since the gene product (preproinsulin) undergoes substantial post-translational modification in pancreatic β cells before it becomes biologically active. The aim of the present study was to develop recombinant canine single chain insulin (SCI) analogues that could be produced in a prokaryotic expression system and which would require minimal processing. Three recombinant SCI constructs were developed in a prokaryotic expression vector, by replacing the insulin C-peptide sequence with one encoding a synthetic peptide (GGGPGKR), or with one of two insulin-like growth factor (IGF)-2 C-peptide coding sequences (human: SRVSRRSR; canine: SRVTRRSSR). Recombinant proteins were expressed in the periplasmic fraction of Escherichia coli and assessed for their ability to bind to the insulin and IGF-1 receptors, and to stimulate glucose uptake in 3T3-L1 adipocytes. All three recombinant SCI analogues demonstrated preferential binding to the insulin receptor compared to the IGF-1 receptor, with increased binding compared to recombinant canine proinsulin. The recombinant SCI analogues stimulated glucose uptake in 3T3-L1 adipocytes compared to negligible uptake using recombinant canine proinsulin, with the canine insulin/cIGF-2 chimaeric SCI analogue demonstrating the greatest effect. Thus, biologically-active recombinant canine SCI analogues can be produced relatively easily in bacteria, which could potentially be used for treatment of diabetic dogs

Emotionality and fact orientation in environmental activism: A comparative content analysis of the communication of Fridays for Future and the Last Generation on Instagram

The increasing threat of climate change is particularly characterized in public discourse in Germany by the environmental activism of Fridays For Future and Letzte Generation (trans-lated: Last Generation). Both environmental movements mobilize by communicating the causes of problems, approaches to solutions and possible ways to motivate people. Although emotionalized and fact-oriented communication suggests a significant influence on this, the topic has not yet been researched. A comparative content analysis of 283 image and video posts from both climate protest movements on Instagram from 2023 revealed the predominant role of anger, determination, despair and regret in the diagnostic and prognostic attribution of both movements. Determination, hope and gratitude could be identified in the motivational function of the communication of both activist groups. While Letzte Generation often makes negative emotional statements, Fridays For Future is characterized by a more positive emotionality. Particularly with regard to the attribution of problems, both movements consistently refer to facts, whereby Letzte Generation in particular explicitly refers to sources. The work provides insights into the role of emotionality and fact orientation in a comparison of the two currently most relevant German climate protest movements. It thus offers a starting point for further research into the mobilization communication of environmental movements