Extracellular and intracellular NO detection in plants by diaminofluoresceins

Many assays focus on determining NO content within plant tissues to assess the actual concentration that impacts on cellular processes. Diaminofluorescein fluorescent dyes (DAFs) have been very widely used by plant scientists to reveal likely sites of NO production inside and outside cells. In general, DAFs dyes react with N2O3, a byproduct of NO oxidation, resulting in fluorescence. It is initially available in the form of diacetate (DAF-2DA), which allowed the ready absorption by the cells. The diacetate group is removed by cell esterases leaving the membrane impermeable to DAF-2 and available for N2O3 nitration to generate the highly fluorescent triazole (DAF-2T). Here, we describe two methods for detection of NO by fluorescence, one for NO extracellular detection by DAF-2 and the other one for NO intracellular detection, in this case using DAF-2DA1990103108CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPNão tem2012/19167-0; 2015/21546-7N.M.S. acknowledges the São Paulo Research Foundation (Fapesp, Brazil) (Grant no. 2012/19167-0 and 2015/21546-7), and R.V.R. and E.C.M. acknowledge the fellowship granted by the National Council for Scientific and Technological Development (CNPq, Brazil). The authors also acknowledge the Centre for Research in Biosciences at University of the West of England (UWE

Spatial co-localisation of firms and intrepreneurial dynamics The combinatory system view

In this conceptual paper, I propose an interpretation of business dynamics in terms of the spatial co-localisation of firms—considered as an intelligent cognitive system—in a circumscribed area in order to form clusters of various types. I interpret clustering by adopting the methodology of multi-agent combinatory systems: that is, systems formed by collectivities of agents (firms) in which an internal feedback recombines the micro behaviours of the agents (localisation) in order to produce a macro effect (cluster) which, in turn, modifies the attractiveness of the area and conditions the subsequent localisations. I also present the idea that if a cluster has fitness advantages for new firms, then usually new entrepreneurs are formed within it and the cluster widens, due to the endogenous genesis of new firms