Nucleic acid-based fluorescent probes and their analytical potential

It is well known that nucleic acids play an essential role in living organisms because they store and transmit genetic information and use that information to direct the synthesis of proteins. However, less is known about the ability of nucleic acids to bind specific ligands and the application of oligonucleotides as molecular probes or biosensors. Oligonucleotide probes are single-stranded nucleic acid fragments that can be tailored to have high specificity and affinity for different targets including nucleic acids, proteins, small molecules, and ions. One can divide oligonucleotide-based probes into two main categories: hybridization probes that are based on the formation of complementary base-pairs, and aptamer probes that exploit selective recognition of nonnucleic acid analytes and may be compared with immunosensors. Design and construction of hybridization and aptamer probes are similar. Typically, oligonucleotide (DNA, RNA) with predefined base sequence and length is modified by covalent attachment of reporter groups (one or more fluorophores in fluorescence-based probes). The fluorescent labels act as transducers that transform biorecognition (hybridization, ligand binding) into a fluorescence signal. Fluorescent labels have several advantages, for example high sensitivity and multiple transduction approaches (fluorescence quenching or enhancement, fluorescence anisotropy, fluorescence lifetime, fluorescence resonance energy transfer (FRET), and excimer-monomer light switching). These multiple signaling options combined with the design flexibility of the recognition element (DNA, RNA, PNA, LNA) and various labeling strategies contribute to development of numerous selective and sensitive bioassays. This review covers fundamentals of the design and engineering of oligonucleotide probes, describes typical construction approaches, and discusses examples of probes used both in hybridization studies and in aptamer-based assays

Participatory adaptation planning and costing. Applications in agricultural adaptation in Western Kenya

Adaptation to climate change is an important theme in the strategy and policy of institutions around the world. Billions of dollars are allocated every year, based on cost estimates of actions to cope with, or benefit from the impacts of climate change. Costing adaptation, however, is complex, involving multiple actors with differing values and a spectrum of possible adaptation strategies and pathways. Currently, expert driven, top-down approaches dominate adaptation costing in practice. These approaches are subject to misallocation, with global funds not always reaching vulnerable communities in most need. This paper introduces an analytical framework called Participatory Social Return on Investment (PSROI), which provides a structured framework for multi-stakeholder planning, selection and valuation of appropriate methods of adaptation. The broader economic, social and environmental impacts of these adaptation actions are explored and valued through a participatory process. PSROI is strength-based, building local capacity and generating stakeholder buy-in. The financial valuation generated provides an additional tool for examining and prioritizing adaptation actions based on their impact. Results from a pilot of the PSROI framework in a smallholder farming community in Western Kenya provide empirical evidence for the difference between expert driven desk-based and ground-based cost estimates that involve local communities. There was an approximate 70 % reduction in the valuation of an agroforestry intervention, selected by the local community, when compared between the desk-based valuation and that of the local community, using primary field data. This reduced expectation of the desk-based PSROI is justified by coherent explanations such as lack of knowledge about the intervention, misconception about the potential costs and benefits, and the risk-averse nature of the farmers. These and other important insights are fundamental for planning and decision-making, as well as appropriate targeting and delivery of funding for adaptation.Abrar S. Chaudhury, Ariella Helfgott, Thomas F. Thornton, Chase Sov