A new selective fluorescent probe based on tamoxifen

© 2016 Elsevier Ltd. Developing targeted validation probes that can interrogate biology is of interest for both chemists and biologists. The synthesis of suitable compounds provides a means for avoiding the costly labeling of cells with specific antibodies and the bias associated with the interpretation of biological validation experiments. The chemotherapeutic agent, tamoxifen has been routinely used in the treatment of breast cancer for decades. Once metabolized, the active form of tamoxifen (4-hydroxytamoxifen) competes with the binding of estrogens to the estrogen receptors (ER). Its selectivity in ER modulation makes it an ideal candidate for the development of materials to be used as chemical probes. Here we report the synthesis of a fluorescent BODIPY®FL conjugate of tamoxifen linked through an ethylene glycol moiety, and present proof-of-principle results in ER positive and ER negative cell lines. Optical microscopy indicates that the fluorescent probe binds selectively to tamoxifen sensitive breast cancer cell lines. The compound showed no affinity for the tamoxifen resistant breast cancer lines. The specificity of the new compound make it a valuable addition to the chemical probe tool kit for estrogen receptors

Aging blow fly pupae using hyperspectral imaging

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Lipidomic features of honey bee and colony health during limited supplementary feeding

Honey bee nutritional health depends on nectar and pollen, which provide the main source of carbohydrates, proteins and lipids to individual bees. During malnutrition, insect metabolism accesses fat body reserves. However, this process in bees and its repercussions at the colony level are poorly understood. Using untargeted lipidomics and gene expression analysis, we examined the effects of different feeding treatments (starvation, sugar feeding and sugar + pollen feeding) on bees and correlated them with colony health indicators. We found that nutritional stress led to an increase in unsaturated triacylglycerols and diacylglycerols, as well as a decrease in free fatty acids in the bee fat body. Here, we hypothesise that stored lipids are made available through a process where unsaturations change lipid\u27s structure. Increased gene expression of three lipid desaturases in response to malnutrition supports this hypothesis, as these desaturases may be involved in releasing fatty acyl chains for lipolysis. Although nutritional stress was evident in starving and sugar-fed bees at the colony and physiological level, only starved colonies presented long-term effects in honey production

LATERAL BRANCHING OXIDOREDUCTASE acts in the final stages of strigolactone biosynthesis inArabidopsis

Strigolactones are a group of plant compounds of diverse but related chemical structures. They have similar bioactivity across a broad range of plant species, act to optimize plant growth and development, and promote soil microbe interactions. Carlactone, a common precursor to strigolactones, is produced by conserved enzymes found in a number of diverse species. Versions of the MORE AXILLARY GROWTH1 (MAX1) cytochrome P450 from rice and Arabidopsis thaliana make specific subsets of strigolactones from carlactone. However, the diversity of natural strigolactones suggests that additional enzymes are involved and remain to be discovered. Here, we use an innovative method that has revealed a missing enzyme involved in strigolactone metabolism. By using a transcriptomics approach involving a range of treatments that modify strigolactone biosynthesis gene expression coupled with reverse genetics, we identified LATERAL BRANCHING OXIDOREDUCTASE (LBO), a gene encoding an oxidoreductase-like enzyme of the 2-oxoglutarate and Fe(II)-dependent dioxygenase superfamily. Arabidopsis lbo mutants exhibited increased shoot branching, but the lbo mutation did not enhance the max mutant phenotype. Grafting indicated that LBO is required for a graft-transmissible signal that, in turn, requires a product of MAX1. Mutant lbo backgrounds showed reduced responses to carlactone, the substrate of MAX1, and methyl carlactonoate (MeCLA), a product downstream of MAX1. Furthermore, lbo mutants contained increased amounts of these compounds, and the LBO protein specifically converts MeCLA to an unidentified strigolactone-like compound. Thus, LBO function may be important in the later steps of strigolactone biosynthesis to inhibit shoot branching in Arabidopsis and other seed plants

Rethinking soil water repellency and its management

Soil water repellency (SWR) is a widespread challenge to plant establishment and growth. Despite considerable research, it remains a recalcitrant problem for which few alleviation technologies or solutions have been developed. Previous research has focused on SWR as a problem to be overcome, however, it is an inherent feature of many native ecosystems where it contributes to ecosystem functions. Therefore, we propose a shift in the way SWR is perceived in agriculture and in ecological restoration, from a problem to be solved, to an opportunity to be harnessed. A new focus on potential ecological benefits of SWR is particularly timely given increasing incidence, frequency and severity of hotter droughts in many regions of the world. Our new way of conceptualising SWR seeks to understand how SWR can be temporarily alleviated at a micro-scale to successfully establish plants, and then harnessed in the longer term and at larger spatial scales to enhance soil water storage to act as a “drought-proofing” tool for plant survival in water-limited soils. For this to occur, we suggest research focusing on the alignment of physico-chemical and microbial properties and dynamics of SWR and, based on this mechanistic understanding, create products and interventions to improve success of plant establishment in agriculture, restoration and conservation contexts. In this paper, we outline the rationale for a new way of conceptualising SWR, and the research priorities needed to fill critical knowledge gaps in order to harness the ecological benefits from managing SWR

Production of the seed germination stimulant karrikinolide from combustion of simple carbohydrates

The naturally occurring seed germination stimulant karrikinolide is formed from the combustion of plant material including cellulose. It has previously been reported that combustion of simple carbohydrates such as d-glucose does not produce extracts containing karrikinolide. Moreover, it was reported that extracts with germination-promoting ability could be obtained only by combustion of simple carbohydrates in the presence of amino acids such as l-glycine. By employing a 13C-labeled karrikinolide to physically quantify natural karrikinolide, we now show that it is produced from combustion of simple carbohydrates in similar amounts regardless of whether l-glycine is present or not. The addition of l-glycine appears to be beneficial in reducing the inhibitory effect of smoke extracts and provides a greater concentration range for effective germination-promoting activity

Regulation of seed germination and seedling growth by chemical signals from burning vegetation

It is well known that burning of vegetation stimulates new plant growth and landscape regeneration. The discovery that char and smoke from such fires promote seed germination in many species indicates the presence of chemical stimulants. Nitrogen oxides stimulate seed germination, but their importance in post-fire germination has been questioned. Cyanohydrins have been recently identified in aqueous smoke solutions and shown to stimulate germination of some species through the slow release of cyanide. However, the most information is available for karrikins, a family of butenolides related to 3-methyl-2H- furolsqb2,3-crsqbpyran-2-one. Karrikins stimulate seed germination and influence seedling growth. They are active in species not normally associated with fire, and in Arabidopsis they require the F-box protein MAX2, which also controls responses to strigolactone hormones. We hypothesize that chemical similarity between karrikins and strigolactones provided the opportunity for plants to employ a common signal transduction pathway to respond to both types of compound, while tailoring specific developmental responses to these distinct environmental signals

Structure-activity relationship of karrikin germination stimulants

Karrikins (2H-furo[2,3-c]pyran-2-ones) are potent smoke-derived germination promoters for a diverse range of plant species but, to date, their mode of action remains unknown. This paper reports the structure-activity relationship of numerous karrikin analogues to increase understanding of the key structural features of the molecule that are required for biological activity. The results demonstrate that modification at the C5 position is preferred over modification at the C3, C4, or C7 positions for retaining the highest bioactivity. © 2010 American Chemical Society

The synthesis and biological evaluation of labelled karrikinolides for the elucidation of the mode of action of the seed germination stimulant

Karrikins are a novel class of naturally occurring plant growth regulators that promote seed germination among a diverse range of species. Currently little is known about the mechanism by which these compounds overcome seed dormancy and initiate germination. The preparation of various karrikinolide derivatives provides an opportunity to investigate the karrikinolide mode of action at the cellular and molecular level. The first synthesis and biological evaluation of analogues suitable for use in metabolic labelling, affinity chromatography, photoaffinity labelling and NanoSIMS experiments are reported