Colour development of immunogold-labelled antibodies for light microscopy

We describe an improvement of the immunogold technique, which is based on the colour development of silver-intensified immunogold signals. This method (referred to as the coloured-SIG technique) was found to be as sensitive as the silver-intensified immunogold method and more sensitive (in two of the three tested systems) than immunoenzymatic procedures, such as the peroxidase/antiperoxidase technique or the avidin-biotin system. The coloured SIG-method results in either a magenta-red or a cyan-blue final reaction product. Therefore, this new improvement of the immunogold technique should be useful in double-staining methods, immunoblot methods and conventional immunostaining methods.     

Global mapping of cost‐effective microalgal biofuel production areas with minimal environmental impact

Sustainable alternatives to fossil fuels are urgently needed to avoid severe climate impacts and further environmental degradation. Microalgae are one of the most productive crops globally and do not need to compete for arable land or freshwater resources. Hence, they may become a promising, more sustainable cultivation alternative for the large‐scale production of biofuels provided that substantial reductions are achieved in their production costs. In this study, we identify the most suitable areas globally for siting microalgal farms for biodiesel production that maximize profitability and minimize direct competition with food production and direct impacts on biodiversity, based on a spatially explicit multiple‐criteria decision analysis. We further explore the relationships between microalgal production, agricultural value, and biodiversity, and propose several solutions for siting microalgal production farms, based on current and future targets in energy production using integer linear programming. If using seawater for microalgal cultivation, biodiesel production could reach 5.85 × 10¹¹ L/year based on top suitable lands (i.e., between 13% and 16% of total transport energy demands in 2030) without directly competing with food production and areas of high biodiversity value. These areas are particularly abundant in the dry coasts of North and East Africa, the Middle East, and western South America. This is the first global analysis that incorporates economic and environmental feasibility for microalgal production sites. Our results can guide the selection of best locations for biofuel production using microalgae while minimizing conflicts with food production and biodiversity conservation.     

CYR61/CCN1 and WISP3/CCN6 are chemoattractive ligands for human multipotent mesenchymal stroma cells

Background:  

CCN-proteins are known to be involved in development, homeostasis and repair of mesenchymal tissues. Since these processes implicate recruitment of cells with the potential to be committed to various phenotypes, we studied the effect of CYR61/CCN1 and WISP3/CCN6 on migration of human bone marrow derived mesenchymal stroma cells (MSCs) in comparison to in vitro osteogenic differentiated MSCs using a modified Boyden chamber assay.     

Promiscuity comes at a price: Catalytic versatility vs efficiency in different metal ion derivatives of the potential bioremediator GpdQ

The glycerophosphodiesterase from Enterobacter aerogenes (GpdQ) is a highly promiscuous dinuclear metallohydrolase with respect to both substrate specificity and metal ion composition. While this promiscuity may adversely affect the enzyme's catalytic efficiency its ability to hydrolyse some organophosphates (OPs) and by-products of OP degradation have turned GpdQ into a promising candidate for bioremedial applications. Here, we investigated both metal ion binding and the effect of the metal ion composition on catalysis. The prevalent in vivo metal ion composition for GpdQ is proposed to be of the type Fe(II)Zn(II), a reflection of natural abundance rather than catalytic optimisation. The Fe(II) appears to have lower binding affinity than other divalent metal ions, and the catalytic efficiency of this mixed metal center is considerably smaller than that of Mn(II), Co(II) or Cd(II)-containing derivatives of GpdQ. Interestingly, metal ion replacements do not only affect catalytic efficiency but also the optimal pH range for the reaction, suggesting that different metal ion combinations may employ different mechanistic strategies. These metal ion-triggered modulations are likely to be mediated via an extensive hydrogen bond network that links the two metal ion binding sites via residues in the substrate binding pocket. The observed functional diversity may be the cause for the modest catalytic efficiency of wild-type GpdQ but may also be essential to enable the enzyme to evolve rapidly to alter substrate specificity and enhance k_cat values, as has recently been demonstrated in a directed evolution experiment. This article is part of a Special Issue entitled: Chemistry and mechanism of phosphatases, diesterases and triesterases.