Cocoa pod husk (CPH) is the main by-product (ca. 70-75% weight of whole fruit) of the cocoa harvest, an important and economic crop in developing countries. It is a rich source of minerals (particularly potassium), fibre (including lignin, cellulose, hemicellulose and pectin) and antioxidants (e.g. phenolic acids). An existing practise is the return of CPH to soil with potential benefits (or disadvantages) for cocoa productivity and soil sustainability that have not been fully characterised. Currently, alternative low-value applications of CPH include its use as animal feed, as a starting material for soap making and activated carbon. Other biotechnological valorisation potentials for CPH and its fractions include the production of bio-fuels and their incorporation in food systems. Physical, chemical or biological pretreatment approaches are needed in order to achieve desirable fractions in a cost-effective and sustainable manner for novel applications in food and non-food sectors.
Using Xenopus tropicalis, we present the first analysis of the developmental effects that result from knocking down the function of the three Cdx genes present in the typical vertebrate genome. Knockdowns of individual Cdx genes lead to a similar range of posterior defects; compound Cdx knockdowns result in increasingly severe posterior truncations, accompanied by posterior shifts and reduction of 5′ Hox gene expression. We provide evidence that Cdx and Wnt3A genes are components of a positive feedback loop operating in the posterior axis. We show that Cdx function is required during later, but not early stages of development, for correct regional specification of the endoderm and morphogenesis of the gut. Our results support the hypothesis that during amphibian development the overall landscape of Cdx activity in the embryo is more important than the specific function of individual Cdx proteins. Developmental Dynamics 238:835–852, 2009. © 2009 Wiley-Liss, Inc.
BackgroundFGF signaling has multiple roles in regulating processes in animal development, including the specification and patterning of the mesoderm. In addition, FGF signaling supports self renewal of human embryonic stem cells and is required for differentiation of murine embryonic stem cells into a number of lineages.Methodology/Principal FindingsGiven the importance of FGF signaling in regulating development and stem cell behaviour, we aimed to identify the transcriptional targets of FGF signalling during early development in the vertebrate model Xenopus laevis. We analysed the effects on gene expression in embryos in which FGF signaling was inhibited by dominant negative FGF receptors. 67 genes positively regulated by FGF signaling and 16 genes negatively regulated by FGF signaling were identified. FGF target genes are expressed in distinct waves during the late blastula to early gastrula phase. Many of these genes are expressed in the early mesoderm and dorsal ectoderm. A widespread requirement for FGF in regulating genes expressed in the Spemann organizer is revealed. The FGF targets MKP1 and DUSP5 are shown to be negative regulators of FGF signaling in early Xenopus tissues. FoxD3 and Lin28, which are involved in regulating pluripotency in ES cells are shown to be down regulated when FGF signaling is blocked.ConclusionsWe have undertaken a detailed analysis of FGF target genes which has generated a robust, well validated data set. We have found a widespread role for FGF signaling in regulating the expression of genes mediating the function of the Spemann organizer. In addition, we have found that the FGF targets MKP1 and DUSP5 are likely to contribute to the complex feedback loops involved in modulating responses to FGF signaling. We also find a link between FGF signaling and the expression of known regulators of pluripotency.
SUMMARYLin28 family proteins share a unique structure, with both zinc knuckle and cold shock RNA-binding domains, and were originally identified as regulators of developmental timing in Caenorhabditis elegans. They have since been implicated as regulators of pluripotency in mammalian stem cells in culture. Using Xenopus tropicalis, we have undertaken the first analysis of the effects on the early development of a vertebrate embryo resulting from global inhibition of the Lin28 family. The Xenopus genome contains two Lin28-related genes, lin28a and lin28b. lin28a is expressed zygotically, whereas lin28b is expressed both zygotically and maternally. Both lin28a and lin28b are expressed in pluripotent cells of the Xenopus embryo and are enriched in cells that respond to mesoderminducing signals. The development of axial and paraxial mesoderm is severely abnormal in lin28 knockdown (morphant) embryos. In culture, the ability of pluripotent cells from the embryo to respond to the FGF and activin/nodal-like mesoderm-inducing pathways is compromised following inhibition of lin28 function. Furthermore, there are complex effects on the temporal regulation of, and the responses to, mesoderm-inducing signals in lin28 morphant embryos. We provide evidence that Xenopus lin28 proteins play a key role in choreographing the responses of pluripotent cells in the early embryo to the signals that regulate germ layer specification, and that this early function is probably independent of the recognised role of Lin28 proteins in negatively regulating let-7 miRNA biogenesis.KEY WORDS: lin28a, lin28b, Xenopus, Mesoderm, miRNA, let-7, Pluripotency, Germ layer, FGF, Activin, Nodal Lin28 proteins are required for germ layer specification in Xenopus Laura Faas, Fiona C. Warrander, Richard Maguire, Simon A. Ramsbottom, Diana Quinn, Paul Genever and Harry V. Isaacs* DEVELOPMENT 977 RESEARCH ARTICLE Lin28 function in Xenopus mesoderm specification. This represents the first evidence that Lin28 family genes play a key role in regulating the timing of, and responses to, growth factor signalling in the early development of a vertebrate embryo.A key finding of this study is that, although amphibian lin28 proteins exhibit let-7 miRNA-binding activity, no significant effects on the abundance of let-7a, let-7f and let-7g miRNAs are detected in lin28 knockdown embryos, at the stage when germ layer specification occurs. Our data indicate that the requirement for lin28 function in the initial specification of the mesoderm is likely to be independent of its role in let-7 family biogenesis. By contrast, lin28 function seems to be required to regulate let-7 levels at later stages, after germ layer specification, indicating differential roles for lin28 in amphibian development: a let-7-independent early role and a let-7-dependent later role. MATERIALS AND METHODS Embryo methodsX. tropicalis embryos were produced as previously described (Khokha et al., 2005;Winterbottom et al., 2010). Embryos were injected at the 2-or 4-cell stage and cultured at 22°C. Anima...
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