cAMP in budding yeast: Also a messenger for sucrose metabolism?

S. cerevisiae (or budding yeast) is an important micro-organism for sucrose-based fermentation in biotechnology. Yet, it is largely unknown how budding yeast adapts to sucrose transitions. Sucrose can only be metabolized when the invertase or the maltose machinery are expressed and we propose that the Gpr1p receptor signals extracellular sucrose availability via the cAMP peak to adapt cells accordingly. A transition to sucrose or glucose gave a transient cAMP peak which was maximally induced for sucrose. When transitioned to sucrose, cAMP signalling mutants showed an impaired cAMP peak together with a lower growth rate, a longer lag phase and a higher final OD600 compared to a glucose transition. These effects were not caused by altered activity or expression of enzymes involved in sucrose metabolism and imply a more general metabolic adaptation defect. Basal cAMP levels were comparable among the mutant strains, suggesting that the transient cAMP peak is required to adapt cells correctly to sucrose. We propose that the short-term dynamics of the cAMP signalling cascade detects long-term extracellular sucrose availability and speculate that its function is to maintain a fermentative phenotype at continuously low glucose and fructose concentrations.

Chemicals of concern in plastic toys.

We present a list of Chemicals of Concern (CoCs) in plastic toys. We started from available studies reporting chemical composition of toys to group plastic materials, as well as to gather mass fractions and function of chemicals in these materials. Chemical emissions from plastic toys and subsequent human exposures were then estimated using a series of models and a coupled near-field and far-field exposure assessment framework. Comparing human doses with reference doses shows high Hazard Quotients of up to 387 and cancer risk calculated using cancer slope factors of up to 0.0005. Plasticizers in soft plastic materials show the highest risk, with 31 out of the 126 chemicals identified as CoCs, with sum of Hazard Quotients >1 or child cancer risk >10-6. Our results indicate that a relevant amount of chemicals used in plastic toy materials may pose a non-negligible health risk to children, calling for more refined investigations and more human- and eco-friendly alternatives. The 126 chemicals identified as CoCs were compared with other existing regulatory prioritization lists. While some of our chemicals appear in other lists, we also identified additional priority chemicals that are not yet covered elsewhere and thus require further attention. We finally derive for all considered chemicals the maximum Acceptable Chemical Content (ACC) in the grouped toy plastic materials as powerful green chemistry tool to check whether chemical alternatives could create substantial risks.

How Well Does LCA Model Land Use Impacts on Biodiversity?--A Comparison with Approaches from Ecology and Conservation.

The modeling of land use impacts on biodiversity is considered a priority in life cycle assessment (LCA). Many diverging approaches have been proposed in an expanding literature on the topic. The UNEP/SETAC Life Cycle Initiative is engaged in building consensus on a shared modeling framework to highlight best-practice and guide model application by practitioners. In this paper, we evaluated the performance of 31 models from both the LCA and the ecology/conservation literature (20 from LCA, 11 from non-LCA fields) according to a set of criteria reflecting (i) model completeness, (ii) biodiversity representation, (iii) impact pathway coverage, (iv) scientific quality, and (v) stakeholder acceptance. We show that LCA models tend to perform worse than those from ecology and conservation (although not significantly), implying room for improvement. We identify seven best-practice recommendations that can be implemented immediately to improve LCA models based on existing approaches in the literature. We further propose building a "consensus model" through weighted averaging of existing information, to complement future development. While our research focuses on conceptual model design, further quantitative comparison of promising models in shared case studies is an essential prerequisite for future informed model choice.

Spatial patterns of agricultural expansion determine impacts on biodiversity and carbon storage.

The agricultural expansion and intensification required to meet growing food and agri-based product demand present important challenges to future levels and management of biodiversity and ecosystem services. Influential actors such as corporations, governments, and multilateral organizations have made commitments to meeting future agricultural demand sustainably and preserving critical ecosystems. Current approaches to predicting the impacts of agricultural expansion involve calculation of total land conversion and assessment of the impacts on biodiversity or ecosystem services on a per-area basis, generally assuming a linear relationship between impact and land area. However, the impacts of continuing land development are often not linear and can vary considerably with spatial configuration. We demonstrate what could be gained by spatially explicit analysis of agricultural expansion at a large scale compared with the simple measure of total area converted, with a focus on the impacts on biodiversity and carbon storage. Using simple modeling approaches for two regions of Brazil, we find that for the same amount of land conversion, the declines in biodiversity and carbon storage can vary two- to fourfold depending on the spatial pattern of conversion. Impacts increase most rapidly in the earliest stages of agricultural expansion and are more pronounced in scenarios where conversion occurs in forest interiors compared with expansion into forests from their edges. This study reveals the importance of spatially explicit information in the assessment of land-use change impacts and for future land management and conservation.

Emerging approaches, challenges and opportunities in life cycle assessment.

In the modern economy, international value chains--production, use, and disposal of goods--have global environmental impacts. Life Cycle Assessment (LCA) aims to track these impacts and assess them from a systems perspective, identifying strategies for improvement without burden shifting. We review recent developments in LCA, including existing and emerging applications aimed at supporting environmentally informed decisions in policy-making, product development and procurement, and consumer choices. LCA constitutes a viable screening tool that can pinpoint environmental hotspots in complex value chains, but we also caution that completeness in scope comes at the price of simplifications and uncertainties. Future advances of LCA in enhancing regional detail and accuracy as well as broadening the assessment to economic and social aspects will make it more relevant for producers and consumers alike.

Regulation of two germin-like protein genes during plum fruit development.

Germin-like proteins (GLPs) have several proposed roles in plant development and defence. Two novel genes (Ps-GLP1 and 2) encoding germin-like protein were isolated from plum (Prunus salicina). Their regulation was studied throughout fruit development and during ripening of early and late cultivars. These two genes exhibited similar expression patterns throughout the various stages of fruit development excluding two important stages, pit hardening (S2) and fruit ripening (S4). During fruit development until the ripening phase, the accumulation of both Ps-GLPs is related to the evolution of auxin. However, during the S2 stage only Ps-GLP1 is induced and this could putatively be in a H(2)O(2)-dependent manner. On the other hand, the diversity in the Ps-GLPs accumulation profile during the ripening process seems to be putatively due to the variability of endogenous auxin levels among the two plum cultivars, which consequently change the levels of autocatalytic ethylene available for the fruit to co-ordinate ripening. The effect of auxin on stimulating ethylene production and in regulating Ps-GLPs transcripts was also investigated. These data, supported by their localization in the extracellular matrix, suggest that auxin is somehow involved in the regulation of both transcripts throughout fruit development and ripening.

Molecular characterization of seven genes encoding ethylene-responsive transcriptional factors during plum fruit development and ripening.

Seven ERF cDNAs were cloned from two Japanese plum (Prunus salicina L.) cultivars, 'Early Golden' (EG) and 'Shiro' (SH). Based on the sequence characterization, these Ps-ERFs could be classified into three of the four known ERF families. Their predicted amino acid sequences exhibited similarities to ERFs from other plant species. Functional nuclear localization signal analyses of two Ps-ERF proteins (Ps-ERF1a and -1b) were carried out using confocal microscopy. Expression analyses of Ps-ERF mRNAs were studied in the two plum cultivars in order to determine the role of this gene family in fruit development and ripening. The seven Ps-ERFs displayed differential expression pattern and levels throughout the various stages of flower and fruit development. The diversity in Ps-ERFs accumulation was largely due to the differences in their responses to the levels of ethylene production. However, other plant hormones such as cytokinin and auxin, which accumulate strongly throughout the various developmental stages, also influence the Ps-ERFs expression. The effect of the plant hormones, gibberellin, cytokinin, auxin, and ethylene in regulating the different Ps-ERF transcripts was investigated. A model was proposed in which the role played by the plant hormone auxin is as important as that of ethylene in initiating and determining the date and rate of ripening in Japanese plums.

Comparing domestic versus imported apples: a focus on energy use.

GOAL, SCOPE AND BACKGROUND: The issue of whether food miles are a relevant indicator for the environmental impacts associated with foods has received significant attention in recent years. It is suggested here that issues other than the distance travelled need to be considered. The argument is presented by illustrating the case for the provision of apples. MATERIALS AND METHODS: The effects of variability in primary energy requirements for apple cultivation and for other life cycle stages, seasonality (timing of consumption) and loss of produce during storage are studied in this paper, by comparing apples from different supplier countries for consumption in Europe. RESULTS: Data sources for primary energy use (PEU) of apple production are identified ranging from 0.4-3.8 MJ/kg apples for European and Southern American countries and 0.4-0.7 MJ/kg for New Zealand. This variability is related to different yields and producer management practices in the different countries. Storage loss may range from 5% to 40% for storage periods between 4 and 10 months, and this has a significant effect on the results (e.g. increasing the total PEU by 8-16% when stored for 5-9 months in Europe as compared with a no loss and no storage situation). The storage periods and related storage losses change markedly through the year for imported (i.e. non-European) versus European apples. DISCUSSION: The timing of consumption and related storage losses need to be included in the assessment, as this affects the order of preference for locally sourced versus imported apples. The variability in energy requirements in different life cycle stages, but particularly for the fruit production stage, is also significant in this comparative analysis. CONCLUSIONS: Overall, it seems that there are similarities in the total PEU ranges for European and New Zealand apples during the Southern Hemisphere's apple season (European spring and summer). However, during the European autumn and winter (Northern Hemisphere apple season) PEU values are generally higher for apples imported from the Southern Hemisphere compared with European apples consumed in Europe. However, this latter observation may not hold true where apples for consumption in one European country are imported from another European country, because energy use for road transportation has a significant influence on the result. RECOMMENDATIONS AND PERSPECTIVES: Future studies comparing alternative sources of fresh produce need to account for ranges of data for the fruit production and storage stages, which reflect the seasonality of production.

Transport of proanthocyanidin dimer, trimer, and polymer across monolayers of human intestinal epithelial Caco-2 cells.

The gut absorption of proanthocyanidins (PAs) and of the related (+)-catechin monomer was investigated with colonic carcinoma (Caco-2) cells of a human origin, grown in monolayers on permeable filters. Permeability of various radiolabeled PAs differing in their molecular weight was compared with that of the radiolabeled (+)-catechin. No toxicity was observed at PA concentrations up to the physiological concentration of 1 mM. (+)-Catechin and PA dimer and trimer had similar permeability coefficients (P(app) = 0.9-2.0 x 10(-6) cm s(-1)) close to that of mannitol, a marker of paracellular transport. Paracellular transport was also indicated by the increase of absorption after reduction of the transepithelial electric resistance through calcium ion removal. In contrast, permeability of a PA polymer with an average polymerization degree of 6 (molecular weight 1,740) was approximately 10 times lower (P(app) = 0.10 +/- 0.04 x 10(-6) cm s(-1)). PAs, particularly the most astringent PA polymer, were also adsorbed on the epithelial cells. These results suggest that PA dimers and trimers could be absorbed in vivo and that polymer bioavailability is limited to the gut lumen.

Polymeric proanthocyanidins are catabolized by human colonic microflora into low-molecular-weight phenolic acids.

Polymeric proanthocyanidins are common constituents of many foods and beverages. Their fate in the human body remains largely unknown. Their metabolism by human colonic microflora incubated in vitro in anoxic conditions has been investigated using nonlabeled and (14)C-labeled purified proanthocyanidin polymers. Polymers were almost totally degraded after 48 h of incubation. Phenylacetic, phenylpropionic and phenylvaleric acids, monohydroxylated mainly in the meta or para position, were identified as metabolites by gas chromatography coupled to mass spectrometry (GC-MS). Yields were similar to those previously reported for flavonoid monomers. These results provide the first evidence of degradation of dietary phenolic polymers into low-molecular-weight aromatic compounds. To understand the nutritional properties of proanthocyanidins, it is therefore essential to consider the biological properties of these metabolites.

Lignification in transgenic poplars with extremely reduced caffeic acid O-methyltransferase activity.

Transgenic poplars (Populus tremula x Populus alba) were obtained by introduction of a sense homologous transgene encoding caffeic acid O-methyltransferase (COMT) under the control either of the cauliflower mosaic virus double 35S promoter or of the eucalyptus cinnamyl alcohol dehydrogenase promoter. Although these constructs conferred a moderate overexpression of COMT in some lines, a transgenic line with the double 35S promoter was found where COMT activity in woody tissues was close to zero due to a gene-silencing phenomenon. For the first time in COMT down-regulated trees, this alteration substantially reduced lignin level in 6-month-old trees (17% decrease). Lignin structure was found to be strongly altered, with a two times higher content in condensed bonds, an almost complete lack of syringyl units, and the incorporation of 5-hydroxyguaiacyl units to the most remarkable extent reported so far. Consistent with the higher cellulose content and with the higher condensation degree of the lignin, the impact of the transformation on the kraft-pulping performances of the poplar trees positively affected the pulp yield (10% relative increase), but made lignins less amenable to industrial degradations.

Proanthocyanidins and human health: systemic effects and local effects in the gut.

Proanthocyanidins share common properties with other polyphenols, in particular their reducing capacity and ability to chelate metal ions. However, their polymeric nature clearly makes them different. They have a high affinity for proteins and their absorption through the gut barrier is likely limited to the molecules of low polymerization degree and to the metabolites formed by the colonic microflora, as suggested by in vitro experiments. The nutritional significance of proanthocyanidins is discussed in relation to their physico-chemical properties and bioavailability.

Carbon-14 biolabeling of (+)-catechin and proanthocyanidin oligomers in willow tree cuttings.

Proanthocyanidin polymers, oligomers, and the structurally related monomer (+)-catechin were labeled by incorporation of radioactive precursors in shoots of willow tree (Salix caprea L.). [1-(14)C]-Acetate and [U-(14)C]-phenylalanine precursors were fed through the cut stems or petioles of leaves. Optimization of several parameters such as the nature and origin of the plant material, leaf maturity, nature, and quantity of radioactive precursor applied and the duration of metabolism led to incorporation yields of 3.2% and to specific activities of 500 microCi/g. Detailed characterization of the products (polymerization degree, procyanidin/prodelphinidin ratio, specific activities) and purification by chromatography are reported. Some sugars bound to radiolabeled proanthocyanidin polymers were removed by enzymic treatment with a mixture of glycosidases. A radioactive purity close to 100% and specific activities suitable for bioavailability studies were obtained.