Quantitative Analysis of Protein Expression to Study Lineage Specification in Mouse Preimplantation Embryos

This protocol presents a method to perform quantitative, single-cell in situ analyses of protein expression to study lineage specificationin mouse preimplantation embryos. The procedures necessary for embryo collection, immunofluorescence, imaging on a confocal microscope, and image segmentation and analysis are described. This method allows quantitation of the expression of multiple nuclear markers and the spatial (XYZ) coordinates of all cells in the embryo. It takes advantage of MINS, an image segmentation software tool specifically developed for the analysis of confocal images of preimplantation embryos and embryonic stem cell (ESC) colonies. MINS carries out unsupervised nuclear segmentation across the X, Y and Z dimensions, and produces information on cell position in three-dimensional space, as well as nuclear fluorescence levels for all channels with minimal user input. While this protocol has been optimized for the analysis of images of preimplantation stage mouse embryos, it can easily be adapted to the analysis of any other samples exhibiting a good signal-to-noise ratio and where high nuclear density poses a hurdle to image segmentation (e.g., expression analysis of embryonic stem cell (ESC) colonies, differentiating cells in culture, embryos of other species or stages, etc.).     

用模块化分子元件调控和编程生物系统

合成生物学的目标包括“通过合成来理解生命”以及用现代工程学方法设计合成复杂生物系统．其工程学目标的实现依赖于可集成、可调控、可重用、功能多样的蛋白质、RNA、DNA等基本分子元件．以分子机制为基础,合理设计与实验室进化相结合,改造和创建生物分子的相互作用特异性、调控方式、定量活性等,是实现生物系统人工调控与编程的重要策略,同时为自下而上设计合成日益复杂的人工生物系统奠定基础．     

Unravelling the peculiarities of island life: vicariance, dispersal and the diversification of the extinct and extant giant Galápagos tortoises

In isolated oceanic islands, colonization patterns are often interpreted as resulting from dispersal rather than vicariant events. Such inferences may not be appropriate when island associations change over time and new islands do not form in a simple linear trend. Further complexity in the phylogeography of ocean islands arises when dealing with endangered taxa as extinctions, uncertainty on the number of evolutionary ‘units’, and human activities can obscure the progression of colonization events. Here, we address these issues through a reconstruction of the evolutionary history of giant Galápagos tortoises, integrating DNA data from extinct and extant species with information on recent human activities and newly available geological data. Our results show that only three of the five extinct or nearly extinct species should be considered independent evolutionary units. Dispersal from mainland South America started at approximately 3.2 Ma after the emergence of the two oldest islands of San Cristobal and Española. Dispersal from older to younger islands began approximately 1.74 Ma and was followed by multiple colonizations from different sources within the archipelago. Vicariant events, spurred by island formation, coalescence, and separation, contributed to lineage diversifications on Pinzón and Floreana dating from 1.26 and 0.85 Ma. This work provides an example of how to reconstruct the history of endangered taxa in spite of extinctions and human‐mediated dispersal events and highlights the need to take into account both vicariance and dispersal when dealing with organisms from islands whose associations are not simply explained by a linear emergence model.     

Visual extrapolation under risk: human observers estimate and compensate for exogenous uncertainty

Humans commonly face choices between multiple options with uncertain outcomes. Such situations occur in many contexts, from purely financial decisions (which shares should I buy?) to perceptuo-motor decisions between different actions (where should I aim my shot at goal?). Regardless of context, successful decision-making requires that the uncertainty at the heart of the decision-making problem is taken into account. Here, we ask whether humans can recover an estimate of exogenous uncertainty and then use it to make good decisions. Observers viewed a small dot that moved erratically until it disappeared behind an occluder. We varied the size of the occluder and the unpredictability of the dot''s path. The observer attempted to capture the dot as it emerged from behind the occluded region by setting the location and extent of a ‘catcher’ along the edge of the occluder. The reward for successfully catching the dot was reduced as the size of the catcher increased. We compared human performance with that of an agent maximizing expected gain and found that observers consistently selected catcher size close to this theoretical solution. These results suggest that humans are finely tuned to exogenous uncertainty information and can exploit it to guide action.     

Enhanced Phenolic Diterpenes Antioxidant Levels Through Non-transgenic Approaches

Plant-derived antioxidants are essential in our diet, and antioxidant composition is a key determinant of the quality of plant extracts of interest to the pharmaceutical and food industries. By using carnosic acid as an example of a key antioxidant constituent of rosemary and sage extracts, we discuss the importance of studying non-transgenic approaches to enhancing antioxidant levels in plants and improving the antioxidant composition of plant extracts. In contrast to other terpenoids or phenolic compounds, carnosic acid has only been found in some Labiatae species, such as rosemary and sage. Carnosic acid has medicinal properties; it is a potent antioxidant and protects skin cells against UV-A radiation and cancer. Furthermore, it has been used as a preservative in food and non-food products, displaying important antimicrobial effects. However, the key steps involved in its biosynthesis remain largely unknown, and thus non-transgenic approaches are required to increase its level in plant extracts. Dried rosemary or sage leaves can contain between 0.1% and 7% carnosic acid, depending on the species and variety, but also on plant growth conditions, sample treatment and mode of extract preparation. Furthermore, leaf age, salinity and ionic interactions can also have a significant effect on biosynthesis and therefore have a strong impact on the total antioxidant potential of rosemary and sage extracts. Non-transgenic approaches, used in these or other species, can significantly increase antioxidant levels and therefore provide very significant improvements in the quality of several botanical extracts used in industry, and can be applied as either an alternative or a complement to transgenic approaches.

Enhanced Phenolic Diterpenes Antioxidant Levels Through Non-transgenic Approaches

All authors

T. Tounekti &; S. Munné-Bosch

https://doi.org/10.1080/07352689.2012.696457

Published online:

26 October 2012

FIG. 1 Chemical structure of synthetic and natural plant-derived antioxidants. Note that the hydroxyl groups in the ortho-position at C₁₁ and C₁₂ of the carnosic acid molecule provide high antioxidant properties (color figure available online).     

70.

New activities launched in Warsaw and consultations on emerging ideas     

Jim Fava  Sonia Valdivia  Bas de Leeuw  Andreas Ciroth  Allan A. Jensen  Guido Sonnemann 《The International Journal of Life Cycle Assessment》2008,13(5):371-373

     

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