KH domains with impaired nucleic acid binding as a tool for functional analysis

In eukaryotes, RNA-binding proteins that contain multiple K homology (KH) domains play a key role in coordinating the different steps of RNA synthesis, metabolism and localization. Understanding how the different KH modules participate in the recognition of the RNA targets is necessary to dissect the way these proteins operate. We have designed a KH mutant with impaired RNA-binding capability for general use in exploring the role of individual KH domains in the combinatorial functional recognition of RNA targets. A double mutation in the hallmark GxxG loop (GxxG-to-GDDG) impairs nucleic acid binding without compromising the stability of the domain. We analysed the impact of the GDDG mutations in individual KH domains on the functional properties of KSRP as a prototype of multiple KH domain-containing proteins. We show how the GDDG mutant can be used to directly link biophysical information on the sequence specificity of the different KH domains of KSRP and their role in mRNA recognition and decay. This work defines a general molecular biology tool for the investigation of the function of individual KH domains in nucleic acid binding proteins

Influencia de los métodos de conservación en las actividades enzimáticas de suelos forestales mediterráneos

Soil storage method may alter enzymatic activity being storage conditions of the soil samples prior to analysis decisive for the results. Studies made on freshly collected soils are generally preferred. However it is always not possible due to practical reasons since for example sampling is often restricted to short period of the year or because a great quantity of microbiological analyses must be made on time and by few people. On this context, soil storage methods are needed, being cold at 4°C the most widely used although sometimes alternative storage methods are also utilized. The aim of this study is to evaluate the effect of two alternative storage methods of soil samples (freezing at –20°C and air drying conservation methods) in comparison to cold at 4°C on the enzymatic activities (dehydrogenase, phosphatase, β-glucosidase and urease soil enzymes). Samples of two forest ecosystems (pine and holm oak forest stand) were taken in two different season of the year (winter and spring 2009). Results showed that enzymatic activities differed when freezing or air drying conservation methods were used in comparison with cold soil samples. Generally, alternative soil storage methods presented lower enzymatic activity than cold at 4°C. However, these changes depend on season and sampling location.El método de conservación del suelo utilizado puede alterar la actividad enzimática, siendo decisivas para los resultados, las condiciones de conservación previas a los análisis. Para los estudios es preferible realizar los análisis en muestras frescas y recién cogidas. Sin embargo, esto no es siempre posible por razones prácticas ya que las muestras se tienen que recoger en un corto periodo del año o porque los análisis los debe hacer poca gente y siempre en un determinado tiempo. En este contexto, se necesita un método de conservación, siendo el mantenimiento en el frigorífico a 4°C el método más usado, aunque existen otros métodos alternativos. El objetivo de este trabajo es el de evaluar el efecto de dos métodos alternativos de conservación (congelado a –20°C y secado a temperatura ambiente) sobre las actividades enzimáticas (deshidrogenasa, fosfatasa, β-glucosidasa y ureasa), en comparación con el mantenido en el frigorífico a 4°C. Las muestras se obtuvieron de dos zonas forestales diferentes (pinar y encinar) y en dos épocas distintas (primavera y verano de 2009). Los resultados mostraron diferencias en las actividades enzimáticas cuando se usó el congelado o secado al aire de las muestras de suelo en comparación con el mantenimiento en el frigorífico. De forma general, los métodos de conservación alternativos mostraron una menor actividad enzimática en las muestras de suelo analizadas. Sin embargo, estos cambios dependen de la época del año y la zona de muestreo

Polar Smectic Films

We report on a new experimental procedure for forming and studying polar smectic liquid crystal films. A free standing smectic film is put in contact with a liquid drop, so that the film has one liquid crystal/liquid interface and one liquid crystal/air interface. This polar environment results in changes in the textures observed in the film, including a boojum texture and a previously unobserved spiral texture in which the winding direction of the spiral reverses at a finite radius from its center. Some aspects of these textures are explained by the presence of a Ksb term in the bulk elastic free energy density that favors a combination of splay and bend deformations.Comment: 4 pages, REVTeX, 3 figures, submitted to PR

Time-scarcity, ready-meals, ill-health and the obesity epidemic

In this 3-part paper, we firstly review the interaction of time-scarcity with food-choices, specifically ready-meals, and potential health consequences from their consumption. Secondly we review declared nutrients, in relation to the standard 30% of Guideline Daily Amounts, concluding that popular ready-meals from major UK supermarkets are currently nutritionally haphazard. Thirdly, we present a simple scheme to establish standards for nutritional composition of ready-meals: unless otherwise specified, any meal (the smallest unit of nutrition) as recommended to be eaten or as offered should provide 30%+10% of GDA for energy and pro rata for key nutrients (e.g. sodium, sat fat, vitamin C)

Promoting Essential Laminations

We show that every co--orientable taut foliation F of an orientable, atoroidal 3-manifold admits a transverse essential lamination. If this transverse lamination is a foliation G, the pair F,G are the unstable and stable foliation respectively of an Anosov flow. Otherwise, F admits a pair of transverse very full genuine laminations. In the second case, M satisfies the weak geometrization conjecture - either its fundamental group contains Z+Z or it is word-hyperbolic. Moreover, if M is atoroidal, the mapping class group of M is finite, and any automorphism homotopic to the identity is isotopic to the identity.Comment: 56 pages, 11 figures; version 3: final version, incorporates referee's suggestion