Insertional mutagenesis and marker rescue in a protozoan parasite: cloning of the uracil phosphoribosyltransferase locus from Toxoplasma gondii.

Nonhomologous integration vectors have been used to demonstrate the feasibility of insertional mutagenesis in haploid tachyzoites of the protozoan parasite Toxoplasma gondii. Mutant clones resistant to 5-fluorouracil were identified at a frequency of approximately 10(-6) (approximately 2 x 10(-5) of the stable transformants). Four independent mutants were isolated, all of which were shown to lack uracil phosphoribosyl-transferase (UPRT) activity and harbor transgenes integrated at closely linked loci, suggesting inactivation of the UPRT-encoding gene. Genomic DNA flanking the insertion point (along with the integrated vector) was readily recovered by bacterial transformation with restriction-digested, self-ligated total genomic DNA. Screening of genomic libraries with the recovered fragment identified sequences exhibiting high homology to known UPRT-encoding genes from other species, and cDNA clones were isolated that contain a single open reading frame predicted to encode the 244-amino acid enzyme. Homologous recombination vectors were exploited to create genetic knock-outs at the UPRT locus, which are deficient in enzyme activity but can be complemented by transient transformation with wild-type sequences--formally confirming identification of the functional UPRT gene. Mapping of transgene insertion points indicates that multiple independent mutants arose from integration at distinct sites within the UPRT gene, suggesting that nonhomologous integration is sufficiently random to permit tagging of the entire parasite genome in a single transformation.

Regulation of excitatory transmission at hippocampal synapses by calbindin D28k.

Distinct subpopulations of neurons in the brain contain one or more of the Ca(2+)-binding proteins calbindin D28k, calretinin, and parvalbumin. Although it has been shown that these high-affinity Ca(2+)-binding proteins can increase neuronal Ca2+ buffering capacity, it is not clear which aspects of neuronal physiology they normally regulate. To investigate this problem, we used a recently developed method for expressing calbindin D28k in the somatic and synaptic regions of cultured hippocampal pyramidal neurons. Ninety-six hours after infection with a replication-defective adenovirus containing the calbindin D28k gene, essentially all cultured hippocampal pyramidal neurons robustly expressed calbindin D28k. Our results demonstrate that while calbindin D28k does not alter evoked neurotransmitter release at excitatory pyramidal cell synapses, this protein has a profound effect on synaptic plasticity. In particular, we show that calbindin D28k expression suppresses posttetanic potentiation.

A cura pelo poder

Sarney revela em livro de memórias que conseguiu controlar seus medos com o exercício da presidência. Destaca a saga do cruzado e se diz ressentido com acusações de que o planp foi eleitoreiro.

Modelos urbanos y su proyección literaria en la novela criminal

La novela criminal experimenta en la actualidad un momento de esplendor en las distintas literaturas que se desarrollan en el Estado español, debido al aumento en el número de autores, lectores y obras publicadas. No ha sido un camino fácil, pues este género se ha adaptado tardíamente a la realidad española. De hecho, hasta la década de 1970, cuando la Dictadura franquista llegaba a su fin, no hubo una verdadera tradición de novela criminal urbana en España. En 1974, Manuel Vázquez Montalbán publicó la obra Tatuaje, primera en la que el personaje de Pepe Carvalho aparecía como detective e inicio de la saga más relevante en España. El desarrollo del género fue dispar en las diferentes literaturas del Estado, y mientras Cataluña ya contaba con un respetable bagaje –autores como Manuel de Pedrolo, Rafael Tasis, Jaume Fuster…–, en el País Vasco comenzaba a emplearse el género, y en Galicia y Canarias habría que esperar hasta las décadas de 1980 y de 1990, respectivamente. Esta desigual adaptación de la novela criminal propicia la aparición de diferentes modelos urbanos en las distintas obras y literaturas a las que se adscriben. La ciudad es un elemento característico del género, pues su nacimiento está directamente relacionado con la Revolución Industrial: es cuando surgen las urbes y las sociedades modernas y una burguesía con gran poder adquisitivo y aumenta la población alfabetizada. Es en las ciudades donde se desarrollan las tramas criminales y violentas, pues estas fomentan el anonimato de los asesinos, la creación de cuerpos policiales y la presencia de detectives privados...

Causation and Harm in a Multicomponent World

On September 17, 2015, the Federal Circuit issued another decision in the epic Apple v. Samsung smartphone war. This was the fourth court decision in the ongoing saga to deal with injunctions. Apple IV explained the level of proof necessary to satisfy the "causal nexus" requirement. This requirement had emerged as a response to patent litigations involving products with thousands of features, the vast majority of which are unrelated to the asserted patent. To prove a causal nexus, patentees seeking an injunction have to do more than just show that the infringing product caused the patentee irreparable harm. The harm must be specifically attributable to the infringing feature. In Apple IV, the Federal Circuit noted that proving causation was "nearly impossible" in these multicomponent cases. So it decided to water down the causal nexus requirement saying that it was enough for Apple to show that the infringing features were "important"and customer sought these particular features. This lower standard is an ill-advised mistake that leaves multicomponent product manufacturers more susceptible to patent holdup. My critique takes two parts. First, I argue that a single infringing feature rarely, if ever, "causes" consumers to buy the infringer’s multicomponent products. The minor features at issue in Apple IV illustrate this point vividly. Thus, the new causal nexus standard does not accurately reflect how causation and harm operate in a multicomponent world. Second, I explain why the court was so willing to accept such little evidence of real injury. It improperly applied notions of traditional property law to patents. Specifically, the court viewed patent infringement as harmful regardless of any concrete consequences. This view may resonate for other forms of property where an owner's rights are paramount and a trespass is considered offensive in and of itself. But the same concepts do not apply to patent law where the Supreme Court has consistently said that private interests must take a back seat to the public good. Based on these principles, the courts should restore the "causal nexus" requirement and not presume causation.

Del cómic a la narración transmedia en la formación universitaria. Mapa transmedia de Los Vengadores

Dentro de las nuevas perspectivas educativas, el multimedia y la hipertextualidad son elementos imprescindibles en la formación del alumnado universitario que debe navegar en Internet de una manera efectiva. Entre los múltiples conceptos, el de la narración transmedia (Henry Jenkins 2003; Carlos Scolari 2013) ha asumido una especial relevancia, ya que supone la construcción de un objeto artístico y comercial a través de múltiples medios, digitales y tradicionales, con especial relevancia para distintas áreas de conocimiento. Con el reciente éxito de las adaptaciones cinematográficas del Universo Marvel, el cómic ha resurgido como un elemento vertebrador de muchas de estas obras transmedia. La relación entre texto e imagen, intrínseca en la concepción de la narración gráfica, hace que en los principales ejemplos transmedia se utilice el cómic como un elemento más. Esta investigación es una propuesta para explicar las características principales de la narración transmedia al alumnado universitario a través de los múltiples elementos que nos ofrece la saga de Los Vengadores. Cómic, cine, series, dibujos animados, videojuegos, fanfiction y publicidad son elementos que integran este complejo universo y aportan nuevos significados a la narración, ofreciéndonos nuevas estrategias metodológicas en la difusión del conocimiento.

Post-Summit Commentary – Greece on the brink. EPC Commentary, 29 June 2015

The developments of recent days have been dramatic – the saga of the Greek crisis has probably opened its decisive chapter. Negotiations between Athens and its creditors failed after the Greek government decided to leave the negotiating table and hold a referendum on 5 July. The future of the country in the common currency and the potential consequences for the EU and the euro are uncertain. There are clear signs of fatigue, everywhere. But there is still time to avert the worst, if there is the political will on all sides to work on a new perspective for Greece and for the future of the Economic and Monetary Union (EMU).

How to reconcile the EU border paradox? The concurrence of refugee reception and deterrence. IES Policy Brief 2015/4, November 2015

The refugee crisis that unfolded in Europe in the summer of 2015 questions the effectiveness of European border and refugee policies. The breakdown of the Dublin and Schengen rules due to chaotic situations at the borders in the Balkans marks a critical juncture for the EU. We consider this breakdown as a consequence of a long-lasting co-operation crisis among EU Member States. The most recent Council decision responds to this co-operation crisis (Council Decision 12098/15). This Policy Brief analyses EU policy and politics and argues that plans for refugee relocation and reception centres as well as the use of qualified majority voting in the Council can unfold a dynamic that helps to solve the co-operation crisis. However, underlying the problems of co-operation and effectiveness is the EU’s border paradox: while EU border policy works towards refugee deterrence, EU asylum policy aims at refugee protection. The EU’s approach in regulating borders and asylum can be understood in terms of ‘organised hypocrisy’ (Brunsson, 1993). Reconciling the paradox calls for overcoming such hypocrisy.

By way of introduction; a book list for young people,

Mode of access: Internet.

Recreational reading for young people,

Mode of access: Internet.

The cases of polygamy, concubinage, adultery, divorce, &c. seriously and learnedly discussed. : Being a compleat collection of all the remarkable tryals and tracts which have been written on those important subjects. /

Reissued 1736 with title: Select and curious cases of polygamy.

Of William Morris and his work /

Illustrations of mss. pages of the translated Eyrbyggja saga and Bandamanna saga on p. 29-30.

Seynt Graal, or The Sank Ryal.: The history of the Holy Graal, partly in English verse,

Vol. I, Rev. John Stuart Hippisley Horner's copy (name in red in list of members).

Guta-lagh, das ist : Der insel Gothland altes rechtsbuch : In der ursprache und einer wiederaufgefundenen altdeutschen uebersetzung hrsg. /

"Alte erzạhlung [i.e. Guta saga]": p. [106]-115.