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Gpn3 is required for RNA polymerase II (RNAPII) nuclear targeting. Here, we investigated the effect of a cancer-associated Q279* nonsense mutation in Gpn3 cellular function. Employing RNAi, we replaced endogenous Gpn3 by wt or Q279* RNAi-resistant Gpn3R in epithelial model cells. RNAPII nuclear accumulation and transcriptional activity were markedly decreased in cells expressing only Gpn3R Q279*. Wild-type Gpn3R localized to the cytoplasm but a fraction of Gpn3R Q279* entered the cell nucleus and inhibited Gpn1-EYFP nuclear export. This property and the transcriptional deficit in Gpn3R Q279*-expressing cells required a PDZ-binding motif generated by the Q279* mutation. We conclude that an acquired PDZ-binding motif in Gpn3 Q279* caused Gpn3 nuclear entry, and inhibited Gpn1 nuclear export and Gpn3-mediated RNAPII nuclear targeting.

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Gpn1 and Gpn3 are GTPases individually required for nuclear targeting of RNA polymerase II. Here we show that whereas Gpn3-EYFP distributed between the cytoplasm and cell nucleus, it was mainly cytoplasmic when coexpressed with Gpn1-Flag. Gpn3-Flag retained Gpn1-EYFP in the cytoplasm. However, Gpn3-EYFP/Gpn1-Flag nucleocytoplasmic shuttling was revealed after inhibiting nuclear export with leptomycin B. All Gpn3-EYFP coimmunoprecipitated with Gpn1-Flag, and all Gpn1-EYFP with Gpn3-Flag. Importantly, most endogenous Gpn1 and Gpn3 also associate. Gpn1-Gpn3 interaction was essential to maintain steady-state protein levels of both GTPases. We propose that most Gpn1 and Gpn3 associate, are mobilized, and function as a protein complex.

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The study of biochemical activity of extracts obtained from marine organisms is gaining interest as some have proved to have efficient health or industrial applications. To evaluate lethality and antimicrobial activities, some chemical tests were performed on crude extracts of the octocorals Eunicea sp., Muricea sp. and Pseudopterogorgia acerosa and the mollusks Pteria colymbus, Phyllonotus pomum and Chicoreus brevifrons, collected in Venezuelan waters. The presence of secondary metabolites like alkaloids, unsaturated sterols and pentacyclic triterpenes in all invertebrates, was evidenced. Additionally, sesquiterpenlactones, saponins, tannins, cyanogenic and cardiotonic glycosides were also detected in some octocoral extracts, suggesting that biosynthesis of these metabolites is typical in this group. From the lethality bioassays, all extracts resulted lethal to Artemia salina (LC50<1000 microg/ml) with an increased of lethal activity with exposition time. P. pomum extract showed the highest lethality rate (LC50=46.8 microg/ml). Compared to the octocorals, mollusks extracts displayed more activity and a greater action spectrum against different bacterial strains, whereas octocorals also inhibited some fungi strains growth. Staphylococcus aureus was the most susceptible to the antimicrobial power of the extracts (66.7%), whereas Pseudomonas aeruginosa, Candida albicans and Aspergillus niger were not affected. The antibiosis shown by marine organisms extracts indicates that some of their biosynthesized metabolites are physiologically active, and may have possible cytotoxic potential or as a source of antibiotic components.

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Secondary metabolites, lethality and antimicrobial activity of extracts from three corals and three marine mollusks from Sucre, Venezuela. The study of biochemical activity of extracts obtained from marine organisms is gaining interest as some have proved to have efficient health or industrial applications. To evaluate lethality and antimicrobial activities, some chemical tests were performed on crude extracts of the octocorals Eunicea sp., Muricea sp. and Pseudopterogorgia acerosa and the mollusks Pteria colymbus, Phyllonotus pomum and Chicoreus brevifrons, collected in Venezuelan waters. The presence of secondary metabolites like alkaloids, unsaturated sterols and pentacyclic triterpenes in all invertebrates, was evidenced. Additionally, sesquiterpenlactones, saponins, tannins, cyanogenic and cardiotonic glycosides were also detected in some octocoral extracts, suggesting that biosynthesis of these metabolites is typical in this group. From the lethality bioassays, all extracts resulted lethal to Artemia salina (LC50<1.000μg/ml) with an increased of lethal activity with exposition time. P. pomum extract showed the highest lethality rate (LC50=46.8μg/ml). Compared to the octocorals, mollusks extracts displayed more activity and a greater action spectrum against different bacterial strains, whereas octocorals also inhibited some fungi strains growth. Staphylococcus aureus was the most susceptible to the antimicrobial power of the extracts (66.7%), whereas Pseudomonas aeruginosa, Candida albicans and Aspergillus niger were not affected. The antibiosis shown by marine organisms extracts indicates that some of their biosynthesized metabolites are physiologically active, and may have possible cytotoxic potential or as a source of antibiotic components. Rev. Biol. Trop. 58 (2): 677-688. Epub 2010 June 02.

A los extractos crudos de los octocorales Eunicea sp., Muricea sp. y Pseudopterogorgia acerosa y de los moluscos Pteria colymbus, Phylonotus pomum y Chicoreus brevifrons, se les realizaron pruebas químicas, las cuales evidenciaron en todos ellos, la presencia de metabolitos secundarios como alcaloides, esteroles insaturados y triterpenos pentacíclicos. Sólo se detectaron sesquiterpenlactonas, saponinas, taninos, glicósidos cianogénicos y glicósidos cardiotónicos en algunos de los extractos de los octocorales, lo cual sugiere que la biosíntesis de estos metabolitos es propia de este grupo de organismos. Asimismo, se evaluó la actividad letal y antimicrobiana de los extractos de los octocorales y moluscos. En el bioensayo de letalidad, todos los extractos resultaron letales frente al crustáceo Artemia salina (CL50<1.000μg/ml). La actividad letal incrementσ con el tiempo de exposiciσn. El extracto de P. pomum presentó la mayor actividad letal (CL50=46.8μg/ml). En los ensayos de actividad antimicrobiana, los extractos orgαnicos de los moluscos presentaron una mayor actividad y un espectro de acción mayor contra diferentes cepas de bacterias, respecto a los octocorales; aunque estos últimos también inhibieron el crecimiento de algunas cepas de hongos. Staphylococcus aureus, fue la bacteria más susceptible al poder antimicrobiano de los extractos (66.7%), mientras que la bacteria Pseudomonas aeruginosa y los hongos Candida albicans y Aspergillus niger, no fueron afectados. La antibiosis mostrada por los extractos de los diferentes invertebrados marinos estudiados, indica que algunos de los metabolitos biosintetizados por éstos son fisiológicamente activos con posible potencial citotóxico y/o antibiótico.

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With the intention of bridging the 'digital divide' many programmes have been launched to provide computers for educational institutions, ranging from refurbishing second hand computers to delivering low cost new computers. The fast and economical provision of large quantities of equipment is one of the many challenges faced by such programmes. If an increase is to be achieved in the sustainability of computer supplies for schools, not only must equipment be provided, but also suitable training and maintenance delivered. Furthermore, appropriate recycling has to be ensured, so that end-of-life equipment can be dealt with properly. This study has evaluated the suitability of three computer supply scenarios to schools in Colombia: (i) 'Colombian refurbishment', -refurbishment of computers donated in Colombia, (ii) 'Overseas refurbishment', -import of computers which were donated and refurbished abroad, and (iii) 'XO Laptop', -purchase of low cost computers manufactured in Korea. The methods applied were: Material Flow Assessment, -to assess the quantities-, Life Cycle Assessment, -to assess the environmental impacts, and the application of the Multiple Attribute Utility Theory, -to analyse, evaluate and compare different scenarios. The most sustainable solution proved to be the local refurbishment of second hand computers of Colombian origin to an appropriate technical standard. The environmental impacts of such practices need to be evaluated carefully, as second hand appliances have to be maintained, require spare parts and sometimes use more energy than newer equipment. Providing schools with second hand computers from overseas and through programmes such as 'One Laptop Per Child' has the disadvantage that the potential for social improvements - such as creation of jobs and local industry involvement - is very low.

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