Permanent cell line expressing human factor VIII-related antigen established by hybridization.

A permanent human cell line, EA . hy 926, has been established that expresses at least one highly differentiated function of vascular endothelium, factor VIII-related antigen. This line was derived by fusing human umbilical vein endothelial cells with the permanent human cell line A549. Hybrid cells that survived in selective medium had more chromosomes than either progenitor cell type and included a marker chromosome from the A549 line. Factor VIII-related antigen can be identified intracellularly in the hybrids by immunofluorescence and accumulates in the culture fluid. Expression of factor VIII-related antigen by these hybrid cells has been maintained for more than 100 cumulative population doublings, including more than 50 passages and three cloning steps. This is evidence that EA . hy 926 represents a permanent line

Tourist spaces and tourism policy in Spain and Portugal

Advances in Cultura, Tourism and Hospitality Research;10, 235-249This study analyses the relationship between the development of the tourism policy of Spain and Portugal and their effects on regional imbalances. Despite the proximity of the two countries and their specialisation in tourism, there are few comparative studies on tourism of the two Iberian countries. The study focuses on the two major phases of tourism policy: the period of mass tourism and post-Fordist stage. In the conclusions we refer the debate on the existence of a model of development based on tourism to the Latin countries of Southern Europe and we note the export process of the Spanish low-cost tourism model to other countries.Financiado por el Gobierno de España, Programa Fundamental de Investigación, Proyecto de I+D (CSO2012-30840) "Geografías de la crisis: análisis de los territorios urbanos y turísticos de las Islas Baleares, Costa del Sol y principales destinos del Caribe y América Central"

The complete nucleotide sequence of a beta-globin-like structure, beta h2, from the [Hbb] d mouse BALB/c.

We have determined the complete nucleotide sequence of beta h2, a pseudogene in the mouse beta-globin gene complex. The structure of beta h2 is analogous to that of a normal beta-globin gene, and its nucleotide sequence shares 72% homology with the coding regions of a reference mouse adult beta-globin gene. A frame shift occurs in the first coding region for which a compensatory splicing scheme can be devised. The reading frame is not otherwise disrupted. All of the recognized transcription, translation, and splicing signals in beta h2 are intact, with the exception of the " CCAAT box," which has been altered to GTAAC . We compared the predicted amino acid sequence of beta h2 with other beta-globin sequences. Evidence for a period of divergence without selection in the history of beta h2 was found in a set of codons that are usually highly conserved in productive beta-globin genes. An evolutionary tree constructed from nucleotide sequence suggests that beta h2 originated from the adult genes at least 60 million years ago. After some period as a productive gene, beta h2 was inactivated and has subsequently diverged without selection. Hybridization experiments demonstrated that beta h2 and the surrounding region occur without major alteration in other rodent species. The sequence ( AGCCA - 4n - GTGT ) occurs 5' of the CCAAT box in beta h2 and in many productive globin genes

Critical perspectives on ‘consumer involvement’ in health research: epistemological dissonance and the know-do gap

Researchers in the area of health and social care (both in Australia and internationally) are encouraged to involve consumers throughout the research process, often on ethical, political and methodological grounds, or simply as ‘good practice’. This paper presents findings from a qualitative study in the UK of researchers’ experiences and views of consumer involvement in health research. Two main themes are presented in the paper. Firstly, we explore the ‘know-do gap’ which relates to the tensions between researchers’ perceptions of the potential benefits of, and their actual practices in relation to, consumer involvement. Secondly, we focus on one of the reasons for this ‘know-do gap’, namely epistemological dissonance. Findings are linked to issues around consumerism in research, lay/professional knowledges, the (re)production of professional and consumer identities and the maintenance of boundaries between consumers and researchers

Stability of the thrombin-thrombomodulin complex on the surface of endothelial cells from human saphenous vein or from the cell line EA.hy 926

Protein C activation by alpha-thrombin on the surface of endothelial cells depends on an essential membrane-glycoprotein cofactor, thrombomodulin. In the present study we have monitored the activity of thrombin-thrombomodulin complexes on human saphenous-vein endothelial cells (HSVEC) or on the endothelial cell line EA.hy 926. Cell monolayers were exposed for 5 min to 8.5 nM human alpha-thrombin and then washed to remove unbound thrombin. The cells were then incubated at 37 degrees C for 5-180 min. At the end of the respective incubation periods, purified human protein C (120 nM) was added in order to assay the activity of the thrombin-thrombomodulin complexes present on the cell surface. HSVEC pre-exposed to thrombin retained their full capacity to promote protein C activation up to 90 min after free thrombin was removed. This capacity then decreased slowly to reach 56% of control value after 180 min of incubation. Original activity was 3.8 +/- 0.9 pmol of activated protein C formed/min per ml per 10(6) cells (mean +/- S.E.M., n = 5). The capacity of protein C activation of EA.hy 926 cells remained constant for 120 min after free thrombin was removed, then decreased to 76% of control after 180 min. Original activity was 2.0 +/- 0.4 pmol of activated protein C formed/min per ml per 10(6) cells (mean +/- S.E.M., n = 3). Similar results were obtained with cells fixed with 3% paraformaldehyde. However, during the 5-180 min incubation period, non-fixed cells of both types were capable of significantly internalizing fluorescent acetylated low-density lipoprotein. In the experimental protocol used here, an eventual inhibition of thrombin internalization by protein C can be excluded, as protein C is only added at the end of the incubation period. We conclude that there is no evidence of rapid internalization of thrombin-thrombomodulin complexes on HSVEC or the EA.hy 926 cell line, as assessed by the ability of membrane-bound thrombin to activate protein C