The high- and low-activity forms of human plasma phospholipid transfer protein (PLTP)

Plasma phospholipid transfer protein (PLTP) plays a crucial role in high-density lipoprotein (HDL) metabolism and reverse cholesterol transport (RCT). It mediates the generation of pre-beta-HDL particles, enhances the cholesterol efflux from peripheral cells to pre-beta-HDL, and metabolically maintains the plasma HDL levels by facilitating the transfer of post-lipolytic surface remnants of triglyceride-rich lipoproteins to HDL. In addition to the antiatherogenic properties, recent findings indicate that PLTP has also proatherogenic characteristics, and that these opposite characteristics of PLTP are dependent on the site of PLTP expression and action. In human plasma, PLTP exists in a high-activity (HA-PLTP) and a low-activity form (LA-PLTP), which are associated with macromolecular complexes of different size and composition. The aims of this thesis were to isolate the two PLTP forms from human plasma, to characterize the molecular complexes in which the HA- and LA-PLTP reside, and to study the interactions of the PLTP forms with apolipoproteins (apo) and the ability of apolipoproteins to regulate PLTP activity. In addition, we aimed to study the distribution of the two PLTP forms in a Finnish population sample as well as to find possible regulatory factors for PLTP by investigating the influence of lipid and glucose metabolism on the balance between the HA- and LA-PLTP. For these purposes, an enzyme-linked immunosorbent assay (ELISA) capable of determining the serum total PLTP concentration and quantitating the two PLTP forms separately was developed. In this thesis, it was demonstrated that the HA-PLTP isolated from human plasma copurified with apoE, whereas the LA-PLTP formed a complex with apoA-I. The separation of these two PLTP forms was carried out by a dextran sulfate (DxSO4)-CaCl2 precipitation of plasma samples before the mass determination. A similar immunoreactivity of the two PLTP forms in the ELISA could be reached after a partial sample denaturation by SDS. Among normolipidemic Finnish individuals, the mean PLTP mass was 6.6 +/- 1.5 mg/l and the mean PLTP activity 6.6 +/- 1.7 umol/ml/h. Of the serum PLTP concentration, almost 50% represented HA-PLTP. The results indicate that plasma HDL levels could regulate PLTP concentration, while PLTP activity could be regulated by plasma triglyceride-rich very low-density lipoprotein (VLDL) concentration. Furthermore, new evidence is presented that PLTP could also play a role in glucose metabolism. Finally, both PLTP forms were found to interact with apoA-I, apoA-IV, and apoE. In addition, both apoE and apoA-IV, but not apoA-I, were capable of activating the LA-PLTP. These findings suggest that the distribution of the HA- and LA-PLTP in human plasma is subject to dynamic regulation by apolipoproteins.

Structure, function and intracellular dynamics of alphavirus replication complexes

Intracellular membrane alterations are hallmarks of positive-sense RNA (+RNA) virus replication. Strong evidence indicates that within these exotic compartments, viral replicase proteins engage in RNA genome replication and transcription. To date, fundamental questions such as the origin of altered membranes, mechanisms of membrane deformation and topological distribution and function of viral components, are still waiting for comprehensive answers. This study addressed some of the above mentioned questions for the membrane alterations induced during Semliki Forest virus (SFV) infection of mammalian cells. With the aid of electron and fluorescence microscopy coupled with radioactive labelling and immuno-cytochemistry techniques, our group and others showed that few hours after infection the four non structural proteins (nsP1-4) and newly synthesized RNAs of SFV colocalized in close proximity of small membrane invaginations, designated as spherules . These 50-70 nm structures were mainly detected in the perinuclear area, at the limiting membrane of modified endosomes and lysosomes, named CPV-I (cytopathic vacuoles type I). More rarely, spherules were also found at the plasma membrane (PM). In the first part of this study I present the first three-dimensional reconstruction of the CPV-I and the spherules, obtained by electron tomography after chemical or cryo-fixation. Different approaches for imaging these macromolecular assemblies to obtain better structure preservation and higher resolution are presented as unpublished data. This study provides insights into spherule organization and distribution of viral components. The results of this and other experiments presented in this thesis will challenge currently accepted models for virus replication complex formation and function. In a revisitation of our previous models, the second part of this work provides the first complete description of the biogenesis of the CPV-I. The results demonstrate that these virus-induced vacuoles, where hundreds of spherules accumulate at late stages during infection, represent the final phase of a journey initiated at the PM, which apparently serves as a platform for spherule formation. From the PM spherules were internalized by an endocytic event that required the activity of the class I PI3K, caveolin-1, cellular cholesterol and functional actin-myosin network. The resulting neutral endocytic carrier vesicle delivered the spherules to the membrane of pre-existing acidic endosomes via multiple fusion events. Microtubule based transport supported the vectorial transfer of these intermediates to the pericentriolar area where further fusions generated the CPV-I. A signal for spherule internalization was identified in one of the replicase proteins, nsP3. Infections of cells with viruses harbouring a deletion in a highly phosphorylated region of nsP3 did not result in the formation of CPV-Is. Instead, thousands of spherules remained at the PM throughout the infection cycle. Finally, the role of the replicase protein nsP2 during viral RNA replication and transcription was investigated. Three enzymatic activities, protease, NTPase and RNA-triphosphatase were studied with the aid of temperature sensitive mutants in vitro and, when possible, in vivo. The results highlighted the interplay of the different nsP2 functions during different steps of RNA replication and sub-genomic promoter regulation, and suggest that the protein could have different activities when participating in the replication complex or as a free enzyme.

¿Feminismo indígena? : Un análisis crítico del discurso sobre los textos de la mujer en el movimiento zapatista 1994-2009

This work analyses texts on indigenous women´s participation in the Mexican Zapatista Army, Ejército Zapatista de Liberación Nacional. The EZLN came to public attention after ten years of clandestine organization in 1994 in Chiapas, a southern state of Mexico neighboring Guatemala. Along the invasion of various municipalities in Chiapas, the Zapatista Army published their own Revolutionary Laws, directed to the Mexican government that included a section on women´s own laws. The indigenous women´s participation in a guerrilla movement in the economically poorest area of Mexico raised many questions among Mexican feminists and some of them fiercely criticized the laws for not being liberating or feminist at all. The question is, did the indigenous women want the laws to be feminist? To answer the main research question How is the position of women constructed in the Zapatista discourse? I analyze texts by various actors in the discourse within the theoretical framework of critical discourse analysis and the feminist theories of intersectionality. The connecting point in this interdisciplinary framework is the question of power and hegemony. The actors in the discourse are the women commanders themselves, the men commanders, the Zapatista spokesperson, subcomandante Marcos and the Mexican feminists. The texts analyzed are the letters of the EZLN to the media and discourses in public reunions, first published in Mexican newspapers and international discussion lists on the Internet and after 2005, on the Zapatista´s own webpage. The results show that instead of discussing whether the Zapatista women´s participation is feminist or not, the action itself provoked such wide discussion of the diversity within the feminist movement that it is a contribution itself. The work also shows that the use of language can be one tool in the quite recent paradigm of intersectionality in feminist theories.

Le Grand Tour Du reste de l’Europe jusqu’à la Laponie – et retour

Rencontre entre le Sud et le Nord Depuis des siècles, les pays méditerranéens ont entretenu des relations actives avec le Nord de l’Europe. Les regions du Nord ont été conquises et converties, manipulées et colonisées, mais aussi intégrées dans les réseaux commerciaux, scientifi ques, culturels et touristiques. Les voyageurs du Sud de l’Europe ont peu à peu découvert les regions plurielles du Nord de la Finlande, de la Suède, de la Norvège et de la presqu’île de Kola. L’attraction focntionnait aussi dans l’autre sens. Pour les gens du Nord, scientifi ques ou artistes, le Sud de l’Europe constituait une destination aussi exotique aue la Laponie pour ceux du Sud. Dans ce recueil, nous proposons dix points de vue sur ces échanges et réseaux entre la Sud et le Nord. Le premier chapitre présente Tornio, connue très tôt comme la porte de la Laponie, les premières relations scientifi ques entre le Nord et le Sud, ainsi que les exhibitions de Samis dans les métropoles européennes. Le deuxième chapitre propose une analyse des experiences et des impressions de voyageurs italiens, espagnols et français. Le dernier chapitre est consacré aux voyages d’artistes et écrivains fi nlandais en France, principalement à Paris. Ces rencontres entre le Sud et le Nord qui participèrent à la remise en cause, la relativisation et la construction des identities nationals et ethniques, permirent aussi de créer une conscience européenne commune.

La música en la fiesta del dios Xipe Totec

In Aztec mythology and religion, Xipe Totec ("our lord the flayed one") was a life-death-rebirth deity, god of agriculture, vegetation, the east, disease, spring, goldsmiths, silversmiths and the seasons. The music used for his worship has been described by a diversity of documents (carved stones, depicted codices, and chronicles made by the first European friars arrived to Mexico). This article explores the symbolic content of these descriptions, especially in association with Xochipilli Macuilxochitl, master of music and poetry.

El teponaztli en la tradición musical mexica

Lecture about the teponaztli (a specific type of slit drum), traditionally used by the Aztec (or Mexica) people, and its associated repertoire. Special attention is paid to the usage of this instrument in the Cantares Mexicanos (a song compilation from the 16th century) accompaniment.