Detection of Envelope Glycoprotein Assembly from Old-World Hantaviruses in the Golgi Apparatus of Living Cells.

Hantaviruses are emerging pathogens that occasionally cause deadly outbreaks in the human population. While the structure of the viral envelope has been characterized with high precision, protein-protein interactions leading to the formation of new virions in infected cells are not fully understood yet. We use quantitative fluorescence microscopy (i.e., Number&Brightness analysis and fluorescence fluctuation spectroscopy) to monitor the interactions that lead to oligomeric spike complex formation in the physiological context of living cells. To this aim, we quantified protein-protein interactions for the glycoproteins Gn and Gc from Puumala and Hantaan orthohantaviruses in several cellular models. The oligomerization of each protein was analyzed in relation to subcellular localization, concentration, and the concentration of its interaction partner. Our results indicate that when expressed separately, Gn and Gc form respectively homo-tetrameric and homo-dimeric complexes, in a concentration-dependent manner. Site-directed mutations or deletion mutants showed the specificity of their homotypic interactions. When both glycoproteins were co-expressed, we observed in the Golgi apparatus clear indication of Gn-Gc interactions and the formation of Gn-Gc multimeric protein complexes of different sizes, while using various labeling schemes to minimize the influence of the fluorescent tags. Such large glycoprotein multimers may be identified as multiple Gn viral spikes interconnected via Gc-Gc contacts. This observation provides a possible first evidence for the initial assembly steps of the viral envelope, within this organelle, directly in living cells.IMPORTANCE In this work, we investigate protein-protein interactions that drive the assembly of the hantaviruses envelope. These emerging pathogens have the potential to cause deadly outbreaks in the human population. Therefore, it is important to improve our quantitative understanding of the viral assembly process in infected cells, from a molecular point of view. By applying advanced fluorescence microscopy methods, we monitored the formation of viral spike complexes in different cell types. Our data support a model for hantavirus assembly according to which viral spikes are formed via the clustering of hetero-dimers of the two viral glycoproteins Gn and Gc. Furthermore, the observation of large Gn-Gc hetero-multimers provide a possible first evidence for the initial assembly steps of the viral envelope, directly in the Golgi apparatus of living cells.

Relation between mast cells concentration and serotonin expression in chagasic megacolon development.

Chagas' disease is still reaching about 10 million people in the world. In South America, one of the most severe forms of this disease is the megacolon, characterized by severe constipation, dilated sigmoid colon and rectum and severe malnutrition. Previous data suggested that mast cells and serotonin (5-hydroxytryptamine [5-HT]) expression could be involved in intestinal homeostasis control, avoiding the chagasic megacolon development. The aim at this study was to characterize the presence of mast cells and expression of serotonin in chagasic patients with and without megacolon and evaluate the relation between mast cells, serotonin and megacolon development. Our results demonstrated that patients without megacolon feature a large amount of serotonin and few mast cells, while patients with megacolon feature low serotonin expression and a lot of mast cells. We believe that serotonin may be involved in the inflammatory process control, triggered by mast cells, and the presence of this substance in large quantities of the intestine could represent a mechanism of megacolon prevention.

Measurement and Shaping of Biphoton Spectral Wave Functions.

In this work we present a simple method to reconstruct the complex spectral wave function of a biphoton, and hence gain complete information about the spectral and temporal properties of a photon pair. The technique, which relies on quantum interference, is applicable to biphoton states produced with a monochromatic pump when a shift of the pump frequency produces a shift in the relative frequencies contributing to the biphoton. We demonstrate an example of such a situation in type-II parametric down conversion allowing arbitrary paraxial spatial pump and detection modes. Moreover, our test cases demonstrate the possibility to shape the spectral wave function. This is achieved by choosing the spatial mode of the pump and of the detection modes, and takes advantage of spatiotemporal correlations.

Work and defensive processes in small groups: effects of leader gender and authority position.

The effects of leader gender and degree of authority on work activity and group-as-a-whole defensive processes were examined in two Tavistock-model group relations conferences. The small groups in each conference had two leaders with different degrees of explicit authority, a consultant and associate consultant. In one conference, males were designated as consultants and females as associate consultants. In the second conference these role assignments were reversed. Segments of group interaction following leaders' interventions were scored for primary group activity (dependency, fight/flight, pairing or work) and individual member work. High authority leaders elicited more fight/flight than leaders with secondary authority, who in turn evoked more pairing. High authority female leaders tended to evoke more fight/flight activity than the males paired with them. Groups periodically responded with dependency regardless of leader gender or authority position. Group-as-a-whole work occurred infrequently. Individual work occurred, with no significant difference in frequency, in response to all leaders and in the midst of each group-as-a-whole defensive process.