Cell functional enviromics: applications to Pichia pastoris

The present PhD thesis develops the cell functional enviromics (CFE) method to investigate the relationship between environment and cellular physiology. CFE may be defined as the envirome-wide cellular function reconstruction through the collection and systems-level analysis of dynamic envirome data. Throughout the thesis, CFE is illustrated by two main applications to cultures of a constitutive P. pastoris X33 strain expressing a scFv antibody fragment. The first application addresses the challenge of culture media development. A dataset was built from 26 shake flask experiments, with variations in trace elements concentrations and basal medium dilution based on the standard BSM+PTM1. Protein yield showed high sensitivity to culture medium variations, while biomass was essentially determined by BSM dilution. High scFv yield was associated with high overall metabolic fluxes through central carbon pathways concomitantly with a relative shift of carbon flux from biosynthetic towards energy-generating pathways. CFE identified three cellular functions (growth, energy generation and by-product formation) that together described 98.8% of the variance in observed fluxes. Analyses of how medium factors relate to identified cellular functions showed iron and manganese at concentrations close to PTM1 inhibit overall metabolic activity. The second application addresses bioreactor operation. Pilot 50 L fed-batch cultivations, followed by 1H-NMR exometabolite profiling, allowed the acquisition of data for 21 environmental factors over time. CFE identified five major metabolic pathway groups that are frequently activated by the environment. The resulting functional enviromics map may serve as template for future optimization of media composition and feeding strategies for Pichia pastoris. The present PhD thesis is a step forward towards establishing the foundations of CFE that is still at its infancy. The methods developed herein are a contribution for changing the culture media and process development paradigm towards a holistic and systematic discipline in the future.

Enhancing fan experience during live sports broadcasts through second screen applications

When sports fans attend live sports events, they usually engage in social experiences with friends, family members and other fans at the venue sharing the same affiliation. However, fans watching the same event through a live television broadcast end up not feeling so emotionally connected with the athletes and other fans as they would if they were watching it live, together with thousands of other fans. With this in mind, we seek to create mobile applications that deliver engaging social experiences involving remote fans watching live broadcasted sports events. Taking into account the growing use of mobile devices when watching TV broadcasts, these mobile applications explore the second screen concept, which allows users to interact with content that complements the TV broadcast. Within this context, we present a set of second screen application prototypes developed to test our concepts, the corresponding user studies and results, as well as suggestions on how to apply the prototypes’ concepts not only in different sports, but also during TV shows and electronic sports. Finally, we also present the challenges we faced and the guidelines we followed during the development and evaluation phases, which may give a considerable contribution to the development of future second screen applications for live broadcasted events.