Selected Reaction Monitoring (SRM) frente a Western Blot

We propose the SRM technology as a complementary method to the Western Blot for the detection and quantification of proteins in a sample. The technique Western Blot has its own limitations: i) only a protein-of-choice is detected, ignoring any non-relevant proteins, ii) the sensitivity of the technique depends on the specificity of the antibody and iii) Western Blot is expensive and time-consuming. The advantages of SRM with respect Western Blot are remarkable: i) you can detect up to hundreds of different proteins in a sample, ii) SRM is more sensitive, because just 50 copies of the target protein per cell are enough for the detection and iii) once it has been made an investment in the necessary machinery to develop this technique, the detection of proteins in a sample turns into a cheaper, faster, more specific and full-quantitative procedure, without the need of using antibodies. First of all, SRM requires the identification of little peptides, obtained by tryptic digestion, whose sequence must be unique for a single protein or isoform. There is software for that aim. Then, it’s necessary to create isotope-labeled peptides of that identified for acting as internal standards. That sample is introduced in a triple quadrupole mass spectrometer: it passes through a first quadrupole, which functions as a filter, where the fragments are selected, previously ionized, attending to the mass/charge (m/z) relation that correspond to that unique fragments of the protein of interest. In this first selection may be other peptides from other proteins, with the same m/z but with different sequence. To select those that are exclusive from the target protein, the fragments are moved to a second quadrupole, where they are fragmented again with a physical method, and so new smaller fragments are generated. All the new fragments are conduced to the third quadrupole, where just those which come from the protein of interest are selected, attending at their m/z again. The target peptide concentration is determined by measuring the observed signal response for the target peptide relative to that of the isotopic-labeled peptide, the concentration of which is calculated from a pre-determined calibration-response curve. Calibration curves have to be generated for each target peptide in the sample. Because SRM technology is increasing its use, there have been developed databases where the scientific community upload information about protocols and standards for each protein with the aim to facilitate the work to other researchers.

Mapping the Liminal Experience of Eastern Europe in Selected American Works, 1965-2002

The purpose of this dissertation is to study literary representations of Eastern Europe in the works of celebrated and less-known American authors, who visited and narrated the region between the mid-1960s and early 2000s. The main critical body focuses on Eastern Europe before 1989 and encompasses three major voices of American literature: John Updike, Joyce Carol Oates, and Philip Roth. However, in the last chapter I also explore American literary perceptions of the area following the collapse of communism. Importantly, the term “Eastern Europe” as used in this dissertation is charged with significance. I approach it not only as a space on the map or the geopolitical construct which emerged in the aftermath of the Second World War, but rather as a conceptual category and a repository of meanings built out of fact and fantasy: specific historical, political and cultural realities interlaced with subjective worldviews, preconceptions, and mental images. The critical framework of this dissertation is twofold. I reach for the concept of liminality to elucidate the indeterminacy and malleability which lies at the heart of the object of study—the idea, image, and experience of Eastern Europe. Bearing in mind the nature of the works under analysis, all of which were inspired by actual visits behind the Iron Curtain, I propose to interpret these transatlantic literary journeys in terms of generative experience, where Eastern Europe is mapped as a liminal space of possibility; a contact zone between cultures and, potentially, the locus of self-discovery and individual transformation. If liminality is the metaphor or a lens that I employ in order to account for the nature of the analyzed works and the complex terrain they map, imagology, whose purpose is to study the processes of constructing selfhood and otherness in literature, provides me with the method and the critical vocabulary for analyzing selected literary representations. The dissertation is divided into six chapters, the last of which serves as coda to the previous discussion. The first two chapters constitute the critical foundation of this work. Then, in chapters 3, 4, and 5 I study American images of Eastern Europe in the works written by John Updike, Joyce Carol Oates, and Philip Roth, respectively. The last, sixth chapter of this dissertation is divided into two parts. In the first one, I discuss new critical perspectives and avenues of research in the study of Eastern Europe following the collapse of communism. Then, I carry out a joint analysis of four works written after 1989 by Eva Hoffman, Arthur Phillips, John Beckman, and Gary Shteyngart. The dissertation ends with conclusions in which I summarize my findings and reflections, and suggest implications for future research. As this dissertation seeks to demonstrate, Eastern Europe portrayed in the analyzed works oscillates between contradictory representations which are contingent upon a number of factors, most importantly who maps it and in what context. Even though each experience of Eastern Europe is distinct and fueled by the profiles, identities, and interests of the characters and their creators, I have found out that certain patterns of othering are present in all the works. Thus, my research seems to suggest that there is something of a recurrent literary image of Eastern Europe, which goes beyond the context of the Cold War. Accordingly, while this dissertation hopes to be a valid contribution to the study of literary and cultural mappings of Eastern Europe, it also generates new questions regarding the current, post-communist representation of the area and its relationship to the national tropes explored in my work.