Translating Cards against Humanity for Finnish Users : What Should Be Taken into Consideration?

Tämän tutkielman tavoitteena oli kääntää poliittisesti epäkorrekti korttipeli nimeltä Cards against Humanity siten, että suomenkielinen versio olisi mahdollisimman käytettävä ja hauska. Tämän tavoitteen saavuttamiseksi tutkielmassa hyödynnettiin Tytti Suojasen, Kaisa Koskisen ja Tiina Tuomisen luomaa käyttäjäkeskeinen kääntämisen teoriaa (UCT) ja sen tarjoamia työkaluja, kuten mentaalimalleja ja heuristista arviointia. Lisäksi hyödynnettiin lingvistiikan sekä huumorin ja kulttuurin kääntämisen tutkimuskirjallisuutta. Tutkielman hypoteesi oli, että mitä paremmin käännöstyössä huomioidaan pelin tulevat käyttäjät ja tutkimuskirjallisuus, sitä parempi käännöksestä tulee. Käytettävyyden ja hauskuuden arvioimiseksi tehtiin käytettävyystutkimus, joka suoritettiin testipelien ja kyselylomakkeiden avulla. Tutkimukseen osallistui 42 henkilöä, jotka pelattuaan suomennettua peliä vastasivat taustatietokysymyksiin ja arvioivat mm. korttien käytettävyyttä ja ymmärrettävyyttä, kieltä, hauskuutta, loukkaavuutta ja kokonaisuutta. Kyselylomakkeiden tuottama aineisto analysoitiin kvantitatiivisesti ja kvalitatiivisesti, ja lopuksi aineisto arvoitiin heuristisesti. Tutkimuksen mukaan käännös onnistui käytettävyyden ja hauskuuden tavoitteissaan, ja ennakko-oletuksen mukaisesti tulevien käyttäjien profiloinnilla ja tutkimuskirjallisuuteen tukeutumisella oli positiivinen vaikutus käännöksen laatuun. Käännöksen suurimmat ongelmat liittyivät korttien kokoon ja määrään, sekä sanojen tunnistukseen ja korttien yhteensopivuuteen. Nämä ongelmat olisi olleet todennäköisesti vältettävissä seuraamalla UCT-teoriaa kokonaisvaltaisemmin

The Inducible Intein-Mediated Self-Cleaving Tag (IIST) System: A Novel Purification and Amidation System for Peptides and Proteins

An efficient self-cleavable purification tag could be a powerful tool for purifying recombinant proteins and peptides without additional proteolytic processes using specific proteases. Thus, the intein-mediated self-cleavage tag was developed and has been commercially available as the IMPACT™ system. However, uncontrolled cleavages of the purification tag by the inteins in the IMPACT™ system have been reported, thereby reducing final yields. Therefore, controlling the protein-splicing activity of inteins has become critical. Here we utilized conditional protein splicing by salt conditions. We developed the inducible intein-mediated self-cleaving tag (IIST) system based on salt-inducible protein splicing of the MCM2 intein from the extremely halophilic archaeon, Halorhabdus utahensis and applied it to small peptides. Moreover, we described a method for the amidation using the same IIST system and demonstrated 15N-labeling of the C-terminal amide group of a single domain antibody (VHH)

Lifting the curtain on contemporary office: Expected versus actual practices of virtual multispace office

Objective of the study: The production process of the 21st century in Western countries is increasingly concentrated on creating and sharing knowledge, while the production of physical goods is outsourced to lowercost economies. This so-called knowledge work often takes place outside the traditional temporal and spatial employment spheres as work can often be conducted anytime and anywhere with a laptop and an interned connection. To support these new flexible ways of working, there has been a recent trend in office design towards virtual multispace office design. The purpose of this study is to understand the expectations of managers and designers versus the actual practices of employees as to the use of virtual multispace office space and how the office design has consequence on the everyday work practices of the organizational members. Research method: Earlier research on office spaces in organizational studies has mainly taken the perspective of seeing them as abstract macro-level organizational systems, making assumptions on how organizations adapt to their changing environments through office design. In order to broaden our understanding of organizational life and its consequences of office design, this thesis will adopt a practice perspective in studying organizations - one that sees social and material as mutually entangled or imbricated. The research method utilized in data collection follows the principles of practice theory with fieldwork comprised of direct observations and interviews and informal discussion. Findings: The empirical data revealed how the office space is not a stable box that could be imposed with expected ways of using the space, assuming that the employees would act according to the intentions but that both space and materiality emerge as dynamic and generative forces shaping the individuals appropriating the office space in an on-going manner. This finding became evident through the interplay of identified accommodating practices that were aligned as well as through the resisting practices that were misaligned to the intended ways of using the virtual multispace office. Furthermore, a paradox between the managerial and design discourse on ways of using the virtual multispace office and the realized materiality of the office space was identified

The Inducible Intein-Mediated Self-Cleaving Tag (IIST) System: A Novel Purification and Amidation System for Peptides and Proteins

An efficient self-cleavable purification tag could be a powerful tool for purifying recombinant proteins and peptides without additional proteolytic processes using specific proteases. Thus, the intein-mediated self-cleavage tag was developed and has been commercially available as the IMPACT™ system. However, uncontrolled cleavages of the purification tag by the inteins in the IMPACT™ system have been reported, thereby reducing final yields. Therefore, controlling the protein-splicing activity of inteins has become critical. Here we utilized conditional protein splicing by salt conditions. We developed the inducible intein-mediated self-cleaving tag (IIST) system based on salt-inducible protein splicing of the MCM2 intein from the extremely halophilic archaeon, Halorhabdus utahensis and applied it to small peptides. Moreover, we described a method for the amidation using the same IIST system and demonstrated 15N-labeling of the C-terminal amide group of a single domain antibody (VHH)

Psychosocial difficulties and treatment retention in inpatient detoxification programmes

Aims: Treatment retention is associated with addiction treatment outcomes. Research regarding predictors of retention at inpatient detoxification treatment is limited. The aim of this study was to investigate whether psychosocial difficulties (PSDs) are associated with treatment retention among Finnish inpatients undergoing detoxification treatment. Design: This register-based study included real-life data on detoxification treatment episodes (n = 2,752) between February of 2016 and May of 2019 from several inpatient treatment units in Finland. The PARADISE24fin instrument was used to assess PSDs. Socio-demographic and substance use related variables, as well as PSDs, were analysed with regard to treatment retention. Multiple logistic regression models were used to identify predictors of treatment incompletion. Results: Of the 2,752 detoxification treatment episodes, 80.3% (n = 2,209) were completed. Men and women differed with regard to the variables associated with treatment retention. After adjusting for confounders, younger age (Peer reviewe

The NMR structure of the engineered halophilic DnaE intein for segmental isotopic labeling using conditional protein splicing

Protein trans-splicing catalyzed by split inteins has been used for segmental isotopic labeling of proteins for alleviating the complexity of NMR signals. Whereas inteins spontaneously trigger protein splicing upon protein folding, inteins from extremely halophilic organisms require a high salinity condition to induce protein splicing. We designed and created a salt-inducible intein from the widely used DnaE intein from Nostoc punctiforme by introducing 29 mutations, which required a lower salt concentration than naturally occurring halo-obligate inteins. We determined the NMR solution structure of the engineered salt-inducible DnaE intein in 2 M NaCl, showing the essentially identical three-dimensional structure to the original one, albeit it unfolds without salts. The NMR structure of a halo-obligate intein under high salinity suggests that the stabilization of the active folded conformation is not a mere result of various intramolecular interactions but the subtle energy balance from the complex interactions, including the solvation energy, which involve waters, ions, co-solutes, and protein polypeptide chains. (C) 2022 The Authors. Published by Elsevier Inc.Peer reviewe

Intermolecular protein splicing and its use in biotechnological applications

Inteins are selfish but harmless autocatalytic proteins that perform a post-translational modification, termed protein splicing. In protein splicing an intein excises itself off from the precursor protein and simultaneously ligates the flanking proteins together with a peptide bond. Inteins are found sporadically distributed in unicellular organisms, but their biological functions remain obscure. Importanly, inteins that are split into two can remain active and perform protein ligation by protein trans-splicing (PTS). In principle, PTS allows ligation of any two protein-sequences, with the only requirement being Ser, Thr, or Cys as the first residue downstream of the intein. This has inspired development of numerous biotechnological applications including protein semisynthesis, segmental isotopic labeling, and cyclization. Protein ligation by split inteins is, however, limited by the lengths, substrate specificity, orthogonality, and the reaction yields of the split inteins. The objective of this thesis was to advance the development of protein splicing as a protein-ligation tool. First, the split site of a natively split DnaE intein was shifted in order to engineer a split intein with shorter C-intein that could be easily chemically synthetized. The newly engineered split intein could perform protein ligation in high yields and was demonstrated to be in certain cases even better than the natively split intein. Encouraged by this, 21 more split inteins were engineered starting from four different inteins, guided by the three dimensional structures of these inteins. Split inteins were systematically tested for activity and orthogonality to evaluate their potential for biotechnological applications. Next, the scope was widened to bacterial intein-like (BIL) domains. BIL domains belong to the same superfamily with inteins but are distinct by their distribution and functions and have a wider variety of residues at the downstream junction. The first structure of a BIL domain was solved. It highlighted their homology to inteins as well as allowed engineering of split BIL domains. The split BIL domains could perform protein ligation also with Ala at the downstream splicing junction, although in minute yields, which could be the first step towards nucleophile-free protein ligation. Finally, discovery of a previously not reported intermolecular protein-splicing reaction, termed intein-mediated protein alternative splicing (iPAS), was described. Structural studies revealed that three-dimensional domain swapping is the underlying mechanisms of iPAS. iPAS makes it possible to increase diversity at protein level, without altering the genetic code, and could be used to control protein functions in concentration and expression-order dependent manner. Discovery of this new phenomenon could allow protein interference and is opening new insights into the possible biological functions of inteins.Proteiinien silmukointi työkaluna proteiinien muokkauksessa ja aktiivisuuden säätelyssä Tässä työssä on tutkittu proteiinien silmukointia sekä mahdollisuuksia hyödyntää sitä bioteknologian sovelluksissa. Työssä kehitettiin olemassa olevia proteiinien silmukointiin perustuvia ligaatio-menetelmiä sekä kuvataan uudenlainen intermolekulaarinen proteiinien silmukointi -mekanismi, joka mahdollistaa proteiinien aktiivisuuden säätelyn proteiinitasolla. Proteiinien silmukointi -domeenit voidaan jakaa kolmeen eri luokkaan, joista kahta, inteiinejä ja bakteerien inteiinin-kaltaisia (bacterial intein-like, BIL) domeeneja, on tutkittu tässä työssä. Inteiinit ovat yksisoluisista eliöistä löytyviä autokatalyyttisiä proteiini-domeeneja, jotka silmukoituvat irti esiproteiinista liittäen samalla vapautuvat päät yhteen peptidi-sidoksella. Inteiineistä ei tiedetä olevan hyötyä, muttei haittaakaan, isäntäorganismeilleen. Niitä voidaan kuitenkin hyödyntää useissa bioteknologian, biokemian ja biolääketieteen sovelluksissa, sillä ne mahdollistavat post-translationaalisen proteiinin muokkauksen. Suuri osa sovelluksista perustuu proteiinien trans-silmukointiin, joka on halkaistujen inteiinien välittämä proteiinien ligaatio -reaktio. BIL-domeenit voivat myös silmukoitua mutta niiden pääasiallinen tehtävä on katkaista viereinen proteiini irti. Päinvastoin kuin inteiinit, BIL-domeenien uskotaan olevan tärkeitä isäntäorganismeilleen mutta niille ei ole kehitetty bioteknologisia sovelluksia. Tässä työssä muokattiin uusia halkaistuja inteiinejä sekä tutkittiin näiden bioteknologisten sovellusten kannalta tärkeitä ominaisuuksia in vivo sekä in vitro. Työssä kuvataan uusia aktiivisia halkaistuja inteiinejä, joista muutamalla on sovellusten kannalta erityisen lupaavia ominaisuuksia. Lisäksi työ lisäsi tietämystä halkaistuja inteiinejä kehittäessä huomioitavista seikoista. Työssä ratkaistiin ensimmäinen BIL-domeenin kolmiulotteinen rakenne NMR-spektroskopian avulla. Tämä vahvisti näiden rakenteellisen homologian inteiinien kanssa sekä auttoi suunnittelemaan proteiinien ligaatioon soveltuvia halkaistuja BIL-domeeneja. Työssä osoitettiin, että halkaistujen BIL domeenien avulla on teoriassa mahdollista ratkaista inteiinien reaktiomekanismista juontuva rajoitus kohdeproteiineille, vaikka reaktion tehokkuutta tulisikin parantaa. Kolmantena osuutena työssä kuvataan inteiini-välitteinen vaihtoehtoinen proteiinien silmukointi (intein-mediated protein alternative splicing, iPAS). Rakennetutkimukset osoittivat reaktion johtuvan kolmiulotteisesta domeenien uudelleenjärjestäytymisestä. Ilmiö voitiin toistaa useilla eri inteiineillä sekä eri kohdeproteiineilla, mukaan lukien eräs luonnollisista isäntäproteiineista. Lisäksi iPAS:n osoitettiin mahdollistavan proteiinien aktiivisuuden häirinnän sekä pelastamisen. Näitä havaintoja voidaan hyödyntää uusien työkalujen kehittämisessä ja niillä voi olla merkitystä inteiinien biologisen merkityksen kannalta

Substrate specificities of inteins investigated by QuickDrop-cassette mutagenesis

Inteins catalyze self-excision from host precursor proteins while concomitantly ligating the flanking substrates (exteins) with a peptide bond. Noncatalytic extein residues near the splice junctions, such as the residues at the -1 and +2 positions, often strongly influence the protein-splicing efficiency. The substrate specificities of inteins have not been studied for many inteins. We developed a convenient mutagenesis platform termed "QuickDrop"-cassette mutagenesis for investigating the influences of 20 amino acid types at the -1 and +2 positions of different inteins. We elucidated 17 different profiles of the 20 amino acid dependencies across different inteins. The substrate specificities will accelerate our understanding of the structure-function relationship at the splicing junctions for broader applications of inteins in biotechnology and molecular biosciences.Peer reviewe

Recombinant Spider Silk Protein and Delignified Wood Form a Strong Adhesive System

For developing novel fully biological materials, a central question is how we can utilize natural components in combination with biomimetic strategies in ways that both allow feasible processing and high performance. Within this development, adhesives play a central role. Here, we have combined two of nature's excellent materials, silk and cellulose, to function as an adhesive system. As an initial step in processing, wood was delignified. Without lignin, the essential microstructure and alignment of the wood remain, giving a strong scaffold that is versatile to process further. A recombinant spider silk protein was used as a fully biological and water-based adhesive. The adhesive strength was excellent with an average value of 6.7 MPa, with a maximum value of up to 10 MPa. Samples of different strengths showed characteristic features, with high tear-outs for weaker samples and only little tear-out for strong samples. As references, bovine serum albumin and starch were used. Based on the combined data, we propose an overall model for the system and highlight how multiple variables affect performance. Adhesives, in particular, biobased ones, must be developed to be compatible with the overall adherend system for suitable infiltration and so that their mechanical properties match the adherend. The engineering of proteins gives an unmatched potential for designing adhesive systems that additionally have desired properties such as being fully water-based, biologically produced, and renewable.Peer reviewe

In vivo and in vitro protein ligation by naturally occurring and engineered split DnaE inteins

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