Continuous Flow Biocatalytic Reductive Amination by Co-Entrapping Dehydrogenases with Agarose Gel in a 3D-Printed Mould Reactor

Herein, we show how the merge of biocatalysis with flow chemistry aided by 3D-printing technologies can facilitate organic synthesis. This concept was exemplified for the reductive amination of benzaldehyde catalysed by co-immobilised amine dehydrogenase and formate dehydrogenase in a continuous flow micro-reactor. For this purpose, we investigated enzyme co-immobilisation by covalent binding, or ion-affinity binding, or entrapment. Entrapment in an agarose hydrogel turned out to be the most promising solution for this biocatalytic reaction. Therefore, we developed a scalable and customisable approach whereby an agarose hydrogel containing the co-entrapped dehydrogenases was cast in a 3D-printed mould. The reactor was applied to the reductive amination of benzaldehyde in continuous flow over 120 h and afforded 47 % analytical yield and a space-time yield of 7.4 g L day−1 using 0.03 mol% biocatalysts loading. This work also exemplifies how rapid prototyping of enzymatic reactions in flow can be achieved through 3D-printing technology

FOXN1 forms higher-order nuclear condensates displaced by mutations causing immunodeficiency

The transcription factor FOXN1 is a master regulator of thymic epithelial cell (TEC) development and function. Here, we demonstrate that FOXN1 expression is differentially regulated during organogenesis and participates in multimolecular nuclear condensates essential for the factor’s transcriptional activity. FOXN1’s C-terminal sequence regulates the diffusion velocity within these aggregates and modulates the binding to proximal gene regulatory regions. These dynamics are altered in a patient with a mutant FOXN1 that is modified in its C-terminal sequence. This mutant is transcriptionally inactive and acts as a dominant negative factor displacing wild-type FOXN1 from condensates and causing athymia and severe lymphopenia in heterozygotes. Expression of the mutated mouse ortholog selectively impairs mouse TEC differentiation, revealing a gene dose dependency for individual TEC subtypes. We have therefore identified the cause for a primary immunodeficiency disease and determined the mechanism by which this FOXN1 gain-of-function mutant mediates its dominant negative effect

The additional value of patient-reported health status in predicting 1-year mortality after invasive coronary procedures: A report from the Euro Heart Survey on Coronary Revascularisation

Objective: Self-perceived health status may be helpful in identifying patients at high risk for adverse outcomes. The Euro Heart Survey on Coronary Revascularization (EHS-CR) provided an opportunity to explore whether impaired health status was a predictor of 1-year mortality in patients with coronary artery disease (CAD) undergoing angiographic procedures. Methods: Data from the EHS-CR that included 5619 patients from 31 member countries of the European Society of Cardiology were used. Inclusion criteria for the current study were completion of a self-report measure of health status, the EuroQol Questionnaire (EQ-5D) at discharge and information on 1-year follow-up, resulting in a study population of 3786 patients. Results: The 1-year mortality was 3.2% (n = 120). Survivors reported fewer problems on the five dimensions of the EQ-5D as compared with non-survivors. A broad range of potential confounders were adjusted for, which reached a p<0.10 in the unadjusted analyses. In the adjusted analyses, problems with self-care (OR 3.45; 95% CI 2.14 to 5.59) and a low rating (≤ 60) on health status (OR 2.41; 95% CI 1.47 to 3.94) were the most powerful independent predictors of mortality, among the 22 clinical variables included in the analysis. Furthermore, patients who reported no problems on all five dimensions had significantly lower 1-year mortality rates (OR 0.47; 95% CI 0.28 to 0.81). Conclusions: This analysis shows that impaired health status is associated with a 2-3-fold increased risk of all-cause mortality in patients with CAD, independent of other conventional risk factors. These results highlight the importance of including patients' subjective experience of their own health status in the evaluation strategy to optimise risk stratification and management in clinical practice

Design and fabrication through additive manufacturing of devices for multidimensional LC based on computational insights

One-dimensional column-based liquid chromatography systems do not offer sufficient separation power for the analysis of complex samples. Two-dimensional systems are more powerful. However, they suffer from longer analysis times. Spatial multi-dimensional liquid chromatography may prove an efficient solution, as the peak capacity of the system is ideally the product of those of the individual dimensions, yet the analysis time remains relatively short due to parallel separations. The main aspects we need to investigate for such devices include flow uniformity, analyte transfer between dimensions, band broadening, and confinement of the flow in each dimension. The objectives of this thesis were to study the effects of design, fabrication and operation factors on the performance of the device. For this purpose, both simulations and experiments were conducted. Three-dimensional computational fluid dynamics (CFD) was employed to calculate flow and mass transfer and injections of a mixture of dye and water were simulated. Prototypes were fabricated using 3D-printing, more specifically, stereolithography. Flow confinement, analyte transfer, band broadening and pressure resistance of devices were studied. Feasible designs were proposed for two- and three-dimensional spatial LC devices, using either passive flow confinement based on permeability differences or active flow confinement based on an approach that was patented during this work. Various other applications of 3D-printing in analytical sciences were also studied

Επίδραση του συνδυασμού τιμών των κυριότερων παραμέτρων του κτιριακού κελύφους στις συνθήκες θερμικής άνεσης κατά τους χειμερινούς μήνες

234 σ.Στην παρούσα διπλωματική εργασία εξετάζεται η επίδραση του συνδυασμού τιμών των κυριότερων παραμέτρων του κτιριακού κελύφους στις συνθήκες θερμικής άνεσης κατά τους χειμερινούς μήνες. Αρχικά στο βιβλιογραφικό πρώτο μέρος περιγράφονται τα δομικά στοιχεία του κτιριακού κελύφους ,τα βασικά χαρακτηριστικά τους και ο τρόπος που αυτά επιδρούν στην ενεργειακή κατανάλωση του κτιρίου. Επιπλέον αναπτύσσονται οι έννοιες που είναι απαραίτητες για την κατανόηση της θερμικής συμπεριφοράς του κτιρίου καθώς και το θεσμικό πλαίσιο που εφαρμόζεται για τη μείωση και τον έλεγχο της ενεργειακής κατανάλωσης του κτιριακού τομέα. Στο δεύτερο μέρος, υπολογίζεται η ενεργειακή κατανάλωση συγκεκριμένου κτιρίου αναφοράς κατά τους χειμερινούς μήνες και μελετάται η διακύμανση της ενεργειακής κατανάλωσης του κτιρίου λόγω της επίδρασης του συνδυασμού των κυριότερων παραμέτρων της μόνωσης και των υαλοπινάκων. Στη συνέχεια παρουσιάζονται οι καμπύλες ενεργειακής κατανάλωσης του κτιρίου αναφοράς για διάφορους συνδυασμούς των παραμέτρων και τέλος προσεγγίζονται οι τιμές των παραμέτρων μόνωσης και υαλοπινάκων με τις οποίες επιτυγχάνεται ψευδοαδιαβατικό κέλυφος για κάθε χειμερινό μήνα.The present thesis examines the effect of the combinations of values of the main envelope parameters on building’s thermal comfort during winter. Initially in the bibliographic first part the elements of the building envelope, their basic characteristics and their effect on the energy consumption of the building are described. Moreover, a review is developed about the necessary terms for understanding the thermal behavior of a building and the state laws applied to reduce and control the energy consumption of the building sector. In the second part, the energy consumption of a typical reference building during the winter months is calculated and the heating energy consumption’s variation of the building due to the effect of the combination of the most important parameters of insulation and fenestration is examined. Following are the contours of the daily heating energy consumption of the reference building for various combinations of parameters, and finally there is an approach of the values of insulation and fenestration parameters for which a qausi-adiabatic envelope is achieved for each winter month.Ελένη Θ. Αδαμοπούλο