Proceedings of the 4th MIT Portugal Program Conference by MIT Portugal Program

Proceedings of the 4th MIT Portugal Conference

NEW FRONTIERS

CONTENTS Forward

Conference Program

Doctoral Projects

Bioengineering Systems

Engineering Design and Advanced Manufacturing

Sustainable Energy Systems

Transportation Systems

135

FORWARD Dear Conference participants: The 2014 MIT Portugal Program Conference was conceived as a new, additional occasion for the scientific community to meet with industry, and discuss the role of knowledge transfer in innovation fostering and technological progress. Special emphasis is given to entrepreneurship, with several moments where it will be the main theme of debate. It is nowadays considered one important instrument of transforming ideas into social progress, through new and more effective solutions to problems, helping to create more qualified jobs and to raise social well being. The University of Coimbra has a long track record on innovation fostering and on incubating new business ideas into real companies, now also filling an additional stage, that of accelerating companies development. The Conference is, therefore, a good combination between a spot and a motto: a spot where consistently new businesses and value are encouraged and supported and a Conference motto loaded with hope for the future. This is the first MIT Portugal Conference taking place after the beginning of the second phase of the Program. It is therefore, also, an important occasion for exchanging ideas and do networking, aiming at stronger bonds between all the institutions and industry involved, scientists, students, engineers, industry leaders. In this context, the compilation of the abstracts submitted by PhD students is a token of the creative and dynamic environment that the MIT Portugal Program has been able to create. It is my pleasure to welcome you and invite you to explore it.

Ant贸nio Gomes Martins

Conference chairman

University of Coimbra

CONFERENCE PROGRAM JUNE 27th 2014

8h30 REGISTRATION

13h00 LUNCH

9h00 WELCOME SESSION

14h00 - 16h00 NEW FRONTIERS IN SCIENCE AND ENGINEERING

António Gomes Martins / Conference Chairman Luis Godinho / Faculty of Science and Technology, University of Coimbra João Gabriel Silva / Rector of University of Coimbra António Cunha / Rector of University of Minho, Council of Rectors of Portuguese Universities (CRUP) representative at the MIT Portugal program PGC

9h30 - 11h30 ENTREPRENEURSHIP AT MIT PORTUGAL PROGRAM 9h30 - 10h30 Keynote Speaker: Charles Cooney / MIT “Exploring New Frontiers by Translating Ideas into Commercial Impact “ Introduction to Keynote Speaker: Dava Newman / MIT Portugal Program Director

10h30 - 11h30 DISCUSSION PANEL Diana Fernandes / E3 Forum Gonçalo Amorim / Director of the MIT Portugal Innovation & Entrepreneurship Initiative and Building Global Innovators João Nuno Moreira / University of Coimbra, Treat-U co-founder José Costa / University of Coimbra, WSBP co-founder Joseph Shamon / Independent Board Advisor at MIT Portugal Program Innovation & Entrepreneurship Initiative, Building Global Innovators Teresa Mendes / President, Instituto Pedro Nunes, IPN Moderator: Jorge Figueira / Industry Liason Officer, University of Coimbra

11h30 STUDENTS' RESEARCH HIGHLIGHTS – POSTER SESSION

14h00 - 14h50 NEW FRONTIERS IN SUSTAINABLE CITIES António Gomes Martins / University of Coimbra Claúdia Sousa / MPP student Luís Seca / MPP Student Miguel Moreira da Silva / REN & MPP alumnus

14h50 - 15h25 NEW FRONTIERS IN STEM CELL ENGINEERING & REGENERATIVE MEDICINE Bruce Tidor / MIT Francisco Santos / Cell2B Chief Scientific Officer and Co-Founder Sezin Aday / MPP Student

15h25 - 16h00 NEW FRONTIERS IN DESIGN AND MANUFACTURING APPROACHES Alexandre Silva / Universidade do Minho Dava Newman / MIT Portugal Program Director José Valente / Managing Director Iber-Oleff Nuno Valverde / MPP student

16h00 COFFEE BREAK 16h30 BEST POSTER AWARD 16h45 GRADUATION CEREMONY 17h30 CLOSING REMARKS MIT Portugal Program Directors: Dava Newman, Paulo Ferrão

Doctoral Projects

Analysis of the gene regulatory network underlying pluripotenciality in mouse embryonic stem cells Ana Marisa Mendes Gonçalves Vinhais Guedes MIT Portugal Bioengineering Systems, Instituto de Medicina Molecular and Instituto de Histologia e Biologia do Desenvolvimento, Faculdade de Medicina da Universidade de Lisboa aguedes@medicina.ulisboa.pt Background: Master’s in Molecular Biology and Genetics. Faculdade de Ciências da Universidade de Lisboa Starting year: 2011 Research team: Domingos Henrique1,2; Elsa Abranches1,2 Supervisors: Domingos Henrique1,2; Elsa Abranches1,2; Claudia L. da Silva3 1 Instituto de Medicina Molecular and Instituto de Histologia e Biologia do Desenvolvimento, Faculdade de Medicina da Universidade de Lisboa 2 Champalimaud Neuroscience Programme, Champalimaud Centre for the Unknown 3 Department of Bioengineering and IBB – Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa

OBJECTIVES Dissect the gene regulatory network underlying pluripotenciality in mouse Embryonic Stem cells (mESCs) by investigating the functional role of single-cell fluctuations in NANOG expression and FGF/ ERK signaling, and how this interplay regulates the responsiveness of mESCs to differentiation cues. WORK PLAN Take advantage of a mESC line (Nd) expressing a fluorescent reporter (VNP) under the control of Nanog regulatory regions to: (i) assess NANOG heterogeneity in different culture conditions; (ii) quantify mRNA expression in single-cells of key pluripotency transcription factors by single molecule FISH; and (iii) analyze lineage-affiliated gene expression.

RESULTS We have shown that Nd mESCs cultured in different media conditions display very different NANOG expression levels (Fig. 1 A-C) correlating with differences in cell morphology (Fig. 1 D-F). This heterogeneity is also present at the mRNA level and can be observed when analyzing Oct4 (Fig. 2 D-F) and Nanog (Fig. 2 A-C) mRNA expression in single cells. Correlation analysis between Oct4 and Nanog mRNA expression in individual cells shows that in Serum and SerumLIF cells with low number of Nanog mRNA molecules express varying levels of Oct4 (Fig. 2 G-H). In 2iLIF the correlation between Nanog and Oct4 expression is higher with very few cells showing low levels of expression of either Nanog or Oct4 (Fig. 2 I).

11 FIGURE 1 Nanog heterogeneity in mESCs cultured in different conditions. (A-C) mESC morphology in different culture conditions (Serum, SerumLIF or 2iLIF). (D-F) FACS histograms of Nd cells, grown in Serum, SerumLIF or 2iLIF showing differences in the percentage of Nanog:VNP positive cells. The negative control cells (E14tg2a) are represented in gray.

Further work is now being performed regarding the expression of lineageaffiliated genes, in order to correlate Nanog heterogeneity with the potential of cells to enter specific differentiation paths in different

environments. This analysis should also shed light into how different differentiation programs are wired in mESCs.

FIGURE 2 Single cell analysis of Nanog and Oct4 expression in mESCs grown in different conditions. (A-C) Histograms showing the distribution of Nanog mRNA molecules per cell. The mean, median and standard deviation are shown for each condition, as well as the number of cells analyzed. (D-F) Same as (A-C) for Oct4 mRNA. (G-I) Dotplots showing the correlation between Nanog and Oct4 expression in each cell for the different conditions. The number of cells analyzed is shown, as well as the Pearson (r(P)) and Spearman (r(S)) correlation coefficients.

Generation of cord blood-derived induced pluripotent stem cells using a non-viral reprogramming strategy Ana Lima MIT Portugal Bioengineering Systems, Faculty of Sciences, University Nova de Lisboa anafranlima@gmail.com Background: Degree in Biomedical Sciences/ University of Aveiro/Portugal Master in Molecular and Cell Biology/ University of Coimbra/ Portugal Starting year: 2012 Research team: SandraPinto1; Rajeev Gupta2 Supervisors: Ricardo Neves1; Tariq Enver2 1 Biomaterials and Stem Cell-Based Therapeutics Lab/ CNC, University of Coimbra 2 Cancer Research Institute/ University College of London

OBJECTIVES The aim of this project is to develop a strategy by which human induced pluripotent stem cells (hiPSCs) can be efficiently generated from human umbilical cord blood (UCB) derived hematopoietic stem/ progenitor cells avoiding viral manipulation and DNA integration. The generation of a biodegradable nanotechnology platform that enables a more efficient reprogramming process and avoiding the use of virus would be a great step in terms of a future use of hiPSCs in the clinical context. WORK PLAN During the course of this work we will specifically: (i) identify the compartments of UCB from which hiPSC can be reprogrammed with greatest efficiency; (ii) develop a non-viral nanoparticle based system which can deliver reprogramming proteins and/or microRNAs directly to these cells. Within the first phase that comprises the identification of the hematopoietic stem cells (CD34+) compartments of UCB from which iPSC can be generated

with greatest efficiency, the general strategy would involve the selection of CD34+ specific cell subpopulations by fluorescent activated cell sorting. Regarding the second phase that comprises the optimization of the nanoparticle delivery system to deliver of reprogramming factors to the UCB subpopulation selected in the previous phase, it would involve the adjustment of the physicochemical properties of the nanoparticles to provide a good biocompatibility and efficiency of delivery the different cargo. RESULTS The UCB sorting was optimized and CD133 was the selected marker that provides a sufficient number of cells to proceed with the reprogramming process. It was also possible to optimize some reprogramming conditions within the UCB context. This is shown in figure 1 where an alkaline phosphatase staining shows significant differences within different chemical contexts of reprogramming after infecting UCB cells with a lentiviral vector containing Sox2, Oct4, Klf-4, c-Myc.

FIGURE 1 Different reprogramming efficiencies outputs from UCB cells in the presence of different chemical modulations.

Superhydrophobic surfaces as a platform for encapsulation of cells and drugs for tissue engineering purposes Ana Catarina Lima MIT Portugal Bioengineering Systems, 3B´s Research Group, University of Minho catarina.lima@dep.uminho.pt Background: Starting year: Supervisors:

MSc. in Biomedical Engineering/ University of Minho/Portugal 2010 João F. Mano1 1 3B´s Research Group, University of Minho

OBJECTIVES The aim of this work was to use biomimetic low adhesive superhydrohobic surfaces to produce polymeric-based systems for encapsulation of drugs and cells. A variety of natural polymers (e.g. dextran, chitosan and alginate), crosslinking processes (ionic gelation and photocrosslinking) and particle structures were explored (simple or multilayered systems).

encapsulating drugs and/or cells in mild conditions (fig. 2). Using this methodology the particles were obtained in a dry environment, just involved by air and avoiding the migration of the cells or drugs for the surrounded medium. This allowed a very high encapsulation efficiency, known a priori. In addition the particles were obtained individually (no aggregation). This technique did not required complex and expensive apparatus as well as organic phases.

WORK PLAN Polystyrene and copper superhydrophobic surfaces were obtained by a phase separation and by a chemical-based deposition method, respectively. The spherical-shaped particles were obtained by dispensing, with controlled volumes, aqueous precursor solutions containing polymer monomers, drugs or cells and some other agent intervenient in the crosslinking process (fig. 1). According to the polymer used, the most appropriated crosslinking procedure was applied to harden the particles. Depending on the application aimed for each developed system, the appropriated in vitro and a few in vivo tests were performed to characterize de particle: optical, confocal and scanning electron microscopy, particle size, water uptake capacity, drug release experiments, cell viability and proliferation, implants in rats and others. RESULTS The obtained results demonstrated that biomimetic low adhesive superhydrophobic surfaces were suitable platforms to produce polymer-based particles

14 FIGURE 1 Schematic representation of particles preparation.

FIGURE 2 (A) Alginate beads encapsulating fibronectin; (B) Chitosan beads encapsulating fibroblast cells; (C) Multilayered dextran particles encapsulating 3 fluorescent molecules as model drugs and (D) coreshell particles composed by dextran and alginate encapsulating fibroblast cells in the shell.

Aerosolized gold-nanodevices for lung diseases theragnosis Ana Sofia Silva MIT Portugal Bioengineering Systems, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa asm.silva@campus.fct.unl.pt Background: Masters Degree in Biomedical Sciences, Universidade da Beira Interior, Covilhã, Portugal Starting year: 2011 Research team: Ana-Aguiar Ricardo1; Ilídio Correia2 Supervisors: Ana-Aguiar Ricardo1; Ilídio Correia2 1 Group of Polymer Synthesis and Processing in Supercritical Carbon Dioxide; REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa; 2 Biomaterials and Tissue Engineering Group, Health Sciences Research Center, Faculdade de Ciências da Saúde, Universidade da Beira Interior.

OBJECTIVES Although the role of inflammatory signalling and oxidative stress in lung diseases may be apparent, the lack of efficient drug delivery and real time diagnosis may result in improper treatment leading to fatal lung pathophysiology. Therefore, there is an urgent need to develop new targeted carriers that can provide sustained delivery of potent therapeutics and molecular probes through airways for real time treatment and diagnosis (theragnosis). The project herein presented describes the synthesis of gold nanodevices, their grafting with fluorescent polymers synthesized in supercritical CO2 and subsequent functionalization with specific peptide sequences. Moreover, such nanodevices loaded with specific therapeutics were encapsulated into chitosan microcarriers using supercritical assisted atomization (SAA) for future pulmonary delivery. WORK PLAN • Assembling of biocompatible, targeted and fluorescent gold-nanodevices (GND) and their structural, morphological and physico-chemical characterizations;

• Microencapsulation of GND into chitosan based-dry powder formulations using SAA and their structural, morphological, physico-chemical, pharmacokinetic and aerodynamic performance characterizations; • In vitro and in vivo evaluation of the produced nano-in-micro formulations. RESULTS New nano-in-micro formulations for the delivery of biotech products via pulmonary inhalation were successfully obtained by co-atomization of different functionalized GND with chitosan. The nanocarriers herein synthetized have the adequate hydrodynamic diameter for intracellular targeting and a rapid translocation to lymph nodes and blood which is an added value for pulmonary delivery. GND were efficiently encapsulated into chitosan carriers as corroborated by CLSM. Furthermore, the Andersen Cascade Impactor data shows that these new microcarriers appear to present favorable aerodynamic properties in order to reach the deep lung, where the release of therapeutics is expected to occur.

Acknowledgments:The authors are grateful to financial support from Fundação Calouste Gulbenkian, Fundação para a Ciência e a Tecnologia (FC&T), through the projects PEst-C/EQB/

LA0006/2013, PTDC/EQU-EQU/116097/2009, SFRH/BD/51584/ 2011 and MIT-Portugal Program Bioengineering.

Development of targeted drug carriers for breast cancer therapy Andrea Costa e Silva MIT Portugal Bioengineering Systems, Centre of Biological Engineering, University of Minho andrea.c.esilva@gmail.com Background: Biological Engineering, University of Minho, Portugal Starting year: 2012 Supervisor: LĂgia Rodrigues1 Co-Supervisors: Natalie Artzi2; Leon Kluskens1 1 University of Minho, Portugal 2 Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, USA and the Department of Anesthesiology, Brigham and Womenâ&#x20AC;&#x2122;s Hospital, Harvard Medical School

OBJECTIVES AND WORK PLAN Current cancer treatments include surgical intervention, radiation and chemotherapeutic drugs; however these therapies are non-specific resulting in the destruction of both tumor and healthy cells. In this sense, the development of a targeted drug delivery system is highly desirable. This approach consists essentially in designing smart new anticancer therapeutics meant to specifically target some unique aspects of tumor biology. Such drug delivery systems should enable improved biodistribution, tissue uptake and pharmacokinetics of therapeutic agents. Nanocarrier-based drug delivery offers several advantages such as an increase of drug stability and bioavailability, by preventing its early degradation, the ability to deliver therapeutic agents, and thus reduce non-specific toxicity. The aim of this PhD project is to develop a targeted therapeutic nanocarrier for the treatment of cancer. The isolation of novel targeting moieties â&#x20AC;&#x201C; aptamers - for cancer cells shall be achieved through the SELEX technique. This methodology consists in the use of a random library of single-stranded DNA or RNA, flanked by two primer-binding regions, and uses an iterative process that specifically amplifies sequences that have high binding affinity to a given target. In several selection rounds, binders are amplified

and nonspecific binders are removed in a partitioning step. Furthermore, a rationally designed gene silencing siRNA will be used as a therapeutic agent, directed to targets in relevant pathways involved in the development and proliferation of breast cancer cells. The siRNA will follow conjugation to previously isolated aptamers. This may be achieved either directly or to an encapsulated form of the therapeutic agent in a nanocarrier. In the case of nanocarrier encapsulation, these will enhance the stability, bioavailability and selectivity of these constructs. But more importantly, the ability to design materials with controlled crosslinking density determining material fate would enable to program the delivery rate of the therapeutic agents and prolonging its duration in vivo. The delivery and internalization of the conjugated aptamer-siRNA or nanocarrier will be studied and ultimately, the optimization of the delivery system is predicted. These studies will primarily undergo in vitro validation using various cancer cell models. Eventually, in vivo studies shall also be conducted. RESULTS Ongoing cycles of SELEX. Culture optimization of different cell lines and establishment of procedures for siRNA transfection and protein expression.

Surface modifications to control drug release from therapeutic ophthalmic lenses Andreia Pimenta MIT Portugal Bioengineering Systems, Centro de Química Estrutural at Técnico Lisboa, University of Lisbon andreia.pimenta@tecnico.ulisboa.pt Background: Starting year: Supervisors:

MSc in Biomedical Engineering/Universidade NOVA de Lisboa/Portugal 2012 Rogério Colaço1; Ana Paula Serro1 1 Centro de Química Estrutural at Técnico Lisboa, University of Lisbon

OBJECTIVES The aim of the work is to develop and optimize surface modified/coated drug-releasing hydrogels to be used in therapeutic ophthalmic lenses (soft contact lenses and intraocular lenses). For that, different surface coating/modification strategies will be tested in order to create drug diffusion barriers, which lead to a continuous and gradual dispense of medication to the eye, at adjustable rates, and for extended periods. The drug release kinetics will be investigated through in vitro tests by using equipment specially conceived to simulate the eye. Appropriate physical-chemical and biological experimentation will be performed in view of future (pre)clinical validation.

RESULTS So far, minimal inhibitory concentrations of two antibiotics for three different bacteria that are associated with endophthalmitis (Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus pneumonia) were determined by agar diffusion bioassays. Release studies of these antibiotics from commercial intraocular lenses (loaded with drug by soaking) were performed using the infinite sink method and in hydrodynamic conditions of the aqueous humor through a microfluidic device.

WORK PLAN • Preparation and characterization of materials for ophthalmic lenses; • Drug loading & Surface modifications and/or coating of the ophthalmic material; • In vitro release tests; • Numerical modelling and optimization of drug release profiles.

FIGURE 1 Top: Microfluidic device for simulation of the aqueous humor hydrodynamic conditions. Bottom: Release kinetics of moxifloxacin hydrochloride in hydrodynamic conditions and in sink conditions.

Mesenchymal stem cells as a means to expand and select regulatory T cells for subsequent clinical translation António Soure MIT Portugal Bioengineering Systems, Instituto Superior Técnico, University of Lisbon antoniosoure@tecnico.ulisboa.pt Background: Starting year: Supervisors:

Biochemistry/Faculdade de Ciências, Universidade Lisboa/ Portugal Biotechnology/Instituto Superior Técnico, Universidade de Lisboa/ Portugal 2012 Joaquim Sampaio Cabral1; Cláudia Lobato da Silva1; João Lacerda2 1 Instituto Superior Técnico, Universidade de Lisboa 2 Instituto de Medicina Molecular, Universidade de Lisboa

OBJECTIVES Test the ability of mesenchymal stem cells (MSC) from different sources, including bone marrow (BM) and adipose tissue (AT) to recruit and expand regulatory T cells.

of expanded/induced cells to suppress the proliferation of activated conventional T cells (Tcon). RESULTS

WORK PLAN MSC have been obtained from BM and cultured in vitro until confluent. PBMC were co-cultured with MSC in order to examine the ability of MSC to induce and expand Treg in vitro. Specifically, five different conditions have been tested thus far: • PBMC co-cultured with MSC at a 5:1 ratio • PBMC co-cultured with MSC at a 3:1 ratio • PBMC co-cultured with MSC at a 5:1 ratio, with the addition of 10U/mL of IL-2. • PBMC alone • PBMC alone + IL-2 (10U/ml) In the future, to determine if the mechanism by which MSC induce/expand Treg is due to cell-cell contact or due to the release of soluble factors, transwell experiments will be performed. The stability of Foxp3 expression will be assessed by DNA demethylation analysis. Functionality assays will also be performed in order to determine the ability

FIGURE 1 Phenotypical analysis of the condition in which PBMC were co-cultured with MSC and in which IL-2 (10U/ml) was added (day 7 of experiment). Briefly, lymphocytes were defined as being CD3+ cells (not shown). The gate of CD25bright Foxp3+ cells was defined according to the expression of CD25 and Foxp3 in both the CD4 vs CD25 and CD4 vs Foxp3 dot plots, respectively.

FIGURE 2 The effect of BM MSC on the expansion and recruitment of regulatory T cells. At days 3 and 7 of the experiment, the percentage of both (a) CD25+Foxp3+ cells and (b) CD25+Foxp3+CD127+ was assessed. Results show that PBMC co-cultured with MSC seem to yield a higher enrichment of both CD25+Foxp3+ cells and of CD25+Foxp3+CD127- cells, when compared to PBMC cultured alone, with or without the addition of IL-2. Importantly, the addition of IL-2 to PBMC-MSC co-cultures seems to have a beneficial effect on the enrichment of Tregs, especially on CD25+Foxp3+ cells.

Filtration-based strategies for the downstream processing of human mesenchymal stem cells Bárbara Cunha1,2 MIT Portugal Bioengineering Systems, Universidade Nova de Lisboa bcunha@itqb.unl.pt Background: Starting year: Research team: Supervisors:

MSc. Degree in Biotechnology / Universidade Nova de Lisboa - Faculdade de Ciências e Tecnologia / Portugal 2012 Margarida Serra1,2; Cristina Peixoto1,2; Marta Silva1,2; Manuel Carrondo2,3; Paula M. Alves1,2,3 Margarida Serra (Animal Cell Technology Unit, iBET/ITQB-UNL); Paula M. Alves (co-supervisor, Animal Cell Technology Unit, iBET/ ITQB-UNL) 1 Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal 2 iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal 3 Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Monte de Caparica, Portugal

OBJECTIVES The main aim of this work was to develop an efficient strategy for the downstream processing (DSP) of human mesenchymal stem cells (hMSC). WORK PLAN After cell expansion on microcarriers using stirred tank bioreactor systems, different filtration-based approaches were evaluated for the DSP of hMSC. We have focused on the i) clarification step for microcarrier removal using dead end filtration, where the influence of different filter materials and pore sizes were evaluated. For the ii) cell concentration/volume reduction step, tangential flow filtration technology was investigated, aiming at concentrating hMSC up to a factor of ten. For this purpose, a wide range of operating conditions was tested; examples presented here include the impact of initial cell concentration, membrane material and pore size on cell recovery yields and quality attributes.

RESULTS After hMSC expansion, cells were harvested and separated from the microcarriers using filters with two types of materials (nylon and polypropylene) with pores sizes of 30, 80 and 100 μm. Our results show that both materials have the ability to remove microcarriers without compromising cells’ viability (> 97%). In addition, high pore sizes (> 80 μm) demonstrated to be more suitable for the clarification step, allowing to recover 94% of viable cells (Fig 1A). Regarding the concentration step, our results show that hMSC can be successfully concentrated up to a factor of ten, recovering 80% of viable cells. Initial cell concentrations higher than 2 x 105 cell/mL (Fig 1B), and polysulfone membranes have showed to be key parameters to perform the volume reduction step. Furthermore, at the end of DSP, hMSC have conserved their characteristics, presenting high cell viability (over 90%) and maintaining their immunophenotype, metabolic activity and multilineage differentiation potential (Fig 1C).

20 FIGURE 1 Downstream processing of hMSC. A - Clarification of microcarriers from the cell bulk suspension. B - Impact of the initial cell concentration on the viable cell recovery yield. C - Multipotent differentiation potential of hMSC after processing.

Design and fabrication of zonal cartilage constructs Carla Moura MIT Portugal Bioengineering Systems, CDRsp, Polytechnic Institute of Leiria carla.moura@ipleiria.pt Background: Starting year: Research team: Supervisors:

Master degree /Polytechnic Institute of Leiria/ Portugal 2010 Frederico Castelo Ferreira1; Paulo Jorge Bártolo2,3 Frederico Castelo Ferreira1; Paulo Jorge Bártolo2,3 1 Bioengineering systems, MPP, Investigador FCT, Institute of Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa 2 Centre for Rapid and Sustainable Product Development, Polytechnic Institute of Leiria 3 Manchester Biomanufacturing Centre, School of Mechanical and Aerospace and Civil Engineering & Manchester Institute of Biotechnology, University of Manchester, UK

OBJECTIVES Articular cartilage lacks of a sufficient intrinsic repair response and hence the zonal cartilage structure and function are commonly and irreversibly lost following trauma or cartilage related disease, such as with rheumatoid arthritis and osteoarthritis. Osteoarthritis is characterised by a loss of cartilage, typically progressing from superficial fibrillation to complete erosion until the subchondral bone. This condition affects a large number of patients (9.6% on men and 18% on women), causing pain and debilitation and hence severe loss in their quality of life. This work aims to develop a cartilage substitute based on the combination of 3D high-interconnected porous scaffolds seeded with mesenchymal stem cells (MSC), which are first expanded in the scaffold to larger numbers and then differentiate into chondrocytes. These constructs are expected to facilitate regeneration of articular cartilage. In order to achieve a successful procedure, it is necessary to select the best strategy concerning material properties as well as conditions for cell seeding, culture and differentiation. WORK PLAN The work consists on the manipulation of the Bioextruder parameters to manufacture extruded

layer-by-layer poly(ɛ-caprolactone) scaffolds with different characteristics concerning pore sizes and fibre alignments. The scaffolds obtained were assessed under different conditions for cell seeding, expansion and differentiation. Expansion and differentiation were performed using two atmospheric conditions, normoxia and hypoxia (21% and 5% oxygen, respectively).

FIGURE 1 Culture strategy used for seeding of scaffolds with high chondrocytes densities.

21 FIGURE 2 SEM image of scaffolds.

RESULTS Results obtained showed that pore size does not pose a limitation for cell adhesion and proliferation within the range of pore sizes studied (190-390ฮผm). Fibre alignment (0-45ยบ and 0-90ยบ) also does not influence cell adhesion and proliferation, however regarding the differentiation it influences the dimension of the aggregates that are formed. This study also shows that the best approach towards cartilage regeneration involved the use of hypoxia condition, performing cell expansion in the PCL scaffold before differentiation. Acknowledge: This research is sponsored by the Portuguese Foundation for Science and Technology through MIT Portugal Program, Bioengineering Systems Focus Area, a PhD grant SFRH/BD/73970/2010 and an Investigator FCT research contract IF/00442/2012.

FIGURE 3 Dapi (blue) and Phalloidin (red) staining of the cells adhered to the scaffolds.

Light-activatable polymeric nanoparticles for intracellular delivery in cancer stem cells Carlos Boto MIT Portugal Bioengineering Systems, Center for Neurosciences and Cell Biology (CNC), University of Coimbra, Portugal csmboto@biocant.pt Background: MSc. in Materials Engineering, University of Coimbra, Portugal Starting year: 2009 Research team: Ricardo Neves1; Alberto Martin-Lorenzo2; Isidro Sanchez-Garcia2; Rajeev Gupta3; Tariq Enver3 Supervisors: Lino Ferreira1 1 Center for Neurosciences and Cell Biology (CNC), University of Coimbra, Portugal 2 Institute of Biomedical Research of Salamanca (IBSAL), University of Salamanca,Spain 3 UCL Cancer Institute, University College London, London, United Kingdom

OBJECTIVES Nanoparticles (NPs) may be used as vehicles for the intracellular delivery of biomolecules and ultimately to modulate the activity of cancer cells. Here we present a novel lightdissociable polymeric NP formulation that has approximately 120 nm in diameter, positive net charge, and disassemble when exposed to UV light (365 nm) or blue light (405 nm) releasing nile red (NR) and retinoic acid (RA) (Fig. 1). These NPs can be taken up rapidly (4 h) by acute myeloid leukemia (AML) cells (K562, NB4, U937 and AML stem cells). Importantly, our results show that the light activation of NPs contribute for the differentiation of these cells at levels not observed using formulations that release passively RA or by conventional chemotherapy. This study highlights the importance of drug spatial positioning and concentration to treat leukemia. WORK PLAN Diameter, counts and zeta potential of NPs in response to light over time were recorded. The in vitro effects of retinoic acid (RA)-loaded NPs on: (i) the erythroid differentiation of K562 at day 9

was assessed by cytochemical staining with benzidine solution, (ii) the granulocytic differentiation at day 6 and 3 for human NB4 and U937 (PLZF/RARA), respectively, was assessed by flow cytometry using CD11b as a marker, (iii) reduction of stem cell colonies in AML stem cells was assessed by colony-forming cell (CFC, 2 weeks) and long-term culture initiating cell assays (LTC-IC, 7 weeks). Three independent runs were done for each experimental group and for each run three technical replicates were performed. RESULTS The activation of NPs promotes rapid and efficient RA delivery (80% in 10 min). Lightactivated RA -NPs induce 1.92 (±0.17) times higher levels of erythroid differentiation in K562, 1.19 (±0.01) times of granulocytic differentiation in NB4 and 1.45 (±0.03) times of granulocytic differentiation in U937 cells as compared to non-activated NPs. In addition, light-activated RA -NPs induce 1.83 (±0.17) times higher levels of erythroid differentiation in K562, 1.12 (±0.01) times of granulocytic differentiation in NB4 and 1.44 (±0.04) times of granulocytic differentiation

FIGURE 1 Photo-disassembly of NPs. (A) A suspension of NPs was exposed to UV light (365 nm, 100 Watts) for up to 10 min. At each time, the size, zeta potential and number of NPs (kcps) in the suspension was evaluated by dynamic light scattering. (B) Normalized fluorescence vs. time for the same NR-loaded NP formulation when exposed or not to UV light (365 nm, 100 Watts), showing the increase in the release rate with UV exposure. Nile Red in aqueous solution exposed to UV light is presented to demonstrate its photostability.

in U937 cells as compared to 1µM RA in solution (Fig. 2). It should be noted that RA -NPs contain 10 times less RA than the one used in solution. This experimental result is particularly important in the RA-low sensitive cell line U937-PLZF/RARA where the high RA intracellular release is able to overcome the low sensitivity of the cell. Finally, AML stem cells treated with light-activated RA -NPs showed 69.6±9.2 % and 61.8±10.1 % less colonies, in CFC and LTC-IC colony assays, respectively (Fig. 3). Next we evaluated the in vivo effect of our light-activatable RA+ NPs in human AML cells. Cells treated with light-activated NPs (blue bar, Fig. 4) present higher differentiation

FIGURE 2 Effect of RA+ NPs on human leukemia cells. Schematic representation of the methodology. (A) Myelocytic differentiation (CD11b+ cells) of human Zn-induced U937-B412 cells cultured with light-activated NPs or soluble RA. Zn-induced U937-B412 cells were cultured for 3 days. (B) Percentage of CD11b+ cells in Zn-induced U937-B412 cell cultures after 1 day of culture. Cells cultured with 10 -7 M of vitamin D3 for 1 day were used as positive control. Results are expressed as Mean ± SEM (n =3). *P<0.05, * *P<0.01, * **P<0.001.

marker expression than cells treated with RA+ NPs without light activation. CONCLUSION Our results show that our light-activatable NPs are able to: (i) Quickly release retinoic acid when exposed to light; (ii) Photo-disassemble within cells; (iii) Successfully induce differentiation of a retinoic acid-resistant-leukemia cell line; (iv) Decrease the clonogenic potential of leukemia stem cells from human AML bone marrow; (v) Be activated in vivo by a blue laser and differentiate successfully into myelocytic lineage.

FIGURE 3 Differentiation of AML stem cells cultured with light-activated NPs or soluble RA. Cell differentiation was evaluated by a colony forming unit assay at day 14. (A) AML stem cells were cultured for 14 days with RA in medium (102 - 105 nM) or RA+ NPs (0.01 - 10 µg/mL) or blank NPs (10 µg/ mL), exposed or not to UV light. Cell differentiation was evaluated by a colony forming unit assay at day 14. (B) Long-term culture-initiating cell assay results. AML stem cells were cultured on feeder layers for 5 weeks and then on methylcellulose medium for 14 days with blank NPs (10 µg/mL) or RA+ NPs (10 µg/mL) exposed or not to UV light. Results are expressed as a mean percentage of control plates containing only AML cells. Results are expressed as Mean ± SEM (n = 3). * P<0.05, ** P<0.01, *** P<0.001.

FIGURE 4 In vivo differentiation of NB4 cells exposed to light-activatable RA+ NPs. (A) Schematic representation of the in vivo experimental set up. Cells were treated with blank or RA+ NPs (10 µg/mL) for 4 h, washed, and then activated or not with a blue optical fiber (405 nm, 80 mW) for 5 min. Cells were then resuspended in a 1:1 (v/v) Matrigel solution and subcutaneously injected in a PDMS cylinder construct implanted in the dorsal region of mice. After 5 days, cells were removed from the construct and characterized by FACS, for CD11b expression. (B) Percentage of CD11b+ cells in human leukemia NB4 cells collected 5 days after subcutaneously injection. Results are expressed as Mean SEM (n = 4). * P<0.05, * *P<0.01, ***P<0.001.

Delivery of antimicrobial peptides for the treatment of mycobacteriosis César Oliveira Costa MIT Portugal Bioengineering Systems, Centre of Biological Engineering & Life and Health Sciences Research Institute, University of Minho cesaroliveira.costa@gmail.com Background: MSc. Bioengineering / Faculty of Engineering - University of Porto / Portugal Starting year: 2012/2013 Research team: João Pedro Silva1; Jeremy Sousa2; António Gil Castro2; Jorge Pedrosa2; Rui Appelberg3; Miguel Gama1 External collaborators: Fernando Martinez Estrada4; Subhankar Mukhopadhyay4; Siamon Gordon4; Erik de Vries5 Supervisors: Miguel Gama1; Rui Appelberg3; Jorge Pedrosa2 1 Centre of Biological Engineering, University of Minho, Portugal 2 Life and Health Sciences Research Institute, University of Minho, Portugal 3 Institute for Molecular and Cell Biology, University of Porto, Portugal 4 University of Oxford, England 5 University of Groningen, The Netherlands

BACKGROUND AND OBJECTIVES Mycobacterium tuberculosis, which resides inside macrophages, has always been recognized as one of the most “successful” pathogens. Standard treatments have already been used for decades and, therefore, resistances to the first-line medicines are increasing. Additionally, poor patient compliance with stringent therapies is often pointed out as a major reason leading to treatment failure. Antimicrobial peptides (AMPs), a promising new class of broad spectrum antibiotics, are less prone to result in pathogen resistances due to their target (cellular membranes) and rapid action. In our laboratory we search for AMPs with potent activity against mycobacteria and try to develop efficient delivery systems based on self-assembled colloidal nanocarriers. Additionally, this systems are expected to reduce peptide toxicity and enhance selective uptake on infected cells. Finally, the use of encapsulated drugs in mycobacterial therapy may help reducing drug administration schedules which would ultimately improve patient compliance. WORK PLAN 1. Optimization of therapeutic formulations for AMPs controlled delivery. 2. In vitro and in vivo delivery of AMPs to infected macrophages; Analysis of mechanisms of therapeutic action and potential side effects. 3. Study binding, internalization mechanisms and intracellular fate of the nanocarriers (University of Oxford). 4. In vivo biodistribution of the nanocarrier (University of Groningen). 5. Study the global pharmacoeconomic impact of AMPs as new therapeutic agents to fight tuberculosis.

RESULTS We have used encapsulated AMPs (Fig. 1) to tackle an infection promoted by M. avium in mouse macrophages in vitro. During 7 days the peptide has killed more than 99% of initial bacterial burden. The bactericidal effect has already been confirmed using infected mice in vivo, by applying the therapeutic formulation via the pulmonary route (Fig. 2). Currently, we are moving for more potent mycobacteria, such as M. tuberculosis, which shows identical susceptibility to the antimicrobial formulation.

FIGURE 1 Schematic representation of the peptide nanocarrier.

FIGURE 2 Intratracheal drug delivery using a MicroSprayer® aerosolizer in mice.

Towards scalable production of functional iPSC-derived cardiomyocytes Cláudia Correia MIT Portugal Bioengineering Systems, Universidade Nova de Lisboa ccorreia@itqb.unl.pt Background: MSc. Degree in Biotecnology / Universidade Nova de Lisboa - Faculdade de Ciências e Tecnologia / Portugal Starting year: 2011 Research team: Margarida Serra1,2; Nuno Espinha1,2; Marcos Sousa1,2; Catarina Brito1,2; Karsten Burkert 3; Yunjie Zheng3; Jürgen Hescheler3; Manuel J. T. Carrondo2,4; Tomo Saric3; Paula M. Alves1,2 Supervisors: Paula M. Alves1,2, Margarida Serra1,2 (co-supervisor), 1 ITQB-UNL, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal 2 iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal 3 Institute for Neurophysiology University of Cologne, Cologne, Germany 4 Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Monte de Caparica, Portugal

OBJECTIVES The main aim of this study was to develop a scalable and robust bioprocess for the production of pure and functional induced pluripotent stem cells (iPSC)-derived cardiomyocytes (CMs). WORK PLAN Our strategy consisted in designing integrated bioprocesses for CM differentiation and purification by culturing iPSCs as 3D aggregates in environmentally controlled bioreactors, where the necessary conditions to control stem cell fate are thoroughly tuned. Distinct bioreactor systems (stirred tank and Wave bioreactors) were used and different bioprocess parameters were tested. In particular, we explored the impact of the dissolved oxygen (DO) and hydrodynamic forces on CM differentiation.

16 days of culture. Moreover, this study shows for the first time that wave-induced agitation enhances further CM differentiation of iPSCs, increasing CM differentiation yields up to 60 (Fig. 2), while reducing the time needed for differentiation process. In this strategy, an earlier and higher expression of mesorderm and cardiac specific genes was detected when compared to stirred tank bioreactor cultures. In an 11-day differentiation protocol, 2.3±0.1 x106 CM/ mL were obtained with high purity (97%) and quality in 1 L bioreactor run. Morphological and functional characterization analysis confirmed that CMs presented typical cardiac morphology (Fig. 3), calcium transients, electrophysiological profiles and drug responsiveness.

RESULTS Our results showed that DO and agitation type/mode are key parameters in the bioprocessing of iPSC, affecting cell differentiation towards functional CMs. By combining a hypoxia culture (4% O2 tension) with an intermittent agitation profile in stirred tank bioreactors, we were able to improve about 1000-fold the differentiation yields (up to 43 CM per initial iPSC) and achieve high CM purities (>97%, Fig. 1) after FIGURE 2 Final CM differentiation yield (CMs produced per initial iPSC) achieved with different bioreactor culture strategies.

26 FIGURE 1 Confocal microscopy of iPSC-derived cardiospheres. The frequency and distribution of α-MHC-positive (green) cells is shown. Nuclei are labeled with DAPI (blue). Scale bars: 100 µm.

FIGURE 3 Structural characterization of iPSC-derived CMs produced in fully controlled bioreactors. Cells stained positive for the CM specific markers: α-MHC (green) and titin (red). Nuclei were counterstained with DAPI (blue). Scale bars: 50 µm.

Neurogenesis in the subventricular zone is modulated by MicroRNA-124 loaded nanoparticles Cláudia Saraiva MIT Portugal Bioengineering Systems, Faculty of Science and Technology, Universidade Nova de Lisboa; Health Sciences Research Centre, University of Beira Interior c.m.m.saraiva@gmail.com Background: Starting year: Research team: Supervisors:

Master in Biochemistry/ University of Coimbra/ Portugal 2012 Cláudia Saraiva1,2; José Paiva3; Lino Ferreira3,4; Liliana Bernardino2 Liliana Bernardino2 1 Faculty of Science and Technology, Universidade Nova de Lisboa 2 Health Sciences Research Centre, University of Beira Interior 3 Biocant - Center of Innovation in Biotechnology, Cantanhede 4 Center for Neuroscience and Cell Biology, University of Coimbra

OBJECTIVE The subventricular zone (SVZ) lining the lateral ventricles comprises the largest population of neural stem cells (NSCs) in the adult mammalian brain. NSCs are multipotent and can give rise to neurons and glia cells. MicroRNA (miR)-124 has been recently described to trigger neuron commitment of NSCs. However, current strategies to deliver miRs into cells or tissues are not efficient. Thus, identifying new platforms to deliver proneurogenic molecules such as miR-124 is crucial to boost neurogenesis of endogenous NSCs. The main goal of this work is to study the inductive effect of miR-124-loaded nanoparticles (miR-124 NPs) in the differentiation of NSCs into new neurons. WORK PLAN P1-3 C57BL/6 mice were used to obtain stem/progenitors cell cultures from the SVZ. The resulting neurospheres were seeded on coverslips and the cell monolayer obtained was then transfected with several concentrations (1, 10 and 20μg/mL) of NPs complexed with 200nM of miR-124.

RESULTS We found that 1μg/mL of NPs did not interfere with cell toxicity (propidium iodide assay), proliferation (BrdU assay) or oligodendrocyte commitment (olig2-immunoreactivity) of SVZ cells. Interestingly, 1μg/mL of miR-124 NPs was able to increase the differentiation into neurons (NeuN-immunoreactivity) in about 25% compared with controls. The relative mRNA amount of two validated miR-124 targets, sox9 and jagged1, was also assessed by qPCR. As expected, miR-124 NPs reduced the expression of both genes as compared with controls. Moreover, a significant decrease in the number of sox9-and jagged1-immunoreactive cells was also obtained. Taken together, our results showed that the presence of miR-124 delivered by NPs increase the neuronal commitment of SVZ cells, being the 1μg/mL of the miR-124 NPs the most suitable formulation. These results provide clear evidences to support the use of miR-124 NPs as a new therapeutic approach to boost brain repair endogenous mechanisms in the setting of neurodegenerative diseases.

Towards improved predictability in pre-clinical research: Human dopaminergic 3D in vitro model for development of gene delivery strategies Daniel Simão1,2 MIT Portugal Bioengineering Systems, Universidade Nova de Lisboa dsimao@itqb.unl.pt Background: MSc. Molecular Genetics and Biomedicine; Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa; Portugal Starting year: 2011 Research team: Catarina Pinto1,2; Margarida Serra1,2; Ana P. Teixeira1,2; Isabella Saggio3,4; Giampietro Schiavo5; Eric J. Kremer 6; Paula M. Alves1,2; Catarina Brito1,2 1 ITQB-UNL – Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal 2 BET – Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal 3 Biomedical Science Park San Raffaele, Rome, Italy 4 Department of Genetics and Molecular Biology, Sapienza University of Rome, Rome, Italy 5 Cancer Research UK, London, United Kingdom 6 Institut de Génétique Moléculaire de Montpellier, Montpellier, France

OBJECTIVES Development of a human 3D neural in vitro model making use of human midbrain-derived neural progenitor cells (hmNPC) and evaluation of hd-CAV-2 efficacy and safety. WORK PLAN Dynamic culture systems strategies were adopted for hmNPC aggregation and 3D differentiation. Physical and chemical parameters, such as hydrodynamics, process duration, media composition and oxygen levels, were optimized to improve differentiation efficiency. The generated 3D model was explored for characterization and better understanding of hd-CAV-2 particular features in human neural cells, enabling to evaluate its efficacy and safety for gene delivery strategies.

RESULTS Differentiation of hmNPC neurospheres resulted on a significant enrichment in neurons, astrocytes and oligodendrocytes. Global gene expression analysis revealed the activation of neurogenesis developmental pathways during 3D differentiation. Differentiated cells presented dense filopodia, critical in axonal development, guidance and synapse formation. Moreover, an increase in the expression of the dopaminergic (DA) markers along with an increase in synaptic functionality and ability to synthesize and release dopamine, strongly indicated the generation of DA neurons. Under optimized transduction conditions hd-CAV2-GFP vector transduction led to stable long-term transgene expression with low toxicity. Assessment of hd-CAV-2 specific tropism in comparison with hAd serotype 5 (hAd5), showed that hd-CAV-2 have increased neuronal tropism in contrast to

higher transduction of the glial cell population with hAd5. The 3D model developed herein, along with a comprehensive toolbox of characterization methods, represents a powerful complementary tool, applicable in preclinical research for assessing promising genebased therapies and new compounds. Data generated on hd-CAV-2 transduction in a physiologically relevant 3D cell context, will allow for a better understanding of the impact of viral vectors on human neural cells and complement the in vivo studies on CNS transduction to improve predictability of clinical trial outcomes.

FIGURE 1 3D neural differentiation strategy based on dynamic culture systems. (A) Use of stirred-tank bioreactors enables control over the main culture parameters. (B) This strategy is amenable to optimization for multiple human stem cell sources. (C) The 3D differentiation process is based on sequential steps (expansion, aggregation, differentiation and maturation), where media composition is the key variable, by adding or removing growth, differentiation or neurotrophic factors.

Modulation of hematopoietic stem cell (HSC) activity Emanuel Quartin Costa MIT Portugal Bioengineering Systems, UC Biotech, Center for Neurosciences and Cell Biology, University of Coimbra, Portugal emanuelquartin@gmail.com Background: Starting year: Research team: Supervisors:

Msc in Human Biology and Enviornment, FCUL, Portugal 2012 Ricardo Neves1; Lino Ferreira1; Carlos Boto2 Ricardo Neves1; Lino Ferreira1 1 Center for Neurosciences and Cell Biology, University of Coimbra 2 Universidade de coimbra

OBJECTIVES Methodologies to increase the number or engraftment efficiency of hematopoietic stem cells (HSCs) could provide substantial clinical benefit. We hypothesize that the transient ectopic delivery of HOXB4 in transplanted HSCs may potentiate their number and final engraftment. The main goal of this project is to study the effect of HOXB4 in terms of HSC renewal and engraftment into an animal model. RESULTS As there is no commercial available active HOXB4, using a prokaryotic protein expression system we produced and purified a histidine tagged TAT-HOXB4 fusion proteins. Using a BL21(DE3)pLysS E.coli strain we produced TAT-HOXB4 that was later purified using a Histrap column, where we tested different concentrations of imidazole to obtain more pure fractions. Also different desalting/renaturation methods were tested. The purity and concentration of the TAT-HoxB4 and TAT-mutHoxB4 fusion proteins were determined

FIGURE 1 HOXB4 activity confirmation by binding to the DNA probe. 1:HOXB4; 2:Random DNA probe; 3: HOXB4 DNA probe; 4: HOXB4 + DNA probe; 5- HOXB4 + DNA probe + DTT

by SDS-PAGE and detected with Coomassie Blue or SYPRO Ruby staining After having the protein purified, we assessed protein activity by electrophoretic mobility shift assays (EMSA) and by cell cycle studies (G0 induction) in a murine pro-B cell line. For the EMSA, different concentrations of HOXB4 were incubated with or without DTT and with a DNA probe containing the HOXB4 consensus binding sequence the samples were loaded in a non-denaturing gel later stained with SYBR green (fig.1). For the cell cycle studies, cells were treated with different concentrations of HOXB4 and at different time points. Results showed a nearly 8-fold induction of G0 at and 48h, after of HOXB4 treatment (fig.2).

FIGURE 2 Cell Cycle study after 48h treatment of BaF3 cells treated with produced HOXB4 and comercial HOXB4. A- Effect of HOXB4 in the different phases of the cell; B- G0 fold increase, normalized with the mock treatments.

Scalable expansion of human mesenchymal stem/stromal cells as 3-D aggregates towards an improved cellular product Francisco Moreira MIT Portugal Bioengineering Systems, BERG, Instituto Superior Técnico francisco.moreira@ist.utl.pt Background: Starting year: Supervisors:

Masters degree in Biotechnology in Instituto Superior Técnico, Universidade Técnica de Lisboa Bachelor degree in Biochemistry in Faculdade de Ciências, Universidade de Lisboa 2012 Joaquim Sampaio Cabral1; Cláudia Lobato da Silva1 1 BioEngineering Research Group, Instituto Superior Técnico, Universiddade de Lisboa

OBJECTIVES Isolation and cultivation of human MSC from different sources as 3-D spheroids under xeno-free conditions aims to define optimal culture conditions for MSC culture as aggregates. Considering the clinical application of MSC, the use of fetal bovine serum arises additional problems, mainly focused on product safety. Although BM is the most commonly used source of MSC, in the last decade the therapeutic potential of MSC isolated from a variety of tissues have been also demonstrated, among which AT and UCM are the best studied. MSC will be cultured as spheroids (3-D) and compared with adherent monolayers (2-D). The main two objectives for this first part of the work are the optimization of static culture conditions for

human MSC (derived from 3 sources - bone marrow (BM), adipose tissue (AT) and umbilical cord matrix (UCM)) cultivation as 3-D spheroids, under xenofree conditions; and the in vitro characterization of human MSC cultured as spheroids. WORK PLAN Human MSC were isolated in the laboratory and the spheroids were formed by aggrewellTM. Three different sources were tested: BM, UCM and AT. These were cultured under two different media, one containing FBS and another XenoFree. Further more, from each type of cells two sizes of aggregates were formed: 100 cells/spheroid and 1000 cells/spheroid. RESULTS

FIGURE 1 Comparison between the diameters of the spheroids formed from BM M69A07 donor, in the two different media (Blue – DMEM; Red – XF). A) 100cells/ sph present similar values of diameter for either type of media. B) 1000cells/sph seems to show a tendency of higher diameter for DMEM cultured spheroids, however the error associated in the measurements prevents to draw a definite conclusions.

30 FIGURE 2 Comparison between the diameters of the spheroids formed from AT L09602 donor, in the two different media (Blue – DMEM; Red – XF). A) 100cells/ sph present similar values of diameter for either type of media. B) 1000cells/sph seems to show a tendency of higher diameter for XF cultured spheroids, however the error associated in the measurements prevents to draw a definite conclusions.

One-step transformation of microorganisms Gabriel Mendes1 MIT Portugal Bioengineering Systems, Department of Biological Engineering, University of Minho pintomendes.gabriel@deb.uminho.pt Background: Starting year: Research team: Supervisors:

PgD in Micro Nanotehcnology/ University of Minho/ Portugal 2010 Gabriel Mendes1; Pedro Viera2; Manuel Mota1 ; Leon Kluskens1 ; Senentxu lanceros-Mendez3; Alan Hatton4 Manuel Mota1; Leon Kluskens1; Senentxu lanceros-Mendez3 1 Department of Biological Engineering, University of Minho 2 Department of Biological Engineering, University 3 Department of Physics, University of Minho 4 Department of Chemical Engineering, Massachusetts Institute of Technology

OBJECTIVES This project aims to develop products that simplify the laboratory methods used for the genetic improvement of microorganisms with commercial potential that can be applied in biotechnology, in areas such as food industry, environment or energy. These products will constitute a new methodological paradigm through its simplicity and will substitute classical procedures that are more time-consuming, more expensive and technically more demanding. Transformation of microorganisms (introducing DNA into microorganisms for a genetic improvement) is a cumbersome procedure normally needing three main steps, namely: (1) preparation of microorganisms (weakening of natural barriers to receive foreign DNA); (2) a shock step (normally through an electric pulse, jump in temperature or by using ultrasounds); and finally the (3) a recovery step in which the microorganisms are grown in rich medium to recover from the previous step. With our technology we envisage attaining transformation in a single step through spreading microorganisms and foreign DNA on novel transformation petri-dishes. WORK PLAN Synthesis of superparamagnetic monodisperse magnetic nanoparticles (with diameters of less than 10 nm) and functionalization with cationic molecules such as dopamine, 4-aminobenzoic acid and 2-pyrrolidinone. Synthesis and functionalization of magnetic nanoneedles. Use external magnetic field or friction forces to induce membrane permeabilization. Synthesized or natural nanomaterials are used to facilitate entrance of DNA in microorganisms such as Escherichia coli, Bacillus subtilis and Saccharomyces cerevisiae, all of them GRAS microorganisms. The microorganisms mixed with plasmids are spread on culture petri dishes in the

Poster

presence of nanomaterials and with addition of adequate markers (for instance ampicinin 100µg/ mL, IPTG 1mM and X-Gal 40µg/mL for Escherichia coli.) transformed colonies are produced (Yoshida & Sato, 2009). RESULTS Nanomaterials were synthesized and functionalized with cationic molecules. First attempts to transform non-competent JM109 Escherichia coli with pUC19 plasmid and intact BY4741 Saccharomyces cerevisiae with pyes2- isc1 plasmid have succeeded by using nanoneedles. Maximum transformation efficiencies obtained for Escherichia coli are on the order 1.6x105 CFU/µg DNA and 5.8x103 CFU/ µg DNA for Saccharomyces cerevisiae. Essays on Bacillus subtilis are in progress.

FIGURE 1 Synthesis by thermal decomposition of mono-dispersed magnetic nanoparticles with diameters of about 5nm (S. Sun et al., 2004). Nanoparticles are capped by oleic acid and a ligand exchange reaction is performed with small cationic molecules such as dopamine and 4-aminobenxoic acid to promote dispersion in water and affinity with plasmids. Scale bar is 5 nm.

FIGURE 2 Magnetization loop for magnetic nanoparticles capped with oleic acid, dopamine and 4-aminobenzoic acid. All types of magnetic nanoparticles are superparamagnetic at room temperature. The cationic molecules were used to cap the nanoparticles through ligand exchange (Lattuada & Hatton, 2007). References: 1. Lattuada, M., & Hatton, T. A. (2007). Functionalization of monodisperse magnetic nanoparticles. Langmuir, 23(4), 2158–2168. doi:Doi 10.1021/La062092x. 2. Sun, H., Chen, B., Jiao, X., Jiang, Z., Qin, Z., & Chen, D. (2012). Solvothermal Synthesis of Tunable Electroactive Magnetite Nanorods by Controlling the Side Reaction. The Journal of Physical Chemistry C, 116(9), 5476–5481. doi:10.1021/jp211986a. 3. Sun, S., Zeng, H., Robinson, D. B., Raoux, S., Rice, P. M., Wang, S. X., & Li, G. (2004). Monodisperse MFe2O4 (M =

FIGURE 3 Synthesis of magnetic nanoneedles by a solvathermal process (H. Sun et al., 2012). Nanoneedles are capped with oleic acid and a ligand exchange reaction is performed to improve dispersion in water and affinity for plasmids. Scale bar is 100 nm. Fe, Co, Mn) nanoparticles. Journal of the American Chemical Society, 126(1), 273–9. doi:10.1021/ja0380852. 4. Yoshida, N., & Sato, M. (2009). Plasmid uptake by bacteria: a comparison of methods and efficiencies. Applied Microbiology and Biotechnology, 83(5), 791–8. doi:10.1007/ s00253-009-2042-4.

Peroxiredoxin/thioredoxin system: Forecasting the peril Gianluca Selvaggio MIT Portugal Bioengineering Systems, Faculty of Science and Technology, New University of Lisbon, Portugal gianluca.selvaggio@gmail.com Background: Starting year: Research team: Supervisors:

MSc. in Biomedical Engineering, University of Bologna, Italy 2011 Pedro Coelho1 Armindo Salvador1 1 Centre for Neuroscience and Cell Biology – University of Coimbra

OBJECTIVES Cells occasionally have to cope with high H2O2 concentrations for long periods, due to their involvement in catalysis and folding maintenance, the irreversible oxidation of reactive thiols is deleterious for organisms. Here we hypothesize that the Peroxiredoxin/ Thioredoxin/Thioredoxin Reductase/Protein-Dithiol System (PTRTD) drives anticipatory blocking control (ABC) of dithiols as disulfides thus protecting them. We examined the design requirements for such a system to operate effectively and we compared these requirements to the actual design in human erythrocytes.

must be lower than the maximum flux of peroxiredoxin disulfide reduction and formation. Additionally, we identified a trade-off between the robustness of signal transduction and the NADPH expenditure in the process. A comparison of experimental data to the theoretical predictions above indicates that the design of the PTRTD system in human erythrocytes accomplishes effective integration between anticipatory blocking, antioxidant protection and redox signaling.

WORK PLAN We applied the approach described in (1; 2) by: i) scaling the system to reduce the number of parameters, ii) decomposing the design space into phenotypical regions, iii) determining the steady state behaviors, iv) grading them using a set of quantitative performance criteria derived from the ideal system properties, v) seeking for the principles that warrant the criteria satisfaction vi) comparing the required design principles with the observed phenotype. RESULTS The operating regimes were graded, based on the following performance criteria: a) efficient scavenging capacity, b) low NADPH consumption, c) effective signal propagation and d) effective ABC. The design principles were as follows, for human erythrocytes: i) the equilibrium constant for thiol-disulfide exchange between thioredoxin and the protein dithiol should be in the range 0.03<K<21 to allow the protein full protection; ii) the maximum flux of thioredoxin reduction

FIGURE 1 PTRTD Scheme.

References 1. Savageau, M. A., “Design principles for elementary gene circuits: Elements, methods, and examples,” Chaos: An Interdisciplinary Journal of Nonlinear Science, Vol. 11, No. 1, 2001/03/01, pp. 142–159. 2. Pedro M. B. M. Coelho, Armindo Salvador, M. A. S., “Quantifying Global Tolerance of Biochemical Systems: Design Implications for Moiety–Transfer Cycles,” PLoS Comput Biol, Vol. 5, No. 3, March 20, 2009, pp. e1000319.

Personalized 3D vascular constructs for toxicity/drug screening assessment Inês Honório MIT Portugal Bioengineering Systems, Biomaterials and Stem Cell-Based Therapeutic Laboratory, UC Biotech - Technologial Park of Cantanhede, FCT-UNL, Portugal honorio.ines@gmail.com Background: Starting year: Research team: Supervisors:

Msc. Molecular and Genetics Biology, Faculdade de Ciências da Universidade de Lisboa, Portugal 2011 Helena Vazão1, Mário Grãos2 Lino Ferreira3 1 UC Biotech - Technologial Park of Cantanhede 2 Biocant 3 Universidade de Coimbra

WORK PLAN The development of new tools for the rapid toxicological/drug screening assays of chemical substances, at cellular levels, is of high interest, providing patient-specific information about the toxicity/therapeutic effect of a pharmaceutical compound. Since vascular cells control the permeability of blood vessels, regulate inflammation, immunity and cell growth, among other key functions, it is of utmost importance to assess vascular toxicity and develop new drugs able to modulate the activity of endothelial cells. The overall aim of this project is the development of personalized “vascular kits” for risk assessment of chemicals and pharmaceutical compounds that can be used for high-throughput screening. Pluripotent stem cells such as human embryonic stem cells (hESCs) or inducible pluripotent stem cells (iPSCs) represent a potential source of vascular cells for the development of vascular kits. Recently, it was isolated in our lab a population of vascular progenitor cells from hESCs that can give rise to endothelial and smooth muscle cells and form microvessels

when injected in vivo. In this project we would like to extend our differentiation protocol to iPSCs that are very promising for personalized kits given that they can be generated from somatic cells isolated from patients using a seminal technology developed by Yamanaka and collaborators in 2006. For the development of the kits, the cells should form a vessel where the medium can flow. Recently one of us has engineered constructs with perfusable microvasculature networks, which will be used for the development of the vascular kits. RESULTS iPSCs has been differentiated in smooth muscle and endothelial like cells using diferentiation protocols previously established in the lab for the human embryonic stem cells. Micro-channels were produced in order to evaluate the capacity of smooth muscle and endothelial cells to adhere and react to physiological shear stress conditions. The same as been done using iPSC derived cells.

Urinary rhabdosphincter bioengineering – A decellularized matrix for modeling stress urinary incontinence in vitro Irina Neves Simões1 MIT Portugal Bioengineering Systems, Department of Bioengineering and IBB – Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Portugal irinansimoes@tecnico.ulisboa.pt Background: Starting year: Research team: Supervisors:

Master’s in Biotechnology, IST-UL, Portugal 2011 Paulo Vale3; Shay Soker2; Anthony Atala2; Rute M. Noiva4; Belmira M. Carrapiço4; M. Conceição Peleteiro 4; Joaquim M. S. Cabral1; Cláudia L. da Silva1; Pedro M. Baptista2,5,6 Cláudia L. da Silva1; Joaquim M. S. Cabral1; Pedro M. Baptista2,5,6 1 Department of Bioengineering and IBB – Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Portugal 2 Wake Forest Institute for Regenerative Medicine, Winston-Salem, NC, USA 3 Serviço de Urologia, Hospital Garcia de Orta, Almada, Portugal 4 Interdisciplinary Centre of Research in Animal Health, Faculty of Veterinary Medicine, Universidade de Lisboa, Portugal 5 IIS Aragón, CIBERehd., Zaragoza, Spain 6 Aragon Health Sciences Institute (IACS), Zaragoza, Spain

OBJECTIVES Establish an in vitro urethral model based on acellular urethral matrices for the screening of new drugs, therapies and the study of stress urinary incontinence (SUI). WORK PLAN Piglet urethras were decellularized using different concentrations of Triton X-100 and SDS and a perfusion- and agitation-based system. Cell removal was assessed by H&E staining and confirmed by DNA extraction. The presence of specific ECM proteins was evaluated by immunofluorescent stainings. Total collagen, elastin and glycosaminoglycans were quantified by colorimetric assays. 300μm thick acellular scaffold sections were repopulated with human skeletal muscle myoblasts (hSkM) and further differentiated. Cell presence and distribution was assessed by H&E and immunohistochemistry. RESULTS SDS solutions at 0.5% and 1% concentrations successfully decellularized the urethra, as shown by the absence of purple basophilic staining on H&E (Fig.1).

FIGURE 1 H&E staining of native (a) and acellular (b and c) bioscaffolds (0.5% SDS, b; 1% SDS, c).

The 0.5% SDS removed 93.4±2.6% of DNA material, while 1% SDS removed 96.1±0.0%. Immunofluorescent staining and colorimetric quantification indicated no major loss or modification in the localization of ECM proteins. Still, elastin was significantly decreased (~19%) in the acellular ECM. hSkM were

able to repopulate the acellular scaffold and form myotubes under the appropriate conditions (Fig. 2).

FIGURE 2 H&E staining (a-c) and immunohistochemistry staining (d-h) of the scaffolds recellularized with hSkM. The presence desmin (d), Myosin Heavy Chain (e), Actin (f) MyoD (g), and Pax3 (h) was evaluated.

hSkM deposition was mainly observed in the peripheral areas of the scaffold, due to intrinsic limitations of this cell type. Fibroblasts and bone marrow mesenchymal stem cells (BM-MSCs) were able to penetrate the scaffold to a greater extent (Fig. 3).

FIGURE 3 H&E staining recellularized bioscaffolds using hskMyob (a), BM-MSCs (b) and L929 fibroblasts (c).

Future studies include the optimization of the recellularization process, using a dynamic perfusion system, assessing cell adhesion, proliferation and function of the bioengineered urethral rhabdosphincter.

Affinity ligands for phosphoproteomics Íris Luz Batalha MIT Portugal Bioengineering Systems, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa irisbatalha@gmail.com Background: Starting year: Supervisors:

BSc in Applied Chemistry and MSc in Biotechnology, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, Portugal 2009 Ana Cecília Roque1; Christopher Robin Lowe2; Olga Iranzo3 1 Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa 2 Department of Chemical Engineering and Biotechnology, University of Cambridge 3 Instituto de Tecnologia Química e Biológica, Universidade NOVA de Lisboa

OBJECTIVES Protein phosphorylation has been vastly associated with highly incident human diseases, such as cancer and Alzheimer's. However, it is difficult to identify and quantify phosphorylated proteins and peptides by Mass Spectrometry (MS) due to their low stoichiometry and abundance in plasma and serum samples. Current methodologies used to overcome these issues are either unspecific or costly. The aim of this project is to develop novel synthetic affinity ligands using a high-throughput platform, which are both target-oriented and inexpensive. WORK PLAN Three combinatorial libraries of ligands have been synthesized in 96-well blocks using agarose as solid support. Amine, carboxylic acid, and aldehyde components have been selected in order to mimic key amino acids involved in phosphate recognition, such as Arg, Lys, Tyr, Ser, His, Gly, and Asn. After several steps of optimization, two lead ligands were selected. These ligands were further synthesized in magnetic nanoparticles, and screened against a semi-complex mixture of tryptic digested α-casein, β-casein and bovine serum albumin (BSA). Samples were analysed using MALDI-TOF MS. RESULTS 232 tailor-made synthetic ligands have been synthesized using three different chemistries. Lead ligands

FIGURE 1 Schematic representation of the research strategy followed in the present work.

presented selective binding towards phosphorylated species at acidic pH, high binding capacities and up to 70% recovery yields upon elution. Ligands immobilized on magnetic nanoparticles present higher binding capacities (100x), but at the cost of lower selectivity. A mixture of trypsin-digested α-casein, β-casein, and BSA was efficiently enriched in monoand multi-phosphorylated peptides, comparable to the results obtained when employing a conventional protocol with Ti4+-IMAC.

Human hematopoietic stem cells treated with lysophosphatidic acid have enhanced survival in ischemic conditions and preserve cardiac tissue after myocardial infarction Ivana Kostic1,2,3 MIT Portugal Bioengineering Systems, Instituto Superior Técnico, Universidade de Lisboa, Portugal ivana_k@mit.edu Background: Starting year: Research team: Supervisors:

MsC in Human and Animal Physiology/University of Belgrade/Serbia 2010 Isabel Carvalho1*; Helena Vazão1,2; Sezin Aday1,2; Tiago Carvalheiro3; Mário Grãos1,2; António Duarte 4; Lino Gonçalves 4; Artur Paiva3; Lino Ferreira1,2 Lino Ferreira2,3, lino@biocant.pt; Jeffrey Karp5, jeffkarp@mit.edu 1 Biocant, Technology Transfer Association, Cantanhede, Portugal 2 CNC-Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal 3 Histocompatibility Center 4 Department of Cardiology, University Hospital of Coimbra, Coimbra, Portugal 5 Brigham and Women’s Hospital, Harvard Medical School, USA *Authors contributed equally.

OBJECTIVES For clinical efficacy, it is imperative that stem cells survive and engraft into the host tissue. Unfortunately, most of the cells die a few days after delivery, mainly due to ischemia. Some methodologies have been proposed to augment cell survival, but they have shown limited effectiveness. Work plan: Our approach was to augment cell survival by preconditioning CD34+ cells isolated from umbilical cord blood(1) with several pharmacological agents(2). Lysophosphatidic acid (LPA) treatment under hypoxia and serum-deprived conditions showed the most promising results. RESULTS The pro-survival effect of LPA is concentration dependent (from 1-100 μM), statistically significant as compared to the control (untreated cells) at 1 μM of LPA. The pro-survival effect of LPA decreases as a function of hypoxia time. The percentage of viable cells in LPA-treated CD34+ cells decreased from 78% at day 1 to 40% at day 3. To identify the receptor that is mediating the pro-survival effect of LPA (100 μM), we used several antagonists and inhibitors

where PPARg had the highest effect. It decreased significantly the number of viable cells induced by LPA from 77% to 53%. To determine whether LPA can induce the proliferation of CD34+ cells[3], a suspension of untreated or LPA-treated cells was exposed to hypoxia for 24 h and then to normoxia for 6 additional days. LPA-treated cells increased their number approximately 3-fold over the 7 days period while untreated cells decreased to half their initial number. After 1 day of hypoxia, both untreated or LPA-treated CD34+ cells started to differentiate into mast cells (14-17%) and neutrophils (5-8%). In the in vivo studies, the median of the differences in the fractional shortenings was 7.5 and 2.6 in LPA-treated cells + gel and sham groups, respectively, being the differences statistically significant. No statistical differences were observed between hearts treated with CD34+ cells and sham. References: 1. Haider, H.; Ashraf, M. J Mol Cell Cardiol 2008, 45, (4), 554-66. 2. Pedroso, D.C., et al., PLoS One, 2011. 6(1): p. e16114. 3. van Corven, E.J., et al., Cell, 1989. 59(1): p. 45-54an.

Engineering umbilical cord blood-derived hematopoietic stem/progenitor cells’ microenvironment toward efficient megakaryo/thrombopoiesis Javad Hatami MIT Portugal Bioengineering Systems, Department of Bioengineering and IBB - Institute for Biotechnology and Bioengineering – Instituto Superior Técnico (IST), Lisboa, Portugal Javad.hatami@ist.utl.pt Background: Starting year: Supervisors:

MSc, Biomedical Engineering, Amirkabir University of Technology -Tehran Polytechnic, Tehran, Iran 2010 Frederico Castelo Ferreira1; Claudia Lobato da Silva1 1 Department of Bioengineering and IBB - Institute for Biotechnology and Bioengineering – Instituto Superior Técnico , Universidade de Lisboa, Portugal

OBJECTIVES To study the ex-vivo expansion and differentiation of umbilical cord blood hematopoietic stem/progenitor cells (HSC) toward Megacaryocyctic lineage. The main idea explored herein is to integrate an optimize HSC expansion stage with an efficient Megakaryocyte differentiation stage and then maturation into blood platelets. I intend to design a system that allows a certain degree of independent optimal conditions for each stage, achieving high yields of megakaryo/thrombopoiesis. RESULTS Co-infusion of ex-vivo generated megakaryocytic progenitors with hematopoietic stem/progenitor cells (HSC/HPC) may contribute to a faster platelet recovery upon umbilical cord blood (UCB) transplantation. A two stage protocol containing cell expansion and megakaryocyte (Mk) differentiation was established using human UCB CD34+-enriched cells. The expansion stage used a pre-established protocol supported by a human bone marrow mesenchymal stem cells (MSC) feeder layer and the differentiation stage used TPO (100 ng/mL) and IL-3 (10 ng/mL). 18% of culture-derived Mks had higher DNA content (>4N) and were able to produce platelet-like particles. The proliferation extent of CD34+ cells obtained in the expansion stage (FI-CD34+), rather than expansion duration, determined as a key parameter for efficient megakaryocytic differentiation. A maximum efficiency yield (EY) of 48±7.7 Mks/input

CD34+ cells was obtained for a FI-CD34+ of 17±2.5, where a higher FI-CD34+ of 42±13 resulted in a less efficient megakaryocytic differentiation (EY of 22±6.7 and 19±4.6 %CD41).

FIGURE 1 Effect of UCB CD34+-enriched cell’s expansion stage, co-cultured with a feeder layer, on Megakaryocytic differentiation: EY (A) and %CD41 (C) were evaluated with respect to FI-CD34+ (EY and FI-CD34+ were significantly different among all groups, p<0.05). EY (B) and %CD41 (D) were evaluated with respect to the expansion duration (no significant differences between groups, p>0.3). %CD41 was significantly different between G2 and either G1 or G3, p<0.05), n≥4 for each group.

Heterologous production of curcuminoids in E. coli through an artificial biosynthetic pathway Joana L. Rodrigues MIT Portugal Bioengineering Systems, Department of Biological Engineering, School of Engineering, University of Minho, Braga, Portugal joana.joanalucia@deb.uminho.pt Background: Starting year: Supervisors:

Master in Biological Engineering/ University of Minho/ Portugal 2010 LĂgia L. Rodrigues1; Kristala L. J. Prather2; Leon Kluskens1 1 Department of Biological Engineering, University of Minho 2 Department of Chemical Engineering, Massachusetts Institute of Technology

ABSTRACT Curcuminoids are natural pigments from plants that have been reported as potential cancer-fighting drugs. Nevertheless, they have a poor bioavailability. Cellular uptake is low, and they are quickly metabolized once inside the cell, requiring repetitive oral doses to achieve sufficient concentration inside the cell for therapeutic activity. The aim of this work is to engineer an artificial biosynthetic pathway for the production of curcuminoids by Escherichia coli. Starting from the substrate tyrosine, the curcumin pathway involves several enzymatic steps: conversion of tyrosine to p-coumaric acid; conversion of p-coumaric acid to caffeic acid; production of caffeoyl-CoA from caffeic acid; production of feruloyl-CoA from caffeoyl-CoA; and finally the production of curcumin from feruloyl-CoA and possibly other curcuminoids, due to enzyme promiscuity. The enzymes involved in the two first enzymatic steps are tyrosine ammonia lyase from Rhodotorula glutinis, P450 CYP199A2 from Rhodopseudomonas palustris, and the redox partners pdr from Pseudomonas putida and pux from

R. palustris. Coumaric acid and caffeic acid were successfully produced. A coumaroyl-CoA ligase from Arabidopsis thaliana is being explored for the conversion of the different carboxylic acids into their corresponding CoA esters. Different combinations of this enzyme and caffeoyl-CoA 3-methyl transferase may lead to the production of different curcuminoids. For the last step of the pathway two approaches are being studied: the use of diketide-CoA synthase and curcuminoid synthase from Curcuma longa, and curcumin synthase from Oryza sativa that itself catalyzes both steps. Curcumin and bisdemethoxycurcumin were produced using both approaches and their production was confirmed by HPLC analysis, as well as by the yellow color of the culture supernatant. Successful construction of the curcuminoids biosynthetic pathway would mark a significant step forward in the in situ production of these poorly soluble, anti-carcinogenic compounds. Acknowledgments: The author thanks the Master student Rafael AraĂşjo for the support given in the laboratory.

hPin1 WW domain - From production to engineering applications Margarida Dias MIT Portugal Bioengineering Systems, Chemistry Department, Faculty of Science and Technology - Universidade Nova de Lisboa amd18816@campus.fct.unl.pt Background: Starting year: Supervisors:

MSc in Biotechnology and BSc in Applied Chemistry at FCT-UNL , Portugal 2010 Olga Iranzo1; Ana Cecília A. Roque2 1 Institut des Sciences Mol.culaires de Marseille, France 2 Chemistry Department, Faculty of Science and Technology - Universidade Nova de Lisboa

OBJECTIVES Development of an affinity support for capture phosphorylated peptides and proteins based on Human Pin1 WW domain [3]. WORK PLAN In this work was explored the potential application of a native binding domain to phosphorylated peptides for purification of proteins. We consider a WW domain - Pin1, which was produced chemically and purified, it was immobilized in a solid support and the affinity studies were preformed. RESULTS Human Pin1 (hPin1), a mitotic peptidyl-prolyl isomerase (PPIase) has been identified as a mitosis cell cycle regulator [1,2,3]. The WW domain region of this protein specifically recognizes Pro-rich ligands and phosphorylated in Ser and Thr residues[4]. In this work, the stability of a point mutated version of hPin1 at the N and C-terminals (hPin1_mutated) was studies using Molecular Dynamics (MD) studies. During MD simulations, hPin1_mutated maintains its characteristic WW folding (hairpin β-sheet motif), in

40 FIGURE 1 Best Docking solution between hPin1 and peptide VPRpTPV. In yellow are represented the residues in the “X-P groove”.

addition the stability of β-sheets increased. Furthermore, automated molecular docking studies were performed to predict the affinity of hPin1_mutated towards different phosphorylated and non-phosphorylated peptides, including VPR(pT)PV and PPPPYP. The results demonstrated a higher affinity for phosphorylated peptides. The Ka determined in silico is comparable to what is described in the literature for the native ligand [4] , and the best conformation is represented in Fig.1 [5-6]. hPin1_mutated was produced by solid-phase peptide synthesis using an automated microwave synthesizer and Fmoc-chemistry with incorporation of Pseudoproline dipeptide units to facilitate the synthesis and hPin1_mutated was obtained with high purity. The peptide was characterized Mass Spectrometry (ESI-MS) and Circular Dichroism (CD). The CD spectra showed the characteristic signal of a WW domain folding, as well as the stability expected for this small domain. hPin1_WWmutated was immobilized successfully in solid support and affinity studies were conducted with peptides and model proteins phosphorylated. The experimental results demonstrated affinity towards the phosphorylated peptide and less affinity for non-phosphorylated version. Furthermore, it was possible to capture a model protein phosphorylated in Ser and Tyr, and the same protein without phosphorylation was not capture. In addition, the protein elution was also tested and it was observed protein recovery. These results indicate that hPin1 WW domain can be used in biotechnology applications for recognition and possible purification of phosphorylated peptides and proteins. References: 1. Sudol, M. and T. Hunter, . 103,1001-1004,2000. 2. Macias, M.J., et al., Nat. Struct. Biol.,7, 375-379, 2000 3. Lu, P.J., et al., Science, 283, 1325-1328, 1999 4. Verdecia, M.A., et al., Nat Struct Mol Biol, 7,(8), 639-643, 2000 5. Zarrinpar, A. and W.A. Lim, Nat Struct Mol Biol, 7(8), 611-613,2000 6. Zhang, M., et al.,. ACS Chemical Biology, 7 (8), 1462-1470, 2012

Membrane reactors in process intensification of mannosylerythritol lipids production from sugars Marisa Santos MIT Portugal Bioengineering Systems, Department of Bioengineering and Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa; Laboratório Nacional de Energia e Geologia, I.P., Unidade de Bioenergia Background: Starting year: Research team: Supervisors:

Integrated Master in Biological Engineering/Universidade do Algarve/ Portugal 2011 Nuno Faria1,2; César Fonseca2; Frederico Castelo Ferreira1 Frederico Castelo Ferreira1; César Fonseca2 1 Department of Bioengineering and Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa 2 Laboratório Nacional de Energia e Geologia, I.P., Unidade de Bioenergia

OBJECTIVES Mannosylerythritol lipids (MELs) are glycolipids with surface active properties produced by Pseudozyma sp. strains. These biosurfactants are usually produced from vegetable oils. The use of vegetable oils in industrial processes will hardly be sustainable, due to competition with food and feed supply chain and their increasing prices. Moreover, the use of vegetable oils for MEL production has the additional disadvantage of difficult downstream separation of the glycolipid produced from the remaining substrate. The work herein presents the first attempt to use yeast cell recycling in fed-batch reactors for MEL production. This process targets the conceptual use of lignocellulosic wastes. WORK PLAN There is a call to obtain sustainable products, nondependent from fossil fuels and not competing with food supply chain. Therefore, the development of a sustainable process for MELs production requires the use of new carbon sources, acquired at low cost. This work targets the use of fermentation processes coupled to ultrafiltration membranes for retention of yeasts and/or polysaccharides.

FIGURE 1 Scheme for a Fed-batch bioreactor with yeast cell recycling through an ultrafiltration membrane unit.

RESULTS Using a membrane system, after 10 days of fermentation we were able to attain higher MEL concentration than we usually achieve after 16 days using a pulse system. Also, in the continuous feeding process, it was observed a gradual increase in MEL production during entire time of culture, contrary to what was observed in the pulse feeding process, where MEL concentration remains constant (or fluctuating) during time. Fatty acids (FA) decreased initially to reach a plateau, presumable a steady state between production fluxes from carbon source consumption and FA used for MEL production. A fed-batch and continuous reactor coupled to an ultrafiltration membrane unit has the potential to contribute to improve the production of MELs using sugar-based substrates, namely from lignocellulosic wastes, overcoming the need of using vegetable oils as carbon source. Acknowledgements: This work was co-funded by Fundação para a Ciência e a Tecnologia (FCT) through the following initiatives, MIT-Portugal Program - Bioengineering Systems Focus Area; NTF, PhD grant (SFRH/BD/33724/2009); MS, PhD grant (SFRH/BD/78420/2011), FCF, “Investigador FCT” Program (IF/00442/2012); CF, “Ciência 2008” Program. Moreover, this work was co-funded by the National Exploratory Project “TAKEOFF” through FEDER “Programa Operacional Factores de Competitividade” - COMPETE (FCOMP-01-0124-FEDER-041191) and national (FCT) funding (EXPL/AAG-TEC/0610/2013). The authors acknowledge the Portuguese Yeast Culture Collection (PYCC), CREM-FCT/UNL, for providing the yeast strains.

Chemical and physical engineering of MSC microenvironment to tune their secretory profile Marta Costa MIT Portugal Bioengineering Systems, Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa marta.costa@tecnico.ulisboa.pt Background: Starting year: Supervisors:

Biochemistry/FCUL/Portugal; Bioengineering and Nanosystems/IST/Portugal 2012 Frederico Ferreira1; Cláudia Lobato da Silva1; Todd McDevitt 2 1 Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa 2 Stem Cell Engineering Center, Georgia Tech

OBJECTIVES The beneficial effects of mesenchymal stem cells (MSC) on regenerative medicine are not only related to their differentiation capacity but also to the bioactive molecules secreted by these cells. Microenvironmental stimuli can be explored to modulate MSC secretory profile and increase their survival, namely through cells encapsulation and delivery of small molecules and/or growth factors (GF).

FIGURE 1 Encapsulation of hBM MSC in solid (A) and liquid (B) core alginate beads.

WORK PLAN • Chemical and physical engineering of MSC microenvironment to tune their secretory profile. • Development of an encapsulation system capable of increasing MSC survival. • Tailor the angiogenic potential of DMOGtreated MSC.

RESULTS Initial attempts to encapsulate hMSC in solid core-alginate beads showed inhibition of cells proliferation and, overtime, formation of MSC aggregates with uncontrollable sizes. To better control MSC size and promote increased nutrient exchange, encapsulation of hMSC in liquid core-alginate beads can be performed (Figure 1). Delivery of small molecules (CHIR99021, BIO, p.e.) and/or GF (FGF2) was addressed to improve the survival/proliferation rates of hMSC. Additionally, the use of heparin MPs was explored to more efficiently deliver FGF2 to MSC. Over 80% of FGF2 adsorbs electrostatically to heparin MP, highlighting the potential of this material to gather important GF for MSC survival. Regarding small molecules potential effect on MSC secretory profile, treatment of MSC with DMOG, a small molecule that can activate the expression of HIF-1α in cells at normal oxygen tension, was shown to enhance VEGF secretion. Its angiogenic activity might have a positive impact on MSC survival once transplanted into a diseased model.

FIGURE 2 Delivery of FGF2-conjugated heparin MP to monolayer MSC (A). A higher number of heparin MP seems to hinder MSC proliferation, potentially due to GF gathering from the microenvironment (B).

FIGURE 3 VEGF secreted by hMSC treated with DMOG (0 and 500 μM) in both serum and serum-free media.

Patient’s access to breakthrough innovation: A regulatory/technology perspective on clinical translation cases of cell therapy medicinal products Miguel Amador MIT Portugal Bioengineering Systems, IN+, Instituto Superior Técnico, University of Lisbon, Portugal miguel.amador@tecnico.ulisboa.pt Background: Starting year: Supervisors:

Msc. in Biomedical Engineering, Instituto Superior Técnico, University of Lisbon, Portugal 2012 Manuel Heitor1; Frederico Ferreira2; Miguel Amaral1; Kenneth Oye3 1 IN+, Instituto Superior T´ecnico, University of Lisbon 2 IBB, Instituto Superior Técnico, University of Lisbon 3 Political Science and Engineering Systems, Massachusets Institute of Technology

OBJECTIVES Cell Therapy Medicinal Products (CTMP) are an emerging Regenerative Medicine field technology, which holds promising new therapies, for areas where significant clinical need still exist, but are yet far from their promising impact, with stakeholders facing challenges with its inherent complex dynamic therapeutic action. While access to breakthrough therapies is a matter of public interest, it is the responsibility of stakeholders to guarantee their safety and efficacy, expressed in their regulatory environment. Seen as one of the hurdles taking CTMP to a broader usage poses a challenge of risk governance and development of public policies. This research work aims to contribute with evidences to the debate regarding the urgency to provide adequate early access to CTMP to patients by: understanding the suitability of regulatory frameworks for the clinical translation of CTMP to provide patients access to therapies, in a timely manner, and in acceptable levels of efficacy and safety; defining CTMP stakeholders (i.e. promoters, regulators, patients) capability to provide acceptable levels of efficacy and safety in a sustainable way.

WORK PLAN To study the challenges on clinical translation of CTMP, this work is focused on developing case studies, involving different stakeholders, in order to: 1) Provide evidence of the urgency of patient early access to breakthrough CTMP; 2) Provide evidences of the capability to give proper access to CTMP to patients under current and alternative regulatory frameworks; 3) Understand how technologies are enabling new therapies and an adequate regulatory evaluation.

FIGURE 2 Towards new Regulations: fast-track approval path? It is possible without compromising safety and efficacy?

FIGURE 1 Manufacturing cell based products: zero risks at high costs. Is it possible to improve?

RESULTS On this research work, it was elaborated a report, gathering multiple Portuguese stakeholders, highlighting the complexity of addressing regulatory issues in health care, which goes beyond the changes at the regulatory level and the analysis at a local level. Case studies of CTMP clinical translation are being developed in Europe and planned to be extended during a research period in Boston.

Yeast mannosylerythritol lipids from lignocellulose: A novel strategy for the production of jet biofuel Nuno Faria MIT Portugal Bioengineering Systems, Department of Bioengineering, IBB-Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa nunosek@gmail.com Background: Starting year: Research team: Supervisors:

Master degree/ University of Minho/ Portugal 2009 Marisa Santos1; Bruce Tidor2; César Fonseca3; Frederico Castelo Ferreira1 Frederico Castelo Ferreira1; César Fonseca3; Bruce Tidor2 1 Department of Bioengineering, IBB-Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa 2 Biological Engineering and Computer Science, Massachusetts Institute of Technology 3 Laboratório Nacional de Energia e Geologia, I.P., Unidade de Bioenergia

OBJECTIVES Biofuels and bio-based chemicals have gained attention as renewable alternatives to liquid transportation fuels and chemicals derived from petroleum. This work aimed to explore the natural ability of non-conventional yeast, Pseudozyma spp., to convert lignocellulose sugars into mannosylerythritol lipids (MEL), towards the sustainable production of a biosurfactant, at low substrate cost and direct product recovery. Moreover, MEL is explored as a novel biological precursor for the sustainable production of jet biofuel, exploiting the characteristic size of their lipidic chains. WORK PLAN To accomplish the goals, the work consisted in exploration of process configurations to produce MEL from lignocellulose or lignocellulosic derived sugars and/ or co-production of other interesting molecules (e.g. enzymes), exploration of yeast metabolism and characterization of physical-chemical properties of blends of conventional fuel and MEL-derived molecules. RESULTS The performed work demonstrated the capacity of Pseudozyma spp. in converting pentoses, D-xylose/D-glucose mixtures, cellulose and xylan into MEL. While different process configurations were

developed for MEL production from cellulosic substrates combining commercial enzyme cocktails and Pseudozyma spp., the ability to produce own xylanolytic enzymes allowed the direct conversion of xylan into MEL. Dynamic mathematical models describing the metabolic pathways for the conversion of lignocellulosic sugars into MEL were developed, as tools for improving bioprocesses and for future metabolic engineering and synthetic biology approaches. The experimental design for 13C-NMR studies was established, allowing the determination of flux distribution at branch points of metabolism. Finally, the utilization of MEL as a biofuel precursor was assessed through the characterization of physicochemical properties of products obtained from the lipidic chains of lignocellulosic MEL, and in their blend with conventional jet fuel, against specifications for jet fuels. Acknowledgements: This work was co-funded by Fundação para a Ciência e a Tecnologia (FCT) through the following initiatives, MIT-Portugal Program - Bioengineering Systems Focus Area; NTF, PhD grant (SFRH/BD/33724/2009); MS, PhD grant (SFRH/BD/78420/2011), FCF, “Investigador FCT” Program (IF/00442/2012); CF, “Ciência 2008” Program. Moreover, this work was co-funded by the National Exploratory Project “TAKE-OFF” through FEDER “Programa Operacional Factores de Competitividade” - COMPETE (FCOMP-01-0124-FEDER-041191) and national (FCT) funding (EXPL/AAG-TEC/0610/2013). The authors acknowledge the Portuguese Yeast Culture Collection (PYCC), CREM-FCT/UNL.

Development of hydrogel-based dressings for skin wound regeneration Patrícia Morgado MIT Portugal Bioengineering Systems, Departamento de Química, REQUIMTE, Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa (FCT-UNL), Campus de Caparica, Portugal patriciaicmorgado@gmail.com Background: Starting year: Research team: Supervisors:

MSc in Biomedical Sciences, Faculdade de Ciências da Saúde da Universidade da Beira Interior (FCS-UBI), Av. Infante D. Henrique, 6200-506 Covilhã, Portugal. January 2012 Ana Aguiar-Ricardo1; Ilídio J. Correia2 Ana Aguiar-Ricardo1; Ilídio J. Correia2 1 Departamento de Qu.mica, REQUIMTE, Faculdade de Ci.ncias e Tecnologia da Universidade Nova de Lisboa (FCT-UNL), Campus de Caparica, Portugal; 2 Centro de Investigação em Ciências da Saúde (CICS), Faculdade de Ciências da Saúde da Universidade da Beira Interior (FCS-UBI), Covilhã. Portugal.

OBJECTIVES Among the many diseases that affect the skin, burns are responsible for thousands of deaths every year worldwide. To overcome this health problem, this work plan aims to develop hydrogel-based matrices able to perform a controlled release of biologic active agents essential for restoring skin barrier and functions. WORK PLAN: 1. Development hydrogel-based wound dressings (WD) loaded with bioactive nano/microcarriers. 2. Characterization of the carriers and WD: morphology; mechanical properties; water uptake and contact angle analysis; water vapour permeability; in vitro release studies. 3. Evaluation of the suitability of the systems for the biomedical application proposed, through in vitro and in vivo studies.

RESULTS Asymmetric membranes have been reported as ideal WD for skin regeneration. Thus in this study supercritical carbon dioxide (scCO2) phase inversion technique was applied to prepare poly(vinyl alcohol)/ chitosan (PVA/CS) asymmetric membranes. The PVA/CS dressings produced are recovered in a dry state but, due to their high swelling capacity, they can be used to produce hydrogels able to maintain the moisturized environment needed for wound healing (WH). The ibuprofen (IBP) loaded membranes showed a fast release profile, which is a huge drawback for its application in the WH process. An ideal WD should allow drug delivery over 12h, since the peak of the inflammatory phase only occurs after this period. To overcome this handicap, microcarriers (poly(glyceroldimethacrylate) (PGDMA) and β-cyclodextrins (B-CD)) loaded with IBP were introduced into the matrices. The drug release was extended when B-CD were added to membranes. Furthermore, the loading of the carriers did not change membranes’ properties, making them suitable to be used as WD in skin wound regeneration. Acknowledgements: The authors are grateful to financial support from Fundação para a Ciência e a Tecnologia (FC&T), through the projects PEst-C/EQB/LA0006/2013, SFRH/BD/80648/2011 and MIT-Portugal Program Bioengineering.

FIGURE 1 Schematic representation of the two main approaches followed to prepare the hydrogel-based wound dressings.

Vascular cells derived from HutchinsonGilford progeria syndrome (HGPS) inducible pluripotent stem cells Patricia R. Pitrez MIT Portugal Bioengineering Systems, Biological Engineering, IBB Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Portugal pitrezpatricia@gmail.com Background: Starting year: Research team: Supervisors:

MSc. in Biological Engineering, University of Minho, Portugal 2010 Helena Vazão1 , Xavier Nissan2 Lino Ferreira1, Isabel Rocha3 1 CNC - Center of Neurosciences and Cell Biology, University of Coimbra, Coimbra, Portugal 2 CECS, I-STEM, AFM, Institute for Stem cell Therapy and Exploration of Monogenic diseases, Evry cedex, France 3 IBB – Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Braga, Portugal

OBJECTIVES To study the vulnerability of smooth muscle cells (SMCs) in Hutchinson-Gilford Progeria Syndrome (HGPS). WORK PLAN Hutchinson-Gilford Progeria Syndrome is a rare, progressive premature aging disease in children that leads to vascular SMCs degeneration and premature atherosclerosis. Death occurs at a mean age of 13 years, usually from heart attack or stroke. One of the hallmarks of the disease is the dramatic loss of vascular SMCs in large arteries. In consequence, the arteries undergo vascular remodeling and calcification. So far, it is unclear the reason underlining the sensitivity of SMCs in the context of the disease. To clarify the mechanisms behind SMCs loss, it’s important to have human HGPSSMC; however, these cells are difficult to obtain from Progeria patients.

Therefore, we derived HGPS induced pluripotent stem cells (iPSCs) from skin fibroblasts and then differentiate them into SMCs. RESULTS In this work, iPSCs obtained from HGPS fibroblast patients were successfully differentiated into HGPS-SMCs. These HGPS-SMCs showed impaired maturation and an upregulation of progeria markers when comparing with SMCs differentiated from healthy iPSCs. HGPS-SMCs shared similar features observed on progerin-expressing cells such as activation of several effectors of NOTCH signaling pathway and response to farnesyltransferase inhibitors. Therefore we have developed a cell model that is suitable for drug screening and to study the mechanism of SMC loss. This finding opens new opportunities for the treatment of HGPS disease and diseases related to vascular ageing.

A model-based approach to correct drop foot using electrical stimulation P. L. Melo MIT Portugal Bioengineering Systems, IDMEC, Mechanical Engineering, Instituto Superior Técnico, University of Lisbon, Portugal, Portugal paulo.de.melo@tecnico.ulisboa.pt Background: Starting year: Research team: Supervisors:

MSc. in Biomedical Engineering, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Portugal 2008 Miguel Tavares da Silva1; Dava Newman2; Jorge Martins1 Miguel Tavares da Silva1; Dava Newman2; Jorge Martins1 1 Instituto Superior Técnico, University of Lisbon, Portugal 2 Massachusetts Institute of Technology, Cambridge, MA

OBJECTIVES Drop foot patients are among one of the groups that has largely benefited from FES over the last decades, with several successful generations of the peroneal nerve stimulator currently available (1). These systems are often based in the use of a sensor, such as a foot switch or a tilt sensor, which times the swing phase and delivers a preset stimulation envelope to the dorsiflexors, enabling toe clearance (2). Within this scope, a new method to model the dynamics of the electrically stimulated dorsiflexors is proposed, to help chart the way towards, even more integrated, patient-specific model-based approaches to correct drop foot. WORK PLAN A novel model-based approach to correct drop foot is implemented as a two-stage process: two experimental trials tailored to address specific gait phases affected by drop foot; and a black-box modeling procedure, using the trials’ input-output data sets, to model the dynamics of the electrically stimulated muscles. These, easy to implement clinically, mimic the two phases mostly affected by drop foot, swing and controlled plantarflexion. Using system identification techniques, linear autoregressive models (specifically the Output-Error, OE, structure) are identified for each phase using different types of excitation inputs. RESULTS First evidences showed high prediction rates using OE linear models, but also that the excitation inputs of triangular nature have the highest accuracy for these types of movements and are therefore recommended for the development of patient-specific modelbased controllers to be used under the clinical

rehabilitation setting, where cycling motions are often important. On the other hand, pseudo-random multi-level excitation inputs were the most robust across all conditions, and still maintained a good prediction performance, making these types of inputs the best candidates for the development of gait neuroprostheses (see Figure 1).

FIGURE 1 Left upper plot: Identification result using a FRN data set (model orders: nb= 2, nf = 2, nd = 1). Right upper plot: validation result using the previous model on another FRN data set. Lower left plot: identification result using a STR data set (model orders: nb = 4, nf = 4, nd = 1). Right lower plot: validation result using the previous model on another STR data set.

References 1. Peckham, P., Functional electrical stimulation for neuromuscular applications, Vol. 7, Annual Review of Biomedical Engineering, 2005. 2. Cameron, M., “The walkaide R functional electrical stimulation system – A novel therapeutic approach for foot drop in central nervous system disorders,” European Neurological Review, Vol. 5, No. 2, 2010, pp. 18–20.

Investigating regulatory T cells by single cell RNA-sequencing Ricardo Júdice Miragaia MIT Portugal Bioengineering Systems, Department of Biological Engineering, University of Minho miragaia@ebi.ac.uk Background: Starting year: Supervisors:

Master’s degree in Genetics and Molecular Biology, Faculty of Sciences of the University of Lisbon, Portugal 2012 Sarah Teichmann1; Eugénio Ferreira2 1 European Bioinformatics Institute (EMBL-EBI)/Wellcome Trust Institute, University of Cambridge 2 Department of Biological Engineering, University of Minho)

OBJECTIVES Single-cell transcriptomics is an emergent and powerful approach to understand the complexity inherent in many cell populations. Its relevance has been growing as increasing evidence supports the existence of marked heterogeneity within cell populations previously considered as homogeneous. This is particularly important for many immune cell types, where single cell RNA-sequencing has the potential to provide profound new insights into subpopulation structure. This project focuses on a particular immune lineage - Regulatory T (Treg) cells – which play a central role in peripheral tolerance to self- and non-self-antigens. By single-cell mRNA sequencing of Treg cells, we intend to address their remarkable gene expression plasticity, determine Treg identity, define subsets, and investigate their functions, surface markers and key features. Results from this work will provide a more comprehensive overview of how immune homeostasis is regulated. WORK PLAN The initial phase of this project consists of characterizing Treg identity at the single-cell level. As a first step, we are focusing on Treg isolation protocols from diverse tissues in collaboration with research groups with previous expertise on the field. The resulting single cell suspensions are then run on the C1™ microfluidics systems (Fluidigm®) for single cell capture, reverse transcription and cDNA amplification. Next, we barcode and prepare the Illumina libraries for sequencing. Using this methodology, we plan to get single-cell transcriptomics data from immunologically relevant tissues. Extensive bioinformatics

analysis on these data will provide new insights into Treg biology. In a second stage, the most interesting features of Treg biology will be extracted and prioritised for further study. A combination of adoptive transfers of newly discovered Treg subsets and manipulations targeting newly identified Treg genes will be used as validation assays.

FIGURE 1 Project Workflow. (A) Regulatory T (Treg) cell suspensions are obtained from mouse tissues. (B) Single cells are then captured in separate compartments on C1™ chips and lysed. The C1™ system is responsible for the reverse-transcription of the mRNA released from each cell, as well as the amplification of the resultant cDNA. After appropriate Library Prep, (C) the cDNA is sequenced on the Illumina® platforms. (D) The single-cell transcriptomics data obtained presents itself as a brand new open window to Treg biology. After thourough analysis, interesting findings will be tested in vivo and/or in vitro.

Novel methodology to predict septic shock outcome using fuzzy modeling Rúben Duarte Pereira MIT Portugal Bioengineering Systems, Institute of Mechanical Engineering, CIS/IDMEC-LAETA, Instituto Superior Técnico, Universidade de Lisboa rubens.dmap@gmail.com Background: Starting year: Supervisors:

MSc in Biomedical Engineering / Instituto Superior Técnico and Faculdade de Medicina, Universidade de Lisboa / Portugal 2009 João Miguel da Costa Sousa1; Stan N. Finkelstein2 1 Institute of Mechanical Engineering, CIS/IDMEC-LAETA, Instituto Superior T.cnico, Universidade de Lisboa 2 Engineering Systems Division, Massachusetts Institute of Technology

OBJECTIVES Severe sepsis and septic shock constitute major clinical and economic healthcare problems, and early therapy is suggested to influence outcome of the patients in the intensive care unit (ICU). Though, medical scores for risk assessment in the ICU are complex and sparsely calculated. The aim of this work is to use computer algorithms that are able to model relations in data to improve the outcome of septic shock patients in the ICU. WORK PLAN A retrospective study using a cohort of adult septic shock patients from a publicly available database was performed. Only patients comprising data for a pre-determined set of physiological variables considered were included. From a total of 583 patients that underwent septic shock, 117 met inclusion criteria for this study. Takagi-Sugeno fuzzy modeling was used to model predictors of septic shock outcome (deceased or survived). RESULTS We developed a classifier for septic shock outcome based only on 1 hour of measurements of 4 physiological variables (“BPpHT” set): Arterial blood pressures, blood pH and thrombocytes levels.

The model correlates more than 0.79 (p < 0.05) with at least one of the medical risk scores studied along the ICU stay of each patient, see Figures 1 and 2, and achieves 81% accuracy when compared to medical decision, see Figure 3. The model provides accurate hourly risk of death in septic shock patients and enables highly dynamic interpretation of the clinical situation.

FIGURE 2 Variation of the medical scores for the patients in Figure 1 during the ICU stay (the X-axis, time, is common to both graphics).

FIGURE 3 Comparison of the classification of risk between the classifier developed and the medical decision model. Value “1” means “risk of decease” and value “0” means “not at risk”.

49 FIGURE 1 Variation of the output of the classification model developed, purple, for one of the patients during the ICU stay (outcome is “deceased”). Variations of the physiological variables used are also shown.

Bulk fluorescence long-term monitoring of serotonergic neuronal activity in behaving mice Sara Matias MIT Portugal Bioengineering Systems, Champalimaud Neuroscience Programme, Champalimaud Centre for the Unknown sara.matias@neuro.fchampalimaud.org Background: Starting year: Research team: Supervisors:

Biomedical Engineering/ Instituto Superior Técnico/ Portugal 2008 Eran Lottem1; Guillaume P. Dugué2 ; Zachary F. Mainen1 Zachary F. Mainen1; Manuel Nunes da Ponte3 1 Champalimaud Neuroscience Programme, Champalimaud Centre for the Unknown 2 CNRS UMR, INSERM, Paris, France 3 REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa

OBJECTIVES Serotonin (5-HT) is a neuromodulator involved in a wide range of brain functions and diseases. We want to investigate the dynamics of 5-HT neuronal activity in the dorsal raphe nucleus (DRN) during behavior. WORK PLAN We used viral infection of the Cre-dependent genetically encoded calcium indicator GCaMP6s in specific neurons using cre-expressing mouse lines and implanted single multimode optical fibers in the targeted areas. Fluorescence emission was collected using a PMT. We validated the setup using EYFP expressed in 5-HT neurons. We found that we could measure protein expression levels over a period of weeks. Subsequently, we monitored 5-HT activity in the dorsal raphe nucleus of anesthetized mice in response to electrical stimulation and found rapid and reliable calcium transients that scaled in magnitude with the number of stimulation pulses. RESULTS In behaving mice, we found robust calcium signals in the dorsal striatum of D1-cre (n=4) and of D2-Cre (n=4) mice that corresponded to contraversive turning (D1-Cre: 4% deltaF/F; D2-Cre: 2% deltaF/F) but not to ispiversive turning, which was not observed in EYFP controls. To identify the stimuli or events

50 FIGURE 1 Schematics of fiber optic- based setup used for fluorescence measurement of neuronal activity.

that activate 5-HT neurons we designed an olfactory-guided decision-making task in which head-fixed mice have to discriminate between various odors in order to decide whether to lick a water spout to receive reward or not to lick to avoid air-puff punishment. Besides GCaMP6s we are now also expressing tdTomato in 5-HT cells to control for movement artifacts. Using these two approaches we are studying specifically the function of the serotonergic neurons of the DRN during sensory, motor and reward events.

FIGURE 2 Both the D1 and the D2 pathways are active during contraversive turning. a) and c) Representation of the contraversive (green) and ipsiversive (blue) turns in freely moving mice for the D1 (a) and the D2 (c) pathways. b) and d) Fluorescence measurement of neuronal activity during the turns represented in a) and c) respectively; insets: YFP controls.

FIGURE 3 Serotonergic activity during a classical pavlovian task in behaving mice. Neural activity was measured with the setup shown in figure 1 during a an olfactory-guided decision-making task. Serotonergic activity during the conditioned stimulus seems to be correlated with the value of the outcome.

Design of cell-instructive interfaces resorting to marine-origin resources and human platelets Sara M. Oliveira MIT Portugal Bioengineering Systems, 3B’s Research Group, University of Minho oliveiramsara@gmail.com Background: Starting year: Research team: Supervisors:

Biomedical Engineering/University of Minho/Portugal 2010 Sara M. Oliveira1,2,3; Rogério R. Pirraco2,3; Tiago H. Silva2,3; Vitor E. Santo2,3; Manuela E. Gomes2,3; Alexandra P. Marques 2,3; Rui L. Reis2,3; João F. Mano2,3 João F. Mano2; Rui L. Reis2 1 MIT Portugal Bioengineering Systems, 3B’s Research Group, University of Minho 2 3B’s Research Group – Biomaterials, Biodegradables and Biomimetics, AvePark, Zona Industrial da Gandra S. Claúdio do Barco, 4806-909 Caldas das Taipas, Guimarães, Portugal 3 ICVS/3B’s – PT Government Associate Laboratory, Braga/Guimarães, Portugal

OBJECTIVES The main objective was to develop tunable cost-effective cell instructive interfaces resorting to marine polysaccharides/polyelectrolytes (PE’s) and autologous bioactive factors from human platelets lysate (PL) through layer-by-layer (LbL) assembling, a bottom-up approach for 2D/3D structures modification. The main objective was to verify whether the PL could be incorporated in the multilayered structures, and investigate how to tune the effect of the interface on several cell phenotypes. WORK PLAN Several 2D and 3D multilayered nano/sub-micron structures containing PL and marine origin PE’ such: κ-, ι-, λ-carrageenan (Car), chitosan and alginate, among other LbL parameters varied, were developed. Coatings were characterized and cell-inductive potential assessed. RESULTS Briefly, some of the major achievements are stated in this section. Mitogenesis. The nanocoating affected morphology and increased by 2-fold the proliferation of adipose derived stem cells (hASCs) – Fig.1.

FIGURE 1 hASCs morphology.

Angiogenesis. The nanocoatings could induce endothelial cells to form tube-like structures mediated

by the interaction of VEGF/FGF cell receptors and the growth factors incorporated – Fig.2.

FIGURE 2 Cell morphology, in presence and absence of VEGF/FGF receptors inhibitor.

Osteogenesis. In absence of the most common osteogenic inducer (dexamethasone, Dex), the multilayered structures could induce osteogenesis of hASC which effect varied with the assembling parameters – Fig. 3.

FIGURE 3 Calcium deposited (red) after 32 days.

Overall, results indicate that PL can be incorporated and the final cell instruction is tunable, which is an important feature to reduce variability associated with patient platelets and the own cells regeneration potential. With this methodology highly bioactive cell interfaces for several TERM applications can be developed. Provisional Patent Request n°20141000042365:“Layer-by-Layer with Platelet’s Derivatives and Uses Thereof” 51

A three-dimensional multibody model of the human ankle-foot complex Sérgio Gonçalves MIT Portugal Bioengineering Systems, IDMEC-IST, Instituto Superior Técnico, Universidade de Lisboa sergio.goncalves@tecnico.ulisboa.pt Background: Starting year: Research team: Supervisors:

Biomedical Engineering, IST, Portugal 2011 Tiago Malaquias1 Miguel Tavares da Silva1 1 IDMEC-IST, Instituto Superior Técnico, Universidade de Lisboa

OBJECTIVES The human movement, in particular the human gait, has been studied over time to understand the complex relation between the musculoskeletal and neurologic systems. Some of the most common neuromuscular disorders result in variations of the movement patterns, hindering the performance of daily tasks, such as walking, climbing stairs, etc. Multibody based methodologies have been applied in the study of the kinematics and dynamics of these patterns, enabling the development of new therapeutic approaches and medical devices optimized for each subject and each pathology. This way, a proper ankle-foot complex model is crucial to understand the interactions between the body and the ground and their influence in locomotion. WORK PLAN In this work, a novel three-dimensional foot-leg model with thirteen degrees-of-freedom, which includes four segments (toes, mid-forefoot, rear-foot and leg) modelled using six rigid bodies and five kinematic pairs, is proposed (see Fig. 1). A multibody formulation with natural coordinates is applied to describe the biomechanical system. The required anthropometric parameters were estimated to allow the anatomic modelling of this complex. The kinematic

description of the ankle considers the inclusion of the non-intersecting axes of the talocrural and talocalcaneal joints; a massless link formulation is applied to depict the offset between both joints, avoiding the modelling of the talus bone. RESULTS The reliability of the model was tested by applying it in the inverse dynamics study of the gait patterns of a male subject. The kinematic and dynamic outcome of these simulations are presented in Figs. 2a) and 2b). The obtained results are consistent with the literature, attesting the applicability and reliability of the model. The introduction of the Midtarsal joint allowed the analysis of other DOFs, which are usually not considered in the current models. The obtained patterns for this joint evidenced that the motion at this level is significant, as well as they enable the study of the foot spring behavior.[1]–[3]

FIGURE 2 a) Joint angular displacement (Left); b) Moment of force (Right).

References:

52 FIGURE 1 Schematic representation of the axes of rotation (red) and the DOFs of the model and the corresponding multibody formulation (blue and black).

[1] T. M. Malaquias, S. B. Gonçalves, and M. T. Silva, “A Three-dimensional Multibody Model of the Human Ankle-Foot Complex,” in EUCOMES - 5th European Conference on Mechanism Science, 2014, pp. 1–8. [2] T. M. Malaquias, S. B. Gonçalves, and M. T. Silva, “Development of a Three-dimensional Multibody Model of the Human Leg and Foot for application in Movement Analysis,” in IMSD - 3rd Joint International Conference on Multibody System Dynamics, 2014, pp. 1–10. [3] S. B. Gonçalves, T. M. Malaquias, and M. T. Silva, “Development of a Three-dimensional Multibody Model of the Human Leg and Foot for application in Movement Analysis (in preparation),” 2014.

Development of 3D in vitro models for prediction of hepatic metabolism and toxicity Sofia Rebelo1,2 MIT Portugal Degree Program, Faculdade de Ciências, Universidade Nova de Lisboa sofiarebelo@itqb.unl.pt Background: Starting year: Research team: Supervisors:

MSc/ University of Lisbon/Portugal 2010 Rita Costa1,2; Catarina Brito1,2; Paula Alves1,2 Paula M. Alves1,2; Catarina Brito1,2 1 Instituto de Tecnologia Química e Biológica (ITQB) – Universidade Nova de Lisboa (UNL), Oeiras, Portugal; 2 Instituto de Biologia Experimental e Tecnológica (IBET), Oeiras, Portugal

OBJECTIVES In vitro recapitulation of liver function is a valuable tool for the pharmaceutical industry, for toxicology testing, drug screening and target validation at early stages of the drug development. Moreover, humanbased in vitro systems play an important role in the clinics, for disease modeling and external support in patients with hepatic failure. The goal of my thesis was to develop physiologically relevant in vitro models of hepatic function, combining three-dimensional (3D) configuration with stirred tank bioreactors to attain a controlled, in-vivo like culture system.

Additionally, isolation of human hepatocytes obtained from ressected liver tissue of hepatectomized patients was optimized. Freshly isolated HH were cultured as spheroids in stirred tank bioreactors under physiological conditions (low oxygen and perfusion operation mode). As spheroids were formed, human Mesenchymal Stem Cells (hMSCs) were inoculated and its effects on hepatocytic function were assessed (Fig. 2).

WORK PLAN Two human hepatic models were used: HepaRG, a hepatoma-derived cell line, and primary cultures of human hepatocytes (HH). HepaRG is a bipotent line which differentiation is achieved after DMSO supplementation, despite its inherent effects on modulation of mechanisms of hepatic function. A DMSO-free strategy was developed for the differentiation of HepaRG, based on aggregation, alginate microencapsulation and culture in stirred conditions (Fig. 1).

RESULTS HepaRG microencapsulated spheroids present polarized hepatocyte-like cells and interconnected biliary network, while maintaining drug metabolism activity and biosynthetic functions. HH co-cultures with hMSC exhibit higher viability and maintenance of hepatocyte-specific functions. The work herein developed delivers systems with enhanced/sustained hepatic functions, suitable to predict hepatic metabolism. These newly developed 3D in vitro models contribute to the collection of tools available for preclinical drug development, increasing its efficiency and accelerating drug development pipelines.

FIGURE 1 Schematic representation of the strategy applied to develop a 3D model of HepaRG cell line.

FIGURE 2 Schematic representation of the strategy for the co-culture 3D model of human hepatocytes and hMSC.

Design of magnetic nanoparticles for cancer diagnostics and therapy Susana Palma MIT Portugal Bioengineering Systems, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa s.palma@campus.fct.unl.pt Background: Starting year: Supervisors:

MSc. Biomedical Engineering, Instituto Superior Técnico, Portugal 2010 A. Cecília Roque1 1 REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa

OBJECTIVES Production and characterization of iron oxide superparamagnetic nanoparticles (MNP) for biomedical applications such as cell labeling and cancer-theranostics with Magnetic Resonance Imaging (MRI). WORK PLAN Synthesis of MNP by the thermal decomposition of Fe(acac)3 in organic solvent followed by ligand exchange reaction to render the MNP hydrophilic; Functionalization of hydrophilic MNP with biocompatible and bioactive molecules; Characterization of the physiochemical, magnetic and relaxometric properties of the developed systems using transmission electron microscopy (TEM), dynamic light scattering (DLS), Fourier Transform Infrared spectroscopy (FT-IR), Thermogravimetric Analysis (TGA), Vibrating Sample Magnetometry (VSM), magnetic relaxometry and MRI phantom images; In-vitro evaluation of cell-particle interactions using the MTT assay, Prussian Blue staining, and characterization of the cell-labeling efficiency by Inductively Coupled Plasma and in-vitro MRI.

RESULTS Two natural hydrophilic polysaccharides were explored as stabilizing agents for MNP: gum arabic (GA) and a fucose-rich exopolysaccharide (EPS) produced by a in-house isolated bacterial strain (Enterobacter A47 (DSM 23139)). The resulting nanosystems consist of several 8 nm magnetic cores entrapped in a network of biopolymer, totalizing a hydrodynamic diameter of 235 nm for MNP-DMSA-GA and 160 nm for MNP-DMSA-EPS. These particles are efficient T2 MRI contrast agents (Fig. 1B) and possess a high r2/r1 value (350 for MNP-DMSA-GA and 132 for MNP-DMSA-EPS). In vitro experiments with HCT116 cell line show the effect of the MNP in cell viability (Fig. 1C) and yielded IC50 values of 110, 60 and 80 µg/(Fe)/ml for MNP-DMSA, MNP-DMSA-GA and MNP-DMSA-EPS, respectively. MNP-DMSA-GA are the most internalized particles, inducing a Fe concentration-dependent variation of contrast in in vitro MRI images.

FIGURE 1 (A) MNP production routes; (B) MRI phantom images of water dispersions of MNP-DMSA-GA (a) and MNP-DMSA-EPS (b) at increasing iron concentrations; (C) Evaluation of MNPs citotoxicity by MTT assay.

Rational design of bacteriophages as a platform for cancer therapy Tânia Mendes MIT Portugal Bioengineering Systems, Centre of Biological Engineering, University of Minho taniamendes@ceb.uminho.pt Background: MSc in Biological Engineering/ School of Engineering, Department of Biological Engineering, University of Minho / Portugal Starting year: 2012 Supervisors: Lígia Rodrigues1; Leon Kluskens1 1 Centre of Biological Engineering, University of Minho

OBJECTIVES The aim of this work is to engineer a bacteriophage-based platform to specifically target, invade and control cancer. WORK PLAN Firstly, a targeted drug-carrying bacteriophage will be constructed. Genetic manipulation to endow phages with cancer-specificity-conferring ligands, as well as chemical conjugation to load the targeted phages with cytotoxic drugs will be performed. Secondly, novel ligand peptides will be screened and selected to inhibit cell proliferation and induce cell death when coupled with drug-carrying phages. Phage display and iPhage technology to screen surface- and internalizing-peptide libraries will be applied. Thirdly, a phage-based gene therapy approach will be developed to regulate cancer proliferation and metastasis. Specifically, phages will carry zinc finger nucleases specific for mammalian target of rapamycin (mTOR) and ribosomal S6 kinase (RSK) pathways. Fourthly, the effect of temperature will be studied to induce modifications of portal and protein capsid conformation on phages, triggering nanoparticle disassembly

and cargo delivery. Finally, the mechanisms underlying internalization and trafficking of engineered phages will be elucidated. RESULTS Using molecular biology tools a filamentous phage, M13KE, was engineered to display a target-specific (231 peptide) and cell-penetrating peptides (Penetratin and TAT-HIV) in the pIII minor capsid protein in order to promote specific internalization, thereby facilitating the direct delivery of the drug. Engineered phages were conjugated with doxorubicin, an anti-carcinogenic drug, and incubated with breast cancer (MDA-MB-231) and breast epithelium (MCF10-2A) cell lines to evaluate internalization of phage particles and cytotoxicity. Up to now, there are good indicators that drug-carrying phages equipped with specific peptides for cancer cells can be specifically internalized promoting cell death. Acknowledgement: The author acknowledges the Lipid’NP’Phage project team (PTDC/SAU-BMA/121028/2010) for support given in the laboratory.

Development of a new nanostructured scaffold for neural stem/progenitor cell transplantation Tânia Vieira MIT Portugal Bioengineering Systems, Faculdade de Ciências e Tecnologias (FCT) – Universidade Nova de Lisboa (UNL) tssvieira@gmail.com Background: MSc. Biomedical Sciences, Universidade da Beira Interior, Portugal Starting year: 2012/2013 Supervisors: Célia Henriques1; João Paulo Borges1; Ana Sofia Falcão2 1 CEFITEC , Faculdade de Ciências e Tecnologias,Universidade Nova de Lisboa 2 iMED, Faculadade de Farmácia – Universidade de Lisboa

OBJECTIVES The complexity of the CNS and of its response to a SCI makes recovery of functionalities an unsolved clinical problem. The present investigation is supported by promising reports regarding the potential success of neural stem progenitor cells (NSPC) transplantation assisted by a scaffold. The work will be focused on the synthesis of new biodegradable polyurethanes, which will be used as the raw materials for the production of scaffolds through the electrospinning technique. The produced scaffolds will be evaluated in vitro. WORK PLAN The work will be divided into several tasks including (1) the production and characterization of new polyurethanes extended with natural polymers (chitosan and gelatin), (2) the fabrication and characterization of nanofibrous mats with different morphologies (by electrospinning), and (3) the evaluation of primary NSPCs adhesion, differentiation and viability on the produced fibrous mats. RESULTS Polyurethane based on chitosan (PU-CS) was successfully produced and characterized through 1H NMR (Figure 1), demonstrating the involvement of the CS on the final polyurethane structure. The synthetized PU-CS was used to produce nanofibers matrices through the electrospinning technique. The solvent system used to produce the solutions of PU-CS was tetrahidrofuran/dimethylformamide. Several solution concentrations, from 4 to 8%, were tested to verify their effect on the final fiber morphology (figure 2 A-C). For both concentrations the fibers possess a regular morphology without beads defects. The fibers diameter increased with the increase in solution concentration (figure 2 D). 56

FIGURE 1 1H NMR spectra of the synthetized PU-CS.

FIGURE 2 SEM images of electrospun nanofibrous matrices produced from several solution concentrations: (A) 4 %, (B) 6 % and (C) 8 %. (D) Average fiber diameter of the produced fibers.

Multidisciplinary optimization of wind turbine blade Abdolrasoul Sohouli MIT Portugal EDAM – Leaders for Technical Industries, Mechanical Engineering Department Instituto Superior Técnico, University of Lisbon abdolrasoul.sohouli@ist.utl.pt Background: Starting year: Supervisors:

Mechanical Engineering 2012 Afzal Suleman1 1 Mechanical Engineering Department, Instituto Superior Técnico, University of Lisbon)

OBJECTIVES This research seeks the development of a multidisciplinary optimization design approach by considering variable stiffness composite laminates in wind turbine blade. In this research, the effects of main wind turbine blades uncertainties and risk will be also studied to assure the design is less conservative and more cost-efficient. WORK PLAN The project will be divided in four tasks: Task 1: Development of Multidisciplinary Optimization Design A methodology and computational environment will be developed to obtain multidisciplinary optimization design for wind turbine blade by variable stiffness composites. The design drivers will be: • Design for structural performance (tip deflection, fatigue failure, frequency constraints, and light weight); • Design for manufacturability (Continuity of fiber path and curvature of paths); • Design for Cost. The wind turbine blade will be designed in two distinct levels: • Micro–structure and macro-structure (constituents, ply, laminate); • Sub-structures (spar cap, skin…).

Task 2: Reliability Analysis A set of ultimate designs from the Pareto frontier will be selected and then a reliability assessment will be performed for these designs against different failure modes. The selected designs should not be in highrisk region. Task 3: Cost Model A comprehensive cost model will be developed to analysis and compare with conventional components for making decision. The cost involves material, manufacturing and operating cost. Task 4: Decision Tree A decision tree will be modeled as decision support tool to review possible scenarios to ease judge for decision maker. RESULTS The outcomes of the work will be: 99 Fiber paths: Continues fiber paths of each components of wind turbine blade will be obtained as input for fiber placement machine. 99 A reliability assessment: The probability of failure for the new conceptual design will be calculated. 99 A cost model analysis. 99 Decision Analysis.

Improved methodologies for the design of extrusion forming tools Ananth Rajkumar MIT Portugal EDAM – Leaders for Technical Industries, Institute for Polymers and Composites (IPC), University of Minho rajkananth@gmail.com Background: Starting year: Research team: Supervisors:

European Masters in Material Science (University of Aveiro , Portugal. Aalburg University, Denmark. Hamburg University, Germany) Bachelors in Mechanical Engineering (Kanuya Institute of Technology,TamilNadu, India) 2012 Ananth Rajkumar1; João Miguel Nóbrega1; Olga Sousa Carneiro2 João Miguel Nóbrega1; Olga Sousa Carneiro1 1 Institute for Polymers and Composites, University of Minho 2 Soprefa, Componentes Industriais SA, Portugal

OBJECTIVES The main objective of this work is the establishment of new design procedures for the forming tools employed in the extrusion of thermoplastic profiles, namely the extrusion die and the calibration/cooling system. The usual experimental-based trial-and-error design approach will be improved through the employment of numerical modeling tools in combination with automatic design procedures. These aims require the development of adequate numerical tools to model the processes involved, and will be done in the framework of the open source computational library OpenFOAM®. The work will be undertaken in close cooperation with Soprefa, Componentes Industriais SA, a Portuguese company that has been designing and producing thermoplastic profiles for more than two decades. WORK PLAN The plan is to develop numerical routines based on the Finite Volume Method, to perform multi domain simulations for modelling the flow in the extrusion die and the cooling process in the calibrator. This methodology should handle complex geometries and also to take into account the shape changes occurring

in the extrudate between the extrusion die and the calibrator. This requires the implementation of proper free surface (interface tracking) boundary conditions, which will be done in the OpenFOAM® framework, taking advantage of the built in parallelized computations, and will be then associated with optimization procedures. The developed numerical routines will be assessed by experienced designers from Soprefa, using industrial case studies, in order to validate / improve the developed codes. The whole system will be then tested by less experienced designers, to assess its potential to minimize the resources and time spent on the design process, and to guarantee a better performance for the extruded profile. RESULTS The main zones and the parallel zone cross section of an extrusion die designed to produce T-shaped profile are shown in (Fig 1). The cross section of the profile comprises two areas (Elemental Sections – ES) with different thicknesses. Due to the differential flow restriction inherent to each ES it is difficult to achieve an even flow distribution, which will depend on the flow channel geometry (FCG). In order to facilitate the search of a FCG that promotes

60 FIGURE 1 (a) Lateral view of the flow channel of the profile extrusion die (indication of the main zones), (b) Cross section of the parallel zone of the extrusion die flow channel.

a balanced flow, the geometry was parameterized, and adequate geometry and mesh generators were developed. Having in mind the future implementation of automatic optimization approaches, an objective function (Fobj) is considered. This Fobj has a

minimum value of zero and its value decreases with the increase of the FCG performance. These effects were exemplified in a case study were the effect of the geometrical parameter L1 (shown in Fig 1.a) was illustrated. As shown in Fig. 2 and 3, both the Fobj and the velocity distribution are significantly affected by the value of L1, being the most balanced flow distributions associated with the lowest values of the Fobj.

FIGURE 2 Magnitude of the Fobj for different values of the geometry parameter L1.

FIGURE 3 Velocity contours at the flow channel outlet for different values of the geometry parameter L1.

Development of lightweight and cost-efficient exterior automotive body panels for electric vehicles André Camboa MIT Portugal EDAM – Leaders for Technical Industries, Faculty of Engineering, University of Porto andrecamboa@outlook.com Background: Starting year: Supervisors:

M.Sc. Industrial Design and Technology/ University of Beira Interior/ Covilhã 2010 João Nunes1; Fernando Alves2 1 MIT Portugal EDAM – Leaders for Technical Industries 2 INEGI, Faculty of Engineering of University of Porto

OBJECTIVES Current electric vehicles still present a considerable low autonomy range as major disadvantage, which with higher car price leads to a low public demand and, consequently, to very small production volumes. A problem that might be solved through several improvements, among them, the reduction of vehicle´s weight by recurring to low density materials and cost-efficient manufacturing processes. Relatively to the automotive exterior body panels, such problem narrows the available materials and manufacturing technologies, once those conventional are associated to higher tooling and manufacturing costs. Thus, polymer-metal hybrid (PMH) configurations based on polydicyclopentadiene

(PDCPD) arises as a relevant solution for both weight and cost reduction. WORK PLAN The development of PDCPD based PMH exterior body panels are followed by the early stages of design till their manufacturing. Several designs, mechanical simulations, mechanical tests, manufacturing tests and cost analysis were performed. RESULTS The preliminary cost results shows, for instance, that adhesive bonding is a promising assembly process beyond 5000 till a 35000 units, considering the component geometry analyzed in this research work.

FIGURE 1 Process-based cost model diagram.

62 FIGURE 2 Cost comparison by assembly type (Left); Maximum acceptable production volume by gluing.

Decision tool for sustainable supply chain design and planning Bruna Mota MIT Portugal EDAM – Leaders for Technical Industries, Department of Management and Engineering, Instituto Superior Técnico, University of Lisbon bruna.mota@tecnico.ulisboa.pt Background: Starting year: Supervisors:

Master of Biomedical Engineering from Instituto Superior Técnico, University of Lisbon, Portugal 2012 Ana Paula Barbosa-Póvoa1; Maria Isabel Gomes2 1 Department of Management and Engineering, Instituto Superior Técnico, University of Lisbon 2 Department of Mathematics, Faculty of Science and Technology, Nova University of Lisbon

OBJECTIVES The growing awareness of the limited availability of resources and the continued population growth are pressing governments and companies to work towards sustainable development. Due to the complexity of such problem very little research has been done on both the environmental and social components of sustainability within supply chain design context. This research aims to contribute to filling this gap through the development of a decision tool for the design and planning of supply chains that integrates the three pillars of sustainability: economic, environmental and social. WORK PLAN In a first phase of this research the development of a mathematical optimization model is expected. Due to the socio-economic complexity of the problem, as well as to the number of stakeholders involved, the mathematical optimization model is expected to be complemented with a multicriteria decision analysis. The main phases of this research are: 1. Literature review and preliminary research; 2. Supply chain modeling; 3. Development and integration of sustainability indicators; 4. Model validation through case-studies; 5. Multicriteria decision analysis.

RESULTS Literature review has shown that social indicators, which can be integrated in the mathematical optimization model, are nonexistent. In a preliminary research case study, a Social Benefit indicator which prefers the location of the network facilities in less developed regions was developed and introduced in a supply chain design and planning model as an objective function. A Pareto efficient frontier was established between the social and economic objectives, as shown in Figure 1.

FIGURE 1 Multi-objective approach for the maximization of Social Benefit with Cost limitation.

The results show how much an improvement in the social objective would cost to implement. This type of results is expected to provide important insights to both governments and companies in how to evaluate economic incentives for facility location.

The use of advanced technologies on lifecycle assessment of infrastructures Bruno Gonçalves MIT Portugal EDAM – Leaders for Technical Industries School of Engineering, University of Minho brunosfg@gmail.com Background: Starting year: Supervisors:

Masters in Industrial and Management Engineering, Production and Systems Department, University of Minho, Portugal) 2010 Paulo Lourenço1; José Matos1 1 ISISE – Civil Engineering Department, University of Minho

OBJECTIVES This PhD has 3 main phases/objectives: development of a mobile platform to streamline field inspections and infrastructures assessment; development of a probabilistic condition state index prediction model based on Markovian processes; integrate previous objectives and apply optimization tools for generation of scenarios and decision-making.

inspections classification. The inspection results will be converted into normalized indexes to be used in optimization models (considering the infrastructures performance and constraints) to trace the best scenarios for maintenance/intervention. These scenarios will help managers in the decision-making process, but the final decision will be always responsibility of the managers.

WORK PLAN Infrastructure management must be based in a whole-life costing model, which consists in determining the total cost of any infrastructure from its initial conception to the end of its service life. Whole-life costing models rely on the treatment of infrastructure’s data by application of several methods, analyses and tools. The quality of a WLC model will heavily depend on the quality of available data. Thus the quality of data gathered in field inspections is a key element of WLC models. Although great developments were made by scientific community to provide tools for data gathering, there are still problems with data quality due to subjectivity on field inspections, as it depends from who realizes it. Proper inspections may lead to lower costs in maintenance, repair and rehabilitation during the lifecycle of an infrastructure. In conditions of limited budget a good schedule of high quality inspections is required. An effective inspection system must be able to be operated by different type of users. A platform, for a tablet pc, was developed for inspection tasks in order to expedite the processes of storing and using information. The platform is remotely connected to the main database and presents standard inspection files, combined with the possibility of obtaining technical information of the element being inspected (such as historical data, previous inspections data, theoretical-technical information, etc.) in order to increase the inspections’ objectivity. This platform aims to make information available in real-time and aims to be a step forward to standardize and automatize the

RESULTS The objective of developing a mobile device application to standardize and streamline field inspections was successfully accomplished. The MIP although being a technical tool has a user-friendly interface design. It combines a very clean, simple and easyto-use interface design with technical information gathering and management. The Mobile Inspection Platform (MIP) (Figure 1 and Figure 2) implements a relational information scheme that correlates all the information gathered in field inspections allowing for detailed traceability. It also enables the desired “one-step” data handling being the collected data automatically stored. The MIP provides technical information to inspectors so that they can better assess the infrastructures. It also provides normalized inspection forms that lead to the same inspections type of outputs. The MIP enables inspectors to gather geo-referenced multimedia information (images and videos with GPS information) . The MIP introduces the 5-step process to detailed assess the condition state index for bridges that significantly reduces the degree of objectivity. In fact, all the implementations in the MIP lead to better inspections reducing the degree of objectivity, enabling inspections direct comparison through CS indexes, streamlining the flow of information with a clean, easy-to-use and effective user interface design. A mobile platform has always room for further developments triggered by scientific knowledge advancement and technical equipment advancements. So is the MIP.

FIGURE 1 MIP – “General Description screen” and “Components Selection screen”.

FIGURE 2 MIP – “Condition State Inspection screen” and “Standard Damages Identification screen”.

Integration of engineering, manufacturing and economic issues in design against impact of polymer automotive components Carlos Nuno Veiga Barbosa MIT Portugal EDAM â&#x20AC;&#x201C; Leaders for Technical Industries, Institute for Polymers and Composite, School of Engineering, University of Minho cbnuno@dep.uminho.pt | nnunext@hotmial.com Background: Master in Polymer Engineering/School of Engineering/University of Minho, Portugal Starting year: 2011/2012 Scientific supervisors: Ricardo SimĂľes1; Julio Viana2 Technical supervisors: Markus Franzen3; Thomas Baranowski3 1 Polytechnic Institute of Cavado and Ave 2 Institute for Polymers and Composites, University of Minho 3 Ford Research & Advanced Engineering Europe

OBJECTIVES It is well established that the thermomechanical conditions imposed during injection molding (IM) process affect the morphological development of a plastic part, thus determining its final mechanical properties. Also, the cost of the final part is a function of the processing conditions, geometry and material of the part and tooling. Along these lines, this work aims at developing an integrative methodology to establish relatively faster and accurately the most reliable and cost-effective design alternatives based on functional performance and manufacturing costs of IM plastic components. WORK PLAN The work plan with a timeline and milestones is shown in Figure 1.

FIGURE 1 Project schedule.

RESULTS The IM of the tensile-impact test specimens of two different polypropylene automotive grade materials was completed following a predefined design of experiment (DOE) plan. The mechanical characterization was carried out using state of the art equipment for static and dynamic (high-speed) tensile testing. Regarding the morphological characterization of the moldings, several techniques were applied to assess the main microstructural features: differential scanning calorimetry; polarized light microscopy; x-ray diffraction technique; and Raman spectroscopy. Flow simulations of the tensile-impact test specimens were replicated with the software Autodesk Moldflow Insight (AMI) 2012, according to the predefined DOE. The outputs of those simulations were used to calculate the thermomechanical indices through a dedicated computer application. This developed software offers a predictive capacity in terms of local mechanical properties. Further work encompasses the buildup of a cost model to predict manufacturing costs as well as a decision making approach at part level design. The integration of the flow simulation results with the structural analysis software packages (e.g. Radioss) and the respective experimental validation of the integrative methodology represent the final challenge of this work.

Selection of composite materials considering cost and environmental factors in the early phases of the design process Elçin Calado MIT Portugal EDAM – Leaders for Technical Industries, Mechanical Engineering Department, IST, University of Lisbon elcin.calado@tecnico.ulisboa.pt Background: Starting year: Research team: Supervisors:

Industrial Engineer/ Istanbul Technical University/ Turkey 2012 Arlindo Silva1; Marco Leite1; Tânia Estêvão2; Filipe Maia3; Marta Henriques3; Sara Perez Lopez3; Cristiana Perreira3 Arlindo Silva1; Marco Leite1 1 Mechanical Engineering Department, IST, University of Lisbon 2 CEIIA Prototype & Tests Unit 3 CEIIA Aeronautical Platform

OBJECTIVES The objective of this study is to create a material selection framework that will lead the designer to select the most suitable composite material for the aerospace applications based on the targets related to product performance, economic achievements and environmental impacts over a product’s life-cycle in a multi-disciplinary and multi-objective optimization scenario. WORK PLAN The material selection framework (Figure 1) requires three main tasks to be achieved.

orientation) that are non-dominated with the Pareto front concept. For that, the Direct Multi Search (DMS) [1] optimization is chosen as the solver. PRELIMINARY RESULTS This research study is performed with collaboration between CEIIA and Instituto Superior Técnico. The first and second tasks were done in a preliminary way with simplified methods. The study will continue with real case scenarios from CEIIA’s ongoing projects. A process based cost model(PBCM)[2] was developed specifically for the pre-impregnated composite fibers. After validating the link between FEM and DMS, the process based cost model of composite prototyping will be integrated to the material selection interface. In this way, second task will also be accomplished. The final algorithm of the material selection interface will be as shown below (Figure 2).

FIGURE 1 Material Selection Framework.

The first and second tasks, linking technical performance and economic performance to the material selection interface, have been completed. The third task is to calculate the environmental performance of the design such as EI99, total energy consumption, recyclability during the product’s life cycle. The calculations performed by Eco Audit (TBD) module will be integrated with the material selection interface as well. The framework will therefore analyze, from a database of materials and processes, what are the solutions (material, process, ply thickness and ply

FIGURE 2 Material Selection Interface Algorithm. Bibliography 1. A. Custódio, J. Madeira, A. Vaz, and L. Vicente, “Direct multisearch for multiobjective optimization,” SIAM J. Optim., vol. 21, no. 3, pp. 1109–1140, 2011. 2. M. Johnson and R. Kirchain, “Quantifying the effects of parts consolidation and development costs on material selection decisions: A process-based costing approach,” Int. J. Prod. Econ., vol. 119, no. 1, pp. 174–186, May 2009.

Integrating comfort in air-vents design Inês Costa MIT Portugal EDAM – Leaders for Technical Industries, Instituto Superior Técnico, Universidade de Lisboa ines.b.f.costa@tecnico.ulisboa.pt Background: MSc in Biological Engineering, Instituto Superior Técnico, Universidade de Lisboa, Portugal Starting year: 2011 Supervisors: António Ribeiro1; Mihail Fontil2 1 Mechanical Engineering Department, Instituto Superior Técnico, Universidade de Lisboa 2 Iber-Oleff, Portugal

OBJECTIVES Original equipment manufacturers (OEM) are becoming more interested in the sense of brand and comfort perception of clients, as differentiation strategy. However, these issues are highly subjective and difficult to define, thus the comfort requirements provided to suppliers are generally ill-defined. OEM suppliers usually perform several iterations in a trial and error base, before the approval of the final prototype. The current product development methods are not very systematic, require a lot of resources and do not assure a successful outcome. The main objective of this project is to develop a methodology to assist OEM suppliers in product development projects, leading to a higher rate of design project success and less iterations.

performance parameters. The results are schematically represented in Figure 2. In a second stage, we interviewed a panel of experts (open interview), in order to identify the percepts associated to the actuation feature. (More detailed results are under non-disclosure agreement)

WORK PLAN To achieve the proposed objective, we will create a model of comfort, which will correlate the physical performance of the product under development with the perception of the user. Therefore, the first phase of the work comprised: the identification and measurement of the performance parameters, through physical methods; and the identification and measurement of the percept, through survey methods. We will then use the identified parameters to model the user comfort perception.

FIGURE 1 Actuation feature of the air-vent.

RESULTS In this project, we are following the development of a new air-vent. Air-vent comfort can be divided in airflow (e.g. air dispersion) and actuation features (e.g. haptic feedback). We present here the results of the analysis of an actuation feature (Figure 1). This feature can be rotated and haptic and acoustic feedbacks indicate a specific position. We measured the performance of this actuation feature and we selected the important

FIGURE 2 Performance measurement and parameters identification.

New opportunities in vehicles design and development to improve efficient energy use – An engineering systems approach to improve vehicle’s eco-efficacy Irene Carvalho MIT Portugal EDAM – Leaders for Technical Industries, Mechanical Engineering, Instituto Superior Técnico, Universidade de Lisboa, Portugal irene.carvalho@ist.utl.pt Background: Materials Engineering/ Instituto Superior Técnico/ Portugal Starting year: 2009 Research Team: Arlindo Silva1; Ricardo Simões2,3 Supervisors: Arlindo Silva1; Ricardo Simões2,3 1 EDAM, Instituto de Ciência e Engenharia de Materiais e Superfícies, Instituto Superior Técnico, Universidade de Lisboa, Avenida Rovisco Pais, 1 1049-001 Lisboa, Portugal, arlindo.silva@ist.utl.pt 2 Escola Superior de Tecnologia, Instituto Politécnico do Cávado e do Ave, Campus do IPCA, 4750-810 Barcelos, Portugal, rsimoes@ipca.pt 3 EDAM, Institute for Polymers and Composites – IPC/I3N, University of Minho, 4800-058 Guimarães, Portugal, rsimoes@dep.uminho.pt

OBJECTIVES i. Understand in which extend a modular design of a passenger car plays a role in avoiding fuel consumption. The modular design of a vehicle is defined as a passenger car allowing a flexible removal/inclusion of rear parts of a vehicle, allowing minimum structural mass being used for passengers’ mobility, according to the load factor needs, per trip and during the useful life of the vehicle. ii. Bring new insights to the field of automotive innovation and European legislation aiming at reducing fuel consumption, and CO2 emissions, of passenger cars. iii. Conceive a modular vehicle design for personal mobility for validation. WORK PLAN i. Mathematical modeling: an analytical model has been developed to compute the potential effects in fuel consumption through a modular concept of a vehicle, quantifying their dependence on vehicle design parameters (the vehicle mass), and user behavior parameters (driving patterns of car use and driving cycles); ii. Car use and type choice behaviors: in order to understand if modular designs of a vehicle would be a market need (and type of need) and a want, four focus groups were conducted; a questionnaire is about to be launched; iii. Front-end design of a modular concept of a vehicle, for validation.

RESULTS

FIGURE 1 Variation of fuel consumption as function of use and design dependent parameters, for the use case of high use of the modular car in the lightest mode in urban cycle.

FIGURE 2 Relative values of m+ and m-, in kg, obtained for particular vehicle and user behavior.

Other results may be found in: 1. Carvalho, I., Baier, T., Simoes, R., & Silva, A. (2012). Reducing fuel consumption through modular vehicle architectures. Applied Energy, 93, 556–563. doi:10.1016/j.apenergy.2011.12.004. 2. Carvalho, I., Baier, T., Araújo, A., Kirchain, R., Simoes, R., & Silva, A. (2012). Reengineering the concept of urban cars: A case study. In 15th International Conference on Experimental Mechanics (Vol. 1, pp. 683–684). Porto, Portugal.

New roles for engineering and industrial design in the multidisciplinary development of highly complex products João Filipe Figueiredo MIT Portugal EDAM – Leaders for Technical Industries, Faculty of Engineering, University of Porto figueiredo.joao@fe.up.pt Background: Starting year: Supervisors:

Industrial Design & Technology, University of Beira Interior 2011 Nuno Correia1; Inês Secca Ruivo2 1 INEGI, Faculty of Engineering, University of Porto 2 Visual Arts Department, University of Évora

ABSTRACT The interdisciplinary relationship between industrial design and mechanical engineering has always been sensitive. The present research aims to develop a multidisciplinary methodological understanding of this relationship, taking into account a historical retrospective of engineering, industrial design and technology, in order to build a framework in tight accordance with current technological trends and social paradigms. Several relevant design processes are under detailed study and analysis, as their discussion

and comparison form the basis for the framework. Once this is completed, the work in association with research partners, concerning the ongoing methodology’s application, will begin, within a product development environment. Within this research, technology is proposed as the mediator between engineering and industrial design, in processes that have led to innovation. Moreover, in the conflict between technology and society lied the reform and regulation for design practice, in its broadest sense.

Hybrid CFRP/Steel composite bolted joints João Ribeiro Lopes MIT Portugal EDAM – Leaders for Technical Industries, Instituto Superior Técnico, Universidade de Lisboa joao.ribeiro.lopes@ist.utl.pt Background: Starting year: Supervisors:

MSc in Mechanical Engineering - Instituto Superior Técnico Universidade de Lisboa - Portugal BSc in Mechanical Engineering - Instituto Superior Técnico – Universidade de Lisboa – Portugal Ten years professional experience in aeronautic industry. 2011 Manuel de Freitas1; Pedro Ponces Camanho2 1 Instituto Superior Técnico, Universidade de Lisboa 2 Faculdade de Engenharia da Universidade do Porto

OBJECTIVES Recent research in the space industry has shown a significant increase in bearing strength of composite bolted joints can be achieved by a lay-up of metallic plies and composite plies in bolted areas, or a hybrid CFRP/Steel laminate. The objective of this research is to develop this technology to an aeronautical application where it will have to cope with new problems like fatigue, reparability, inspectability and cost-effective design. WORK PLAN The first part of the research deals with the mechanical adhesion between CFRP and austenitic steel. Two test programs were performed: Inter-Laminar Shear Stress and Single Lap Shear Stress. In both cases numerical models using the Finite Element Method (FEM) were built where the numerical results were compared with the experimental data. The available results show that the shear strength of the CFRP/ Steel interface is very close to the shear strength of the conventional CFRP/CFRP interface. The second part of the research deals with fatigue

tests of hybrid bolted joints. The objective is to compare the fatigue behavior of the hybrid CFRP/Steel with the conventional CFRP specimens. A FEM numerical model will be built and the experimental results will be compared with the numerical simulations. The third part of the research deals with cost. The objective is to measure the impact that this new technology has on manufacturing cost. The Process Based Cost Model (PCBM) will be applied in an aeronautical component in an industrial context. Two cost models will be built: A cost-model with the new hybrid CFRP/Steel joint and a similar cost-model with the conventional technology. RESULTS The overall result is to increase the Technology Readiness Level of this technology by assessing: • The mechanical performance of the hybrid joints, namely the static and fatigue strength when compared with conventional solutions. • The impact on cost of this technology; • Operational issues like inspectability and reparability.

Automatic detection of drusen associated with age-related macular degeneration in optical coherence tomography: A graph-based approach Jorge Miguel Gomes de Oliveira MIT Portugal EDAM – Leaders for Technical Industries, School of Engineering - Industrial Electronics Department, University of Minho, Portugal id4327@alunos.uminho.pt Background: Starting year: Supervisors:

MSc in Biomedical Engineering - Medical Electronics , University of Minho - School of Engineering - Industrial Electronics Department, Portugal 2012 Carlos Silva1, Manuel Ferreira2 1 Centro Algoritmi, University of Minho; 2Enermeter - Sistemas de Medicação, Lda / Centro Algoritmi - Universidade do Minho

OBJECTIVES The main objective of this work is the development of an automatic algorithm for drusen segmentation in Optical Coherence Tomography (OCT) volumes. Drusen are accumulations of materials at the bottom of the retina. Drusen’s volume can give valuable information to the physicians about the progress of the age-related macular degeneration, a hazardous and widespread disease. WORK PLAN This work is divided into four main tasks: literature review; implementation of a denoising method; development of a segmentation algorithm; building of a database. The main objective of the thesis is to develop a segmentation technique, implying that the implementation of the denoising method needs to be swift. Thus, some previous work, present in the literature, will be adapted for this problem. In this work, a graph-based technique will be developed. This technique will be based on graph cuts. The main reasons are: flexibility of the framework (can incorporate soft and hard constraints), immunity to local minima, trivial incorporation of 3D information and provable good results for similar problems. The previous works on graph cuts are not

specifically meant for handling drusen and that will be of one the contributions that this work will aim at. Another contribution is the utilization of new kinds of energy functions that can better model the problem (e.g. non-submodular functions). Additionally, new features might be integrated in the energy function (e.g. shape priors), always with the goal of improving the segmentation. To test the algorithm it will be necessary to construct a database that contains OCT volumes and respective manual segmentations by an expert. The database will be used to test the performance of the algorithm in conditions similar to clinical practice. RESULTS Quite a significant literature review has been performed and some denoising methods have been adapted to this problem (Figure 1). At the moment, options for the segmentation algorithm are being studied, implying that no segmentation results are available. An example of the results of a drusen segmentation algorithm is presented in Figure 2 to provide some insight of the objective of this work.

72 FIGURE 1 OCT image (a) and respective denoised version (b).

FIGURE 2 OCT image with large drusen (a) and the respective automatic segmentation result (b).

Development of 3D chitosan pieces for medical applications Lígia Figueiredo MIT Portugal EDAM – Leaders for Technical Industries, Department of Mechanical Engineering, Instituto Superior Técnico lmjfigueiredo@gmail.com Background: Starting year: Supervisors:

Master in Biomedical Engineering/Instituto Superior Técnico, Portugal 2012 Alexandra Rodrigues1; Frederico Ferreira2; Luís Pinto3 1 Institute of Materials and Surfaces Science and Engineering, Instituto Superior Técnico 2 Department of Bioengineering and IBB, Instituto Superior Técnico 3 Altakitin, S.A.

OBJECTIVES The aim of this research is to produce chitosan-based products to be used as (internal) bioasborbable fixation devices for orthopedic applications. To do so, it is necessary to find out relevant processing methodologies to produce chitosan-based products with predictable mechanical properties, while keeping the chemical functionalities intact. Those devices need to have the right strength, stiffness and hardness, which are necessary to allow the correct implant positioning, the movement and comfort of the patient until the biological tissue is reconstructed. Additionally, the constructed devices must be biocompatible, induce the bone formation and degrade over time while maintaining suitable properties profile.

be their mechanical strength, degradation profile and biocompatibility. Phase 2: To produce bioabsorbable implants based on chitosan, a production process was previously developed. However, it still needs improvements in order to be stable and produce implants with predictable properties. Therefore, methods to reduce the amount of bubbles generated during the dissolution were studied, which included the reduction of viscosity using temperature. Furthermore, the molds were optimized, which involved the usage of cellulose dialysis membranes (Figure 1 to Figure 2). The produced specimens will now start to be tested mechanically and analyzed by SEM to compare with the previous produced.

WORK PLAN This PhD work is divided in four main phases, which are complementary to each other. The Phase 1 consists in the market analysis; Phase 2 consists in the improvement of the current production process; Phase 3 will involve the addition of other materials (blends) to the chitosan; Phase 4 will correspond to the CE approval and Cost-effectiveness analysis of the implant. RESULTS Phase 1: Analyzing the global market of orthopedic implants, it was concluded that it is projected to reach US$ 46.5 billion by 2017 from an estimated US$ 21.1 billion in 2007. The largest segment of the orthopedic implant market is the reconstructive joint segment, where the knee reconstruction is the largest segment. An intensive literature review was made in order to analyze the mechanical and biologic properties of the materials that compose the current bioasbsorbable implants as well as to understand the advantages and limitations of such implants. The information gathered allowed to conclude that there are problems with the performance of the current implants and the main points of improvement should

FIGURE 1 Precipitation of the chitosan solution under an alkaline environment (NaOH).

73 FIGURE 2 Dense chitosan pieces obtained after the precipitation phase.

Reconfiguring facilities and logistics in dynamic supply networks of SMEs Maria Manuela Azevedo MIT Portugal EDAM – Leaders for Technical Industries, Faculdade de Engenharia da Universidade do Porto mmanuazevedo@gmail.com Background: Starting year: Research team: Supervisors:

Production Engineer / University of Minho / Portugal 2009 Jorge Pinho de Sousa1; José Crispim2; Stanley Gershwin3; Jaime Sá4; Paulo Oliveira5 Jorge Pinho de Sousa1; José Crispim2; Stanley Gershwin6 1 EDAM / Transportation System, FEUP 2 School of Economics and Management, University of Minho 3 EDAM/Department of Mechanical Engineering, MIT 4 General Management, Simoldes Plásticos 5 Logistic Management, Simoldes Plásticos 6 Massachusetts Institute of Technology

ABSTRACT In a world with increasing uncertainty and with accelerated changes, companies need advanced and innovative tools to deal with these situations and to respond quickly and efficiently. This is particular critical with SMEs (Small and Medium Enterprises) as they are very vulnerable to changes in normal business environments. Given the high variety of products manufactured simultaneously and the ever decreasing lifetime of these products, the need for adjustments in the layout of the companies also grows proportionally. Quite often machines are big and difficult to move, most of the time these changes do not happen or are continually being postponed. New machines are often located in sites creating difficult material flows, and thus decreasing their efficiency. At the same time it is important to look at the whole supply network, and not only at the company itself, as a way to understand how similar equipments are or can be used by others, or how they can be shared, to be used in an efficient way.

This project aims to develop a methodology or a set of tools to facilitate collaboration and information sharing between different entities (SME), to support the reconfiguration of facilities and logistic systems. This tools will guarantee higher levels of efficiency, flexibility and control of supply networks of SMEs, operating with complex, large added value products, characterized by low volume, large variety and customer centered production. With this project we expect to create a decision support framework capable of analyzing and assessing different policies and solutions in what concerns the reconfiguration of facilities, in particular by the redesigning currently layouts, and logistic systems, taking into account multiple criteria and perspectives. Such framework will help SMEs in the formation and operation of dynamic networks, able to quickly and efficiently respond to emerging market opportunities. Research results will be fully assessed and validated at Simoldes Plásticos Group.

Use of HP-RTM as a new possible manufacturing process for production of continues carbon fibre reinforced polymer Masoud Bodaghi MIT Portugal EDAM â&#x20AC;&#x201C; Leaders for Technical Industries, Department of Mechanical Engineering, University of Porto Masoud.bodaghi@fe.up.pt Background: Starting year: Supervisors:

Master of Material science - school of materials science and engineering / Iran 2011 Nuno Correia1 1 INEGI, University of Porto

OBJECTIVES Use of HP-RTM as a new possible manufacturing process for production of Continues Carbon Fibre Reinforced Polymer: (1)Determine effect of process parameters, such as resin velocity, viscosity, and location of gates and vents on mechanical performance of HP-RTM composite parts, (2) The compaction response of fibrous preforms subjected to the high pressure is of importance to understand this technique. Among other things, the compaction behaviour of reinforcement specifies thickness variation of the composite part, mould deflection and fibre volume fraction as well as the permeability. However, no analytical model of the compacted reinforcement under the high pressure has been published to date. It is expected that having an understanding of compaction behaviour will provide increased awareness of the process definition in order to simplify mould tool design. Furthermore, enhanced finished product quality will be plausible as a result of greater knowledge of micromechanical response of the reinforcement. (3) Among the parameters that influence resin flow, permeability of fibrous preform has a key role due to its importance for optimization of molding and infusion process: A quantitative evaluation of the uncertainty of permeability measurments in varaible thickness fibre reinforcements. (4) HP-RTM is a process that works under high flow rate and low viscosity. Because this is such a novel process no one has yet postulated a model for the description of its flow. One way to deal with the problem is to create a number of piecewise functions that, as a whole, work to describe the phenomena that occur inside the mould.

WORK PLAN

RESULTS

Sources of error resulting to systematic uncertainty in measured permeability

Planning and scheduling optimization in biopharmaceutical facilities Miguel Vieira MIT Portugal EDAM – Leaders for Technical Industries, Instituto Superior Técnico, Universidade de Lisboa migueljvieira@tecnico.ulisboa.pt Background: Starting year: Supervisors:

Masters in Chemical Engineering, Instituto Superior Técnico, Universidade de Lisboa 2011 Tânia Pinto Varela1; Ana Paula Barbosa Póvoa1 1 CEG-IST, Instituto Superior Técnico, Universidade de Lisboa)

OBJECTIVES The optimal scheduling of industrial operations is crucial for improving performance and adjust production flexibility to market demand. The application of modeling optimization to real industrial problems often stumbles either to represent the complex requirements of the manufacturing process, to tackle large temporal horizons or even to address operational parameters’ uncertainty. This research aims to develop efficient modeling techniques to solve large scale scheduling problems, able to identify optimal scheduling of tasks and resources. WORK PLAN The pharmaceutical sector has been dominated by chemically synthesized drugs. However, bio-based pharmaceuticals have been gathering particular attention recently due to its results in the treatment of diseases such as cancer, diabetes or growth disturbances. Biopharmaceuticals processes are mainly composed by two steps refereed as upstream and downstream processing. The mathematical formulation of scheduling problems has to accurately address the specific constraints of these steps, for example, the uncertainty associated with upstream processes parameters, or the complex operating conditions of downstream operations. The work plan consists in the identification of these different biopharmaceutical process constraints and the development of efficient model formulations for scheduling problems. RESULTS The initial study explored the identified gap within the scheduling of multi-stage multi-product biopharmaceutical processes, proposing the development

of a mixed integer linear programing model based on a continuous-time representation of the Resource Task Network formulation. The model addressed some of the main scheduling constraints of bioprocesses, mainly, the traceability of different production lots and the storage constraints of intermediate products regarding inherent shelf-life limitations. Figure 2 exemplifies the optimal model results of a scheduling problem, outlined by the biopharmaceutical production process of Figure 1, for the profit maximization.

FIGURE 1 Example of a two-stages biopharmaceutical production process.

FIGURE 2 Production schedule example.

Applying systems engineering approaches for quality improvement of manufacturing systems Muhammad Arsalan Farooq MIT Portugal EDAM – Leaders for Technical Industries, Department of Mechanical Engineering, University of Porto farooq.arsalan@fe.up.pt Background: Starting year: Research team: Supervisors:

Masters in Industrial Engineering, Erasmus Mundus Exchange Master Program, Germany – Romania – Italy. 2011 Sergio Tavares1 Henriqueta Nóvoa2; António Araújo2; Randolph Kirchain3 1 Institute of Mechanical Engineering and Industrial Management – INEGI, Porto 2 Department of Industrial Engineering and Management, University of Porto 3 Engineering Systems Division Lab, MIT

OBJECTIVES An international consumer goods packaging industry is facing a strong challenge to improve the quality of its products. Although the industry is producing products with a quality already above the international rules and regulations, customers are always in the quest of even higher quality and defect free products. Solving this problem is a big challenge for this company, not only due to the high production rates of assembly lines, but also due to the low cost of the final product. Considering the problem definition, the objective of the PhD research is to develop novel methodology to better understand relations between different manufacturing activities, with the final goal of reducing the risk of product failure as well as improving the final product quality. WORK PLAN Based on the objectives, the research has the following work plan. 1. State-of-the-art review; 2. Internship at aerosol manufacturing industry; a) Develop a new methodology of engineering system approach to quality management; b) Process mapping or modeling; c) Develop a tool based on Design Structure Matrix concepts. 3. Measuring system complexity using components modularity metrics; 4. Quantification of system components for the new tool; 5. Application of quality improvement tools and techniques; 6. Process and/or technology based cost modeling; 7. Thesis preparation. RESULTS In order to reduce system complexity and highlight critical manufacturing process points, a new tool – the Non Conformity Matrix (NCM) - was developed based on Design Structure Matrix (DSM) principles.

A 10-step methodology to apply NCM to industrial problems was proposed as shown in table 1. Also, Figure 1 shows NCM built (a) based on available information in the industry, (b) based on expert’s knowledge and (c) based on systems engineers knowledge. Simultaneously, engineering analysis together with quality improvement tools were applied, which resulted in highlighting a specific region of the system. In order to further explore the specific region pointed out in the previous analysis, Design of Experiments (DoE) was applied. The results from DoE showed significant factors that affect the product quality and recommended combinations of factors that could improve the manufacturing process. i. I dentification and definition of the defect or problem to be analyzed; ii. Identification of all NCs along the production line of a product; iii. Collection and analysis of all relations between NCs with clear explanations about each dependency (including interviews to operators, quality controllers, managers and engineers); iv. Transfer all the data to a NCM, parsed by manufacturing process, and evaluation of the final NCM; v. Apply mathematical operations (e.g. clustering and sequencing algorithms) to the NCM; vi. Evaluate and characterize the final NCM; vii. Apply quality improvement tools based on the previous selection of what are the critical quality characteristics (response variable) under analysis; viii. Improve the manufacturing process, according to the results; ix. Evaluate again the relations of NCs, deleting the NCs that were eliminated; x. Update and evaluate NCM. TABLE 1 10 step methodology for NCM implementation

FIGURE 1 The three NCMs.

Advanced in mould assembling technologies for high precision polymer based optical components Nelson Oliveira MIT Portugal EDAM – Leaders for Technical Industries, University of Minho Background: Starting year: Supervisors:

Master Integrated in Polymer Engineer/ Minho University/ Portugal 2009 António Pontes1 1 Polymer Department, Minho University

OBJECTIVES • Mapping of the technologic development needs and scientific base support in the in-mould assembling technologies; • To study the feasibility of the use of a laser head operating inside a mould for different operations and specifically for welding; • Development of an adequate cost model for in-mould assembling technology; • Development of a prototype mould including in-mould laser operating head and in-mould welding using the injection of a welding ring sealing, and • Tested with optical components for lightening automotive systems.

FIGURE 1 The laser laser head system.

WORK PLAN

Task 1. Mapping of in-mould assembling technologies Task 2. Development of a laser head system Task 3. Development of a prototype mould Task 4. In-mould assembling technologies validation RESULTS Task 2. Development of a laser head system To do the development of the welding process was used an IA20 robot controlled by a control unit NX100 and a diode laser Laser LM100 (Mergenthaler Laser Technology), equipped with a pyrometer LH500. The laser source and the control system of the robot are presented in the following figure 1. With this technology were welded several materials as PMMA with PC (mixed with carbon black). In the following figure 2 is presented the mechanical resistance of the seam.

FIGURE 2 Resistance of the welded PC/PMMA specimens.

Were also welded PP with PP (mixed with carbon black). In the following figure 3 is presented the mechanical resistance of the seam.

FIGURE 3 A new analysis for evaluate the resistance of the PP/PP seam

After the NIR analysis, was decided to study different material combinations. This had the objective of analyzing their implementation on the case study. The first material combination analyzed was the PC/ PMMA. They presented a good seam resistance. Different PPs were also studied. They presented a seam with smaller resistance compared to PC/PMMA. All the materials presented higher standard deviation.

Task 4. In-mould assembling technologies validation The main objective of this thesis was the development of a manufacturing cell with the capability of producing on the same moulding cycle a complete and functional subsystem. To accomplish this objective, several technologies were combined. In the following figure 4 is presented the equipments used in the manufacturing cell.

Task 3. Development of a prototype mould In the design of the mold was considered the laser welding and the in-mold assembling. To do these two processes in the same cycle was necessary to design an assembling system. The system uses a two the rotary plates that had the objective to transport the different parts to the assembling and welding zone. The plates had also the objective to eject the final subsystem. In the following figure 4 is presented the two halves of the mould.

FIGURE 5 Manufacture cell.

The case study produced with this technology is presented in the figure 6.

FIGURE 4 The mould.

FIGURE 6 Final rear lamp.

Transition from Product-focused and Service-focused design methods to a Product-Service Systems design method: using a user-centered design approach in the front-end phase Nina Costa MIT Portugal EDAM – Leaders for Technical Industries, Mechanical Engineering Department, Faculty of Engineering, University of Oporto ninacosta89@hotmail.com Background: Starting year: Supervisors:

Bachelor in Technology and Product Design/ University of Aveiro/ Aveiro/ Portugal. Master in Industrial Design/ Faculty of Engineering, University of Oporto/ Porto/ Portugal 2011 Lia Patrício1 1 Department of Industrial Engineering and Management, Faculty of Engineering University of Oporto

OBJECTIVES • Study and develop a Product-Service System (PSS) method that can support both manufacturing and service companies in moving from traditional product-focused or service-focused methods, to the design of integrated PSS solutions; • Facilitate communication between stakeholders with different backgrounds through visualization tools capable of supporting the co-creation process and definition of requirements for new PSS solutions.

4. R eflection and systematization of the results obtained through the PSS methods and tools tested in real context and creation of a new PSS method; 5. Overall, the above mentioned stages follow a design research approach through in which new methods are developed and validated, through an iterative process of conceptualization, application and reflection.

FIGURE 1 From customer contextual analysis to PSS Requirements and design of Solutions.

RESULTS • Create a framework with existing PSS methods and tools; • Provide a new method applicable for different types of industry, for supporting the co-creation process, definition of requirements and design of new PSS solutions; • Clarify the new designer’s role and value within the co-creation process; • Create a visual toolbox that enhances constructive thinking and enables interaction with customers and stakeholders with different backgrounds.

WORK PLAN 1. Literature review and systematization of existing methods and tools for designing Product-Service Systems (PSS): analyzing their advantages, disadvantages, and potential applications in industry; 2. Development of a new PSS method that incorporates product-design and service-design methods for the design of integrated PSS solutions; 3. Apply the PSS methods developed through case studies in product-focused design and service-focused design companies for the development of concrete solid new PSS solutions;

FIGURE 2 General guideline to design integrates PSS in the front-end phase of the design and development process.

An engineering model to address kinesthetic and acoustic percept requirements in automotive human-machine interfaces design Nuno Miguel Pereira dos Anjos Valverde MIT Portugal EDAM – Leaders for Technical Industries, Instituto Superior Técnico, Universidade de Lisboa nuno.miguel.valverde@tecnico.ulisboa.pt Background: Starting year: Supervisors:

M.Sc. in Mechanical Engineering, IST-UL 2011 António Relógio Ribeiro1; Mihail Fontul2 1 Grupo de Vibrações e Ruído, Instituto Superior Técnico, Universidade de Lisboa 2 IBER-OLEFF, SA

OBJECTIVES In the automotive industry, manufacturers are trending their marketing in the sense of pleasure and emotion. Moreover, automotive OEMs seek to establish their specific brand feeling: not only a perception of quality but individuality in the usage of their cars that distinguishes them from competition. As shown in Figure 1, we are developing a research in the framework of designing Human-Machine Interfaces (HMI) for automotive applications. We aim to achieve an engineering model for assisting the design of automotive dash-board interfaces when psychophysical and subjective perceptual requirements are given. The case-study is a product from a Portuguese manufacturer, supplier to a foreign OEM, where the client has imposed such ill-defined requirements. WORK PLAN We developed a Design of Experiment (DOE), obtaining a set of mechanical push-buttons where design variables have been changed. The sample set has perceptible differences in tactile and acoustic behaviors, but remains visually identical to the user. Measured data and other engineering parameters are measured and paired along subjective evaluations in a Kansei Engineering methodology.

We aim at translating perceptions and feelings into adequate physical properties to the case-study in order to satisfy kinesthetic and acoustic percept requirements for the final product. The knowledge areas and overall workflow are summarized in Figure 2. RESULTS We’ve measured the kinesthetic behavior (force-displacement curves) for actuating the sample push-buttons and recorded their sound in a full anechoic chamber. From this measured data, we identified and obtained values for engineering parameters. Further, we’ve treated acoustical parameters with psychoacoustics, focusing in very short (burst) sounds. We are performing a set of interviews with expert and non-expert population to evaluate the samples. Our further work aims at using adequate data analysis techniques, including adapting and applying evolutionary algorithms, establishing a model that contributes to the design of automotive HMIs.

82 FIGURE 1 Research problem and main research goals.

FIGURE 2 Research study-case, knowledge areas and overall workflow.

On the development of internal legitimacy in intentional innovation networks Ricardo Greenfield MIT Portugal EDAM – Leaders for Technical Industries Mechanical Engineering – Design Studio / CESE, Faculty of Engineering of the University of Porto / INESC-TEC ricardo.greenfield@gmail.com Background: Starting year: Supervisors:

MSc Electrical and Computer Engineering / Faculty of Engineering of the University of Porto/ Portugal 2011 António Lucas Soares1; Ana Barros2 1Department of Informatics Engineering / UESP, FEUP / INESC-TEC 2 UESP, INESC-TEC

OBJECTIVES Due to their positive effects on firm and regional performance, many attempts to create intentional innovation networks have been undertaken by policy makers, groups of SMEs, large hub-firms, universities and research institutes. However, intentional innovation networks tend to have a smaller success rate than innovation networks that emerge naturally, and further insights in how to increase the success of intentional innovation networks are therefore desired but still lacking in both academia and practice. Previous research indicates that the establishment of early internal legitimacy in intentional networks is a good indicator of future network success. Despite this, the development of internal network legitimacy in intentional network formation remains under- researched. This PhD project aims to: 1) – Identify the drivers influencing the development of internal network legitimacy; 2) – Identify in which contexts these drivers can be influenced by human action; 3) – Develop a measurement instrument for the assessment of internal

network legitimacy; and finally 4) – Develop a methodology supporting the application of best practices for the establishment internal network legitimacy. WORK PLAN The methodology employed to accomplish these goals will be that of case studies for theory building, along with a quantitative study to create the internal legitimacy measurement instrument. Two embedded multi-case studies shall be conducted at the network level with networks in the German and Portuguese industrial context. Each of the two embedded multi-case studies shall consist of multiple network cases. RESULTS Findings are expected to generate knowledge regarding how to adjust intentional innovation network operations to increase chance of future network success. Finally the measurement instrument may be used in future as a generally applicable metric for the comparison of different networks.

The user's emotions and experience problem: Guidelines to support the development of new institutions centered on users’ emotions and experiences Rita Rodrigues MIT Portugal EDAM – Leaders for Technical Industries, Mechanical Engineering Department, Faculdade de Engenharia da Universidade do Porto mdi11015@fe.up.pt/dli12002@fe.up.pt Background: Starting year: Supervisors:

Master degree in Industrial Design, Faculdade de Engenharia da Universidade do Porto, Portugal 2012 João Manuel R. S. Tavares1 1 Mechanical Engineering Department, Faculdade de Engenharia da Universidade do Porto

OBJECTIVES The aim of this project is to study the user experience and emotions inside a specific institution, and see their relation with the facility layout and other elements, such as, for example, the signage and informational symbols. The objective is to investigate the factors of the layout and other physical elements that can be improved to provide positive emotions and a better experience to the user. WORK PLAN Literature Review phase – 1st year Collect scientific papers, books and other types of literature, and study those documents in order to define a course of action for the project depending on the gaps identified. Observation and Information collection phase – 2nd year 1st semester The type of institution to be studied will be selected; when access to the institution is granted, the aim will be to get in touch with the users and employees and to collect the information important to this research. Some research methodologies will be used in order to collect their experiences and its relation with the facility’s environment.

Definition and evaluation of guidelines | Thesis Development – 2nd semester of 2nd year and 1st semester of 3rd year The information collected will be used to produce the expected main out coming of this project: a set of guidelines. These guidelines will be established according to the information collected that indicates that there can be improvements in some of the elements of the institution, and then, if possible, the intention is trying to evaluate those guidelines. Thesis Development – 2nd semester of 3rd year This semester will be dedicated to the thesis development and writing. RESULTS The aimed main result of this research will be to define a set of guidelines that can be used by the developers of new institutions, and that can help them on avoiding problems that the existing institutions have. These guidelines will help them to build the best environment in the institution that will lead to a better experience and better emotions felt by the users when using the services provided.

NetWave: Managing dynamic networks through SME collaboration Senay Sadıç MIT Portugal EDAM – Leaders for Technical Industries, Faculdade de Engenharia da Universidade do Porto ssadic@inescporto.pt Background: Starting year: Supervisors:

MSc. in Industrial Engineering / Istanbul Technical University / Turkey 2009 Jorge Manuel Pinho de Sousa1; José António Crispim2 1 Department of Industrial Engineering and Management, University of Porto 2 Department of Management, University of Minho

OBJECTIVES As an emerging business form derived from supply chain collaboration “Dynamic manufacturing networks (DMN)” represents goal-oriented networks of Small and Medium Enterprises (SMEs) that utilize rapid network formation tools to quickly respond immediate business opportunities. Forming DMNs is frequently addressed as a survival and sustainability tool in to operate in global markets. This thesis aims to develop a methodology and several tools to support SME Networks in DMN formation process. Our objective in this work is not to just come up with a DMN formation tool to assist production assignment and lot sizing decisions but more of a Decision Support platform that tracks performances of partner SMEs and maintain a fair, sustainable and profitable collaboration with optimal operational decisions.

Netwave Project plans to add insight into Dynamic Manufacturing Formation and SME Collaboration literature streams, by developing new methodologies, processes and ICT decision support tools. RESULTS • A framework to support order promising process of SME manufacturing networks in and track individual and network-wide performances. • A multi-objective DMN formation and operational planning tool to help SMEs in managing manufacturing and logistics processes. • Preliminary tests of the new methods.

WORK PLAN The thesis basically has 3 main components: Literature Review: • Identifying the current state of art and research opportunities in dispersed SME networks and DMN formation Order Promising Module • Designing an Order Promising Process to Support formation of DMNs. • Supporting performance tracking, rescheduling modules and provide a platform that aligns operational and strategic decisions Dynamic Network Formation and Operations Tracking Module • A Multi Objective MILP Model to support DMN formation and Operational Plans in SME networks

Integrated management of residential energy resources (Energy Box) Ana Soares MIT Portugal Sustainable Energy Systems, University of Coimbra argsoares@inescc.pt Background: M.Sc. Energy for Sustainability/ University of Coimbra/ Portugal and M.Sc. Electrical and Computers Engineering/ University of Coimbra/ Portugal Starting year: 2010 Research Team: The ongoing research, of which some results are presented here, is part of the Energy Box project. This project is a cooperation between the University of Coimbra and the ESD-MIT and aims to develop, redesign and extend the concept of Energy Box initially proposed in Livengood and Larson (“The Energy Box: Locally Automated Optimal Control of Residential Electricity Usage”, Service Science, 1, 1-16, 2009). http://www.uc.pt/en/org/inescc/Projects/ongoing_projects_pub/Energy_Box Supervisors: Carlos Henggeler Antunes1; Álvaro Gomes1 1 Dept. of Electrical and Computer Engineering, University of Coimbra and INESC Coimbra

OBJECTIVES The aim of this research is to design a methodology to be implemented in an energy management system for controlling domestic energy resources in a smart grid scenario in order to minimize domestic end-user’s electricity bill, and maximize end-users’ satisfaction and the integration of local generation. The methodology takes into account the existence of dynamic tariffs, indicative or emergency signals, and quality of service constraints as well as the uncertainty associated with demand and renewable generation in order to obtain robust decisions. WORK PLAN In order to accomplish the aims of this research, the following steps have been defined: • analysis of the characteristics that allow part of the demand to be treated as a manageable and responsive resource; • analysis of technical issues associated with each energy resource (loads, local generation, storage); • categorization of loads to be the target of automated demand response actions and analysis of the characteristics of those actions (short time interruptions, re-set of temperature parameters, postponement or anticipation of working cycles); • characterization of the different input signals (electricity prices, indicative and emergency signals, users’ preferences, etc.) to be considered; • development of models able to reproduce electricity demand, micro-generation and energy storage systems; • design and implementation of management algorithms (e.g. using evolutionary algorithms to cope with the combinatorial and nonlinear features of the models) to schedule the use of diverse end-use resources with economic and/or quality of service advantages to the consumer.

• assessment of the impacts on the load diagram and comfort conditions caused by the implementation of the actions performed by the energy management system. RESULTS The methodology being developed is expected to help users to take advantage of different alternatives of using energy services to reduce the electricity bill while keeping the desirable levels of comfort and the quality of the energy services provided (Figure 1 and Figure 2).

FIGURE 1 Example of a Pareto optimal front obtained using genetic algorithms to solve the multi-objective problem considering energy cost and end-user dissatisfaction as objective functions.

FIGURE 2 Example of a solution displaying the schedule of the operating cycle of loads in a planning period.

A model for an aggregator of energy management systems â&#x20AC;&#x201C; a meta-heuristic approach Andreia M. Carreiro MIT Portugal Sustainable Energy Systems, DEEC-FCT, University of Coimbra andreia.melo.carreiro@gmail.com Background: Starting year: Supervisors:

Biomedical Engineer 2010 Carlos H. Antunes1; Humberto M. Jorge1 1 Dept. of Electrical and Computer Engineering, University of Coimbra and INESC Coimbra

ABSTRACT/OBJECTIVES The increased penetration of renewable generation in the electric power system has been leading to a higher complexity of grid management due to its inherent intermittency, also with impact on the volatility of electricity prices. Setting the adequate operating reserve levels is one of the main concerns of the System Operator (SO), since the integration of a large share of intermittent generation requires an increased amount of reserve that is needed to balance generation and load. At the same time, the energy consumption in households has been steadily growing, representing a significant untapped savings potential due to waste and load flexibility (i.e., the possibility of time deferring the use of some equipment). An aggregator has been designed to operate as an intermediary between individual energy management systems (local energy boxes - LEB) and the SO/Energy Market (EM) capable of facilitating a load follows supply strategy in a Smart Grid context. The aggregator is aimed at using the flexibility provided by each end-user aggregated into clusters, via contracts, of demand-side resources to satisfy system service requirements from the SO, involving lowering or increasing the power requested in each time slot, thus contributing to the balance between load and supply, avoiding peaks in the load diagram, and coping with the intermittency of renewable sources, thus offering an attractive alternative to supply side investments on peak and reserve generation.

For this purpose, an optimization model has been developed to maximize the aggregator benefits, taking into account revenues from the SO/EM and payments to end-user clusters as well as functional and technical constraints. An algorithmic approach based on genetic algorithms and differential evolution meta-heuristics has been developed to deal with the combinatorial and dynamic nature of this model, which is supplied with real input data.

FIGURE 1 Energy Box Aggregator - information flows between the grid (SO/EM) and the end-users (clusters).

Decision support tools for the planning of MSW management systems António Lorena MIT Portugal Sustainable Energy Systems, IN+ Center for Innovation, Technology and Policy Research, Instituto Superior Técnico, Universidade de Lisboa antonio.lorena@ist.utl.pt Background: MSc Environmental Engineering (Instituto Superior Técnico, Universidade de Lisboa / Portugal) Starting year: 2010 Supervisors: Paulo Ferrão1; Samuel Niza1 1 IN+ Center for Innovation, Technology and Policy Research, Instituto Superior Técnico, Universidade de Lisboa

OBJECTIVES The urbanization process has come with extensive environmental and health impacts, a large part due to the improper disposal of municipal solid waste. The waste management sector is the human response to the ever-growing fluxes of discarded materials with harmful impacts to the environment and human health. The significant environmental and socio-economic impacts of the MSW management sector calls for the use of Decision Support Tools (DST) to assist the decision-makers, experts and other stakeholders in the decision analysis and participatory processes. In this work, an integrated decision support tool for MSW management planning is proposed. This tool allows to assess the environmental, socio-economic and energy related impacts of MSW management. WORK PLAN The proposed decision support tool can be described as a Waste Input-Output (WIO) model. This model considers the monetary and physical interactions between the economy, final demand (households) and the MSW management sector. This allows to capture not only direct impacts, but also indirect and avoided impacts along the supply chain of the studied sector. The environmental, socio-economic and energy related impacts for each economic and MSW treatment activity are assessed separately and included as parameters in the model. The model is then used with original forecasting models of MSW production to do scenario analysis. RESULTS Several scenarios are analyzed with the proposed model. The reference scenario is the Portuguese MSW sector in 2012. Other studied scenarios include Portugal 2020, Portugal 2030 and Portugal Zero Treatment, among others. For example, the

Portuguese MSW management sector in 2012 represented 419 M€ in Gross Value Added (GVA) (Figure 1). In 2020, in accordance to the proposed Plano Estratégico para os Resíduos Urbanos, the MSW will represent 563 M€ in GVA. Regarding employment, in 2012 the sector had a global impact of more than 14.000 jobs, growing to more than 17.500 by 2020.

FIGURE 1 Global impacts in GVA (M€) in reference scenario Portugal 2012 and Portugal 2020.

FIGURE 2 Global impacts in number of jobs in reference scenario Portugal 2012 and Portugal 2020.

New approach to sustainable urban retrofit with energy performance simulation strategy (Case Study: Coimbra, City Center) Atefeh Salehi MIT Portugal Sustainable Energy System, Faculty of Sciences and Technology, University of Coimbra salehi.atefeh@student.dem.uc.pt Background: Starting year: Supervisors:

Master degree in Architecture / Tabriz Islamic Art University / Iran) 2012/2013 Maria Isabel Morais Torres1; Álvaro de Oliveira Gomes1; João Paulo Cardielos1 1 Faculty of Sciences and Technology, University of Coimbra

OBJECTIVES Nowadays, with the excess new construction and the desertification of historic centers, the concept of rehabilitation/renovation has found a primordial importance to convert the city centers to “living centers” again. But, the rehabilitation cannot be done in a disorganized way. The rehabilitations in city center are currently regarded as an aspect of sustainable development in the international context. Building retrofit is also a vital part of emission reduction and using energy in a more efficient and sustainable way. There are several key issues like mobility, water, waste, material, population, energy, CO2 emission and land use related to building retrofit and all of these sections have interrelation together and could make a model for sustainable building retrofit. WORK PLAN This research thesis aims to provide a radical change in current knowledge for urban sustainability by technical pathway to develop a framework for shaping the systematic retrofitting on old city centers. It will explore a more holistic approach to retrofitting in order to explore the gap between technical solutions with the theories of sustainable and smart city through identifying the sustainable retrofit techniques in urban scale. So, this research, as a theoretical and practical contribution, will consider the rehabilitation of urban fabric by taking into account the energy performance planning in order to obtain a higher level of quality of life and sustainable environment by integrating the rehabilitation concerns of technical, environmental and political implications.

RESULTS Accordingly, the subject of this research thesis is a multi-scale modelling tool to support environmental and energy performance planning in the holistic level by a case study analysis.

The main goal of this research thesis is sustainable urban retrofit addressing to multi-disciplinary subject from different perspectives and advance using the simulation of buildings energy performance in the holistic systematic design approach that combines understanding of both the building and the system. The renewable energy deployment opportunities at different scales (building- urban block) - from the resource potential (regional, topographical, climatic aspects) to technical feasibility and relation to demands (built environment, obstructions, human factors) using Geographical Information System (GIS) tools will be studied, by this multicriteria approach. As, the historic center of Coimbra was recently considered as a UNESCO world heritage site. Paying attention to the potential future of this heritage is the main reason for choosing it as a case study.

FIGURE 1 Some pictures of the case study.

Blending optimization of biodiesel produced with waste and virgin oils Carla Caldeira MIT Portugal Sustainable Energy Systems, University of Coimbra caldeira.carla@dem.uc.pt Background: Starting year: Research team: Supervisors:

Chemistry and Environmental Sciences, Faculty of Sciences, University of Lisbon, Portugal 2013 Carla Caldeira1,2; Elsa A. Olivetti3; Randolph Kirchain3; LuĂs Dias1; Fausto Freire2 Luis Dias1; Fausto Freire2 1 INESC-C, Faculty of Economics, University of Coimbra 2 ADAI- LAETA, Faculty of Sciences and Technology, University of Coimbra 3 Materials Systems Laboratory, Engineering Systems Division, MIT

OBJECTIVES The main aim is to develop a multiobjective optimization (MO) model to optimize blends of virgin and waste oils (WO, i.e., waste cooking oils and animal fats) for biodiesel production that minimize costs and environmental impacts in a life-cycle perspective, incorporating uncertainty and variability. Analyses based on this model will allow industry stakeholders to obtain blends for biodiesel production in compliance with technical standards that are efficient in terms of cost and environmental impact, and will inform policy makers on the effects of changes to biodiesel regulations. WORK PLAN The initial model is a bi-objective optimization mathematical formulation with the goal to minimize costs and life-cycle Greenhouse Gas (GHG) emissions, focused on the feedstocks currently most used in Portugal for biodiesel production. Prediction models for biodiesel properties, based on the chemical composition of the oils, are used to establish constraints. In the next step, a stochastic version of this model will be developed using Chance-Constrained Programming (CCP). This approach will consider the uncertainty and variability (U&V) existing in the oils chemical composition and will determine the raw materials and quantities to be blend for a given confidence level. The complexity of the problem increases when a raw material with high compositional U&V, such as WO, is considered. RESULTS The initial bi-objective model was solved using the Ć?-constraint method and the resulting Pareto curve

revealed the trade-off between costs and GHG emissions, from which it was possible to calculate GHG abatement costs. Analyzing the blends along the Pareto curve, a reduction in GHG emissions was obtained by progressively replacing rapeseed by soya and reducing the palm share in the blend used for biodiesel production (Fig 1).

FIGURE 1 Pareto curve and blends. Model constrained according the EN 14214.

Ongoing research focuses on implementing the chance-constrained stochastic approach and to include WO together with virgin oils in the blend.

Sustainability assessment of building retrofit: An integrated life-cycle approach Carla Rodrigues MIT Portugal Sustainable Energy Systems, Department of Mechanical Engineering, University of Coimbra carla.rodrigues@dem.uc.pt Background: Starting year: Research team: Supervisors:

Architecture (2008), MSc. Energy for Sustainability (2013), University of Coimbra, Portugal 2012 Fausto Freire and researchers from Center for Industrial Ecology; Jeremy Gregory1 Fausto Freire2 1 Concrete Sustainability Hub and Materials Systems Laboratory, Massachusetts Institute of Technology 2 Faculty of Sciences and Technology, University of Coimbra

OBJECTIVES The main goal of this research is to assess retrofit strategies for improving the environmental and economic life-cycle (LC) of residential and commercial buildings in Mediterranean climates. Major retrofit measures are currently being modelled in two buildings (one residential and one commercial) to perform a critical review and assessment of two building environmental assessment (BEA) methods and certification tools (e.g. BREEAM (UK), LEED (US)). WORK PLAN An integrated cost and environmental life-cycle model combined with thermal dynamic simulation are being developed and implemented to different building envelope retrofit measures for the selected buildings. Final and primary energy, environmental impacts and (equivalent annual) costs are being assessed and trade-offs will be identified. A sensitivity analysis will be incorporated in order to increase the robustness of the results, by analyzing occupancy patterns, energy price, etc.

RESULTS A comprehensive assessment of the building envelope retrofit has already been performed. A LC impact assessment performed for the residential building shows that, for roof insulation thicknesses greater than 80 mm, the reduction in energy consumption during the use phase, due to a further increase of 40 mm, is not significant (< 5%), while there is an increase of 6 to 20% of the environmental impacts associated with the embodied phase (Figure 1). Polyurethane foam and expanded polystyrene present the lowest total LC impacts for the roof and exterior walls, respectively. Insulation placed inside of the exterior walls has lower total LC impacts than placed outside (Figure 2). A sensitivity analysis was performed to assess the influence of the occupancy level in the whole LC of the building. LC impacts decrease about 20% from low to high occupancy level (due to a 40% decrease of energy requirements during the use phase). The marginal benefit of insulating the exterior walls is higher (about 35%) in the high occupancy level than in the low occupancy level (about 20%). This research intends to support building stakeholders (sustainability consultants, users and designers), policy makers and the LCA community.

94 FIGURE 1 Life-cycle environmental and primary energy assessment of the roof insulation options: RW, XPS and PUR (0, 40, 80 and 120 mm); per 1 m2 of living area over a period of 50 years

FIGURE 2 Life-cycle impact assessment of the exterior walls insulation options: no insulation, RW and EPS; inside (40 and 80 mm) and outside (40 mm); per 1 m2 of living area over a period of 50 years.

Large scale impacts of energy and power demand management: The Portuguese case Catarina Sabino MIT Portugal Sustainable Energy Systems, Energy for Sustainability Initiative, University of Coimbra csabino@student.dem.uc.pt Background: Starting year: Research team: Supervisors:

MSc in Energy and Environmental Engineering, Faculty of Sciences of the University of Lisbon, Portugal 2012 António Martins1 António Gomes Martins2 1 Electrical Engineering Department, FCT – University of Coimbra, EMSURE Project 2 Energy for Sustainability Initiative, University of Coimbra

OBJECTIVES The main goal of this work is to analyze the effect, caused by the implementation of Demand-side Management (DSM) measures in the Portuguese residential sector, on energy consumption, on economy and on GHG emissions. The Portuguese electric power system, some economic aspects of the country, and general characteristics of the Portuguese population concerning energy saving behavioral will be considered. The results will help understanding and quantifying the effects and benefits of three DSM measures – peak shaving, load shifting and strategic conservation – and will support those decision makers responsible for implementing and designing energy efficiency measures in planning, choosing and designing instruments for DSM and behavior change programs before implementation. WORK PLAN A country-level analysis will be performed using the Portuguese MARKAL/TIMES model to quantify the energy and economic savings and GHG emissions reduction caused by peak shaving, strategic conservation and load shifting induced by habitual and investment behavior changes. Specific behavioral characteristics of the Portuguese population related to energy savings in the residential sector, and characteristics of the Portuguese electric power system will be linked to the model, as well as issues related to the new concept of smart grid (electric vehicles, micro-generation systems and smart meters). The main stages of the work are: 1. Collect data; 2. Perform a meta-analysis of the collected data; 3. Define and model scenarios, using the MARKAL/TIMES model; 4. Integrate energy consumer’s behavior issues in the model; 5. Integrate smart grids and electric vehicles issues in the model.

RESULTS Four criteria were assessed for 25 different residential equipment: i) the ownership rate of the equipment, ii) its annual energy consumption, iii) its load diagram shape and iv) the availability of general quantitative data. According to these criteria, lighting systems, washing machine, refrigerator, television, computer and dishwasher are the types of equipment with higher potential for actions targeting behavior to be analyzed in this work. A set of common consumers’ behavior, as well as its effect on the electricity consumption, was identified for the equipment. For each one, an average adoption rate was estimated based on data from previous published studies, which reported surveys (launched in Portugal) in one of the phases. Average values were determined weighted by the samples sizes.

FIGURE 1 Common behaviors and their influence on electricity consumption by type of equipment.

All the behaviors were categorized according to the type of loads they represent: 1) investment dependent loads: all the identified equipment except the television and computer, 2) deferrable loads: washing

FIGURE 2 Loads prone to energy conservation actions: adoption rate of behaviors (weighted average of the adoption rates of different studies) and â&#x20AC;&#x153;virtualâ&#x20AC;? sample size (the sum of the samples sizes of each study).

machine and dishwasher, 3) loads responding to changeable parameter settings: refrigerator and 4) loads prone to energy conservation actions: all the identified equipment. Deferrable loads: in Portugal, the dual rate tariff is subscribed by only 15 % of the consumers. Moreover, according to two studies only 21 and 33 % of the washing machines and dishwashers, respectively, have an option to postpone its operation which is actually used by the users. Loads responding to changeable parameter settings: the operating temperature of the majority of the refrigerators (in Portugal and all over the world) is significantly above the ideal (5 Â°C), and thus this type of load does not represent a relevant energy saving potential.

FIGURE 3 Investment dependent loads: typical replacement rate of the equipment, prevailing energy efficiency class (EEC) and percentage of equipment with an EEC inferior to the prevailing class.

Improving methodological development of life cycle sustainability assessment Chongyang Du MIT Portugal Sustainable Energy Systems, Department of Mechanical Engineering, University of Coimbra yangyangeco@gmail.com Background: Starting year: Supervisors:

Master of Environmental Management / Duke University / USA 2012/2013 Fausto Freire1; Luis Dias2 1 ADAI- LAETA, Faculty of Sciences and Technology, University of Coimbra; 2 INESC-UC Faculty of Economics, University of Coimbra;

OBJECTIVES Life Cycle Sustainability Assessment (LCSA) is nowadays the only life-cycle based sustainability assessment tool with the focus on a product throughout its life cycle. A Life Cycle Sustainability Assessment study is composed of three dimensions of the assessment including Environmental Life Cycle Assessment, Social Life Cycle Assessment, and Life Cycle Costing. This doctoral research project aims at contributing on the methodological development of Life Cycle Sustainability Assessment, testing the feasibility of LCSA through case studies, and providing decision support for policy makers according to LCSA results. WORK PLAN • Understanding the status-quo theoretical development of Social Life Cycle Assessment through a comprehensive literature review • Testifying the feasibility, practicability, and limitations of Social Life Cycle Assessment through case study • Contributing on the methodological development of Social Life Cycle Assessment based on the experience from case study • Modifying the case studies by implementing the improved Social Life Cycle Assessment methodology and comparing with the original results • Conducting Life Cycle Costing and Environmental Life Cycle Assessment case study based on the same product system settings • Integrating results from S-LCA, LCC and LCA and providing decision support based on the results through multi-criteria decision analysis RESULTS We have conducted a comprehensive literature review on Social Life Cycle Assessment, and ready to move forward to apply the gained knowledge on conducting our own SLCA case study. Social Life Cycle Assessment (SLCA) is developed as

a methodology to assess the positive or negative social and socio-economic impacts or potential impacts along the product life cycle. Although there have existed a number of tools focused on assessing social aspects of sustainable development, SLCA differs itself from other tools by its objects on products and services, and its scope concerning the entire life cycle. Regarding the young history of SLCA, it has evolved tremendously in the last years with several important documents published by UNEP/SETAC providing a framework and guidance on conducting SLCA. The framework proposed in the Guidelines is in line with the ISO14040 and 14044 standards for Life Cycle Assessment with some adaptations for the social aspects. Therefore, SLCA follows the four main iterative phases in LCA, namely definition of the goal and scope, life cycle inventory analysis, life cycle impact assessment, and life cycle interpretation. A number of questions worth further research in the domain of SLCA such as indicator selection, data sources, data type, impact assessment methods and interpretation of the results.

FIGURE 1 UNEP/SETAC Guidelines (2009) and Methodological Sheet of Social Life Cycle Assessment (2013).

A sustainable approach towards 100% renewable energy for all electric services on islands Christian Wimmler MIT Portugal Sustainable Energy Systems, Faculty of Engineering, University of Porto c.wimmler@fe.up.pt Background: Starting year: Research team: Supervisors:

MSc. / Newcastle University / UK 2011 Golnar Hejazi1 Eduardo de Oliveira Fernandes2; Carlos Moreira3; Stephen Connors 4 1 MIT Portugal Sustainable Energy Systems, Faculty of Engineering, University of Porto 2 Mechanical Engineering Department, Faculty of Engineering, University of Porto 3 Electrical Engineering Department, Faculty of Engineering, University of Porto 4 Massachusetts Institute of Technology

OBJECTIVES Islands often are confronted with severe energy challenges especially those far from the main land which operate as isolated energy systems. In those cases electricity, the queen of the energy vectors, shall be obtained from diverse sources to alleviate the burden of fossil fuels. Thus, the generation of electricity from renewable energy sources in combination with electricity storage becomes an irrecusably challenge in the nearby future. A conceptual methodology was established to support decision makers in defining an appropriate energy mix to cover all electric services from renewables. WORK PLAN A 5-step work plan was defined. Step 1 assesses the current and forecasts the future demand. Load profiles, representing each hour of the year, are created. Thereby, aspects such as the amount of energy to be covered from electricity, energy efficiency and peak shifting are considered to create diverse scenarios. Step 2 pre-selects technologies, whereas only natural conditions and site characteristics are applied to reduce the number technologies for a given site location. Only suitable technologies are evaluated within a sustainability assessment (step 3) where each remaining technologies undergoes multi-criteria-decision making (MCDM). Technical, economic, environmental and social criteria are applied. Step 4 determines the energy mix. Thus, a time-series approach was chosen to identify combinations of renewable energy technologies that require low storage capacities to cover all electricity needs over the year. In step 5 an analysis of the overall system, including technology mix and storage capacity, for each scenario is performed by applying MCDM. Hence, the most appropriate scenario is identified. 98

RESULTS The inevitability for energy carrier shifts towards electricity could be proved. For islands insights into the new energy paradigm could be gained. As offshore technologies emerge and develop, the likelihood of having those technologies covering large shares of the energy mix surge.

FIGURE Example of load profile within new energy paradigm.

FIGURE Example of covering future demand solely on renewable energy resources and storage systems.

Assessment of diffuse radiation models in Azores Clarisse Magarreiro MIT Portugal Sustainable Energy Systems, Departamento de Engenharia Geográfica, Geofísica e Energia, Faculdade de Ciências - Universidade de Lisboa clmagarreiro@fc.ul.pt Background: Starting year: Supervisors:

MsC/ Faculdade de Ciências - Universidade de Lisboa / Portugal 2012 Pedro M. M. Soares1; Miguel C. Brito1 1 Departamento de Engenharia Geográfica, Geofísica e Energia, Faculdade de Ciências -Universidade de Lisboa)

OBJECTIVES The present study explores the applicability of different existing correlation models of diffuse fraction and clearness index or other plain parameters to the Azorean region. WORK PLAN Reliable data provided by the Atmospheric Radiation Measurements (ARM) Climate Research Facility from the Graciosa Island deployment (May 2009 - December 2010) of the ARM Mobile Facility (http://www.arm. gov/sites/amf/grw) was used to perform the analysis. The hourly models used were selected considering a minimum input of meteorological parameters, models’ algorithmic ease of use and reported model performance. A broad range of locations from where models derived was also taken into account. Performance of the studied models was then assessed by means of standard statistical errors (MAE; MAPE; RMSE and MBE) and by probability density functions (PDFs) matching score (S). RESULTS Models only based on the clearness index underestimated diffuse irradiance in their higher values, approximately after 300 Wm-2.

Boland-Ridley-Lauret model (BRL) presented the best results expressed not only in the statistical errors values but also in the PDF matching score. Even so, exhibiting a MAPE of 23%; a RMSE of 35 Wm-2 and BIAS of -10 Wm-2, is still far from being ideal. It is clear that a more suitable model for the Azores region is required.

FIGURE 1 Probability density functions for the tested models and measured data.

99 TABLE 1 Statistical performance of the assessed models.

Thermal storage in isolated minigrids to optimize renewable integration Diana Neves MIT Portugal Sustainable Energy Systems, SESUL, Faculty of Sciences, University of Lisbon dpneves@fc.ul.pt Background: Starting year: Research team: Supervisors:

Master on Energy and Environmental Engineering; Bachelor in Meteorology, Oceanography and Geophysics, both by Faculty of Sciences, University of Lisbon, Portugal 2009/ stand-by 2010-2012/ expected to finish in 2015 Carlos A. Silva1 Carlos A. Silva1; António Vallera2 1 WS Energia Chair, IN+, Instituto Superior Técnico, University of Lisbon 2 SESUL, Faculty of Sciences, University of Lisbon

OBJECTIVES Evaluate if thermal storage by hot water solar systems, can be used to integrate more renewable resources on isolated mini grids, and which decisions of energy management and demand response, optimize best the electric dispatch. Corvo island, in Azores is used as a case-study, and integrates the project Green Islands.

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WORK PLAN • Literature review on micro-communities with hybrid renewable energy systems (HRES): ○○ D. Neves, C. A. Silva, and S. Connors, “Design and implementation of hybrid renewable energy systems on micro-communities: A review on case studies” Renew. Sustain. Energy Rev., vol. 31, pp. 935–946, Mar. 2014. • Analyzing the demand patterns of electricity and modeling the hot water demand with the implementation of Solar thermal systems and heat pumps: ○○ D. Neves and C. A. Silva, “Modeling the impact of integrating Solar Thermal Systems and Heat Pumps for Domestic Hot Water in electric systems - The case study of Corvo,” Renew. Energy, vol. under revi, 2014. • Optimizing domestic hot water electric loads with demand response: ○○ D. Neves and C. A. Silva, “Optimal electric dispatch on isolated mini-grids, using genetic algorithms as demand response strategy,” submission in process, 2014. • Comparing different software in modeling the mini grid of Corvo and integration of renewable supply for electricity. • Test optimization of demand response with renewable resource forecast with genetic algorithms on own model. • Compare which investments, accounting with energy and storage costs, are more valuable towards sustainability of the island.

RESULTS From literature review on stand-alone systems: D. Neves, C. A. Silva, and S. Connors, “Design and implementation of hybrid renewable energy systems on micro-communities: A review on case studies,” Renew. Sustain. Energy Rev., vol. 31, pp. 935–946, Mar. 2014. • Current implementation of HRES is limited by existing systems and end-use energy; • Most common configuration is Diesel + Wind + Solar; • Renewable resource supply is limited by storage systems and hardly go over 80% on total energy delivered. Looking at thermal storage, through solar systems and heat pumps, and its hourly impact in the grid by electric backup, we modeled different demand profiles and assumptions relating to the hour of backup: D. Neves and C. A. Silva, “Modeling the impact of integrating Solar Thermal Systems and Heat Pumps for Domestic Hot Water in electric systems - The case study of Corvo,” Renew. Energy, vol. under revi, 2014.

FIGURE 1 Comparison of off-peak backup model with disperse off-peak model for winter week 10 (winter).

• H eat pumps are responsible for the major increase in load (around 5%) and solar thermal systems for the increase in peak load (during winter days, when solar doesn’t respond completely to demand).

TABLE 2 Quantitative comparison of best models with different formulations. TABLE 1 Final energy and peak load comparison between different models.

Using genetic algorithms to optimize the daily economic dispatch of DHW load, we used different formulations to test which was best: D. Neves and C. A. Silva, “Optimal electric dispatch on isolated mini-grids, using genetic algorithms as demand response strategy,” submission in process, 2014.

FIGURE 2 Qualitative comparison of best daily models with different formulations.

• From DHW backup needs, we could only re-place the solar thermal backup with the GA optimization, since heat pumps work on a continuously cycle; • We used to formulations of power re-placement: in terms of nº of switched on systems (GA systems) and in terms of amount of energy (GA energy), finding the second formulation more sensitive to daily demands smaller than the installed capacity, and that is the case even in winter with only 63 kWh of daily demand; • The model “fill 170kW” was done by optimizing the two working generators for 65% of their capacity, using 170 kW as constant dispatch every time the base load as inferior; • We encountered that, even with big increases in energy demand, the DHW backup, if well placed, can optimize the daily dispatch with none or fewer increase in the costs; • Preferably the DHW load should meet off-peak hours in order to get advantage of the full capacity of working generators, and avoiding to meet the heat pumps backup hours or peak hours; • Storage of hot water can act as demand response on the grid.

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Environmental sustainability assessment of soybean and palm biodiesel systems: A life-cycle approach Érica Geraldes Castanheira MIT Portugal Sustainable Energy Systems, ADAI-LAETA, Center for Industrial Ecology, University of Coimbra erica@dem.uc.pt Background: Starting year: Research team: Supervisors:

MSc in Energy and Environmental Management, University of Aveiro, Portugal 2008/09 Fausto Freire1; João Malça1; Luís Dias2; Randolph R. Kirchain3; Ece Gulsen3; Elsa Olivetti3 Fausto Freire1 1 ADAI-LAETA, Center for Industrial Ecology, University of Coimbra 2 INESC Coimbra, University of Coimbra 3 Engineering Systems Division, MIT

OBJECTIVES In Portugal more than 50% of biodiesel is produced from imported soybean and palm oil. The increasing global consumption of these commodities for bioenergy purposes has been accompanied by a growing concern about their impacts, including the potentially high environmental impacts associated with intensive land-use practices and land-use change (LUC). My PhD thesis presents an environmental sustainability assessment of biodiesel systems. Critical modeling issues were addressed and assessed through Life-Cycle (LC) models applied to different chains, pathways and scenarios for biodiesel produced from soybean and palm cultivated in South America.

calculated based on different approaches and different LC impact assessment methods were adopted. A sensitivity analysis of alternative multifunctionality procedures for dealing with co-products (allocation and substitution) was performed. The GHG savings from replacing diesel with palm and soybean biodiesel were also assessed, with the aim of providing support for Portuguese companies in calculating and meeting the GHG saving criteria for biodiesel presented in the European Renewable Energy Directive (RED). SELECTED RESULTS

WORK PLAN A comprehensive evaluation of the implications of alternative scenarios was carried out. The scenarios addressed were LUC, palm fertilization schemes, soybean cultivation systems, biogas management at palm oil extraction mill and soybean transportation. LUC emissions were also calculated based on the expansion of the actual Colombian palm area and the actual soybean area (in four Brazilian states). Nitrogen field emissions were

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FIGURE 1 GHG intensity of soybean biodiesel (energy allocation), considering 3 pathways, 4 Brazilian soybean origins and the actual expansion (1985 to 2006) of the soybean area.

FIGURE 2 GHG savings from replacing diesel with palm (a) and soybean (b) biodiesel using various multifunctionality approaches: substitution (scenario A and B), energy, price and mass allocation.

Enhancing freedom and ensuring efficiency and security: Improving the electricity market in Brazil via virtual reservoir model Felipe Calabria MIT Portugal Sustainable Energy Systems, Faculty of Engineering, University of Porto (FEUP) felipecalabria@gmail.com Background: Starting year: Supervisors:

Master of Science/ Federal University of Pernambuco (UFPE)/ Brazil 2012 João Paulo Tomé Saraiva1 1 Department of Electrical and Computer Engineering, Faculty of Engineering, University of Porto

OBJECTIVES The main objective is to develop and test a new market design for electricity markets with large penetration of hydros. Through a virtual reservoir model, it is intended to enhance the freedom for participants bear their contracts without compromising the levels of security of supply and efficiency in the use energy resources. WORK PLAN

RESULTS Several countries embarked on a path characterized by market liberalization in order to implement decentralized and competitive structures. Brazil, which presents a power system with strong predominance of hydros with multiple owners and where more

than 75% of the total installed capacity comes from renewable, has made significantly progress towards liberalization. Nevertheless, the short-term market price is calculated by computational models based on dynamic and linear programming and, therefore, it is not the result of market participant interactions. Moreover, the analysis performed on the market completeness shows that the conciliation between market design and physical operation of the power system doesn’t occur smoothly. Generators assume their commercial commitments through long and medium-term contracts. However, the dispatch is centralized and they are not allowed to decide their own generation in order to bear their contracts, which expose them in the short-term market. In addition, the mechanism implemented to share this risk doesn’t allow generators to develop their own strategy and risk perception. The proposed model aims to replace this mechanism by adding an energy right account scheme through a virtual reservoir model. Moreover, through this model the short-term price formation will be influenced by the interaction of participants and it can be designed to produce hourly prices via a day-ahead market.

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Planning generation in liberalising and decarbonising electricity systems: The Portuguese case Filipa Amorim MIT Portugal Sustainable Energy Systems, Instituto Superior Técnico, University of Lisbon, Portugal filipa.amorim@tecnico.ulisboa.pt Background: Starting year: Supervisors:

Graduated in Economics/ FEUC, University of Coimbra, Portugal 2007/2008 Victor Martins1; Jorge Vasconcelos2; Patrícia Pereira da Silva3 1 ISEG Department of Economics, University of Lisbon 2 Instituto Superior Técnico, IN+, University of Lisbon 3 FEUC Department of Economics, University of Coimbra

OBJECTIVES To study the current transitions of electricity systems in EU and understand the rising generation costs of electricity prices. This implies understanding the introduction of free market rules in electricity sectors and the transition to decarbonised electricity systems. Given the complexity and singularities of each case and context, the Portuguese case has been chosen because it exemplifies a liberalising electricity system that already has integrated with success a high penetration of intermittent renewables. In this sense, it well embodies how the diagnosed problems will be generalized in all Europe in the medium-term.

FIGURE 1 Electricity generation contractual categories relative to demand.

WORK PLAN The existing subsidies to generation have been quantified through their whole duration in time after a consistent subsidized producers’ database has been built because there is no public access to this information. A prospective economic analysis of the Portuguese electricity system has been carried up to year 2030, based in the developed producers’ database cost model, and up to year 2050, through a newly developed integrated Portugal-Spain TIMES model (The Integrated Markal-Efom System). RESULTS Potential for a competitive wholesale generation market in Portugal: One has to wait until 2020 to achieve 33% of expected demand to be satisfied in the free market and until 2025 to reach 67% share (Figure 1). There is indeed very limited room for new generation plants, beyond what is already granted or planned, to cover expected average demand growth.

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Generation legacy contracts costs to final consumers: Total generation legacy costs will grow until 2017 (absolute higher peak) and again until 2020 (second relative higher peak) as illustrated in Figure 2.

TABLE 1 Legacy costs scenarios.

Total generation costs to final consumers: In 2020, if the market price is 60 €2011/MWh, total generation costs will ascend to nearly 6,000 M€2011, the case in which legacy generation costs represent more than 75% of total generation costs. If the market price is 80 €2011/MWh, these will ascend to

6,450 Mâ&#x201A;Ź2011 and if market price is 100 â&#x201A;Ź2011/MWh to 6,970 Mâ&#x201A;Ź2011. End-user generation costs balance and future prices trends: Consumers may expect to benefit from State guaranteed prices in the cases of an annual average market prices growth of 3.8% or more (real terms). To note that the estimated natural gas prices increases are in Europe between 3.7% and 4.4% until 2015 (IEA, WEO 2012). Electricity generation prices will continue in ascension.

Benefits from the integration of decarbonising systems: The Iberian Case: Despite Portugal is a small country within an isolated Iberian electricity market, the analysed roadmap to 2050 under almost full decarbonisation supports the conclusion that there is room for improved overall efficiency and lower risk, while further investing in endogenous low carbon generation resources in Iberia and that Portugal becomes a net exporter.

FIGURE 2 Aggregate electricity generation legacy contract costs.

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Increasing energy access for isolated communities Golnar Hejazi MIT Portugal Sustainable Energy Systems, Faculty of Engineering, University of Porto Golnar.Hejazi@fe.up.pt Background: Starting year: Research Team: Supervisors:

MSc. /Newcastle University/UK; BSc./Iran University of Science and Technology/Iran 2011 Christian Wimmler1 Eduardo de Oliveira Fernandes2; Manuel Matos3; Stephen Connors 4 1 MIT Portugal Sustainable Energy Systems, Faculty of Engineering, University of Porto 2 Mechanical Engineering Department, Faculty of Engineering, University of Porto 3 Electrical Engineering Department,Faculty of Engineering, University of Porto 4 Massachusetts Institute of Technology

OBJECTIVES Energy and poverty are comprehensive concerns that affect over 2 billion people. To solve the issues and correspond to the â&#x20AC;&#x153;energy for allâ&#x20AC;? paradigm, solutions to increase energy access and improve the comfort level of life are highly valued. Providing a livable planet for all through the ages shall be guaranteed. While most energy solutions are tailored towards modern energy systems, they are neither replicable nor applicable to respond to the needs of isolated communities in developing countries. Sustainable energy solutions to fulfill the energy needs by considering local resources and dispersed energy generation are sought. Hence, it is intended to develop a methodology appropriate for remote areas far from the grid. A bottom-up approach is defined to identify closest to ideal solutions to cover at least the minimum energy needs. WORK PLAN The initial stage assesses the current energy needs, the potential of renewable sources and designs a hypothetical village. The research proposes a methodology to cover the minimum energy requirements. Hence, several technologies are identified and selected to cover the main useful services within the village. Based on the evaluation with sustainable criteria, a pre-selection of technologies is made. Scenarios based on the combination of pre-selected technologies to cover all services are built. Thus, a mathematical formulation is established. Considerations of attributing energy to all required services, rather than a single one, are made. In the

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FIGURE 1 Conceptual framework for energy access in isolated communities.

further process only scenarios that satisfy the energy access level are designated. Each scenario is evaluated based on different aspect by the use of multi-criteria decision making to identify the most appropriate solutions. RESULTS The initial analysis confirms that most areas are far below minimum energy access. The solutions are presented in a conceptual, systematic and mathematical approach. Indeed, the defined approach promises to be a high impact concept that changes for a better life.

Energy consumption and certification in the Portuguese residential sector Gonçalo Pereira MIT Portugal Sustainable Energy Systems, Center for Innovation, Technology and Policy Research - IN+, Instituto Superior Técnico goncalo.duarte.pereira@tecnico.ulisboa.pt Background: MSc. Mechanical Engineering/Instituto Superior Técnico/Portugal Starting year: 2011 Supervisors: Carlos Silva1; Susana Vieira2 1 Center for Innovation, Technology and Policy Research - IN+, Instituto Superior Técnico, Universidade de Lisboa 2 Center of Intelligent Systems - IDMEC - LAETA, Instituto Superior Técnico, Universidade de Lisboa

OBJECTIVES Evaluate the energy consumption and certification in the Portuguese residential sector; Study energy storage options for the residential sector. WORK PLAN Using a smart metering pilot, evaluate the differences between estimated (from energy certification) and measured consumptions; Evaluate the main drivers for energy consumption for the residential sector; Evaluate energy storage options for different end-uses. RESULTS In Figure 1 is shown the relationships between dwelling features for a smart metering case study in Portugal. The feature grouping gives insight into the importance of dwelling, equipment and behavior related features. In Figure 2 it is shown the relationship between two of the features, previously shown in Figure 1, with natural gas consumption and exterior temperature. Analyzing Figure 2 it is visible that dwellings with the central heating equipment have a quadratic relationship with exterior temperature due to the environment heating function of the central heating. The tankless boiler equipment, since it only has the water heating function, displays a linear relationship with exterior temperature. Figure 2 also shows that only bellow 16°C the dwelling’s inhabitants start to use the environment heating function of the central heating. Since the energy certification process assumes that dwellings are maintained at a constant 20°C during the heating season (i.e. Winter) there is a clear gap between measured and estimated energy consumption. Furthermore, the more efficient boilers used for central heating end up consuming more energy than their tankless counterparts for periods without environment heating needs (i.e. more efficiency ≠ less consumption).

FIGURE 1 Dwelling features relationships.

In Figure 3 is shown the gap for different energy classifications. As is visible in Figure 3, dwellings with higher energy classifications have higher final energy consumptions than it is estimated by the energy certification process. As a final remark, please note that the energy consumption targets established by the energy certification decrease with better energy classifications.

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FIGURE 2 Relationship between natural gas consumption, central heating, tankless boiler and exterior temperature.

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FIGURE 3 Comparing estimated and measured energy needs.

Extended life cycle assessment of low energy houses in the Mediterranean context Helena Monteiro MIT Portugal Sustainable Energy Systems, Mechanical Engineering Department, FCTUC, University of Coimbra helena.monteiro@dem.uc.pt Background: Starting year: Research team: Supervisors:

MSc in Energy for sustainability/UC/ Portugal; Degree in Architecture/UP/Portugal 2009/2010 Fausto Freire1; John Fernandez2 Fausto Freire1; John Fernandez2 1 Mechanical Engineering Department, University of Coimbra; 2 Architecture Department, MIT

OBJECTIVES This research goal is to support the development of low-energy houses in a Mediterranean climate, considering a life cycle (LC) perspective in order to avoid problem-shifting. Thus this study has two objectives: 1) to comparatively assess LC energy and environmental impacts of different passive construction measures for a single family house; 2) to assess the differences among LC results for different Portuguese locations in order to expand the conclusions to a Mediterranean context.

most important LC process for all alternatives studied so far. However, if a more Realistic operational pattern scenario is assumed, the most important LC process depends on the alternative construction adopted: material production becomes the most important process for the scenarios with higher insulation and a lower total ventilation level.

WORK PLAN A LC model has been implemented for a single family house including five LC processes: material production, transport, maintenance, operational heating and cooling. An integrated life-cycle assessment and building dynamic simulation of the dwelling has been performed in order to assess passive construction measures, such as the envelope insulation level, the total ventilation rates, the use of single or double glazing windows, and the influence of different exterior wall solutions. The operational heating and cooling energy loads have been obtained for two alternative operational patterns: a Permanent operational pattern (100%) with permanent interior set-points between 20ยบC and 25ยบC, and a more Realistic operational pattern (25%) assuming a moderate occupancy with a moderate HVAC level. Selecting some key passive option from the parametric analysis performed other passive construction alternatives will be assessed: different window/wall ratios and orientations. Furthermore, the house alternatives will be assessed for different Portuguese (Mediterranean) locations. RESULTS The current results show that combining two simple passive construction measures (a high envelope insulation level, and an air tight envelope) may lead to important LC energy savings. Considering the Permanent operational pattern scenario, heating is the

FIGURE 1 Case study, single-family house located in Coimbra (A): 2nd floor (B); 1st floor (C).

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FIGURE 2 Final Energy (kWh/year) for heating and cooling the house (with double hollow brick and double glazing windows) considering different thermal insulation levels and total ventilation levels.

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FIGURE 3 Life cycle non-renewable primary energy (MJ) results for the realistic operational pattern assumed, considering the alternatives studied (4 total ventilation levels and 5 insulation thicknesses).

Novel Multivariate adaptive self-learning methodology for efficient energy systems in buildings Henrique Pombeiro MIT Portugal Sustainable Energy Systems, Centre for Innovation, Technology and Policy Research – IN+, Instituto Superior Técnico, Lisbon University henrique.pombeiro@tecnico.ulisboa.pt Background: Starting year: Supervisors:

MSc in Environmental Engineering, Instituto Superior Técnico, Univeristy of Lisbon, Portugal 2013 Carlos Silva1 1 Center for Innovation, Technology and Policy Research - IN+, Instituto Superior Técnico, Universidade de Lisboa

OBJECTIVES My PhD will focus on the energy modelling and optimization with a research grant from an EU CIP Project. I am developing self-learning models that can model energy demand in specific locations in the IST Taguspark building through neuro-fuzzy programing. Having the base scenario a prediction model set, I develop optimization models through evolutionary algorithms to optimize consumption across the building. A real application is foreseen. Additionally, I am developing a multivariable analysis that can model energy consumption in different zones with a pilot in 2000 houses, funded by ERSE, in Portugal. The model will comprise specific data and also more general at the level of the region such as CENSOS and GIS. The most effective feedback mechanisms should be achieved and, with that, an effective policy recommendation tool should be developed. I am complementing my MIT Portugal PhD with KIC InnoEnergy PhD, which will help me focusing my Thesis towards the Industry.

standardized linear regression that is accepted for the EU verification standards. Figure 1 displays the mean squared and mean absolute errors for two models: one linear (LR) and another based in feedforward backpropagation neural networks (NN) algorithm for an amphitheater.

FIGURE 1 MSE and MAE for LR and NN.

These preliminary results show an evolution in the energy modeling, which can still be optimized with the inclusion of variables and with the identification of discrete events. Consequently, effective optimization models will be developed.

WORK PLAN I divide my PhD in two main topics: the intelligent management of a service building and the design of energy policies to encourage household consumers to adopt more efficient consumption behaviors, the later beginning at the end of 2014. Both topics are under the framework of two projects which will help me validating results in real life scenarios. RESULTS At the moment, few neuro-fuzzy baseline models were developed with higher performances than the

FIGURE 2 Model of the amphitheater.

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Integrating urban design and sustainable development: Life-cycle energy and greenhouse gas emissions from urban residential patterns Joana Bastos MIT Portugal Sustainable Energy Systems, ADAI-LAETA, Department of Mechanical Engineering, University of Coimbra jbastos@student.dem.uc.pt Background: Starting year: Supervisors:

Architecture, University of Lisbon 2011 Fausto Freire1; Stuart Batterman2 1 ADAI-LAETA, Department of Mechanical Engineering, University of Coimbra 2 Department of Environmental Health Sciences, University of Michigan

OBJECTIVES The overall goal of to research is to contribute to the understanding of the linkages between urban design, primary energy requirements and greenhouse gas (GHG) emissions associated with different residential patterns. The research incorporates a life-cycle perspective and the methodological approach contributes to the improvement and development of life-cycle assessment (LCA) applied to an urban scale. WORK PLAN A comparative assessment of energy requirement and GHG emissions from urban residential patterns is being developed for two case studies based on existing area in the metropolitan area of Lisbon, considering three urban components: i) construction of buildings and infrastructure; ii) operational requirements of buildings and infrastructure; and iii) user transportation (including private and public transport). In addition, scenarios examining future development alternatives will be analyzed in order to identify directions and opportunities for a more environmentally sustainable urban development.

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PRELIMINARY RESULTS Firstly, a comparative life-cycle assessment of three building types in Lisbon was performed [1], comprising construction, retrofit and use phases. Over the 75-year lifespan, the use phase accounted for most (69-83%) of the primary energy requirements and GHG emissions. Larger buildings had lower life-cycle energy requirements and GHG emissions on a square meter basis; however, the pattern was reversed on a per person basis, and larger buildings had higher results. Recently, we compared a city apartment building with a suburban semidetached house [2], integrating user transportation in the assessment, to explore the linkage between location and energy requirement and GHG emissions. Whereas the largest share of impacts in the apartment building was associated with building use (40-45%), in the suburban house transportation accounted for most energy and GHG emissions (65-70%). 1. Bastos, J., Batterman, S. and Freire, F. (2014) Life-cycle energy and greenhouse gas analysis of three building types in a residential area in Lisbon, Energy and Buildings 69: 344-353 2. Bastos, J., Batterman, S. and Freire, F. (2014) Integrating transportation in the life-cycle assessment of buildings: a comparative assessment of two housing typologies in Lisbon (in submission)

The technical and economic feasibility of energy efficiency and renewables integration in schools with an ESCO business model approach Joana Rafael MIT Portugal Sustainable Energy Systems, IST – Instituto Superior Técnico, Universidade de Lisboa joanarafael@tecnico.ulisboa.pt Background: Starting year: Research team: Supervisors:

Degree in Architecture/ FAUTL / Portugal) (Sustainability and Environmental Management / Harvard University / USA 2012 Tiago Brázio1 Paulo Ferrão2 1 MIT Portugal Sustainable Energy Systems, , Instituto Superior Técnico, Universidade de Lisboa 2 Center for Innovation, Technology and Policy Research - IN+, Instituto Superior Técnico, Universidade de Lisboa

OBJECTIVES This paper is about the technical and economic feasibility of introducing renewable energy systems and efficiency measures in the Portuguese schools that were part of the recent government renovation program – using as a case study a newly renovated school in Lisbon, Portugal. The Parque Escolar (PE) interventions aimed at providing higher building energy performance, better learning conditions and better thermal comfort and air quality conditions. However, this renovation implied a significant increase of equipment and installed power in the schools and thus an average increase of electricity consumption of 39% and 253% for natural gas in the school under study. A hypothesis is tested as a possible solution to tackle the very high energy costs burden that schools are facing by proposing an Energy Services Company (ESCO) business model approach to ensure schools life cycle sustainability.

WORK PLAN On this scope, we studied a school refurbished by PE in Lisbon - Escola Secundária Vergílio Ferreira (ESVF) as a representative case study. First, we analyzed its energy consumption before and after the renovation, verified its compliance with the regulatory framework - RCCTE (Regulation for the Thermal Characteristics of Buildings) and RSECE (Regulation of the Acclimatization Energy Systems of Buildings) - and then, quantified the energetic and economic impact of PV integration and the replacement of the existent gas boiler with a heat pump. Lastly, we conducted a project appraisal with an ESCO business model approach. RESULTS The hypothesis tested for an ESCO approach proved to be financially viable according to the Capital Budgeting Model used. Thus, the hypothesis is validated as a possible solution for energy efficiency financing for PE, transferring school’s operational costs along to the private sector while creating a new market demand.

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Optimal control of the indoor climate João Carrilho MIT Portugal Sustainable Energy Systems, ADAI-LAETA, University of Coimbra joao.carrilho@dem.uc.pt Background: Starting year: Supervisors:

MEng in Engineering Acoustics and Vibration, University of Southampton, United Kingdom 2011/2012 Manuel C. Gameiro da Silva1 1 ADAI-LAETA, Department of Mechanical Engineering, University of Coimbra

OBJECTIVES There are several options for improving the energy efficiency of heating, ventilation and cooling systems, but it is often very difficult to determine which will materialize into the best solution. Given the requirements for maintaining a comfortable thermal environment and an acceptable air quality in buildings, despite the ever changing outdoor conditions, developing an energy-efficient strategy to control the indoor climate becomes not only a control problem but also a dynamic, multi-objective, optimization problem. The focus of this research is on using engineering modeling, control and optimization techniques to develop an optimal control strategy for maintaining the indoor climate within requirements, using a minimum of energy. Engineering methods are used in modeling the dynamics of heat transfer and airflow in buildings, along with system identification theory and model predictive control techniques, to develop an innovative control strategy for energy-efficient indoor climate control. This implies quantifying the compromises between conflicting requirements and objectives in order to provide assistance in the selection of the most adequate strategy. Having identified the best realizable strategy, the ultimate purpose is to develop a supervisory control algorithm, capable of taking decisions based on predicted environmental conditions, suitable for deployment both in new buildings and in retrofitting scenarios.

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WORK PLAN 1. Background and context; deﬁnition of main research questions and objectives; literature review and critique: A multidisciplinary literature review is undergoing, since the start of the project, with thorough organization into the diverse topics that are of particular relevance, along with a critical analysis of the main results reported by other researchers. 2. Dynamical modelling and system identiﬁcation of the building system:

This task introduces the main theoretical aspects used in developing a model of the building system dynamics, including the passive subsystems, the active subsystems, the occupants, the exterior environment, etc. System identiﬁcation theory is used in estimating the system parameters. Within this task, it is foreseen to use a climatic chamber facility to experimentally validate the system identiﬁcation algorithms developed. 3. Predictive and optimal control of the indoor climate: This task introduces the main theoretical concepts, based on the developments of the previous task, for the optimal predictive control of the indoor climate. Modern state-space control theory, associated with optimization techniques, will be used in developing a supervisory controller capable of identifying the best course of action for improved energy efficiency with the ventilation and climatization systems available in a building. An experimental validation exercise is also planned for this task, which will possibly overlap with the experimental campaign of task 2. RESULTS Work is in progress on the system identification approach to determine ventilation Air Exchange Rates (AERs) in buildings: 1. Conventional AER assessment from tracer gas decay profiles (fig. 1) 2. Estimating Time Varying Air Exchange Rates (fig. 2) (João Carrilho, Stuart Batterman and Manuel Gameiro da Silva. Paper presented at CLIMA 2013: the 11th REHVA World Congress & 8th International Conference on IAQVEC, held in Prague, Czech Republic, 16-19 June 2013)f 3. Instantaneous Infiltration Rates from Atmospheric CO2 Daily Cycle (fig. 3) (João Carrilho, Mário Mateus, Stuart Batterman and Manuel Gameiro da Silva. Manuscript in preparation for journal publication)

FIGURE 1 Example of the conventional method for assessing ventilation AERs using tracer gas techniques, in this case CO2 produced by the building occupants. The AERs are the slopes of the fitted straight lines to the natural logarithm of the decay of CO2 concentration. Data obtained over a week in a residential flat.

FIGURE 2 Example of time-varying ventilation rates determined from a novel collocation scheme for system identification of AERs from tracer gas time series (solid line: LTV model), in this case CO2 produced by the building occupants. Data obtained over 24 h in a small, naturally ventilated office. Shaded areas identify the periods were the occupants were present, during which time the office windows were left open. The dashed line (LTI model) is the AER obtained from the conventional profile fitting method.

115 FIGURE 3 Example of time-varying ventilation rates determined from a novel system's approach to the time evolution of the outdoors atmospheric CO2 and its correlated response indoors. Infiltration AERs measured over a week in a residential home. Shaded areas identify night periods.

High quality silicon ribbons for solar cells José Mário Pó MIT Portugal Sustainable Energy Systems, Faculty of Sciences University of Lisbon jmpo@fc.ul.pt Background: Starting year: Supervisors:

Energy and Environment Engineering, FCUL 2011/2012 Jorge Maia Alves1 1 Dept. of Geographical Eng., Geofisics and Energy (DEGGE), Faculty of Sciences, University of Lisbon

OBJECTIVES Develop a process for high quality and low cost silicon sheets growth for solar cells. WORK PLAN: This work focuses on the development of a technological solution to decrease material and the energy footprint during Silicon wafer production for Solar Cells, the Silicon over Dust Substrate (SDS) technology. To achieve higher quality (and solar cells with higher conversion efficiency), optimal thermo-mechanical stresses during Silicon crystallization are studied for the SDS geometry. Aided by Computational Fluid Dynamics Finite Element Analysis (CFDFEA), a prototype furnace is designed, built, and the quality of the output samples is evaluated. RESULTS

FIGURE 1 Temperature distributions output of the computational fluid dynamics model, simulated on a 210 mm by 70 mm by 0.3 mm silicon sheet, with a locally induced linear molten zone; Bottom: corresponds to the temperature distribution for the ribbon with no thermal conditioning. Top: the sample is thermally conditioned to remain at 1000ºC.

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FIGURE 2 Proposed prototype for the establishment of an optical linear molten zone in a silicon ribbon by laser stimulation, with thermal conditioning.

FIGURE 3 Prototype assembly and testing.

Economical evaluation of emerging biofuel feedstocks: The microalgae case study Lauro André Ribeiro MIT Portugal Sustainable Energy Systems, University of Coimbra lribeiro@inescc.pt Background: Starting year: Supervisors:

Bachelor and Master degrees in Business Administration (UFRGS-Brazil) in 2005 and 2008. 2009 Patrícia Pereira da Silva1, Stephen Connors2 1 Faculty of Economics, University of Coimbra 2 Massachusetts Institute of Technology (MIT)

OBJECTIVES The proposed study aims to analyze the innovation path of cultivating microalgae with biofuel production and assess the economical, political and technological factors that are critical to the success of this emerging biofuel feedstock. WORK PLAN The purpose of this study is to verify who are the key players in this emerging market, analyze the present economical and political situation, evaluate possible opportunities and weaknesses and forecast ways to enhance the feasibility of large-scale microalgae biofuel industries and compare this emerging feedstock with other options that could replace our oil consumption. The study aims to make the intersection of three major areas of knowledge: the Economic Policies, Processes of Technological Diffusion and Economical Evaluation / Performance analysis of the technology. Applying Delphi and modeling tools, this study examines different market penetration scenarios of microalgae biofuels. RESULTS Technological advances in cultivation and extraction of oil from microalgae are scientifically well known, and should continue to move forward in the coming years with increasing investment in R&D in this area. In order to boost development, public investment in R&D is the most important policy to be adopted by countries. Developing strategies aimed to renewable resources; applying tax incentives and subsidies; and issuing mandatory country objectives are also encouraged.

FIGURE 1 Delphi Survey Preliminary Results “When will it happen?” (Answers of 55 algae experts).

FIGURE 2 Possible future scenario with strong support (adapted from Lee, 2011). Reference: Lee DH Algal biodiesel economy and competition among bio-fuels. Biores. Tech. 2011,102:43-49.

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Managerial flexibilities and competition In renewable energy investment Luciana Barbosa MIT Portugal Sustainable Energy Systems, Instituto Superior Técnico, Universidade de Lisboa, Portugal lu_salles@hotmail.com Background: Starting year: Research team: Supervisors:

Master degree in finance / Lancaster University / UK 2013 Alberto Sardinha1; Monika Tarsalewska2 Paulo Ferrão3; Artur Rodrigues 4 1 INESC-ID and Instituto Superior Técnico, Universidade de Lisboa, Portugal 2 Exeter University, UK 3 Instituto Superior Técnico, Universidade de Lisboa, Portugal 4 School of Economics and Management, Department of Management, University of Minho

OBJECTIVES This paper presents a model that identifies the optimal time to deploy a renewable energy project, and shows how the probability of arriving a competing investor may affect the investment threshold and the value of the project.

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WORK PLAN Deploying a renewable energy project is an important decision that can facilitate sustainability. However, investment in the power sector is characterized by uncertainties in the energy prices, market evolution, and policy intervention that can have a significant impact on the financial performance of a project. For instance, feed-in tariff (FIT) program is a long-term contract with energy producers (e.g., homeowners, business and organizations such as schools and community groups) to incentivize energy generation through renewable energy sources. Furthermore, as economies try to attract investors, one has to take into account the effect of competition in order to value a renewable energy project. Therefore, the high sunk cost in these types of investments significantly increases the risk of the project, especially in dynamic economies where the investor faces competition. In order to value renewable energy project, managers need a valuation technique that can incorporate uncertainties of dynamic economies and take into account managerial flexibilities. However, standard static valuation techniques, such as NPV, underestimate the value of the project as the methods do not take into account the flexibilities that are present in dynamic environments. For example, a valuation technique must consider an option to expand when conditions are favorable or an option to abandon when the project is a failure. Hence, we believe that the real options approach provides a good framework for valuations of renewable energy projects, since managerial flexibilities and uncertainties are incorporated into the model.

This work presents a model to value managerial flexibilities and identify the optimal timing for the investment in a renewable energy project taking into account the effect of competition. Thus, the purpose of the paper is to present a model that values renewable energy investments with embedded options and calculates the optimal time to invest in these projects. In addition, a key contribution of our model is that it analyzes the effect of competition on the value of the option and the time to launch the project. In order to evaluate the value added by the real options, we calculate a strategy that considers managerial flexibilities and competition against another strategy based on a static decision making process. The former strategy considers an investment on renewable energy project where managers have the flexibility to wait for the best moment (i.e., when the market is favorable and due to the fear of competition) to deploy the project; if the market does not become favorable within a finite time period, the managers have also the option to abandon the project. The latter strategy includes the same renewable energy project; however, the project is always launched at time 0. The optimal critical operational revenue, in which the investment in the renewable energy project becomes optimal (also known as optimal stopping threshold), is calculated using the analytic approximation for the American call option. This is an extension of the Barone-Adesi and Whaley (1987) approximation, where we include both the diffusion parts and the jump to the project. Hence, we developed a Matlab code (available upon request) to simulate several strategies incorporating real options where the fear of preemption and the FIT bonus are incorporated. RESULTS The results show that when the probability of the competitor arrival increases, the value of the option decreases and the time to invest accelerates.

Decision-support tool for wind power plants participating in electricity markets Luis Rodrigues Jr MIT Portugal Sustainable Energy Systems, Departamento de Engenharia Geográfica, Geofísica e Energia Departamento de Engenharia Geográfica, Geofísica e Energia, Faculdade de Ciências da Universidade de Lisboa luiscarlosjr@gmail.com Background: Energy and Environment Engineering/ FCUL/ Portugal, Electrical Engineering / UFSC/ Brazil Starting year: 2013 Supervisors: Ana Estanqueiro1; Ricardo Lima1 1 Energy Analysis and Networks Unit , Portuguese National Laboratory for Energy and Geology (LNEG)

OBJECTIVES Wind energy has developed into a prominent energy source in many of the electric power systems worldwide. Nevertheless, the deployment of the technology is slowing down due to most government’s skepticism on continuing encouraging FIT as occurred in the past 20 years. Additionally, the massive installation of wind turbines complicate the operation of a power system, as their production fluctuations and limited capacity credit require additional reserves to keep the balance between production and consumption. In order to end FIT and make wind plants responsible for their integration costs, these plants have been pushed to electricity markets and grouping into clusters in order to compete with conventional generation. This work aims at improving the participation of wind power in electricity markets and exploit wind turbines technological capabilities by developing decision-support tools for wind generator owners. These tools will assist wind energy promoters in taking optimal decisions regarding wind power plants operation and its bidding strategy in the many market frameworks available in modern electricity markets. WORK PLAN Optimal bidding strategies will be investigated considering the actual working mechanisms of electricity markets, the influence of wind power clusters on market clearing price and envisioning trends in market design. Moreover, the wind turbines rich variety of technical capabilities regarding power production modulation

will be modeled and made available for the decision-support tool. In a later phase, an integrated bidding strategy for wind energy producers that own mixed generation portfolio will also be addressed. The mixed generation portfolios might include battery energy storage, hydro and thermal generation assets. Decision strategies should take into account the flexibility offered by the assets in order to reduce imbalance costs caused by the uncertainty on wind and hydro production. RESULTS Expected results of the application of the decision-support tool include a boost in the effectiveness of market participation of wind power producers, reducing integration costs of wind energy and guaranteeing a reasonable profit margin of wind power producers.

FIGURE 1 Information exchange diagram of a decision-support tool for the operational management of a mixed generation portfolio.

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An innovative strategy for power system restoration using utility scale wind parks Luis Seca MIT Portugal Sustainable Energy Systems,Faculdade de Engenharia da Universidade do Porto pds11006@fe.up.pt Background: Starting year: Supervisors:

MSc/ FEUP/ Portugal 2011 Carlos Moreira1; JoĂŁo PeĂ§as Lopes1 1 Department of Electrical and Computer Engineering, Faculdade de Engenharia da Universidade do Porto

OBJECTIVES The objective of this thesis is to present an innovative Power System Restoration (PSR) that exploits the flexibility offered by large offshore wind parks, in particular with HVDC connection with Voltage Source converter (VSC), to support restoration following a black out in a transmission grid. Restoration sequences comparing different strategies in a VHV AC network with conventional generation were implemented for two cases: with and without the participation of a large offshore wind farm in the restoration plan. In order to evaluate its benefit, a typical transmission network with all its models was implemented in EUROSTAG. WORK PLAN Very ambitious targets for renewable energy integration in all EU countries have been established. The large scale integration of renewable based energy sources has however a significant impact in power system operation. In effect, these large shares of variable energy sources replace conventional

120 FIGURE 1 Load pick-up versus time on the system during the PSR.

synchronous units, thus reducing system inertia as well as frequency and voltage regulation capabilities. This fact, together with the need of exploitation the network closer to its technical limits has exposed the power systems to vulnerabilities, being the worst consequence the occurrence of blackouts that may cover large geographical areas. Consequently, a combined approach that uses utility scale wind farms connected to the transmission level for PSR is clearly regarded as an added value. In this research one has developed restoration sequences comparing strategies implemented for two cases: 1. Without the participation of wind generation; 2. With a large offshore WF participating the restoration plan. RESULTS One can observe that the load pick up in the system 2 is faster than in 1 and, as seen in pictures 2 and 3, over voltages are lower in system 2.

FIGURE 2 Behavior of bus voltages during the PSR for system 1).

FIGURE 3 Behavior of bus voltages during the PSR for system 2).

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Energy and urban sustainability: On the thermal assessment in housing architectural design Mariana Abrunhosa Pereira MIT Portugal Sustainable Energy Systems, INEGI/IDMEC-FEUP, Faculty of Engineering, University of Porto pds10001@fe.up.pt Background: Starting year: Supervisors:

Architecture degree, Faculty of Architecture, University of Porto, Portugal 2010 Eduardo Oliveira Fernandes1; Manuel Correia Guedes2 1 Faculty of Engineering, University of Porto 2 Department of Civil Engineering and Architecture, Instituto Superior Técnico, Universidade de Lisboa

OBJECTIVES The aim of this thesis is to define the order of merit or of design priority of the architectural and construction parameters that have more influence in the thermal behavior of housing in Portugal and subsequently, to create a design aid that leverages on the former Portuguese pioneer thermal regulation, RCCTE.

UA and inertia curves as it is illustrated in Figure 1. This will allow the architect to be acquainted with the actual quantification of the options/decisions at an earlier phase of the building design, before all sophisticated simulations could have been undertaken.

WORK PLAN The literature review explored both the scientific knowledge of building physics and good architectural practices in their proper climatic context. It was possible to conclude that although many studies approached the role of some relevant input parameters taking into account design parameters that influence the building thermal behavior (such as site constraints) and, in more than one way, aiming to express the thermal performance (such as of a component or the whole building), it became clear in this research that the fundamental parameters are contained in the net gain methods of which the EN ISO 13790 ‘Thermal performance – calculation of energy use for space heating’ and the Portuguese RCCTE are both representative. The reworking of these parameters in the light of a design aid tool suggests however new approaches such as in the assumptions regarding zoning, and the expression of the glazing needed or due. RESULTS After exploring several avenues such as that of decision trees, it was possible to conclude that a straightforward encapsulation of all essential parameters can be made through normalized relationships between ‘heating/cooling gains’ and ‘energy needs’, expressed in a family of equivalent glazing, global

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FIGURE 1 Nomogram to be used during the architectural building design process for the optimization of the heating and cooling needs for different UA [W/°C] as a function of the glazing area/useful floor area, applied to a given south oriented space, with heavy thermal inertia and without shading devices.

Energy behaviours impact on energy consumption - A case study Marta Lopes MIT Portugal Sustainable Energy Systems, FCT - University of Coimbra, Portugal Dept. of Environment, ESAC - Polytechnic Institute of Coimbra, Portugal; INESC Coimbra, Portugal mlopes@esac.pt Background: MSc in Environmental Management, Materials and Wastes Valorisation, University of Aveiro, Portugal BSc in Environmental Engineering, IST â&#x20AC;&#x201C; Technical University of Lisbon, Portugal Starting year: 2010 Research team: This research is being developed under a multidisciplinary R&D environment on end-use energy efficiency in the scope of projects ENERGYBOX (http://www.uc.pt/en/org/inescc/Projects/energy_box) and EMSURE (http://www.uc.pt/en/efs/ research/emsure). Supervisors: Carlos Henggeler Antunes1; Nelson Martins2 1 Dept. of Electrical Engineering and Computers, University of Coimbra, Portugal 2 Dept. of Mechanical Engineering, University of Aveiro, Portugal

ABSTRACT Energy behaviours represent an important underexploited resource in the context of promoting end-use energy efficiency in the residential sector. However, addressing energy behaviours is a complex task and more effective behaviour change interventions and policies grounded on comprehensive approaches of energy behaviours are required. This work aims at presenting the preliminary results of an integrative approach on energy behaviours influence on energy consumption. A real-world intervention involving 128 Portuguese households was implemented during 2011/13. Energy consumption was monitored with a time step of 15 minutes using a smart monitoring device (http://www.cloogy.com/en/), and qualitative variables were characterised using self-reported questionnaires available â&#x20AC;&#x201C; to households through a web platform (e.g., http://www.ces.uc.pt/inqueritos/ sintra). Qualitative variables included the socio-demographic characteristics of the household, their activities and social practices, the energy resources and services they use, the building characteristics, the physical environment, their energy behaviours profile and personal characteristics. A multivariate statistical approach to energy behaviours influence on energy consumption is presented and discussed. Factor analysis of selected behaviours enabled identifying three main energy behaviour dimensions in the studied sample. Furthermore, multiple regression analysis using the list of selected variables yields solutions accounting for approximately 61% of the variance in energy consumption of the sample. The implications for tailored policy and planning interventions are discussed in relation to energy behaviours.

METHODOLOGY

RESULTS

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Acknowledgement: The author would like to express her gratitude to ISA - Intelligent Sensing Anywhere, AMES - Sintra Municipal Energy Agency, and Lisboa E-Nova - Lisbon Municipal Energy and Environmental Agency for the technical support. This work has been framed under the Energy for Sustainability Initiative of the University of Coimbra and partially supported by the Energy and Mobility for Sustainable Regions Project (CENTRO07-0224-FEDER-002004) and by Fundação para a Ciência e a Tecnologia (FCT) under grant SFRH/BD/51104/2010 and project grants MIT/SET/0018/2009 and PEst-OE/ EEI/UI308/2014.

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Exploratory analysis into resource metabolism of (urban) economies Michal Monit MIT Portugal Sustainable Energy Systems,Instituto Superior Técnico, Universidade de Lisboa michal.monit@singularityu.org Background: Starting year: Research team: Supervisors:

M.Eng. in Energy Engineering, AGH-UST, Poland M.Sc. in Renewable Energy Science, RES, Iceland 2010 Urban metabolism group, IST-UTL Paulo Ferrão1; André Pina1 1 Center for Innovation, Technology and Policy Research - IN+, Instituto Superior Técnico, Universidade de Lisboa

OBJECTIVES • Construct an accessible framework enabling resource metabolism studies based on widely available, top-level data; • Explore driving forces behind resource consumption of economies and interconnections with changes in socio-economic variables; • Extend the country-level method down to urban level; • Test for existence of power law scaling phenomena in resource consumption data, disaggregated by material type, • Improve on the ways urban metabolism data is visualized and communicated. WORK PLAN • Data gathering and processing; • Data gathering and processing, continued; • Construction of a “plug and play” framework for analyzing inputs and outputs of economies,

•

• • •

by material / product type and quantity; based on widely published country statistics (e.g. UN, OECD, Eurostat); Exploratory analysis of evolution of size and structure of resource consumption of economies, at a national level, and its interlink with changes in socio-economic variables; Incorporate economic data and allocate resources to specific economic activities and bring down the scale of analysis down to urban level; Test and compare the framework against published resource metabolism and consumption studies, both at national and urban level; Publish validated results; Prepare ground and discuss possible applications and extensions of this work.

RESULTS (TENTATIVE)

125 FIGURE 1 Resource metabolism of Portugal and the Lisbon Metropolitan Area. Colors represent different material categories. Grays denote different economic activities. Data sources: UN Stats, OECD, Eurostat; Portugal (domestic extraction) – MOSUS; Lisbon Metropolitan Area – work of Samuel Niza and Leonardo Rosado

FIGURE 2 DMC by material type, plotted against urban population, in percent. Thinner end of line corresponds to beginning of time series data (1970-2008), color-coded by country. Data from PNAS paper by Wiedmann, et al. (2013), and World Bank data.

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FIGURE 3 GDP per capita, const. 2005 US dollars, vs. urban population [%]. Thinner end of line corresponds to beginning of time series data (1970-2008), color-coded by country. Data from PNAS paper by Wiedmann, et al. (2013), and World Bank data.

The comprehensive analysis of the effects of additives on working parameters of diesel based engines Mohamamdhossein Ghodsirad MIT Portugal Sustainable Energy Systems, Mechanical Engineering department, University of Coimbra, mh.ghodsirad@gmail.com Background: Starting year: Research team: Supervisors:

Mechanical Engineering/ Ferdowsi University of Mashhad/ Iran 2011 Luis Serrano1,2; Joao Teixeira1 Manuel Gamiero da Silva1; Antonio Portugal3 1 Department of Mechanical Engineering, University of Coimbra 2 IPL, School of Technology and Management, Polytechnic Institute of Leiria, Portugal 3 Department of Chemical Engineering, University of Coimbra

OBJECTIVES The main objective of the reported work was to develop a methodology to assess the fuel consumption and the efficiency of the propulsion system of passenger cars equipped with internal combustion engines in road tests carried out to compare the performance of fuels with different formulations. The second objective is to optimize the efficiency of the working regime of this injectors by the introduction of appropriate additives. WORK PLAN This investigation will be carried out by two different test methods: Engine test bench and Real on-road test. In the road test, the examination has been done out in highway travels. A Peugeot 407, has been equipped with two vehicle data loggers. A real-time fuel consumption metering system is combined with a GPS logger and an OBD II data logger which provides the vehicle and engine information available at the CPU unit, while the pressure sensors are connected to three pressure taps placed on the front of the vehicle to provide information about the relative velocity between the car and the air and the flow angularity. The second phase of the tests, the engine test bench, a direct injection diesel engine will be examined to discriminate between a fuel that produces no measurable deposits and one which produces deposits that cause loss in power under field conditions considered unacceptable by engine manufacturers. RESULTS The presented results in Figure 1 are extremely encouraging by giving the level of correlation between the values obtained by the calculated engine load expressed by the driving force on the wheels and the measured fuel consumption. It can be concluded that the measurement have been made with a high accuracy and all relevant variables have been considered.

FIGURE 1 Temporal evolutions of the driving force and the fuel consumption of the vehicle.

The same data file that gave rise to the figure above are represented Figure 2, all fuel consumption values during the trip correspond to the driving force transmitted from the engine to the wheels at the same time. The representation used in Figure 2 has the advantage of allowing a single aggregate data from different graphic paths. It is possible to identify differences in terms of consumption, for example, comparing the regression lines obtained for two different fuel formulations.

FIGURE 2 Fuel consumption as a function of driving force.

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Decision support methodology for national energy planning in developing countries: An implementation focused approach Nathan Lee MIT Portugal Sustainable Energy Systems, INEGI/IDMEC, Faculty of Engineering, University of Porto nathan.lee@fe.up.pt Background: Starting year: Supervisors:

Advanced Studies Diploma/ Faculty of Engineering, University of Porto/ Portugal Bachelor of Science- Engineering Physics/ Miami University of Ohio/ USA 2011 VĂtor Leal1 1 INEGI/IDMEC, Faculty of Engineering, University of Porto

OBJECTIVES The objective of this thesis is to contribute to the advancement of methodologies supporting national EP practices in developing countries, where the Economic Community of West African States (ECOWAS) was chosen to limit the scope. Specifically this work will address the need for an EP methodology allowing actors to identify objectives specific to the geographic context and to account for them in the construction and multiple-objective evaluation of multiple energy policy plans.

multiple energy policy plans in their achievement of objectives compared to a no-plan future. WP5 consists of drawing conclusions from the EP exercise, especially about the impact of the specific ECOWAS and ECOWAS+ objectives and attributes identified, and writing of the thesis. RESULTS 1. Already Achieved: i) A set of EP objectives and respective quantifiable attributes have been identified for use within the multicriteria evaluation models; ii) The energy demand and supply models have been constructed, and baseline energy demand and supply projections for the case study have been made. 2. Further Expected: i)EP methodology will be made for ECOWAS member states, which is adaptable to other developing countries; ii) An assessment of the impact of accounting for context specific EP objectives will be conducted.

FIGURE 1 Map of ECOWAS region [1].

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WORK PLAN The work is separated into 5 work packs (WP). WP1 consists of a literature review of EP objectives and quantifiable attributes used in developed countries and the ECOWAS and the identification of additional objectives and attributes possibly implicitly referred to in the EP activities of developing countries (ECOWAS+). WP2 consists of the construction of Energy Demand and Supply models for national EP, as well as multicriteria evaluation models. WP3 is comprised of the elaboration of a case study of Ghana and calculation of baseline energy demand projections. WP4 consists of the development and evaluation of

TABLE 1 EP Objectives and respective attributes.

References: 1. European Commission: Eurostat. West Africa Page: ECOWAS Map. 2009. http://epp.eurostat.ec.europa.eu/. *Identifier added to graphic by authors.

Evaluation of key determinants of electricity market integration in a context of high penetration of renewables Nuno Figueiredo MIT Portugal Sustainable Energy Systems, EFS Initiative, University of Coimbra nuno.figueiredo@portugen.com Background: Starting year: Supervisors:

Masters in Mechanical Engineering, University of Coimbra 2011 Patrícia Pereira da Silva Surname1 1 Faculty of Economics, University of Coimbra

OBJECTIVES Assess the influence of the high penetration of RES-E on the level of electricity market integration.

markets), as one could expect by the concept definition of market splitting.

WORK PLAN • Overview of electricity market restructuring in Iberia • Regional electricity market interconnections • Data extraction and processing • Modeling • Analysis of results • Policy recommendations RESULTS The estimated models aim to provide indications about the behaviour of the market splitting mechanism in the Iberian spot electricity markets, considering the wind power penetration in these same markets. We use logit models to express the probability response for market splitting of day-ahead spot electricity prices together with explanatory variables like wind speed, available transmission capacity and electricity demand. In this study the wind speed is the main explanatory variable used in our models, expressing the amount of electricity generated from wind energy sources. The two other explanatory variables used are related with demand of each country and the Available Transfer Capacity (ATC) between both spot electricity markets. Prediction results from the models express that market splitting probability increases when the wind speed is higher, consequently with higher wind power generation. This can be explained by having low marginal cost electricity available to flow across the border. Market splitting probability decreases with increasing ATC (allowing higher flows of electricity between

FIGURE 1 Market splitting probability vs Average Wind Speed.

FIGURE 2 Market splitting probability vs ATC.

The European interconnection capacity target of 10% of the peak demand of the smallest interconnected market has to be reconsidered, in order to keep electricity market integration a reality. Investment in interconnection capacity has to follow the investment and deployment of further wind power capacity, so coordination policies governing both interconnection development and renewable incentives should be designed.

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Development of a climate performance indicator for analysis of low energy buildings Nuno Martins MIT Portugal Sustainable Energy Systems, SESUL, Faculdade de Ciências da Universidade de Lisboa, University of Lisbon nunomrmartins@hotmail.com Background: Starting year: Supervisors:

Energy and Environment Engineering, Faculdade de Ciências da Universidade de Lisboa, Portugal 2012 Guilherme Carrilho da Graça1 1 Centre for Sustainable Energy Systems Gaculty of Sciences, University of Lisbon

OBJECTIVES Development of a climate performance indicator, that focuses on a given building’s energy performance and its relation with local climate WORK PLAN Development of the framework Development of secondary indicators Expansion to other locations, other building typologies and other passive methods

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potential (CI). The best climates and buildings are expected to have CIs and BIs that are close to one. Secondary indicators provide a more detailed analysis, evaluating climate availability and building necessity for particular passive solutions. By comparing these indicators and their impact on the simulation results, empirical relations were determined, allowing an estimation of the impact of a given solution without the need for detailed thermal simulation.

RESULTS Nuno R. Martins, Guilherme Carrilho da Graça, A climate performance indicator for analysis of low energy buildings, Proceedings of BS2013: 13th Conference of International Building Performance Simulation Association, Chambéry, France, August 26-28 2013, Pages 1348-1356 The proposed framework is based on three interrelated energy performance indicators, also called primary indicators. The total building climate performance indicator (BCP) is the product of the climate potential (CI) and building indicator (BI). BI is building dependent and characterizes the capability of a given building to maximize the existing climate

FIGURE 2 SI NV* and decrease in colling load.

FIGURE 1 Average BI T.

FIGURE 3 SI NV+TM and decrease in colling load.

A novel approach for optimal integration of electric vehicles with internal combustion engines vehicles in urban freight transport Parisa Ahani MIT Portugal Sustainable Energy Systems, Institute of Mechanical Engineering, Instituto Superior TĂŠcnico, Universidade de Lisboa prsahani1@gmail.com Background: Starting year: Supervisors:

MSc in Energy for Sustainability/ University of Coimbra/ Portugal 2013/2014 Tiago Farias1; Sandra Melo1 1 Engineering, Instituto Superior TĂŠcnico, Universidade de Lisboa

OBJECTIVES Urban freight transport sector is being pressured to reduce the impact of its operations within urban areas. One of the solutions pointed out by various researches is the use of electric vehicles for delivering goods and parcels to customers. Electric vehicles have various advantages such as low emission and low operation cost per kilometer, which make them a viable option mostly from the standpoint of public stakeholders. On the other hand, their relatively high purchase costs make them, at least currently, not so attractive for private stakeholders operating in urban freight transportation sector. The main goal of this research study is to develop a new model for optimal integration of electric vehicles with internal combustion engine (ICE) vehicles in urban freight transport.

best combination of vehicles (small electric vehicles and diesel vans) for their urban freight transportation services over some planning horizon.

WORK PLAN We propose a mixed-integer programming formulation for determining an economically and environmentally optimal combination of electric vehicles with ICE vehicles in urban freight transport applications over some planning horizon. The developed model takes into account the risk associated to some uncertainties such as fossil fuel price uncertainty, electric vehicles purchasing cost uncertainty, and battery technology uncertainty, which is a novel approach in this regard. We also examine various risk measurements for analyzing the risks associated to such uncertainties. A thorough analysis of the involved risks would lead to more economically and environmentally viable decisions by the concerned private and public policymakers.

FIGURE 1 A sample of expected results.

RESULTS The developed model will then be applied to two case studies (Lisbon and New York City) to determine the

FIGURE 2 Illustration of a network of depot and demand points.

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Network reinforcement planning in a smart grid environment Ricardo Ferreira MIT Portugal Sustainable Energy Systems, Faculty of Engineering of the University of Porto (FEUP) rjcf@inesctec.pt Background: Starting year: Research team: Supervisors:

MSc / FEUP/ Portugal 2011 Ricardo Ferreira1 Manuel Matos1; JoĂŁo PeĂ§as Lopes1 1 Faculty of Engineering of the University of Porto (FEUP)

OBJECTIVES The main purpose of this research is to conceptualize and develop new models, algorithms and methodologies for electricity distribution system planning in a smart grid environment. WORK PLAN Development of mathematical models for: (1) optimal placement and daily operation of distributed storage devices within an electricity distribution system (2012); (2) optimal daily operation of flexible electrical loads and for optimal electricity distribution network reconfiguration (2013). Development of a smart distribution reinforcement planning model that includes all of the aforementioned daily optimization models (2014). RESULTS Optimal placement and daily operation of distributed storage devices in an electricity distribution system. The 14-bus low voltage 400 kV power distribution system shown in Figure 1 was used to evaluate the optimal daily operation algorithm. Generator symbols represent PV sources. Three load levels were considered. The level of PV generation within the second hour was gradually increased until a technical constraint was violated (there is no PV generation at the other two hours). In that situation, the magnitude of nodal voltages V12 and V13 rose above the maximum acceptable limit of 1.05 pu. Then, two 20 kWh devices were assumed to be installed. Using non-linear programming and an evolutionary algorithm, it was possible to obtain the optimal location of the two storage devices: bus 4 and bus 10.

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FIGURE 1 LV grid and low profiles used to evaluate the optimal storage algorithm.

Also, an optimal operation strategy for the storage devices was obtained, as shown in Figure 2. The strategy allowed eliminating the overvoltages at V12 and V13. The evolution of the state-of-charge (SOC), as well as the energy stored (Sin) and subsequently given back by the devices (Sout) throughout the three simulation time intervals is also shown.

FIGURE 2 Optimal strategy of operation for the two storage devices.

Dynamic fleet-based life-cycle assessment of electric mobility pathways: an application to light-duty passenger vehicles in Portugal Rita Garcia MIT Portugal Sustainable Energy Systems, ADAI-LAETA, Department of Mechanical Engineering, University of Coimbra rita.garcia@dem.uc.pt Background: Starting year: Research team: Supervisor:

M.Sc. in Environmental Engineering/ University of Coimbra/ Portugal 2010 Fausto Freire1; Luís Dias2; Jeremy Gregory3; Pedro Marques1 Fausto Freire1 1 ADAI-LAETA, Department of Mechanical Engineering, University of Coimbra 2 Faculty of Economics, University of Coimbra 3 Engineering Systems Division, Materials Systems Laboratory, Massachusetts Institute of Technology

OBJECTIVES The main goal of this research is to assess the conditions under which the introduction of electric vehicles (EVs) in the Portuguese fleet is environmentally beneficial compared to existing vehicle systems (with a focus on greenhouse gas (GHG) emissions). Furthermore, it aims at assessing the major sources of uncertainty associated with these conditions. WORK PLAN This research focuses on the Portuguese light-duty passenger vehicle fleet. The time frame of analysis is 1995-2030, with focus on exploring pathways of EV adoption for reducing impacts over 2010-2030. The work plan encompasses the development of a dynamic fleet-based life-cycle (LC) model by integrating a vehicle stock model of the Portuguese fleet with dynamic vehicle LC models in order to assess fleet LC GHG emissions over time. Three vehicle technologies are modeled: diesel internal combustion engine vehicle (ICEV), gasoline ICEV, and battery EV (BEV), including vehicle manufacturing, fuel production, electricity generation and vehicle operation. Scenario analysis is then used to evaluate the technologies adopted as well as changes in vehicle and contextual characteristics and their effect in the fleet GHG emissions through 2030. Lastly, uncertainty analysis is performed by conducting a probabilistic scenario analysis of key parameters in order to understand the implications of changing assumptions on the outcomes of the study. RESULTS Preliminary results of the scenario analysis are presented. Table 1 shows the four deterministic scenarios evaluated, in contrast with the 2010 fleet, and Figure 1 the LC GHG emissions evolution for each scenario. Total LC GHG emissions of the fleet are expected to increase until 2019 in the “no change”

scenario, due to the combined effect of fleet size and VKT growth. As the fleet size and VKT stabilize, a 6% reduction compared to 2010 is observed. Until 2023, reducing fuel consumption of new ICEVs (“ICEV dominates”) has a larger effect on the LC GHG emissions than the introduction of BEVs in the fleet (“BEV dominates”), since it takes time for BEV share in the fleet to become significant. Nevertheless, a slightly higher reduction in 2010-2030 GHG emissions is obtained in this scenario (36%) than in the “ICEV dominates” scenario (33%). The “combined” scenario allows an extra 5% reduction (41%).

TABLE 1 Scenarios of market share and new vehicle fuel consumption.

FIGURE 1 Total LC GHG emissions of the fleet for the four scenarios in Table 1, from 1995 to 2030.

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Three level energy decoupling: Decoupling at the primary, final and useful level of energy consumption Zeus Guevara MIT Portugal Sustainable Energy Systems, FCUL-IST, University of Lisbon zeusguevara@campus.ul.pt Background: Starting year: Research team: Supervisors:

M.Sc. Sustainable Energy Systems, Technological University of Eindhoven, The Netherlands 2010 João Rodrigues1 Tiago Domingos1; Tânia Sousa1; João Serra2 1 Instituto Superior Técnico de Lisboa, Universidade de Lisboa 2 Faculdade de Ciências da Universidade de Lisboa

OBJECTIVES Reducing the energy intensity of the economy (energy decoupling) is one of the top priorities of the sustainability agenda. As energy is fundamental for every economic activity, any measure to reduce its consumption must be adequately designed so not to negatively affect economic growth. Primary energy intensity, which is the standard indicator for energy decoupling analysis, has a high degree of aggregation and therefore is unable to give enough information about its components. The objective of this thesis is to improve the understanding of energy decoupling by including into the analysis the different levels of energy consumption: Primary, final and useful.

Furthermore, the thesis includes original pieces of work: the first structural decomposition analysis of the Portuguese renewable energy transition, a better approach to track energy efficiency trends of the economy and the first input-output model of useful exergy.

WORK PLAN The work consisted of building a model that disentangled the structure of the energy sector (i.e. Primary to useful energy) from the rest of the economy. To do so several methodological tools were used: Useful work accounting, decomposition techniques, and input-output analysis. RESULTS The main contribution is the decomposition of the primary energy intensity indicator into several self-contained components: Structure and efficiency of primary-to-final conversion; structure and technical efficiency of final-to-useful conversion; useful exergy demand composition and intensity; economic structure; and composition of final demand of non-energy goods and services. Through this model, it is possible to analyze the relative contributions of energy and economic transitions on the overall energy performance of an economy.

134 Eq. 1. Primary-Final-Useful-Economy model of primary energy intensity

FIGURE 1 Structural decomposition analysis of primary energy use in Portugal 1995-2010: Primary – Final – Economy model.

FIGURE 2 Index decomposition analysis of industrial exergy intensity changes in Mexico 1993-2009 by the traditional and the UW-based approaches: Final – Useful – Economy model.

A framework for sustainable pavements implementation applied to portuguese conditions Bahareh Tavallaee MIT Portugal Transportation Systems, Civil Engineering and Architecture Department, Instituto Superior Técnico, University of Lisbon, Portugal tavallaee05@gmail.com Background: MSc. at Pavement Engineering, Shahid Chamran University, Ahwaz, Iran Starting year: 2011 Supervisor: Luís Picado Santos1 Co-supervisor: Jose Manuel Neves1 1 Civil Engineering and Architecture Department, Instituto Superior Técnico, University of Lisbon, Portugal

OBJECTIVES Proposing a comprehensive framework (e.g. MEPDG and its software ME Design) for Portuguese pavement design are demanded by this thesis. This framework includes preparing precise database (to locally calibrate the MEPDG procedure) and adopting a proper behavior model for Portuguese conditions which may result in delivering pavement catalog for Portuguese conditions and developing implementation guidelines for the future adoption of ME Design in Portugal. WORK PLAN In order to approach the defined target, subsequent work areas specified as follow: 1. Literature review on flexible pavement design methods, etc. 2. Data collection for several pavement sections in Portugal (Traffic loading, Material properties, Climate): Selection of pavement sections will be based on some selection criteria such as availability of data, data quality, format of data, etc. 3. Data evaluation (correctness, accuracy, reliability) based on some defined criteria: Check, confirm, and remove outliers. Confirm that selected

pavement sections include values close to the selected performance criteria. 4. Employing data in ME Design in order to verify the quality of database 5. Adopting a proper behavior model for Portuguese conditions and verification of behavior model in ME Design: By analyzing the trends of pavement performance data (rut data, deflections, roughness...) and comparing them with the predicted performance results from the ME Design TM to verify if the performance data is over predicted or under predicted by default empirical (behavior) model for flexible pavements in ME Design. RESULTS The following steps have been accomplished so far: • Writing literature review on Pavement design approaches. • Identifying and organizing required data based on importance for ME Design in separated tables. • Contacting with EP to get and prepare required data (traffic data and material data) for the software. • Collecting and editing the climatic data for required unit and Portuguese conditions.

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Intelligent Traffic Signal Control Cristina Vilarinho MIT Portugal Transportation Systems, Faculty of Engineering of the University of Porto cvilarinho@fe.up.pt Background: Integrated Master in Civil Engineering Faculty of Engineering of the University of Porto, Portugal Starting year: 2012 Supervisors: Jose Pedro Tavares1; Rosaldo J. F. Rossetti2 1 CITTA, Faculty of Engineering of the University of Porto 2 LIACC, Faculty of Engineering of the University of Porto

OBJECTIVES Traffic signal control appeared in response to the need to ensure safety of road users, mainly at intersections. Over the years with the increasing traffic demand, it was realized the impact of traffic lights in terms of efficiency of network operation for the same level of safety. Signal timing offers the opportunity to improve the mobility and contributes to address environmental issues. Nevertheless, the inefficient operation of traffic lights is a common problem certainly experienced on all network settings. Our problem is formulated to perform a real-time signal traffic controller able to find the optimal traffic signal plan including order sequence of signal groups, for local control of traffic lights of isolated intersection. The objectives to be achieved by our traffic control are the following:

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WORK PLAN Literature review concerning existing methods and techniques used in isolated traffic control, as well as other aspects (such as traffic signal constraints, signal control objectives); Develop the methodology and the corresponding implementation algorithms to provide optimal traffic signal control of an intersection; Test and evaluate strategies of traffic signal control in a virtual environment. RESULTS For testing the proposed signal traffic control, it was developed a communication protocol to link our algorithms to the traffic simulator that allows us to get information from the traffic simulation model and modify the its state during the simulation. An integrated traffic control where signal plan design and signal timing are modified at the same time in response to traffic flow fluctuation at a three-arm intersection was developed.

A comprehensive quality indicator for public transport systems evaluation Dimitris Papaioannou MIT Portugal Transportation Systems, Instituto Superior Técnico, University of Lisbon dimitrios.papaioannou@ist.utl.pt Background: Starting year: Supervisors:

Civil Engineering 11’, Democritus University of Thrace, Greece 2011 Luis Miguel Martinez1 1 Civil Engineering Department, Instituto Superior Técnico

OBJECTIVES Development of a Public Transport (PT) quality assessment tool that will be able to evaluate urban PT systems in a comprehensive way. Quality is much more than the elements traditionally studied in the field (information availability, schedule reliability, environmental impacts, etc.). In this work we include and focus on network design and service planning and furthermore want to examine the link between quality on one side and ridership and mode choice on the other side. Current practice shows the difficulty of PT networks in attracting a large number of non-users no matter how much all the traditional quality elements are improved. People are not using public transport either because the system doesn’t cover their needs or because they are captive users of a different mode (mainly car). By taking into consideration all the elements that can affect a person’s choice this research wants to see the effect that each component has and how altering any of them will impact on mode choice and ridership.

WORK PLAN 1. Literature review; 2. Development of indicators in specific elements of urban public transport (accessibility and connectivity) and testing them on existing data; 3. Design and deployment of a survey that will provide all the necessary data for a modelling attempt (4); 4. Development of a quality indicator using S.E.M. and the data gathered from (3); 5. Currently in the deployment of the survey. RESULTS At this stage of the research, the survey is deployed online, disseminated through email lists, social networks and websites, focusing on the cities of Lisbon, Portugal and Thessaloniki, Greece. The aim is to obtain mobility data and preferences regarding PT in these two cities mainly. This data will then be used to develop a SEM model that will rate the performance of PT. Data gathered from other cities will be used for benchmarking and control of the model.

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Understanding and predicting the impact of events on urban mobility Jóni Santos MIT Portugal Transportation Systems, Department of Civil Engineering, University of Coimbra joni@dei.uc.pt Background: Master in Informatics Engineering / University of Coimbra, Portugal Starting year: 2012 Research team: Ana Bastos1; Ana Alves2; Carlos Bento2; Filipe Rodrigues2; Luis Martinez2; João Abreu2 Supervisors: Gonçalo Correia1; Francisco Pereira3 1 Department of Civil Engineering, University of Coimbra 2 Department of Computer Science, University of Coimbra

OBJECTIVES Traffic congestion is increasingly becoming one of the main problems of our cities nowadays. As the world population raises beyond the 7 billion, we realize that more and more individuals want the benefits of urbanization, an increase that our societies, economies and environment, may not be able to sustain much longer. The average commuter spends about 40 hours on traffic congestions per year and this is getting worse every year. Big metropolis need more and more an efficient and dynamic urban planning. This has to be unique to each city and moment in order to provide the best performance in every situation. Ever since the beginning of this century, urban flows and events have been projected online at an increasing rate. Blogs, social networks, detailed digital maps of nearly every place on earth are common realities nowadays. These digital footprints are progressively available and they represent the connection between the physical and the digital part of the city. In our thesis, we intend to take advantage this huge amount of data spread all across the internet (to the city of Lisbon). Moreover, we will perform surveys at events in order to get more detailed information. With these two sources of information, we plan to understand people’s mobility patterns, more specific, mobility regarding to events on urban space.

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We want to understand the impact that events have in a city and then we plan to build models that will predict their impact on the mobility systems. These models will after be used to empower the transportation planners and policy makers with precise information about what will be likely to happen in the next events of the city. In this proposal, we will discuss the methodology that will be used, the survey design and implementation and the problems that we expect to have during the thesis period. This research topic has not been explored much in the past, so there is an opportunity to produce a significant impact on the transportation research state of the art through the completion of this thesis. WORK PLAN • Perform surveys at events in order to get detailed information about people’s mobility to and from events. • Collect all available information about events on online sources. • Build models using this sources of information in order to predict the impact of events on the mobility systems of cities. RESULTS Currently making and gathering data from the surveys and from the internet.

EMS network optimization models with different levels of uncertainty Marco Amorim MIT Portugal Transportation Systems, Civil Engineering Department, Faculdade de Engenharia da Universidade do Porto pdst12005@fe.up.pt Background: Starting year: Supervisors:

Master in Civil Engineering - Transport Infrastructures 2012 António Couto1; Sara Ferreira1 1 CITTA, Faculdade de Engenharia da Universidade do Porto

OBJECTIVES Emergency Medical Response (EMR) systems and its corresponding facilities, vehicle allocation and management have generally been widely and deeply studied subjects. However, most of the studies and efforts have been placed in the mathematical programming – the algorithm itself and the solving technics, mainly heuristics – or in the demand prediction. This has left few concerns to the main objective of the process itself: what are we improving? Of course we can always answer that we are reducing the response time, thus always improving the system. But at which cost and which benefits will we reach? The main objective of this thesis is to measure the social impact of improving the EMR by optimizing the system planning: is a more efficient system also more effective? The research will go through three phases on how to make the system more effective: in the long term (macro-management), in the mid-term (dynamic management), and in the short term (dynamic micro-management), figure 1. The study will comprise all types of EMR calls and the social impact will focus on the road accident victims.

The next step is to study the response system and how to improve it by optimizing the location, number and type of vehicles. This research will approach the three types of management decisions: long, mid and short terms. Finally, the last step will deal with the analysis of the impacts produced by the system optimization. The proposed approaches will be tested in synthetic cities and in the case study of Porto Metropolitan Area using simulation of emergency calls. RESULTS We expect to be able to assess the costs of the EMR system alongside the hospital and social costs of the injuries. This evaluation will involve the analysis the results related to the injury classification MAIS, to be implemented in the EU, the DRG, and the resources required to implement the EMR system.

WORK PLAN The first step is to analyze the severity of the accidents and the benefits of a quick medical response. Using a comprehensive and detailed injury classification, we will assess the impacts of decreasing the medical response in the injury severity. FIGURE 1 Network planning scheme

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Incorporating land-use impacts on BRT and LRT decision making Marcos Schlickmann MIT Portugal Transportation Systems, Faculdade de Engenharia da Universidade do Porto pdst12004@fe.up.pt Background: Master’s degree in Urban Planning - FEUP - Portugal Starting year: 2013 Supervisors: Jorge Pinho de Sousa1; Luis Martínez2 1 NESC Porto - Instituto de Engenharia de Sistemas e Computadores do Porto, Faculdade de Engenharia da Universidade do Porto 2 CESUR, Instituto Superior Técnico, Universidade de Lisboa

OBJECTIVES When governments face the need of improving a transit system they normally have to choose between a set of technological and operational alternatives (e.g., Bus Rapid Transit (BRT), Light Rail Transit (LRT)). In this context, to help correctly evaluate each transport option and its impacts a decision support tool should be used, based for example in a Cost Benefit Analysis or Multiple Criteria Decision Analysis. The objectives of this research are: • To identify, understand and measure the impacts of BRT and LRT systems on accessibility, land uses, land values and densities.

FIGURE 1 BRT

FIGURE 2 LRT

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• To consider those impacts on decision making focusing in sustainability as the major guide to be considered. WORK PLAN To measure those impacts a Hedonic Price Model combined with Geographic Information System (GIS) will be developed, using data collected from the BRT system of Curitiba (Brazil) and the LRT system of Porto (Portugal). After obtaining the results of the analysis, a set of decision-makers, academics and practitioners will be interviewed, in order to understand what level of consideration should be given to those impacts. The final product will be a decision support tool that can help assess more comprehensively why, how and when a BRT or a LRT solution can be more viable. RESULTS The State of the Art and the World shows interesting results: 1. In many cases the final choice are irrational, despite substantial knowledge about the impacts, costs and benefits discussed during the decision making process, favoring one transit solution over another; 2. Many decisions are taken considering the maximum capacity that a solution can deliver instead of the real ridership and demand levels expected for a corridor or for the whole system; 3. Reduction in travel time is still considered as the main criterion to choose a solution, followed by safety concerns, costs and pollution. Land-use, social and economic benefits are sometimes evaluated qualitatively or not considered at all.

Application of datasets from multiple sources for urban and transportation planning: emphasis on cellular network data Merkebe Getachew Demissie MIT Portugal Transportation Systems, Civil Engineering Department, Coimbra University demissie@dei.uc.pt Background: M.Sc. in Transport system - Royal Institute of Technology (KTH) - Sweden Starting year: 2009 Supervisors: Gonรงalo Homem de Almeida Correia1; Carlos Bento2 1 Civil Engineering Department, University of Coimbra 2 Center for Informatics and Systems of the University of Coimbra

ABSTRACT As a result of the increasing number of people, cities face an increasing number of private vehicles and commuters which in turn cause various problems such as traffic congestion, parking difficulties, traffic accidents, loss of space for productive activities, public transport inadequacy and undesirable environmental impacts. In the past, public authorities followed approaches that nowadays are financially unsustainable, focused mainly on expanding the road network to alleviate the problem. However, many analysts argue that the solution for these problems is better addressed through intelligent planning and management of the existing urban and transportation systems. There are various traditional methods for gathering the raw data necessary for urban and transportation planning. Although these methods have the advantage of providing detailed information, their limited coverage and expensive costs of implementation often make them insufficient. More recently, the spread of massive sensoring, namely through the generalized use of cellphone, is producing massive amounts of data with spatio-temporal detail about our daily activities and traveling patterns, which could be important to the planning of urban and transportation systems given their pervasiveness, low cost, and real time nature. In this thesis we explore the use of cellphone data for profiling the dynamics of urban activities and characterizing flows of people for planning of urban and transportation systems in cities. Three types of passive mobile positioning data were used: (1) Call Volume, which is the number of calls; (2) Erlang, which is used to measure the equivalent cellphone traffic

per hour; and (3) Handover, which is the process of transferring an ongoing call from one base station to another without interruption of service. Our observations are based on hourly aggregated cellphone data obtained from a dataset from a telecom company in Lisbon, Portugal. For validation of our results, we collaborated with other data providers in Lisbon to gather different ground truth datasets that could improve our understanding of urban dynamics such as census data, taxi movement, bus movement, traffic count, points of interest, and presence of people. We proposed new approaches to reflect the potential of passive mobile positioning data for urban and transportation planning. Our approach comprises three stages: (1) exploratory data analysis aimed to discover the kind of relationship that emerges between cellular networks data and urban characteristics, activities, and dynamics at a city-scale; (2) use of cellphone data to detect activities associated to the urban areas in what respects to two aspects of activities: spatial patterns of urban activities, and intensities of urban activities along the hours of a day; and (3) extraction of cellular network data for development of models that predict hourly traffic status. Our results confirm that passive mobile positioning data, taking the advantage of its pervasiveness and availability with reasonably less cost, can provide ways to analyse the dynamics of urban activities at a larger scale. In addition, our approach complements traditional urban data collection methods, which are usually made available less frequently to urban and transportation planners, and is especially useful for developing countries where other approaches are too expensive.

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An advanced technological in-vehicle system aiming to reduce the hazardous driving actions Mohammad Mahdi Hajizamani MIT Portugal Transportation Systems - TransportationEngineering, Civil Engineering and Architecture Department, IST, Portugal mhajizamani@yahoo.com Background: MSc. Transportation planning and engineering, Iran Starting year: 2009 Supervisors: Sílvia Costa Shrubsall1 ; José Manuel Viegas1 1 Instituto Superior Técnico, Universidade de Lisboa

OBJECTIVES This project aims to improve road safety by reducing the hazardous driving actions committed by drivers. The main objective of this research is to design an in-vehicle intelligent surveillance and enforcement system (system “X”) which can communicate with the driver, vehicle and environment (including infrastructure, police and motorway operator). This system will warn, help and potentially control the driver to drive in a stable and safe condition. This procedure will lead to less accident and casualties. WORK PLAN 1. Literature review • In-vehicle detection systems and their safety impacts. • In-vehicle action systems (e.g. automatic brake system) and their safety impacts. • Simulation platforms (i.e. traffic module). 2. Data collection • Motorway section specifications and geometric and their associated driving hazard. • Analyzing the trajectories data of the vehicles in a real world traffic flow. 3. Modeling (simulation) • Develop the model in the AnyLogic which is an Agent-Based Modeling (ABM) platform. • Run the simulation model and calibrate the system “X”.

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4. Result analysis • Analyze results from model outputs. • Analyze traffic characteristics (e.g. capacity and level of service) in each scenario. 5. Policy Studies • Discuss about different penetration rates of system “X” and policy strategies to influence penetration. • Political, economic and social analysis. RESULTS Following results are expected by the end of this PhD thesis: • Design and model an in-vehicle system which is able to obtain data from different types of in-vehicle safety systems and sensors to help the driver to have better perception, decision and action in dangerous situations. • Assessing the impact of using this system in the vehicles in different driving conditions. • Discussing legal and economical aspects of the system implementation and necessary requirements. • Finding an optimum point of promoting road safety by increasing the system safety factors while maintaining the proper road capacity.

Operator-based analysis of intercity passenger intermodality Ryan F Allard MIT Portugal Transportation Systems, Department of Civil Engineering and Architecture, IST, University of Lisbon ryan.allard@tecnico.ulisboa.pt Background: Starting year: Research team: Supervisors:

MSc, Transportation Systems, Instituto Superior Técnico; BSc, Aerospace Engineering, MIT 2011/2012 Olga Petrik1; Guineng Chen1; Yu Shen1; Heather Jones1; Marcos Correia1; Maria Pedro1; Filipe Moura2; João Abreu2; Luis Picado Santos2; Luis Miguel Martínez3; Bruno Santos 4; Joseph Sussman5 Filipe Moura2; Joseph Sussman5 1 MIT Portugal Transportation Systems / IST, University of Lisbon; 2 Department of Civil Engineering and Architecture, Instituto Superior Técnico, Universidade de Lisboa; 3 Policy Analyst/ International Transport Forum; 4 Air Transportation and Operations/ Technical University of Delft; 5 Department of Civil and Environmental Engineering and Engineering Systems Division, Massachusetts Institute of Technology

OBJECTIVES Research in intermodality (closely connected transportation systems) has typically been only qualitative, and very little has any measurement of benefits. The importance is briefly highlighted in the illustration of an intercity trip in Figure 1. The connections between the various trip stages of the public transport trip are often so poor (long wait, physical transfer effort, no supporting services) that travellers would rather drive. Additionally transport operators, who would be the ones creating intermodal services, have rarely been the focus of research. This research fills these gaps by attempting to answer the question of “how intermodality may be pursued”. We aim to measure and demonstrate the benefits that could motivate operators to pursue intermodality as an organizational strategy. This would be valuable

for improving the connectedness of long-distance transportation systems worldwide. Research Questions: • What is the value that passengers attach to long-distance intermodal transport services? • How can offering long-distance intermodal services affect transport operators? WORK PLAN For the first research question, we develop a market share model created using discrete choice theory through an intermodal survey for long distance travel in the Iberian Peninsula. We then develop a set of competitive models based on game theory (specifically market estimation/optimization of operator service offerings with competitive and collaborative strategies under a wide variety of

145 FIGURE 1 A Schematic Comparison of the atypical “Most” and “Least” Seamless Intercity Travel Options from the passenger perspective. Note that some elements of seamlessness such as schedule coordination and baggage transfer are omitted.

FIGURE 2 An example of one of the Operator Service Optimization Models used in the Work.

economic and political scenarios). An example is shown in Figure 2. This would allow estimation of the potential effect on operator operations. Policy recommendations will be made based on these results. RESULTS Results will be able to tell us: 1. How much travellers would be willing to pay for reducing travel time and trip connection times, having connection insurance and baggage

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transfer services on intercity trips, and whether these vary by traveller/trip characteristics such as trip purpose or traveller age. 2. What competitive, local, economic and policy environment characteristics make it attractive for operators to provide a connecting service with good connection quality for their travellers, and what policy incentives governments can use to encourage operators to adopt these strategies.