Abstracts

 

The following oral presentations will be given by our members on Saturday 8th July during Session II: Science, Society and Public Engagement

O1: Patricia Bernal. “Bacteria use nano-weapons to protect plants”

Type 6 secretion system (T6SS) is a syringe-shaped molecular weapon that is present in certain types of bacteria. This system is designed to inject toxins (poisons) inside other prokaryotic cells. The bacteria that possess this secretion system or lethal weapon manage to kill many of the other bacteria that coexist in the same environment, and therefore this powerful system of competition confers great evolutionary advantages.
There are bacteria that can protect plants against the attack of organisms that cause diseases. Organisms that cause diseases or infections in plants are known as phytopathogens and the beneficial bacteria that protects them are known as biocontrol agents. Pseudomonas putida is a good example of a biocontrol agent that not only protects plants from the attack of phytopathogens but also promotes their growth.
P. putida KT2440 contains three different T6SS weapons capable of secreting at least 10 different types of toxins. This system is responsible for the death of different phytopathogens very important in agriculture because of the severity of the infections they cause in important crops e.g. potato or cauliflower. The ability of P. putida to kill these pathogens using T6SS is given both in vitro and in plant trials where the plants were protected by the biocontrol agent T6SS. This new mechanism of biocontrol described in bacteria that live in close contact with plants, is a tool that is expected to help us in the near future to combat phytopathogens and thereby protect our crops without the use of chemicals.

O2: Priscila Ramos. “The promise of the pig for mankind, revealing emerging pluripotency in the pig early embryo using single cell transcriptomics” (P15)

The pig is an excellent model for biomedical sciences. Its similarity to humans in size, anatomy and physiology allows it to meet the shortcomings of the mouse model and makes it more suitable for investigating human disorders. Organ shortage is a worldwide problem and a lot of people die every year while waiting for an organ transplantation. Recent advances suggest that this problem could be solved in the future by growing human organs inside animals. In order to generate these chimeric animals, donor human pluripotent cells are injected into pig early embryos to provide them a proper environment to differentiate into multiple tissues, but this process has proven to be very inefficient yet. Cell fate decisions during preimplantation embryo development have been extensively studied in mice and humans, but there is a fundamental gap in our knowledge of early embryogenesis in the pig that is hindering our progress in developing more efficient methods for chimera generation. Here, we set out to obtain the first detailed single cell expression profile of embryonic cells during the emergence of pluripotency in the pig. We present a high-quality single-cell RNA-seq dataset of preimplantation embryos at four different stages. This dataset could be compared to human datasets and to gene expression in human pluripotent cells to determine the precise moment in which these cells could better integrate into pig embryos. Furthermore, we have used this new knowledge to test novel strategies for the isolation and establishment of pig embryonic stem cells.

O3: Alazne Dominguez. “Mammalian Synthetic Biology”

Humans have been manipulating the genetic code of plants and animals for thousands of years by selectively breeding for desirable traits such as cows that produce more milk or beef, the breeding of dogs to be good at hunting. Baker’s yeast has been changed over the years so it ferments to produce the best bread, wine and beer. The difference now is that we can read the genetic code – we better understand what a bit of DNA is actually telling a cell to do. This means we can start to write our own instructions using DNA. So in effect we can ‘reprogramme’ or ‘reengineer’ a cell to do something it wouldn’t naturally do – like make a useful chemical like a medicine or a flavour or fragrance, or even a fuel for your car. T-cell therapy is a working example of synthetic biology in human system. Here a patient’s T cells are removed, synthetically engineered so that they better detect cancer cells and then given back to the patient. But changing one or two traits to help us fight a disease in a specific part of the body is a whole other world from trying to engineer a person’s looks or intelligence. Synthetic biology is not about playing God. It can’t be used to create ‘designer babies’, or to bring back extinct animals such as dinosaurs.

O4: Ibon Santiago. “Active matter: rockets at the nanoscale”

Living matter is a constant source of inspiration for generations of scientists and engineers. One of the features that distinguishes living organisms from inanimate matter is their constant fight to stay away from thermodynamic equilibrium. Just like rockets in space, they achieve this by consuming energy in the environment (or internally stored) to convert it into motion. In particular at small scales, this is achieved by so-called molecular motors. Can we fabricate such tiny motors in the lab?

Progress in nanotechnology has enabled the synthesis of artificial micro and nano devices that can harness chemical energy and translate it into useful motion. These are examples of active matter. I will present the physics behind self-propelled devices and give examples ranging from nanorockets and enzyme powered devices to swarms of bacteria and flocks of birds.

O5: Maria Pin No. “It is just basic”

Are we always looking for a specific application for our research? Sometimes, we do research in Science just to know the answer to unresolved questions without seeking the application of the results. That is what we know as basic research. In some cases, few years later those discoveries are applied in new technology. By re-examining a very specific fundamental research I did in my PhD about cycloisomerisation of 1,6-dienes, I try to show why basic research is important and why we need to keep funding it. First, some well-known examples of fundamental research in Science are first described. Then, I deep into my project by explaining both the good results I obtained and the problems I needed to face. I finish by hypothesising about the future of the project and by reminding that fundamental research is necessary.

O6: Roger Castells Graells. “Generating virus-like particles for potential bionanotechnological applications” (P16)

We are studying virus structures and using synthetic biology approaches to generate new bionanotechnological tools.
Nudaurelia capensis omega virus (NωV) is an insect virus with a pH-controlled maturation mechanism that involves a precise autocatalytic cleavage within the capsid protein. This cleavage converts the full-length protein (α) to the β and γ peptides and is accompanied by a dramatic decrease in the diameter of the particles. The stages in the compaction can be visualized by cryo-EM and the ability to control the movement of the subunits has potential uses in nanobiotechnology. We have produced NωV VLPs in plants and have shown that reduction in pH results in cleavage of the α protein and compaction to mature particles. In order to generate a model of virus maturation we will perform cryo-EM analysis of the process of capsid maturation of NωV VLPs concomitant with pH change.
The production of dynamic virus-like particles as protein-based nanomaterials will allow the development of future applications which require mobile as opposed to static structures, for example, nanomachines with controlled motion.

O7: Maria Cuenca Cambronero. “Forecasting species persistence under climate change conditions”

The decline of water quality due to climate change and land use determined by the use of pesticides and leading to eutrophication is a recognized threat to ecosystem services and to the economy. However, the causes and effects of climate and land use changes occur over many decades and are therefore difficult to measure. Critical for gauging the long-term consequences of climatic change is the reconstruction of evolutionary dynamics over extended time axes. Here, we reconstruct the evolution of fitness traits (mortality, fecundity and growth rate) to climatic change in Daphnia magna populations using animals resurrected from sediment biological archives with known history of eutrophication and temperature changes. Experimental evolution trials are used to measure fitness response of populations sampled across 80 years (1960-present) to the forecasted temperature increase for the upcoming century. In addition, we measure the evolution of fitness traits in response to the combined effect of temperature increase with pesticides and food quality changes. This study will identify fitness traits targets of natural selection and the long-term consequences of environmental stress in metazoan populations.

O8: Mario Gonzalez Jimenez. “Tracking the origin of DNA transcriptional bubbles with OKE spectroscopy” (P28)

The processes important to the biological function of DNA (replication, transcription, denaturation, and molecular intercalation) have in common that they start with the breaking of the hydrogen bonds between the bases of the nucleic acid. Driven by the torsional stress of the molecule, the destabilisation of the weak bonds leads to the splitting of a section of the double helix of DNA into single strands, forming a gap in the nucleic acid known as a transcriptional bubble. In this process, low-frequency phonon-like modes that propagate along the length of the molecule play a key role, expanding and contracting the space between the bases without unstacking them. Despite their importance, the study of these phonon-like modes has been limited by the strong absorption of infrared radiation by water in the terahertz region and only indirect evidence of their existence have been obtained measuring unnatural solid DNA preparations (humidified films, fibres, etc.) with techniques like inelastic x-ray and neutron scattering. We have employed optical Ker-effect (OKE)

The following posters will be presented during the symposium

At the end of Sunday, we will award the prize for the best poster so please do not forget to vote!

Biomedicine

P1: Zuriñe Anton. “Mitochondrial membrane association of human Atg8 proteins during mitophagy”

The phospholipid cardiolipin (CL) has been proposed to play a role in selective mitochondrial autophagy, or mitophagy. CL externalization to the outer mitochondrial membrane would act as a signal for the human Atg8 homologue, LC3. The latter would mediate both mitochondrial recognition and autophagosome formation, ultimately leading to removal of damaged mitochondria. We have applied quantitative biophysical techniques to the study of CL interaction with various Atg8 human homologues, namely LC3B, GABARAPL2 and GABARAP. We have found that LC3B interacts preferentially with CL over other di-anionic lipids, that CL-LC3B binding occurs with positive cooperativity, and that the CL-LC3B interaction relies only partially on electrostatic forces. CL-induced increased membrane fluidity appears also as an important factor helping LC3B to bind CL. The LC3B C-terminus remains exposed to the hydrophilic environment after protein binding to CL-enriched membranes. In intact U87MG human glioblastoma cells rotenone-induced autophagy leads to LC3B translocation to mitochondria and subsequent delivery of mitochondria to lysosomes. We have also observed that GABARAP, but not GABARAPL2, interacts with CL in vitro. However neither GABARAP nor GABARAPL2 were translocated to mitochondria in rotenone-treated U87MG cells. Thus the various human Atg8 homologues might play specific roles in different autophagic processes.

P2: Alba Iglesias Vilches. “Assembly of Combinatorial Synthetic Operons for the Production of Bioactive ”

Bacteria constitute an important source of antibiotics. However, the discovery of new bioactive secondary metabolites is slowing down. The tetronate family of natural products constitutes a structurally and functionally diverse group of secondary metabolites whose potent bioactivities (antimicrobials, antivirals, and anti-cancer agents) makes them attractive targets for clinical and industrial exploitation. The supervisory team have recently elucidated the biosynthesis of the minimal tetronate antibiotic abyssomicin C, solved the crystal structure of key enzymes involved in the biosynthesis, and identified the molecular basis for the first bona fide Diels-Alderase. These findings indicate that the biosynthesis of abyssomicin is highly amenable to re-engineering, as the enzymes involved in the synthesis of the tetronate (AbyA1) and the spiro-tetronate-forming Diels-Alderase (AbyU) are both capable of accepting structurally diverse substrates. One significant challenge in synthetic production of novel tetronate compounds, is the supply of polyketides that vary in their chain length, degree of saturation, and substitution pattern. The aim of this project is to assembly synthetic operons, based on the wild-type abyssomicin pathway originally present in the actinomycete Verrucosispora maris AB-18-032, that will produce diverse polyketides, and thence, novel tetronate compounds for biopharmaceutical development.

P3: Judith Gomez Martinez. “Sperm preparation and egg activation for bovine intracytoplasmic sperm injection (ICSI)”

Bovine ICSI can overcome deficiencies associated with poor sperm viability and motility (e.g. with frozen-thawed sexed semen), but its efficiency is poor. This may be due to suboptimal sperm capacitation, leading to impaired acrosome reaction (AR). Existing bovine ICSI protocols also involve assisted egg activation, which we seek to refine in order to improve ICSI success. Working with frozen-thawed semen from a single sire, in three replicated experiments we found that Lyshophosphatidylcholine (LPC) treatment (100 µg/ml, 15 min) increased (P=0.007) the proportion of AR sperm compared to non-treated controls (0.74±0.019 vs 0.37±0.021). We then assessed dose (10, 40, 80 and 100 µg/ml) and duration of exposure (0, 15, 60 and 120 min) of heparin during sperm preparation followed by LPC treatment. In four replicated experiments,  40 µg/ml heparin for a minimum of 15 min led to the highest (P<0.001) proportion of (0.80) of AR sperm. Next we determined optimal protocols for parthenogenetic activation. In six replicate experiments, a new activation protocol with SrCl2 (20mM) for 6 h plus cytochalasin (CB; 7.5 µg/ml )/cyclohexamide (CHX;10 µg/ml) after a previous incubation in calcium ionophore A23187 (CaI; 5 µg/ml for 5 min), increased (P=0.027) the proportion of blastocysts of activated (0.34±0.06) compare to conventional protocols, including ethanol (7% for 5 min) plus 5 h CHX(0.23±0.05), CaI followed by 5 h in CHX/CB (0.15±0.04) and 6 h in SrCl2/CB after CaI (0.12±0.04). Consequently, ongoing experiments are comparing standard in vitro fertilization with ICSI using the optimal protocols for sperm preparation and egg activation.

P4: Elena Sanchez Heras. “Formation and Function of Membrane Contact Sites between the ER and the phagocytic pathway in the Retina Pigment Epithelia”

We are investigating the formation and regulation of ER membrane contact sites (MCSs) with all stages of the phagocytic pathway in the mammalian retinal pigment epithelium (RPE). The daily renewal of photoreceptor outer segments (POSs) is a process balanced with a disposal mechanism by which the shed POSs undergo acidification along the phagocytic pathway in the RPE, prior to fusion with the lysosomes. When the disposal mechanism is compromised, undigested lysosome remnants of oxidised POSs, termed lipofuscin, remain in the cell and can accumulate with age. Lipofuscin accumulation is a major risk factor implicated in macular degeneration, a degenerative disease of the eye. Here we aim to characterize ER MCSs with the phagocytic pathway, including phago-lysosomes by a combination of electron and super-resolution light microscopy in human macrophages that reproduce the same sequence of phagosome maturation events as the RPE The regulation of ER MCSs with the phagocytic pathway is investigated using siRNA-mediated depletion of candidate tethering proteins in macrophages fed POSs, with findings confirmed in human iPS-RPE cells. The ability to modulate MCSs in all populations of ER-phagosome identified will allow us to further our investigation into the role of MCSs in regulating the maturation of the phagocytic pathway and the processing of POS-derived membrane lipid in the RPE.

P5: Oihane Jaka. “Adipogenic transdifferentiation potential of human primary skeletal muscle-derived fibroblasts is retained after GFP-lentivirus transduction”

Fatty degeneration in skeletal muscle is a hallmark of many myopathies, sarcopenia, obesity and type-2 diabetes. It has been shown that human skeletal muscle fibroblasts (but not the myogenic cells) have the potential for transdifferentiation into adipocytes in culture, suggesting that these cells may be the cause of adipocyte accumulation in muscle. In order to study the transdifferentiation potential of these cells in vivo (xenotransplantation), it is important to genetically label the fibroblasts before transplanting them into the host species to confirm that they still retain their potential for adipogenic transdifferentiation. In this study we have transduced human primary skeletal muscle fibroblasts with a GFP lentivirus to determine if their transdifferentiation potential is retained. The transduction efficiency was 75%, and following treatment with fatty acids (300μM oleic acid and 300μM palmitic acid) for 72 hours, cells were fixed and analysed by immunohistochemistry. The results showed that applying fatty acids to GFP-positive fibroblasts resulted in their transdifferentiation into adipocytes as evidenced by a clear accumulation of Oil Red O positive lipid droplets and increased expression of the adipogenic transcription factors C/EBPα and PPARγ compared to the non-fatty acid treated GFP-positive fibroblasts. This study confirms that GFP lentivirus transduction of human primary skeletal muscle derived fibroblasts does not affect their transdifferentiation potential and can therefore be used to successfully label cells for future in vivo experiments.

This work was funded by the BBSRC.

P6: Sara Alvira de Celis. “Membrane protein insertion and folding by the bacterial holo-translocon”

Maintenance of the proteome integrity is essential for viability. The cell has elaborated parallel and interconnected strategies focused on the synthesis, folding or degradation mechanisms for keeping a homeostatic cell balance. Moreover, many of proteins synthesised by ribosomes must be transported across and into membranes before arriving at their functional destination.

In bacteria, the SecYEG translocon is a protein-conducting membrane channel that can function promoting the transport of secretory proteins or assist the insertion of integral membrane proteins. In this last case, SecYEG associates with YidC and SecDF-YajC to form a functional super-complex called the holo-translocon (HTL) able to insert and fold membrane proteins in a co-translational translocation mechanism universally conserved. In this work, we use classical biochemistry combined with synthetic biology approaches and cryo-electron microscopy to unravel the interactions of a membrane protein with the HTL. We want to address what is the pathway that follows a membrane protein from its synthesis at the ribosome to the passage through the HTL, and elucidate how the HTL folds and promotes the assembling of fundamental proteins within the lipid bilayer. The results of this work will unveil the basis for the potential design of new antibiotics, creating new strategies for fighting against multidrug-resistant bacteria, a current global health emergency.

P7: Nerea Alonso. “Prevalence of low alkaline phosphatase and ALPL mutations in patients with osteoporosis”

Hypophosphatasia a rare inherited skeletal disorder characterised by defective mineralisation, and multiple fractures caused by loss-of-function mutations in ALPL gene, leading to low levels of alkaline phosphatase. The prevalence and clinical significance of low ALP levels in patients with osteoporosis has not been studied. Here we evaluated the prevalence of low ALP levels in a large clinic based population of patients and screened for the presence of ALPL mutations in the subjects compared with controls. From a total cohort of 3285 patients referred to the osteoporosis clinic over a 12-year period we identified 17 (0.52%) in whom ALP levels were low (<40U/L). We selected 30 osteoporotic patients from the same population with normal levels of ALP as controls. Mutations in ALPL were identified in 13/17 patients with low ALP and 0/30 controls (p=0.017). Twelve patients were heterozygous carriers of missense mutations and one had a novel nonsense mutation (c.303c>a, p.Try301X). Of the mutations detected, 9 had previously been reported as causing recessive Hypophosphatasia, but one was novel (c.901a>t, p.Arg301Trp). In silico analysis predicted that p.Arg301Trp is a pathogenic change. Regression analysis showed that ALPL mutation carriers showed a significant reduced number of comorbidities than controls (p=0.001, beta=0.476, 95%CI=[0.28-0.67], r2=23%). In summary, low serum ALP is unusual in osteoporotic patients but a high proportion of patients who have low ALP are carriers of mutations in ALPL. Further studies need to be conducted to address the role of ALPL mutations in osteoporosis.

P8: Maria Ester Quesada del Bosque. “Evaluation the effect of bronchoconstriction on airway epithelial cells responses in asthma”

P9: Lourdes Riquelme Dominguez. “Molecular mechanisms of dopaminergic neurodegeneration in Caenorhabditis elegans”

Neurodegenerative disorders such as Parkinson’s disease affect millions of people and no permanent cure currently exists. Discovery of neuroprotective genes and gene-variants will broaden our understanding of neurodegenerative diseases and suggest new targets for pharmacological or genetic therapies. We have previously established a new model of dopaminergic neurodegeneration in Caenorhabditis elegans. In this model, a dominant (d) mutation in a Transient Receptor Potential channel (TRP), trp-4, results in the progressive and fully penetrant degeneration of dopaminergic neurons. A collection of mutant strains that show neuroprotection from trp-4(d)-induced degeneration was isolated by forward genetic screening. Whole-genome sequencing of one of these mutant strains identified an early stop codon in an ER resident protein. We showed that this mutation was indeed responsible for neuroprotection through three complementary experiments. Based on the molecular identity of this gene and its previously described roles in regulating AMPA receptors, we hypothesize that we have identified a cargo receptor or auxiliary subunit for the TRP-4 channel, and a possible regulator of other members of the TRP channel family. We are currently characterizing its mechanism of function.

P10: Jon Palacios. “In vivo electrophysiological recording of hippocampal cells in head fixed but freely exploring mice”

To fully understand the neuronal network mechanisms that underpin behaviour it is necessary to record from individual neurons in the brain. This is extremely difficult in humans and only possible during procedures requiring the insertion of electrodes for the treatment of certain conditions such as epilepsy. Therefore, animal recordings are required. Extracellular recordings can detect the spiking activity of neurons but to measure the underlying subthreshold membrane dynamics requires intracellular recording. To achieve this in an awake freely moving animal is almost impossible due to mechanical instability. So to circumvent this problem methods to reduce mobility by head fixation have been developed to make recordings in awake mice. One popular technique involves head fixed mice running on a treadmill in a virtual reality environment but in these conditions naturalistic stimuli are limited, in particular the use of the whiskers which are a principle sensory input in mice.
Here we present a novel experimental system where head fixed mice explore a large (~40cm diameter) circular environment that is able to freely move in response to the animals’ movements. At the same time we are able to record neuronal activity from neurons within the brain. The ultimate goal is to be able to study the neuronal responses during decision making tasks based on spatial memory, which will require mice to confidently explore the environment and be free from excessive stress.
Our preliminary data show recordings from principle neurons in the hippocampus in a variety of configurations including extracellular, juxtacellular and whole-cell configurations.

P11: Nuria Casanova Vallve. “The impact of osteoarthritis development on pain, locomotor activity and muscle inflammation and metabolism in a rodent surgical model of osteoarthritis”

Pain, inflammation, inactivity, and muscle atrophy are features of osteoarthritis (OA). Inflammation and inactivity independently induce muscle mass loss and insulin resistance, however relative contribution to muscle decline in OA and the metabolic and molecular mechanisms are unknown. We aimed to quantify the impact of OA progression on pain, activity and muscle inflammation and metabolism in a surgical model of OA. Rats were allocated to 3 groups: meniscus transection (MNX) surgery (n=20) or sham surgery to one limb (n=20), no intervention (naïve, n=20). Pain behaviour and locomotor activity were quantified longitudinally and at 28 and 55 days blood, knee joints and the extensor digitorum longus (EDL) muscle were collected under terminal anaesthesia. Inflammatory cytokines were quantified in blood and muscle metabolites and muscle protein:DNA ratio were determined at day 28 and 55. MNX rats exhibited weight bearing asymmetry (p<0.001) and decreased paw withdrawal thresholds by day 17 (p<0.01), and rearing activity was reduced by day 27 (p<0.05) post-surgery. MNX knee joints had OA pathology on day 28 and 55. No significant levels in blood cytokines were evident on day 55 after OA induction and no difference in muscle metabolites content and muscle protein:DNA ratio between groups at day 28 or 55 were found. Muscle decline and inflammation are not evident in the MNX rat in this study despite changes in pain behaviour and joint pathology. Other factors such as a chronic sedentary life secondary to joint pathology and pain may have a greater role in OA muscle decline.

P12: Pilar Acedo. “Light-based therapies to improve pancreatic cancer outcomes”

Introduction: Pancreatic cancer is one of the 10 most lethal forms of cancer worldwide. Current clinical trials include new combinations of chemotherapy drugs but major limitations remain, therefore new approaches are needed. Our strategy is to target pancreatic cancer using a technique called Photochemical Internalisation (PCI), which can significantly enhance the efficacy of cancer chemotherapy. PCI is a light-triggered intracellular drug delivery method, which combines low dose Photodynamic Therapy (PDT) with chemotherapy to induce efficient cytosolic delivery of therapeutic compounds to their specific subcellular targets.

Aims: To determine the efficacy of PCI in overcoming pancreatic cancer drug resistance compared to conventional monotherapy and to identify the cell death mechanisms induced by these protocols.

Methods: Photoactivatable compounds were co-administered with saporin or gemcitabine, in human pancreatic cancer cell lines and PDX-derived cells. Cell viability was assessed by MTT assays and Trypan Blue & Neutral Red stainings, alongside internalisation and intracellular localisation of photosensitisers (flow cytometry, confocal microscopy).

Results: Despite minimal cell death induced when the drugs or photosensitisers were used in monotherapy, PCI synergistically enhanced saporin and gemcitabine cytotoxicity (p<0.001) using very low concentrations. Moreover, PCI induced endosomal disruption following light excitation and caspases 3/7 activation, in a time dependent fashion.

Conclusion: Our findings demonstrate the potential of PCI to enhance the efficacy of cancer chemotherapy for pancreatic cancer using sub-lethal doses of PDT. To enhance the translational relevance of this Project, we will employ human pancreas 3D-scaffolds, refining dosages before moving on to in vivo experiments.

P13: Isabel Romero Camarero. “Investigating the mechanisms of FoxC1 derepresion in AML”

The transcription factor FOXC1 is derepressed in a tissue-inappropriate manner in human Acute Myeloid Leukaemia with functional consequences and prognostic significance: its mis-expression contributes to a block in monocyte/macrophage differentiation, and enhances clonogenic potential. The mechanisms underlying its mis-expression remain unknown.

CRISPR-Cas9 library screening enables the identification of critical genetic regulators in a variety of biological processes. These pooled approaches require a cellular phenotype that can be assessed by survival, surface markers or fluorescence. We have used the CRISPR/Cas9 technology for generating an mCherry knock-in reporter in the human K562 cell line, in which the FOXC1 expression is silenced. This reporter system is driven by the endogenous FOXC1 promoter. Treatment of cells with inhibitors of the PRC2 complex (known to contribute to FOXC1 repression) leads to increased mCherry fluorescence and upregulation of FOXC1 expression. This cell line tool facilitates studies on transcription of this gene allowing for high-throughput genetic screening to identify genes that maintain FOXC1 repression.

The K562 reporter cell line was transduced with a lentiviral human CRISPR knockout (GeCKO) library containing 122,411 sgRNAs targeting 19,050 protein coding genes and 1864 miRNA precursors. After puromycin selection, the transduced cells were maintained for 4 weeks in culture. mCherry fluorescence was monitored by flow cytometry at different time points and mCherry+ cells were sorted, gDNA was extracted, sgRNA cassettes were PCR amplified and deep sequenced. After successful initial quality assessment of the screen, we have identified promising candidates that we are currently validating using shRNA and pharmacological inhibitors.

P14: Giovanny Rodriguez Blanco. “Tissue proteomics outlines members of the arachidonic acid pathway as markers for prostate cancer”

Prostate cancer (PCa) remains as one of the most common cancer types in men worldwide. We aimed to identify and validate proteins involved in PCa development and progression using nano LC-MS/MS and tissue immunohistochemistry. The proteomics dataset included protein fractions of PCa samples (n=34) and normal adjacent tissue (NAP, n=33), and we used label free quantification (LFQ) to identify differentially expressed proteins between PCa and NAP. Fatty acid synthase (FASN, p=1.05E-07) and Anterior Gradient protein 2 (AGR2, p=7.34E-07) were up-regulated in PCa tissue. In addition, TEBP protein, encoded by PTGES3 gene, was also found highly overexpressed in PCa tissue compared to NAP (p=9.10E-10). Moreover, analysis of 79 arachidonic acid (AA) pathway proteins showed that 15 of them were differentially expressed in PCa compared to NAP (six proteins were down-regulated and nine were up-regulated). Proteins AGR2, FASN and the lipoxygenase LX15B were further validated by immunohistochemistry. LOX5, identified previously as a marker of prostate cancer also part of the AA pathway was also included. Tissue microarray immunohistochemical staining of an independent cohort of patients was performed to study biochemical recurrence (BCR) after radical prostatectomy (RP). It was found that both low percentage of positive tumour cells for AGR2 (HR (95% CI) = 0.61 (0.43-0.93), and low percentage of positive tumour cells for LOX5 expression (HR (95% CI) = 2.53 (1.23-5.22) are predictors of BCR after RP. In conclusion, AGR2, and two proteins of the AA pathway, TEPB and LOX5 can be used as diagnostic and prognostic markers for PCa.

P15: Priscila Ramos. “The promise of the pig for mankind, revealing emerging pluripotency in the pig early embryo using single cell transcriptomics” (O2)

The pig is an excellent model for biomedical sciences. Its similarity to humans in size, anatomy and physiology allows it to meet the shortcomings of the mouse model and makes it more suitable for investigating human disorders. Organ shortage is a worldwide problem and a lot of people die every year while waiting for an organ transplantation. Recent advances suggest that this problem could be solved in the future by growing human organs inside animals. In order to generate these chimeric animals, donor human pluripotent cells are injected into pig early embryos to provide them a proper environment to differentiate into multiple tissues, but this process has proven to be very inefficient yet. Cell fate decisions during preimplantation embryo development have been extensively studied in mice and humans, but there is a fundamental gap in our knowledge of early embryogenesis in the pig that is hindering our progress in developing more efficient methods for chimera generation. Here, we set out to obtain the first detailed single cell expression profile of embryonic cells during the emergence of pluripotency in the pig. We present a high-quality single-cell RNA-seq dataset of preimplantation embryos at four different stages. This dataset could be compared to human datasets and to gene expression in human pluripotent cells to determine the precise moment in which these cells could better integrate into pig embryos. Furthermore, we have used this new knowledge to test novel strategies for the isolation and establishment of pig embryonic stem cells.

P16: Roger Castells Graells. “Generating virus-like particles for potential bionanotechnological applications” (O6)

We are studying virus structures and using synthetic biology approaches to generate new bionanotechnological tools.
Nudaurelia capensis omega virus (NωV) is an insect virus with a pH-controlled maturation mechanism that involves a precise autocatalytic cleavage within the capsid protein. This cleavage converts the full-length protein (α) to the β and γ peptides and is accompanied by a dramatic decrease in the diameter of the particles. The stages in the compaction can be visualized by cryo-EM and the ability to control the movement of the subunits has potential uses in nanobiotechnology. We have produced NωV VLPs in plants and have shown that reduction in pH results in cleavage of the α protein and compaction to mature particles. In order to generate a model of virus maturation we will perform cryo-EM analysis of the process of capsid maturation of NωV VLPs concomitant with pH change.
The production of dynamic virus-like particles as protein-based nanomaterials will allow the development of future applications which require mobile as opposed to static structures, for example, nanomachines with controlled motion.

P17: Alazne Dominguez. “Mammalian Synthetic Biology” (O3)

P18: Elena Moreno Martinez. “Resistance of Saccharomyces cerevisiae to lignocellulose-derived inhibitor”

Production of bioethanol from lignocellulosic hydrolysates is a promising biofuel technology but is subject to inhibition by chemicals released during the pretreatment of biomass. Weak acids, furans and phenols impair yeast growth and fermentation, thus reducing efficiency of the process and increasing the cost. Some of the molecular mechanisms of action and response to these stressors are characterized, but little is known about how uniform such effects are across individual cells within an isogenic cell population. Non-genotypic heterogeneity refers to the expression of different phenotypes within a clonal cell population, despite all individuals being genetically identical. Understanding individual-cell effects of hydrolysate inhibitors could help engineering of population-level resistance. The degree of heterogeneity in resistance traits was specific to particular inhibitors in the fermenting yeast S. cerevisiae. Inclusion of a second inhibitor could alter heterogeneity relative to either inhibitor alone. Short pretreatments with subinhibitory concentrations of the inhibitors indicated adaptive resistance and adaptive heterogeneity responses. RNAi assisted genome evolution was also used to reveal combinations of gene knockdowns that can synergistically increase resistance towards these inhibitors. Findings derived from this research will provide better understanding of the behaviour of yeast in lignocellulosic ethanol fermentations, and might help improve production.

P19: Laura Fort. “Retrovirus reactivation in Motorneuron Disease and Frontotemporal Dementia”

Frontotemporal Dementia (FTD) is the second most prevalent form of dementia, after Alzheimer’s disease, in patients less than 65 years of age. In the last decades, evidence has shown a clinical, genetic and pathologic overlap between FTD and amyotrophic lateral sclerosis (ALS), suggesting a common pathological mechanism. FTD/ALS causing mutations are known to be involved in RNA regulation and endosomal traffic. Retroviruses are found to be abundant in patient tissue post mortem. We have identified mutations in retrotransposon/retrovirus (RT/RV) repressors, BoYb, SoYB and Vretano, via our functional genetic screen in Drosophila for enhancers of FTD-3 caused by a mutant ESCRT-III subunit, CHMP2BIntron5. We also identify an increase in RT/RV copy number in our ALS/FTD model. How endosomal dysfunction and RT/RV de-repression are linked is to be defined and this is the aim of my PhD. A better understanding of the biological mechanism of RT/RV activation in FTD/ALS will help to develop more effective diagnostic and therapeutic strategies for this condition.

 

P20: Margarita Segovia. “Investigating the mechanistic causes of C9orf72 – Related ALS”

Amyotrophic Lateral Sclerosis (ALS) is an adult-onset fatal degenerative disorder characterised by progressive paralysis resulting from relentless degeneration of upper/lower motor neurons (MNs) and death usually within 2-3 years from symptom onset. The C9orf72 GGGGCC-intronic repeat expansion is the most common known genetic cause of ALS accounting for 10% of cases. Our project is focused on identifying the pathophysiological mechanism(s) induced by this hexanucleotide intronic repeat expansion. The development of a reliable cell model is crucial in order to go deeper on the understanding of ALS. We have developed a cellular system consisting in a co-culture of iMNs (motor neurons directly derived from mouse stem cells) and iAstrocytes (astrocytes directly derived from human fibroblasts whose donors are patient carrying the mutation C9orf72). This cell model suggest a great advantage on the generation a proper cell system directly derived from patients, avoiding the use of animal models and saving time regarding the duration of this differentiation (1 month). After the positive molecular characterization obtained, the next step is the electrophysiological characterisation. After the recording of inward and outward currents, our preliminary results show the proper maturation of these iMNs but also some differences in the level of outward currents. Our preliminary data suggests the physiological maturation of this cell model but also the possibility to find a defect related with excitotoxicity. This will be the first step to find new targets and to design new strategies for neuroprotective therapies and identify much-needed biomarkers of this disease.

 

Computer Science and Engineering

P21: Jesus Monge Alvarez. “SMARTCOUGH: Continuos monitoring of cough events based on smartphones”

The potential of telemedicine in respiratory illness has so far failed to live up to expectations, in part because of poor objective measures of symptoms such as cough events, which could lead to early diagnosis or prevention. Considering the burden that these conditions constitute for national health systems, an effort is needed to foster telehealth potential by developing low-cost technology for efficient monitoring and analysis of cough events. Continuous monitoring and analysis of cough events is currently feasible and affordable using generic readily available sensors such as those embedded in current smartphones. This brings along important research challenges in using a smartphone as a medical device, namely the necessity to deal with noisy inputs in mobile environments as well as battery consumption issues related to continuous sensor monitoring and computing. Smartcough implements robust signal processing and efficient machine learning algorithms to overcome those challenges, featuring:
* Continuous smartphone-based cough monitoring (no additional devices required).
* Robust cough detection in noisy environments with low consciousness of the medicalisation of user’s life (e.g., when carrying the phone in pocket/bag).
* Efficient implementation of cough detection algorithms leading to 48h+ autonomy running in background.
* High sensitivity (up to 90.45%) and specificity (up to 99.81%) in a variety of noisy environments (SNR between -15 dB and 15 dB).
* Self-calibration and auto adaptive power threshold to improve accuracy and efficiency.

P22: Carla Gonzalez Soliño. “Enzymatic fuel cells for self-powered healthcare sensors”

Wearable and non-invasive biosensors have appeared as an alternative to the current analytic methods available for health monitoring. They are of paramount interest to patients with chronic diseases, or in medical areas where fast or continuous monitoring is needed.
Amongst these biosensors, enzymatic fuel cells (EFC) are a very promising technology, they convert biomolecules directly into electricity through the action of redox enzymes. Glucose, acetylcholine or lactic acid are suitable fuels for these devices which act as biomarkers for severe diseases such as diabetes or Alzheimer disease. The electrical signal generated is proportional to the concentration of biomolecules. Therefore, EFC can work as amperometric biosensors by choosing specific enzymes that can recognise the desired biomarker.
Despite the advances made in the field, these devices lack of long-term stability as well as power output. This research project thus focusses on developing electrodes with high surface area and conductivity, and on enhancing the electron transfer between the enzyme and the electrode to improve EFC performance. Films based on polypyrrole arise as a suitable material, owing to its good conductivity and porous structure as well as good biocompatibility. Moreover, different types of immobilisation are being studied to improve the enzymes’ stability on the surface. Lastly, we aim to test these devices on non-invasive fluids like saliva or sweat.

P23: Javier Escudero. “Graph regularised tensor factorisation of EEG signals based on network connectivity measures”

A tensor is a multi-way representation of data, or a multidimensional array. Each dimension in the tensor is called a mode or a way. Thus, tensors represent “cubes” of data in 3D or higher dimensions. Tensor analysis can provide a good way to discover the main features of the data and extract the hidden underlying information especially in the case of having big data size. To this end, tensor factorisations represent decomposition methods for high dimensional data and there are primarily used as an extension of singular value decomposition. Tensor factorisation methods have been shown to be powerful for describing signals which in general change in time, frequency, and space in fields such as blind source separation, biomedical signal processing, feature extraction, and classification.

The tensor decompositions can be regularised to alleviate overfitting and small sample size estimation errors by constraining the obtained solution to satisfy some metric. In this work, we provide a novel extension to the theory of graph regularisation for regularising multiple graphs. That is, we make use of prior knowledge about the interrelationships between the data described in the tensor by modelling such relationships as a graph and we employ graph regularised tensor factorisation on an electroencephalogram (EEG) dataset employing brain connectivity networks as the basis of our graphs. Subsequently, we perform graph regularised tensor factorisation on the EEG data to reduce noise and interferences inherent to the EEG. We demonstrate the efficacy of the algorithm theoretically and on some real EEG examples.

P24: Fidel Alfaro Almagro. “UK Biobank Brain Imaging: First 10,000 subjects and new Imaging Derived Phenotypes”

Environmental Sciences

P25: Juan Carlos de la Concepcion. “Natural variation in a rice immune receptor interface extends response to pathogen effectors”

Pathogens deliver an array of molecules, termed effectors, to manipulate host cellular processes for their own benefit. Perception of effectors inside cells largely relies on immune receptors of the NLR (Nucleotide-binding, Leucine-rich Repeat) family, triggering a response to stop infection. This process creates a high selection pressure in the pathogen, driving the emergence of new effector variants that escape recognition. However, plants also evolve receptor alleles with broader recognition for these variants. Here, we unravelled the molecular details of this arms race co-evolution between the rice NLR Pik and the rice blast pathogen effector AVR-Pik.
The rice NLR allele Pikp1 recognizes AVR-PikD through direct binding to an unconventional integrated Heavy Metal Associated (HMA) domain, resulting in disease resistance. Polymorphic effector variants have lower binding affinity for Pikp-HMA in vitro, and evade immune recognition in plants. Another rice NLR allele, Pikm1, recognizes these polymorphic variants, and biochemical characterization of the Pikm-HMA domain has revealed a higher binding affinity for these effectors. Crystal structures of Pikm-HMA complexed with different effector variants provided an atomistic explanation for the extended recognition specificity. This study shows the structural basis of natural variation in pathogen recognition specificities by NLRs, and will allow structure-guided protein engineering to improve disease resistance in crops.

P26: Ana Brugos. “Inactivation of Pectinesterase from Orange Juice Using Thermal Process Generated from Microwave”

Orange juice is one of the most consumed beverages in the world. Only Brazil accounts 85 % of world exports. The short shelf life in natural juice is due to turbidity loss caused by degradation of pectin by enzyme pectinmethylesterase (PME) naturally present in fruit. Turbidity has outstanding importance since it allows the product a freshly extracted juice aspect. Enzymatic inactivation (PME) commercially occur around (95 – 98) °C during (10 to 30) s at the conventional heat treatment. However, recent studies have been performed, in order to test lower temperatures, for shorter periods to prevent over-processing of juice and minimise nutritional loss. In this context, electromagnetic energy has been considered as a distinctive method. In this study, the PME inactivation in orange juice (variety Pêra) was compared through conventional heating and microwaves. Samples with 25 mL were submitted to controlled unifocal microwaves incidence (CEM, star system, USA), at temperature interval from (50 to 70) °C at (0 to 60) s processing times. Temperature-time profiles during the microwave batch processing were acquired using a fibre optic thermometer (Luxtron 812) inserted centrally into the tube. Lethality was performed considering maximum temperature achieved at processes and z-value of 6.5 °C. PME inactivation from (10 to 36) % was achieved for conventional heating, while (18 to 45) % for the microwave process. Isothermal processing time varied from (14 to 69) s at Tref = 50.6 °C to conventional heating, while (16 to 37) s at Tref = 52.8 °C for the microwave process.

Social Sciences and Humanities

P27:  David Gonzalez Alvarez. “Upland Archaeology in the Cantabrian Mountains”

For too long uplands have been regarded as Europe’s last ‘natural landscapes’. However, mountain landscapes are in fact mostly a result of cumulative long-term human interactions with the environment. Recent investigations in the Alps and Pyrenees have emphasized the cultural dynamism and notable anthropogenic impact on European mountains since the Later Prehistory. Meanwhile, the Cantabrian Mountains in the North of Spain are archaeologically under-explored.
This poster summarizes the methods, the aims and the potentialities of my on-going postdoctoral research project, which I am developing one foot at Durham University and the other at the Institute of Heritage Sciences in Santiago de Compostela. Taking inspiration from the above-mentioned research, this project aims to obtain new archaeological information to place the communities who inhabited the Cantabrian Mountains at the forefront of the biography of these cultural landscapes. The nature of human exploitation of the uplands is explored in order to create narratives about the human experiences and the historical processes that mediated in the domestication of upland landscapes. This way, the project provides an archaeological account of the anthropization processes in the the Cantabrian Mountains from the spread of Neolithic (ca. 4800 BC) until the Roman conquest (late first century BC). Historical Ecology and Landscape Archaeology provide the theoretical framework for exploring the available archaeological data and paleoenvironmental sequences in this region, aiming to support a diachronic social interpretation of the cultural landscapes. This is achieved by analyzing the settlement patterns and subsistence systems deployed by the local communities in these mountains.

P28: Amalia Mas Bleda. “Are Altmetrics less biased than citation counts? A comparison of Spanish and UK research”

Research is often assessed with the aid of bibliometric indicators, such as the number of publications in citation-based databases like the Web of Science (WoS) and Scopus, and citation counts. These databases (especially WoS) are biased towards literature in English and towards journal articles. In addition, citation counts take time to accrue and only reflect scientific impacts inside academia. These limitations have led to the development of a variety of alternative indicators, which have the potential to reveal non-scholarly types of impacts and avoid the language biases traditional citation indexes. This study compares Spanish and UK research in eight subject fields using a range of bibliometric and social media indicators. For each field, lists of Spanish and UK journal articles published in the year 2012 and their citation counts were extracted from Scopus. The software Webometric Analyst was then used to extract a range of altmetrics for these articles, including patent citations, online presentation mentions, online course syllabus mentions, Wikipedia mentions and Mendeley reader counts and Altmetric.com was used to extract Twitter mentions. Results show that Mendeley is the altmetric source with the highest coverage, with 80% of sampled articles having one or more Mendeley readers, followed by Twitter (34%). The coverage of the remaining sources was lower than 3%. All of the indicators checked either have too little data or increase the overall difference between Spain and the UK and so none can be suggested as alternatives to reduce the bias against Spain in traditional citation indexes.

Exact Sciences, Physics and Chemistry

P29: Pablo Aparicio Sanchez. “Structural and Electronic Properties of NaVOPO4 polymorphs as Cathode Materials for Na-Ion Batteries”

Many advances in reversible energy storage have been done since the commercialization of the first Li-ion batteries. In the past years, different cations have been studied as an alternative to Li ions. Sodium can be a good alternative to alleviate the concerns of lithium availability, expand the active cathode materials, and improve both rate capabilities and battery life. Sodium-ion batteries (NIBs) have been attracting more attention in the last years due to the abundance of sodium in Earth. Among several different materials used as cathode materials for NIBs, the vanadium-based phosphates have drawn much attention in recent years. Most of these compounds are sodium superionic conductor (NASICON) solids with an open structure for fast sodium transfer. More recently, it has been demonstrated that the vanadyl phosphate VOPO4 with a layered structure is a good candidate as cathode active material. There are several polymorphs of this compound with different structures, such as alpha, alpha-I, alpha-II beta, epsilon, delta, etc. So far only three sodiated products have been identified: alpha, alpha-I and beta. In this work, we have performed density functional theory (DFT) calculations on the alpha, alpha-I and beta NaVOPO4 polymorphs. We have been able to reproduce the experimental structural and cell parameters. We have studied the electronic states of the sodiated and desodiated materials, and compute the band gaps, voltage, and formation energies of the above-mentioned compounds. We have also investigated the Na and vacancy migration barriers using the nudge elastic band (NEB) method, reproducing the experimental observations.

P30: Laura Martinez Maestro. “Crystal phases of MAPbI3 thin films at room and low temperature for solar cell applications”

Organic-inorganic metal-halide perovskite solar cells are emerging as a promising photovoltaic technology to harvest solar energy with efficiencies above 20% reported recently[1] which is comparable to existing thin film photovoltaic technologies based on silicon, cadmium or copper.[2]
The most common perovskite used in the fabrication of efficient perovskite solar cells is methylammonium lead iodide (MA, where MA=CH3NH3PbI3).[3]
There has been a lot of effort to study the morphology and crystallinity of MAPbI3 thin films and their effect in the optoelectrical properties of MAPbI3. However, it has not been until recently when characterization studies addressing MAPbI3 grains within solar cell configuration are beginning to get published. [4]
It is pretty clear than a deeper understanding of the size and interface effects in needed and to do so, Transmission Electron Microscopy (TEM) is a promising technique that would be able to characterize the structural properties of our materials at atomic scale.[5]
In our work, MAPbI3 thin films growth by evaporation method, are investigated due to determine their crystal orientation at low and room temperature using a High resolution STEM microscope.

References.

1. Science 2015, 348, (6240), 1234-1237.
2. Nature Energy 2016, 1, 16081.
4. The journal of physical chemistry letters 2015, 6, (12), 2292-2297.
5. Journal of electron microscopy 2008, 57, (6), 175-179

P31: Maria Crespo Cuadrado. “Nanomaterial risk assessment, a key element for nanotechnology future”

In the last decade, nanotechnology has been confirmed as one of the fastest-growing industries, triggering what some people define as the third industrial revolution. The ability of nanomaterials to improve the properties of everyday products has opened the doors to a wide range of applications from the healthcare to the electronic sector. The key to nanomaterial success is based on the new properties that can be found at the nanoscale. As the volume of the material decreases, quantum mechanics become predominant and surface area is increased enhancing its reactivity. Besides, due to their similar scale, biological elements can be incorporated into nanoparticles, overstepping the limits of traditional medicine and creating a new concept of personalised therapy.

Despite these broad future perspectives, the absence of common international agreements regarding the environmental and health implications of nanomaterials can slow down their progress. Therefore, current efforts are focusing on the development of standard nanomaterial assessments to determine their behaviour through their life cycle and their potential impacts on the ecosystem. The establishment of relevant properties to create safe-by-design nanomaterials and the publication of nanomaterial databases are necessaries to ensure nanotechnology future. However, scientist’s awareness and involvement will be essential to avoid future problems and promote the production of safe nanomaterials from the first stages.

P32: Daniel Escalera Lopez. “Magnetron-sputtered Ni-doped MoS2 nanoclusters for the hydrogen evolution reaction”

Transition metal dichalcogenides (TMDs), and in particular molybdenum disulfide (MoS2), have been reported as promising alternatives as substitutes for precious metals catalysts: for example in the hydrogen evolution reaction (HER). The main limitation of MoS2 lies in the fact that only the Mo-edge sites are catalytically active, leaving both S-edge and basal plane sites inactive. A recent strategy utilized for enhancing MoS2 hydrogen evolution activity is the activation of the S-edge sites by the incorporation of transition metal (TM) atoms (Fe, Co or Ni). TM doping on MoS2 nanoparticles (NPs) is scarcely reported probably due to the difficulty in separating the effects of surface area and morphology changes from the electrocatalytic enhancement To tackle such purpose, we report a novel methodology to prepare TM-doped MoS2 nanoclusters based on a one step, dual target (Ni and MoS2) magnetron sputtering and gas condensation deposition. Aberration-corrected STEM images along with EDX analysis confirm the presence of Ni and MoS2 in the formed nanoclusters. Electrochemical characterization of Ni-doped MoS2 nanoclusters demonstrated a 3-fold increase in exchange current density and a 100 mV shift in the HER onset potential compared with undoped MoS2 nanoclusters, in agreement with results reported previously. Anodic stripping voltammetry experiments on the tested Ni-doped MoS2 nanoclusters reveal distinctive electro-oxidative features related to doped Mo-edge and S-edge sites according to their abundance ratio. Correlation of S-edge doping with the HER catalytic enhancement proves the success of dual target magnetron sputtering deposition for preparing TM-doped TMDs.

P33: Mario Gonzalez Jimenez. “Tracking the origin of DNA transcriptional bubbles with OKE spectroscopy” (O8)

The processes important to the biological function of DNA (replication, transcription, denaturation, and molecular intercalation) have in common that they start with the breaking of the hydrogen bonds between the bases of the nucleic acid. Driven by the torsional stress of the molecule, the destabilisation of the weak bonds leads to the splitting of a section of the double helix of DNA into single strands, forming a gap in the nucleic acid known as a transcriptional bubble. In this process, low-frequency phonon-like modes that propagate along the length of the molecule play a key role, expanding and contracting the space between the bases without unstacking them. Despite their importance, the study of these phonon-like modes has been limited by the strong absorption of infrared radiation by water in the terahertz region and only indirect evidence of their existence have been obtained measuring unnatural solid DNA preparations (humidified films, fibres, etc.) with techniques like inelastic x-ray and neutron scattering. We have employed optical Ker-effect (OKE)