NEU-URV Student Exchange Program · 2020. 1. 13. · take place. Among PEMFCs, we can find hydrogen...

Universitat Rovira i Virgili

Department of Chemical Engineering (http://www.etseq.urv.es/9deq/ca/6-home.html)

Project Proposals

NEU-URV Student Exchange Program

June-July 2020

Total positions offered: 20

http://www.etseq.urv.es/9deq/ca/6-home.html

Department of Chemical Engineering - URV - 1 -

Project #1

Title: WetCAP: Preparation and characterization of biodegradable capsules for oral care products

Supervisors: Marta Giamberini, Bartosz Tylkowski

Contact: [email protected] [email protected]

Research Group: METEOR

Description:

According to Science [1] encapsulation, developed approximately 65 years ago, has been defined as a major interdisciplinary research technology. Encapsulation has been used to deliver almost everything from advanced drugs to unique consumer sensory experiences [2]. Dry mouth (xerostomia) is a common side effect of many prescription and nonprescription drugs, including drugs used to treat depression, anxiety, pain, allergies, obesity, acne, asthma, and others. Moreover, the dry mouth can also be a side effect of muscle relaxants and sedatives. This symptom can also raise a risk of gingivitis (gum disease), tooth decay, and mouth infections, such as thrush [3].

The main objective of the project focuses on preparation and characterization of innovative, biodegradable microcapsules containing wetting/lubricant actives. As a trigger, for a delay encapsulated actives release, we wish to use enzymes which exist in the mouth/saliva. We wish to prepare the capsule shell by applying biodegradable polymers, such as Xiangtan gum or starch. As the encapsulated co-actives we wish to use Cetylpyridinium chloride for sustained antimicrobial effects to get at the therapeutic needs of dry mouth patients. The microcapsules will be prepared by a spray drying method, which is relatively low cost, flexible, and leads to the production of high quality and stable particles. It is the most commonly used encapsulated technique in industry making the WetCAP project easy for scale-up and application in industry. Obtained microcapsules will be characterized by optical and scanning electron microscopes.

Number of positions: 1

1. Zhang, J.; Coulston, R.J.; Jones, S.T.; Geng, J.; Scherman, O.A.; Abell, C. One-step

fabrication of supramolecular microcapsules from microfluidic droplets. Science (New York,

N.Y.) 2012, 335, 690-694.

2. Tylkowski, B.; Pregowska, M.; Jamowska, E.; Garcia-Valls, R.; Giamberini, M. Preparation of a

new lightly cross-linked liquid crystalline polyamide by interfacial polymerization. Application to

the obtainment of microcapsules with photo-triggered release. European Polymer Journal

2009, 45, 1420-1432.

3. Scully, C. 8 - dry mouth (xerostomia and hyposalivation). In Oral and maxillofacial medicine

(third edition), Churchill Livingstone: 2013; pp 91-97.

mailto:[email protected]



Project #2 a & b

Title: Self-assembling polymers for novel proton transport membranes

Supervisors: Dr. Marta Giamberini

Contact: [email protected]

Research Group: METEOR (http://www.urv.cat/recerca_innovacio/en_meteor.html)

Description:

In the last 40 years, there has been a growing interest toward the "hydrogen economy", that is a system where hydrogen is in charge of energy delivery and acts as an energy carrier (like electricity) and not as a primary energy source (like coal or oil)1. This could be a possible solution to the huge energy problems caused by the disappearance of fossil fuels. On the other hand, hydrogen needs to be produced by an energy consuming method, stored and finally used as fuel in a fuel cell to deliver electric energy when needed.

As far as hydrogen production is concerned, the idea of using solar energy to perform water splitting, according to the reaction:

2H2O → 2H2 + O2

is currently under development2. The energy required for this reaction can be supplied by sunlight, but requires the use of a dye-sensitized solar cell, as water does not absorb the electromagnetic radiation emitted by the sun to promote water splitting in a kinetically facile way. This process can be combined with the reduction of carbon dioxide to carbohydrates, and is generally referred to as "artificial photosynthesis". As presented in the international conference “TOWARDS GLOBAL ARTIFICIAL PHOTOSYNTHESIS. Energy, Nanochemistry & Governance”, the artificial photosynthesis project has the category of a global research project comparing or even overtaking the Human Genoma project. The two red-ox half-reactions involved in water splitting must take place in two different compartments so that hydrogen and oxygen gases can be stored in different containers. For this reason, a water-splitting device has to include in a single cell the following components: a light harvesting device, a water-oxidation catalyst, a proton-reduction catalyst and a proton-exchange membrane (PEM). The separation of the anodic and cathodic compartments through a PEM is crucial in order to separate efficiently H2 and O2 as well as to prevent short-circuiting of the cell. Moreover, in such a device the PEM should be capable of resisting quite severe conditions of pH and red-ox potentials.

Fuel cells are a kind of electrochemical device that converts chemical energy directly into electrical energy. They can be divided into five major categories: alkaline, phosphoric acid, solid oxide, molten carbonate and proton exchange membrane. Proton exchange membrane fuel cells (PEMFC) work at relatively low temperature (about 80 ºC), have high power density, can vary their output quickly to meet shifts in power demand and are suitable for automobile applications. In PEMFCs the electrolyte is constituted by a polymer membrane, able to conduct protons, which is sandwiched between two electrodes. They, in turn, contain Pt-based catalysts that allow the oxidation and reduction reactions to take place. Among PEMFCs, we can find hydrogen fuel cells and direct methanol fuel cells (DMFC).

In DMFCs, the fuel is directly methanol; this entails several advantages over fuel cell systems based on hydrogen and compressed natural gas3, as methanol can be simply transported and applied under room temperature and atmospheric pressure by pumping from existing gasoline infrastructures without an external compressor or refrigeration equipment. This makes the volume of DMFCs as small as possible.

Furthermore, methanol has the highest hydrogen-to-carbon ratio among alcohols, thus giving the highest energy generation and the least carbon dioxide emissions; in addition, methanol is miscible with water: as a consequence, DMFCs can be fueled with different concentrations of a methanol aqueous solution. Besides, methanol can be obtained through biomass and is more environmentally friendly than gasoline since it breaks down quickly in the environment. The ideal membrane to be used


http://www.urv.cat/recerca_innovacio/en_meteor.html


in a DMFC should resist to relatively high temperatures, transport protons efficiently, be an insulator to electrons, act as barrier between the anode and cathode, possess good mechanical strength and be inexpensive.

The membranes currently used as PEM are far away to be considered optima with regard to their performance; therefore, the development of new proton exchange membranes is a topic of permanent interest. In this project we tackle this problem by preparing membranes based on self-assembling columnar side-chain liquid-crystalline polymers which lead to the formation of biomimetic ionic channels. These channels contain basic atoms (oxygen or nitrogen) to interact with protons thus allowing their transport across the membrane.

N.A. Kelly, T.L. Gibson, M. Cai, J.A. Spearot, D.B. Ouwerkerk, International Journal of Hydrogen Energy, 35, 892-899 (2010).

T.L. Gibson, N.A. Kelly, International Journal of Hydrogen Energy, 35, 900-911 (2010).

K. Kordesch, G. Simader, Fuel Cells and their Applications, Weinheim, Germany (ISBN 3-527-28579-2), p. 2-8 (1996).

Positions: 2

A. The first one deals with the synthesis and characterization of columnar side-chain liquid-crystalline polymers. This internship is addressed to strongly motivated students with an Organic Chemistry background. We offer the possibility of working with the classical methodology of the organic chemistry synthesis, as well as with techniques such as 1H and 13C NMR, Fourier-transform infra-red spectroscopy (FTIR), polarized optical microscopy (POM), differential scanning calorimetry (DSC), and small and wide-angle X-ray diffraction (XRD).

B. The second one is focused on the preparation and assessment of PEMs based on columnar side-chain liquid-crystalline polymers. In this case, the internship is addresses to strongly motivated students with Applied Chemistry or Chemical Engineering background. The student will have the possibility to work with membrane preparation techniques, microscopic techniques (environmental and conventional scanning electron microscopy -E-SEM, SEM, atomic force microscopy-AFM) and contact angle measurements, as well as with XRD and POM; moreover, permeability tests and electrical characterizations (impedance tests, current-voltage curves, MEA tests) will be eventually performed.


Project #3 a & b

Title: Understanding complex systems using big data

Supervisors: Dr. Roger Guimerà Manrique & Dr. Marta Sales-Pardo

Contact: [email protected] / [email protected]

Research Group: Science and Engineering of Emerging Systems, SEES:lab (http://seeslab.info)

Description:

Technological advances during the last fifteen years have boosted our capacity to generate and store data in many areas of science and business. Indeed, 90% of the world’s stored data has been generated in the last two years, and the availability of such large quantities of data (for example time-resolved geo-location of mobile devices or usage data of social networks) is changing the way we think about crisis response, social mobilization, marketing, and intelligence.

Big data is a particularly relevant opportunity for the study of complex systems such as cells, ecosystems or economies. In complex systems, non-linear interactions among the many components that comprise the system give rise to macroscopic behaviors that cannot be explained by looking at individual components alone. Traditionally, the analysis of complex systems was constrained by the limited information available from the different components (and layers of components) comprising the system (for example, metabolites and proteins in cells, species in ecosystems, and people and institutions in economies). The availability of unprecedented highly detailed data on these systems opens the door to significantly advance our understanding of their behavior, and of how this behavior evolves in time. With large amounts of data we now have the possibility to uncover correlations and patterns that will enable us to build predictive models of these systems.

The goal of our research is to contribute, from the perspective of statistical mechanics, new tools and methods for the analysis and modeling of large-scale datasets.

This project will involve computational work, thus students selecting this project should have an interest in programming.

Positions: 2



http://seeslab.info/


Project #4

Title Transformation of biomass into bioplastics using microwave and microbial fermentation reactors

Supervisors: Dr. Francesc Medina & Dr. Magda Constantí

Contact: [email protected] / [email protected]

Research Group: Heterogeneous Catalysis Group, CATHETER (http://www.etseq.urv.es/catalisi/) and Physical and biological interface Group, INTERFIBIO (http://www.etseq.urv.es/nbg/Interfibio/)

Description:

Nowadays the main industrial use of lignocellulose is direct combustion. Its usage as feedstock for value added compounds has a great potential for replacing petrochemical resources in industry. The chemical structure of lignocellulose obstructs its cleavage into profitable products. Lignocellulose is made of lignin, hemicellulose and cellulose. Its proportion depends on the concrete biomass source, nonetheless cellulose use to be the most abundant one. Thus, finding a successful process to hydrolyse cellulose would lead to a satisfactory usage of biomass as a feedstock to any industrial process. Enzymatic hydrolysis of cellulose is the most common treatment found on bibliography. However, in this work, a new process of hydrolysing cellulose and hemicellulose and taking benefit of it is followed to replace the expensive enzymatic process. Biomass pretreatment is assisted by microwave reactor, and characterized with TOC (total organic carbon), HPLC (high performance liquid chromatography) and XRD (X-ray diffraction). Then the hydrolysed carbohydrates are used in fermentative processes, verifying their aptness for microbial feedstock. Different bacteria are used in order to obtain optically pure lactic acid or PHB; the first one extremely appreciate starting product for polylactic acid (PLA) production, a highly consumed bioplastic. Microwave pretreatment converted 80 % of cellulose and 90% hemicellulose, then bacterial fermentation by L. delbrueckii delbrueckii or B. coagulans produces D-lactic acid 97% optically pure. The growing demand of PLA and PHB together with the capability of explode lignocellulosic biomass potential, place this work as a potential industrial application.

The student will work on the fractionation and characterization of lignocellulose, cellulose and/or hemicellulose materials for their application in microbiological processes.

Positions: 1



http://www.etseq.urv.es/catalisi/


Project #5

Title: Assessment of an electrochemical genosensor array for early detection of individuals at high risk of osteoporosis and osteoporotic fracture.

Supervisors: Ciara O’Sullivan


Research Group: Interfibio Research Group (www.interfibio.com)

Description:

As a consequence of the ageing of the society, osteoporosis (literally “porous bone”) and its complications is becoming more and more prevalent, making the bone disease a health priority in many parts of the world. Usually osteoporosis manifests itself in a drastic manner, i.e. through fracture of the osteoporotic bone in the affected individual. This causes a serious burden for the patient and a high medical care cost for the society. Currently osteoporosis is commonly known as a “silent killer” disease. Up to 20% of patients die in the first year following a hip fracture, mostly due to pre-existing medical conditions. Less than half of those who survive the hip fracture regain their previous level of function [1]. Early detection of individuals at risk of osteoporosis and starting a suitable treatment before fractures and complications arise would drastically improve the current situation. Until now little or no measures for prevention or early detection of osteoporosis are taken, because no simple, yet sufficiently accurate or sensitive tools for early detection are available.

Osteoporosis is a polygenic disorder, determined by the effects of several genes, with each gene separately having only relatively modest effects on bone mass and other determinants of fracture risk [2, 3]. Recent advances in high-throughput “-omics” technologies have greatly increased the scale and scope of genomics research and recent studies have suggested that profiling of genetic variations could not only identify those at high risk of developing osteoporosis but also enhance the predictive accuracy of fracture prognosis. The overall goal of the project is the development point-of-care tool for bone disease (osteoporosis) prevention, detection and treatment.

To achieve this goal an electrode array have been designed and fabricated using screen printing (SP) technology. This screen printed-based chip consists of 64 working electrodes made of gold sharing a common reference and counter electrodes, which will be used in a genetic biosensor for the detection of single nucleotide polymorphisms (SNPs) and microRNAs relevant to osteoporosis diagnosis. The main task of this work will be the assessment of the electrochemical genosensor array, which includes the characterisation of the electrode surface by both electrochemical (cyclic voltammetry (CV) and impedance spectroscopy (EIS)) and microscopic techniques (scanning electron microscope (SEM) and atomic force microscope (AFM). Then, the ability of the electrode surface to support the formation of self-assembled monolayers (SAMs) of DNA and alkanethiols will be evaluated and subsequently, the performance of the screen printed electrode (SPE) array as a biosensor will be assessed for the electrochemical detection of a DNA marker using a model system. Finally, the SPE array will be used for the detection of SNPs and microRNAs for osteoporosis diagnosis.

Literature:

[1] https://www.iofbonehealth.org/facts-statistics: Keene GS, Parker MJ, Pryor GA (1993) Mortality and morbidity after hip fractures. BMJ 307:1248

[2] Williams FM, Spector TD. The genetics of osteoporosis. Acta Reum Port. 2007;32(3):231–40.

[3] Stewart TL, Ralston SH. Role of genetic factors in the pathogenesis of osteoporosis.



www.interfibio.com

https://www.iofbonehealth.org/facts-statistics


Project #6

Title: (Bio)Electrochemical Tools for Integration of Biological Systems and Electronic Components



Research Group: Interfibio Research Group (www.interfibio.com)

Description:

The integration of biological and electronic systems not only excites the imagination but also opens new opportunities and possibilities in human health and rehabilitation. Significant progress has been achieved in receiving signals from living systems and affecting global cell function with external signals. Electrochemistry is the natural method of choice for such interfaces. However, despite the explosion of life science advances, the integration of electronic and biological systems remains woefully underdeveloped. The overall goal of the project is to apply principles of Synthetic Biology to develop biological cells controlled by electronic signals.

The aim is to create synthetic cellular receptors selectively activated by linear peptide ligands. These ligands will be generated by the electrochemical reduction of disulfide bonds in the cyclic precursor peptide. In order to endow the system with the ability to activate a selected ligand:receptor pair in the presence of other disulfide bond‐containing polypeptides, we propose to construct peptide‐selective bio‐electrodes. These electrodes will act essentially as artificial “enzymes” and the engineered synthetic receptors will activate the expression of signal emitting gene products.

To achieve this goal, the first task is to develop a bioelectronic interface on microfabricated electrodes. Initially, the direct electron transfer of the pyrroloquinoline quinone glucose dehydrogenase (PQQ-GDH) system to the electrode surface should be demonstrated using carbon fiber electrodes modified with graphene nanosheets [1]. Graphene provides conductive substrate capable of direct electron transfer but is not easily adapted to microfabrication and patterning. We need therefore to embody PQQ-GDH in a bioelectronic interface based on metals that are common in microfabrication and amenable to selective patterning (i.e. gold). Several solutions will be explored: (i) pattern graphene on photolithographed gold electrodes and (ii) use directional molecular “wires” to link PQQ to gold electrodes facilitating direct electron transfer. The second task is to perform the controlled release of ligands from microfabricated electrodes. The photolithographed electrodes will be modified by pin deposition or by electrochemical patterning with different activating ligands. Then, the ligands will be released by applying potential to individual electrodes exploiting (i) disulphide bond reduction or ester hydrolysis and (ii) host-guest affinity manipulation or ion-chelating capacity of poly-His tags. Finally, the bioelectronic system should be integrated in a compact device. The bioelectronic integrator should be constructed on an electrode array incorporating sensory and ligand-activating electrodes as well as cell-hosting electrodes. The preferred embodiment is to use low cost array-fabrication methods such as screen-printing or printed circuit board (PCB) manufacturing. Once methods are proven, photolithography will be used to achieve the resolution needed.

Literature:

[1] Koushanpour A., Guz N., Gamella M., Katz E., Biofuel Cell Based on Carbon Fiber Electrodes Functionalized with Graphene Nanosheets, ECS Journal of Solid State Science and Technology, 5 (8) M3037-M3040 (2016).



www.interfibio.com


Project #7

Title: Development of sandwich dual aptamer based assay for the detection of specific Mycobacterium Tuberculosis markers.

Supervisors: Ciara O´Sullivan


Research Group: Interfibio Research Group (http://www.etseq.urv.es/nbg/Interfibio/)

Description:

Tuberculosis (TB) remains to be a leading infectious case of mortality worldwide. New diagnostic tools

which will offer ease to use portable devices that do not require laboratory facilities or training of

specialist for the screening of active and latent TB is of high importance1. The detection of TB relies

mainly on microbiological testing of sputum samples 2 and the culture of the Mycobacterium

Tuberculosis itself. Both tests suffer from moderate sensitivity and specificity, smear microscopy can

only provide qualitative results 1 and traditional culture method is time-consuming as requires at least

10-14 days with rapid liquid culture 3.

To diagnose TB accurately, it requires reliable biomarkers. The early secretory antigenic target (ESAT

6) and culture filtrate proteins (CFP10) and (MPT64) are antigens specifically secreted by MbT

identifying pulmonary TB patients. The detection of mycobacterium antigens reported to date use

antibody as recognition element 4–6. An alternative to antibodies that can tolerate extreme

environmental conditions are aptamers. Aptamers are synthetic nucleic acids that specifically bind to a

wide range of targets, from small molecules to whole cells 7–10.

Objective:

The aim of this project is evaluation of selected aptamers against specific tuberculosis targets and the development of sandwich dual aptamer assay for the detection of tuberculosis.

Project outline:

Previously selected aptamers will be truncated in order to reduce size and costs. The secondary structure will be predicted using m-fold and GQRS mapper and the effect of truncation on affinity and specificity will be carried out using Surface Plasmon Resonance (SPR), Apta-PCR Affinity Assay (APAA), Enzyme Linked Aptamer Assay (ELAA). Full-length aptamers and their truncated versions will be evaluated to see which aptamer should be used as reporter and capture molecule and which dual aptamer pairs gives the best performance in terms of affinity and specificity. These studies will be carried out using SPR, Microscale Thermophoresis (MST) and enzyme linked aptamer assay (ELAAs). Using the identified dual aptamer pairs, assays parameters will be optimised by ELAAs. Buffer conditions (e.g. pH, concentration of salts, type of buffer, effect of ions), incubation times, temperatures and concentration of capture and reporter aptamers will be optimised. Optimal assay conditions will be transferred to lateral flow assays (LFAs).


http://www.etseq.urv.es/nbg/Interfibio/


Bibliography:

1. Li, L.; Liu, Z.; Zhang, H.; Yue, W.; Li, C. W.; Yi, C. A point-of-need enzyme linked aptamer assay for Mycobacterium tuberculosis detection using a

smartphone. Sensors Actuators, B Chem. 2018, 254, 337–346 DOI: 10.1016/j.snb.2017.07.074.

2. Monkongdee, P.; McCarthy, K. D.; Cain, K. P.; Tasaneeyapan, T.; Dung, N. H.; Lan, N. T. N.; Yen, N. T. B.; Teeratakulpisarn, N.; Udomsantisuk, N.;

Heilig, C.; et al. Yield of acid-fast smear and mycobacterial culture for tuberculosis diagnosis in people with human immunodeficiency virus. Am. J.

Respir. Crit. Care Med. 2009, 180 (9), 903–908 DOI: 10.1164/rccm.200905-0692OC.

3. Sypabekova, M.; Bekmurzayeva, A.; Wang, R.; Li, Y.; Nogues, C.; Kanayeva, D. Selection, characterization, and application of DNA aptamers for

detection of Mycobacterium tuberculosis secreted protein MPT64. Tuberculosis 2017, 104, 70–78 DOI: 10.1016/j.tube.2017.03.004.

4. Turbawaty, D. K.; Sugianli, A. K.; Soeroto, A. Y.; Setiabudiawan, B.; Parwati, I. Comparison of the Performance of Urinary Mycobacterium

tuberculosis Antigens Cocktail (ESAT6, CFP10, and MPT64) with Culture and Microscopy in Pulmonary Tuberculosis Patients. Int. J. Microbiol.

2017, 2017 DOI: 10.1155/2017/3259329.

5. Sutherland, J. S.; Mendy, J.; Gindeh, A.; Walzl, G.; Togun, T.; Owolabi, O.; Donkor, S.; Ota, M. O.; Kon Fat, E. T.; Ottenhoff, T. H. M.; et al. Use of

lateral flow assays to determine IP-10 and CCL4 levels in pleural effusions and whole blood for TB diagnosis. Tuberculosis 2016, 96, 31–36 DOI:

10.1016/j.tube.2015.10.011.

6. Corstjens, P. L. A. M.; Tjon Kon Fat, E. M.; de Dood, C. J.; van der Ploeg-van Schip, J. J.; Franken, K. L. M. C.; Chegou, N. N.; Sutherland, J. S.;

Howe, R.; Mihret, A.; Kassa, D.; et al. Multi-center evaluation of a user-friendly lateral flow assay to determine IP-10 and CCL4 levels in blood of TB

and non-TB cases in Africa. Clin. Biochem. 2016, 49 (1), 22–31 DOI: 10.1016/j.clinbiochem.2015.08.013.

7. Dausse, E.; Da Rocha Gomes, S.; Toulmé, J. J. Aptamers: a new class of oligonucleotides in the drug discovery pipeline? Current Opinion in

Pharmacology. 2009, pp 602–607.

8. Keefe, A. D.; Pai, S.; Ellington, A. Aptamers as therapeutics. Nature Reviews Drug Discovery. 2010, pp 537–550.

9. Nimjee, S. M.; Rusconi, C. P.; Sullenger, B. A. Aptamers: An Emerging Class of Therapeutics. Annu. Rev. Med. 2005, 56 (1), 555–583 DOI:

10.1146/annurev.med.56.062904.144915.

10. Shamah, S. M.; Healy, J. M.; Cload, S. T. Complex target SELEX. Acc. Chem. Res. 2008, 41 (1), 130–138 DOI: 10.1021/ar700142z.



Project #8

Title: Selection of aptamers for the quantitative detection of mycotoxins in food and feed.

Supervisors: Ciara O´Sullivan



Description:

Mycotoxins are small secondary metabolites of moulds that can cause adverse acute and chronic effects on human and animal health, with side effects affecting the liver, kidney, nervous system, endocrine and immune systems. Some mycotoxins are carcinogenic, some are vasoactive, and often a single mycotoxin can cause more than one type of toxic effect. There are 400 different types of mycotoxins, with 25 of these being the most prevalent. The Food and Agriculture Organisation (FAO) of the United Nations states that nearly 25% of food produced in the world is affected by toxigenic fungi, leading to mycotoxins entering the food chain.

Aptamers are artificial nucleic acids with a specific and complex 3D structure selected by process of SELEX (Systematic Evolution of Ligands by EXponentional enrichment) [1-3]. Aptamers are typically composed of RNA, single-stranded DNA (ssDNA) or a combination of both with unnatural nucleotides, and range in size from approximately 6 to 40 kDa. Due to their three-dimensional structure, characterized by stems, loops, hairpins, bulges, triplexes and quadruplexes, aptamers can bind to a wide variety of targets from single molecules to complex target mixtures or a whole organism. Aptamers are more flexible to different assay formats than their antibody counterparts, allowing them to address the ever-increasing requirements of analytical applications [4-5].

Objective:

The main objective of this project is the selection of aptamer against one of the mycotoxins which could be a posteriori used for the quantitative detection of mycotoxins in food and feed.

Project outline:

The current project focuses on the development of aptamers against mycotoxins. A typical DNA SELEX procedure begins with a nucleic acid library, generally consisting of about 1013 - 1015 different sequences, containing a random region flanked by two primers used for amplification. A randomized ssDNA is incubated directly with the target molecule, which is immobilized on a matrix material (matrix based SELEX) or is free in solution (Capture SELEX). Following incubation, the bound complexes are separated from the unbound and weakly bound oligonucleotides. Target bound oligonucleotides are eluted and amplified by polymerase chain reaction (PCR) and from the resulting double stranded DNA (dsDNA), ssDNA is generated, and this enriched pool of selected oligonucleotides is used in the next SELEX round. Generally, 10-20 rounds of SELEX are required in order to obtain a pool of aptamer candidates with the highest binding affinity for the target. When affinity saturation of the enriched pool of oligonucleotides is achieved, aptamer candidates are sequenced using next generation sequencing platform. Sequence alignments will be used to identify aptamers with homologous sequences and a comparative analysis of the aligned sequences will be performed to determine the consensus motif. Analysis of the conserved regions will be performed for the prediction of the secondary structure of




aptamer candidates as the determined consensus motifs are often located in stem-loop structures, pseudoknot formations, or G-quadruplexes. Knowing the secondary structure of aptamers, truncation studies of selected aptamers will be performed to increase their specificity and affinity, as well as to reduce the length and the cost of the aptamer.

Bibliography:

[1] Ellington AD. And Szostak JW. In vitro selection of RNA molecules that bind specific ligands. Nature. 346(6287): p. 818-

822. (1990)

[2] Robertson D. and Joyce G. Selection in Vitro of an RNA Enzyme that Specifically Cleaves Single-stranded DNA. Nature.

344(6265): 467-468. (1990)

[3] Tuerk C. and Gold L. Systematic evolution of ligands by exponentional enrichment: RNA ligands to bacteiophage T4 DNA

polymerase. Science. 249: 505-510. (1990)

[4] James W. Aptamers. Encyclopedia of Analitical Chemistry, 2000: p. 4848-4871.

[5] Famulok M. and Mayer G. Aptamers as tools in Molecular biology and immunology. Current Topic in Microbiology and

Immunology, 1999. 243: p. 123-136.



Project #9

Title: Electrochemical detection of HPV based on isothermal DNA amplification and copper-catalyzed azide–alkyne click reaction




Description:

Cervical cancer is detectable and preventable in pre-malignant stages through public health interventions such as education, vaccination, screening programs and the early management of pre-malignant lesions. Reliable cervical screening programs have been implemented in most high-income countries and have significantly reduced the cervical cancer incidence and mortality by at least 80%. Although HPV DNA testing is considered an effective and recommended primary screening strategy, it has been associated with variability of results between different tests and increased overscreening/overtreatment of women. Several HPV genotypes have been considered oncogenic or high-risk HPVs by being more frequently associated with cancer development, but natural clearance of infection is also a reality even with such HPVs. Therefore, there is a need to explore new ways to reduce variability in HPV DNA results and to increase specific detection of HPV infections associated with precancer or cancer development. Our group has a wide experience in the development of strategies based on electrochemical DNA detection that can be further implemented in a microfluidic device. The strategy proposed for this project is based on a two-step approach. The first step involves the solid phase amplification of a surface-tethered primer hybridised to a partially complementary ssDNA using alkyne modified dNTPs. The result of this process is a dsDNA containing the enzymatically incorporated alkyne ferrocene, that can be “clicked” to azide-ferrocene in the second step.1,2 Finally the ferrocene modified DNA can be detected by an electrochemical technique (Cyclic Voltammetry, Square Wave Voltammetry or Differential Pulse Voltammetry) The student who chair the project will be trained in PCR, RPA, electrochemistry and in different characterisation techniques.

References

1) Presolski, S. I., Hong, V. P., M.G. Finn, M.G., Curr Protoc Chem Biol. 2011, 3(4), 153–162. 2) Panattoni, A., Pohl, R., Hocek, M., Org. Lett. 2018, 20, 3962−3965





Project #10

Title: Development of an integrated nucleic acid lateral flow device for the early diagnosis of triple-negative breast cancer (TNBC)




Description:

Breast cancer is the most common cancer in women worldwide, being the triple-negative breast cancer

one of the most aggressive subtypes with higher relapse rates and worse prognosis than hormone

receptor positive tumours1. Until the date, self-examination of the breast to mammography screening

are aimed to detect breast cancer. However, the precision of these methods are limited. For this

reason it is crucial to develop a non-invasive method for its early detection based on blood biomarkers.

MicroRNA (miRNA) have been extensively used as biomarkers for the diagnosis of different types of

cancer diseases due to their role in regulating gene expression2–5. Among the most used assays for

the detection of miRNA based on nucleic acid tests6, lateral flow assays present a very simply, rapid

and low-cost platforms that can be applied in the point-of need without requiring trained personnel,

expensive equipment and storage.

The overall objective of this project is the development of an integrated multiplex nucleic acid lateral

flow assay for the detection of miRNAs related with triple-negative breast cancer for its early

diagnostic. To achieve that, we will combine the use of isothermal recombinase polymerase

amplification with tailed primers and fishing strategy with the reported approach of Wee et al., where

two synthetic DNA probes complementary to miRNA sequence are ligated during the hybridization

between DNA-miRNA avoiding the reverse transcriptase step7. Therefore, the final device will integrate

the ligation approach with isothermal amplification using tailed primers and fishing strategy and its final

detection on the strip. We will start as proof-of-concept using two control miRNAs in the blood

samples, one for spike-in approach, since it is not found in human (cel-miR-39-3p)8, and the other for

blood specific approach, since it is expressed in many tissues in humans (hsa-miRNA-103a)9, being

both useful to know the success of the assay in real samples. One time the assay would be validated,

next step will be its applicability in the specific selected miRNAs for the detection of triple-negative

breast cancer.

References

(1) Torre, L. A.; Siegel, R. L.; Ward, E. M.; Jemal, A. Global Cancer Incidence and Mortality Rates

and Trends—An Update. Cancer Epidemiol. Prev. Biomarkers 2016.

(2) Zhu, Y.; Peng, Q.; Lin, Y.; Zou, L.; Shen, P.; Chen, F.; Min, M.; Shen, L.; Chen, J.; Shen, B.

Identification of Biomarker MicroRNAs for Predicting the Response of Colorectal Cancer to

Neoadjuvant Chemoradiotherapy Based on MicroRNA Regulatory Network. Oncotarget 2017, 8

(2), 2233–2248.




(3) Stroese, A. J.; Ullerich, H.; Koehler, G.; Raetzel, V.; Senninger, N.; Dhayat, S. A. Circulating

MicroRNA-99 Family as Liquid Biopsy Marker in Pancreatic Adenocarcinoma. J. Cancer Res.

Clin. Oncol. 2018.

(4) Liu, S. S.; Chan, K. K.; Chu, D. K.; Wei, T. N.; Lau, L. S.; Ngu, S. F.; Chu, M. M.; Tse, K. Y.; Ip,

P. P.; Ng, E. K.; et al. Oncogenic MicroRNA Signature for Early Diagnosis of Cervical

Intraepithelial Neoplasia and Cancer. Mol. Oncol. 2018.

(5) Xu, H.; Wen, Q. MiR31205p Acts as a Diagnostic Biomarker in Nonsmall Cell Lung Cancer and

Promotes Cancer Cell Proliferation and Invasion by Targeting KLF4. Mol. Med. Rep. 2018.

(6) Pimentel, F.; Bonilla, P.; Ravishankar, Y. G.; Contag, A.; Gopal, N.; LaCour, S.; Lee, T.; Niemz,

A. Technology in MicroRNA Profiling: Circulating MicroRNAs as Noninvasive Cancer

Biomarkers in Breast Cancer. J. Lab. Autom. 2015, 20 (5), 574–588.

(7) Wee, E. J. H.; Trau, M. Simple Isothermal Strategy for Multiplexed, Rapid, Sensitive, and

Accurate MiRNA Detection. ACS Sensors 2016, 1 (6), 670–675.

(8) Hara, N.; Kikuchi, M.; Miyashita, A.; Hatsuta, H.; Saito, Y.; Kasuga, K.; Murayama, S.; Ikeuchi,

T.; Kuwano, R. Serum MicroRNA MiR-501-3p as a Potential Biomarker Related to the

Progression of Alzheimer’s Disease. Acta Neuropathol. Commun. 2017, 5, 10.

(9) Wang, W.-X.; Danaher, R. J.; Miller, C. S.; Berger, J. R.; Nubia, V. G.; Wilfred, B. S.; Neltner, J.

H.; Norris, C. M.; Nelson, P. T. Expression of MiR-15/107 Family MicroRNAs in Human Tissues

and Cultured Rat Brain Cells. Genomics. Proteomics Bioinformatics 2014, 12 (1), 19–30.



Project #11

Title: Development of a Chip-SELEX for the selection of aptamers against small molecules




Description:

Aptamers are single stranded oligonucleotides (DNA or RNA) with the ability to fold into stable three-dimensional structures, characterized by stems, loops, hairpins, bulges, triplexes and quadruplexes, that allow them to interact with a wide variety of target molecules from single molecules to complex target mixtures or even a whole organism, with dissociation constants (KD) ranging from picomolar to nanomolar 1. Aptamers offer several advantages over their antibody counterparts for their flexibility to adapt to different assay formats. The main advantages of aptamers are that once they are selected they can be synthesized with high reproducibility and purity from commercial sources and are chemically stable, which offers great flexibility in the design of novel assays with high sensitivity and selectivity 2–5. The process used for the selection and isolation of these aptamers is called systematic evolution of ligands by exponential enrichment (SELEX) characterized by the repetition of successive steps consisting of selection (binding, partition and elution), amplification and conditioning. Classical SELEX is a well-established approach; however, large rounds of SELEX (8-15 rounds) are needed for the selection, which is time-consuming, laborious and sometimes ineffective, particularly as there is no standardized protocol 6,7. Different methodologies have been reported with the aim of reducing the time required to perform the selection of aptamers, such as Capillary Electrophoresis SELEX (CE-SELEX) 8–10. This technique takes advantage of the efficient separation of free nucleic acids from aptamer/target complexes, thus reducing the cycles needed for the isolation of high affinity aptamers to four or sometimes even less. CE-SELEX has also been applied for aptamers against small molecules like N-methyl mesoporphyrin (NMM), which is challenging due to the minimal differences in the mobility shift between aptamer and complex 11. For this project we propose the development of a new method for the selection of aptamers against different small targets based on CE-SELEX technique, but using a portable chip device for the partitioning step.

References

(1) Stoltenburg, R.; Reinemann, C.; Strehlitz, B. SELEX--a (r)Evolutionary Method to Generate

High-Affinity Nucleic Acid Ligands. Biomol. Eng. 2007, 24 (4), 381–403.

(2) Clark, S. L.; Remcho, V. T. Aptamers as Analytical Reagents. Electrophoresis 2002, 23 (9),

1335–1340.

(3) Mairal, T.; Ozalp, V. C.; Lozano Sanchez, P.; Mir, M.; Katakis, I.; O’Sullivan, C. K. Aptamers:

Molecular Tools for Analytical Applications. Anal. Bioanal. Chem. 2008, 390 (4), 989–1007.

(4) Tombelli, S.; Minunni, M.; Mascini, M. Analytical Applications of Aptamers. Biosens.

Bioelectron. 2005, 20 (12), 2424–2434.

(5) Willner, I.; Zayats, M. Electronic Aptamer-Based Sensors. Angew. Chem. Int. Ed. Engl. 2007,




46 (34), 6408–6418.

(6) Lakhin, A. V; Tarantul, V. Z.; Gening, L. V. Aptamers: Problems, Solutions and Prospects. Acta

Naturae 2013, 5 (4), 34–43.

(7) Mencin, N.; Šmuc, T.; Vraničar, M.; Mavri, J.; Hren, M.; Galeša, K.; Krkoč, P.; Ulrich, H.; Šolar,

B. Optimization of SELEX: Comparison of Different Methods for Monitoring the Progress of in

Vitro Selection of Aptamers. J. Pharm. Biomed. Anal. 2014, 91, 151–159.

(8) Mallikaratchy, P.; Stahelin, R. V; Cao, Z.; Cho, W.; Tan, W. Selection of DNA Ligands for

Protein Kinase C-δ. Chem. Commun. 2006, No. 30, 3229–3231.

(9) Mendonsa, S. D.; Bowser, M. T. In Vitro Selection of Aptamers with Affinity for Neuropeptide Y

Using Capillary Electrophoresis. J. Am. Chem. Soc. 2005, 127 (26), 9382–9383.

(10) Mosing, R. K.; Mendonsa, S. D.; Bowser, M. T. Capillary Electrophoresis-SELEX Selection of

Aptamers with Affinity for HIV-1 Reverse Transcriptase. Anal. Chem. 2005, 77 (19), 6107–6112.

(11) Yang, J.; Bowser, M. T. Capillary Electrophoresis-SELEX Selection of Catalytic DNA Aptamers

for a Small-Molecule Porphyrin Target. Anal. Chem. 2013, 85 (3), 1525–1530.



Project #12

Title: Development of an electrochemical lateral flow assay for the ciguatoxins detection Supervisors: Ciara O’Sullivan



Description:

Ciguatera fish poisoning (CFP) is the most commonly reported natural marine toxin-related illness in the world. It is characterised by general, gastrointestinal, cardiac and neurological symptoms. Humans contract CFP by consuming fish that contains ciguatoxins (CTXs), which are marine toxins produced by microalgae of the genera Gambierdiscus and Fukuyoa1. Recently, poisoning cases have been reported in Spain (Canary Islands) and Portugal (Madeira). Additionally, microalgae of the genera Gambierdiscus and/or Fukuyoa have been detected in Balearic Islands, Greece and Cyprus2,3.

Traditional methods for microalgae identification and quantification are time-consuming, laborious and tedious, and require highly trained personnel. Similarly, CTX detection has been hampered by the scarcity, toxicity and chemical complexity of these toxins, and assays developed so far lack proper validation or have raised controversy regarding their performance4–9. Nevertheless, CTX fragments have been rationally synthetized and used to produce convenient monoclonal antibodies to develop immunoassays able to detect CTXs at nano and even sub-nanomolar levels 10–15. Recently, we applied these antibodies for the development of lateral flow assays (LFAs), which provide results in few minutes, with no necessity of trained personnel and specific instrumentations, with low development costs and ease of production. Although, we demonstrated to detect CTXs in extracted fish samples with a limit of detection of 35.66 pg /g, this value is not enough considering the FDA guidance level of 0.01 ppb in fish (10 pg /g). Thus, the main objective of this work is to develop an electrochemical LFA, since much better sensitivity can be achieved using electrochemistry. To address this, LFAs will also be produced with integrated screen-printed electrodes. To further increase sensitivity, the magnetic sandwich assays can be exploited simply adding a planar magnet between the membrane and the backing. The use of different electrochemical techniques will be evaluated for both labelled (e.g. HRP-labelled) and unlabelled formats. The LOD, LOQ, IC50, sensitivity, working range, reproducibility, stability, end-userability, assay time and cost will be evaluated.

References

(1) NAKAJIMA, I.; OSHIMA, Y.; YASUMOTO, T. Toxicity of Benthic Dinoflagellates in Okinawa.

Nippon SUISAN GAKKAISHI 1981, 47 (8), 1029–1033.

(2) Pérez-Arellano, J.-L.; Luzardo, O. P.; Pérez Brito, A.; Hernández Cabrera, M.; Zumbado, M.;

Carranza, C.; Angel-Moreno, A.; Dickey, R. W.; Boada, L. D. Ciguatera Fish Poisoning, Canary

Islands. Emerg. Infect. Dis. 2005, 11 (12), 1981–1982.

(3) Costa, P. R.; Estevez, P.; Castro, D.; Soliño, L.; Gouveia, N.; Santos, C.; Rodrigues, S. M.;

Leao, J. M.; Gago-Martínez, A. New Insights into the Occurrence and Toxin Profile of

Ciguatoxins in Selvagens Islands (Madeira, Portugal). Toxins (Basel). 2018, 10 (12), 524.

(4) Dechraoui, M. Y.; Naar, J.; Pauillac, S.; Legrand, A. M. Ciguatoxins and Brevetoxins,

Neurotoxic Polyether Compounds Active on Sodium Channels. Toxicon 1999, 37 (1), 125–143.




(5) Yasumoto, T.; Fukui, M.; Sasaki, K.; Sugiyama, K. Determinations of Marine Toxins in Foods. J.

AOAC Int. 1995, 78 (2), 574–582.

(6) Lewis, R. J.; Jones, A.; Vernoux, J. P. HPLC/Tandem Electrospray Mass Spectrometry for the

Determination of Sub-Ppb Levels of Pacific and Caribbean Ciguatoxins in Crude Extracts of

Fish. Anal. Chem. 1999, 71 (1), 247–250.

(7) Lewis, R. J.; Yang, A.; Jones, A. Rapid Extraction Combined with LC-Tandem Mass

Spectrometry (CREM-LC/MS/MS) for the Determination of Ciguatoxins in Ciguateric Fish Flesh.

Toxicon 2009, 54 (1), 62–66.

(8) Manger, R. L.; Leja, L. S.; Lee, S. Y.; Hungerford, J. M.; Hokama, Y.; Dickey, R. W.; Granade,

H. R.; Lewis, R.; Yasumoto, T.; Wekell, M. M. Detection of Sodium Channel Toxins: Directed

Cytotoxicity Assays of Purified Ciguatoxins, Brevetoxins, Saxitoxins, and Seafood Extracts. J.

AOAC Int. 1995, 78 (2), 521–527.

(9) Louzao, M. C.; Vieytes, M. R.; Yasumoto, T.; Botana, L. M. Detection of Sodium Channel

Activators by a Rapid Fluorimetric Microplate Assay. Chem. Res. Toxicol. 2004, 17 (4), 572–

578.

(10) Tsumuraya, T.; Sato, T.; Hirama, M.; Fujii, I. Highly Sensitive and Practical Fluorescent

Sandwich ELISA for Ciguatoxins. Anal. Chem. 2018, 90 (12), 7318–7324.

(11) Nagumo, Y.; Oguri, H.; Shindo, Y.; Sasaki, S.; Oishi, T.; Hirama, M.; Tomioka, Y.; Mizugaki, M.;

Tsumuraya, T. Concise Synthesis of Ciguatoxin ABC-Ring Fragments and Surface Plasmon

Resonance Study of the Interaction of Their BSA Conjugates with Monoclonal Antibodies.

Bioorg. Med. Chem. Lett. 2001, 11 (15), 2037–2040.

(12) Nagumo, Y.; Oguri, H.; Tsumoto, K.; Shindo, Y.; Hirama, M.; Tsumuraya, T.; Fujii, I.; Tomioka,

Y.; Mizugaki, M.; Kumagai, I. Phage-Display Selection of Antibodies to the Left End of CTX3C

Using Synthetic Fragments. J. Immunol. Methods 2004, 289 (1–2), 137–146.

(13) Oguri, H.; Hirama, M.; Tsumuraya, T.; Fujii, I.; Maruyama, M.; Uehara, H.; Nagumo, Y.

Synthesis-Based Approach toward Direct Sandwich Immunoassay for Ciguatoxin CTX3C. J.

Am. Chem. Soc. 2003, 125 (25), 7608–7612.

(14) Tsumuraya, T.; Takeuchi, K.; Yamashita, S.; Fujii, I.; Hirama, M. Development of a Monoclonal

Antibody against the Left Wing of Ciguatoxin CTX1B: Thiol Strategy and Detection Using a

Sandwich ELISA. Toxicon 2012, 60 (3), 348–357.

(15) Tsumuraya, T.; Fujii, I.; Hirama, M. Preparation of Anti-Ciguatoxin Monoclonal Antibodies Using

Synthetic Haptens: Sandwich ELISA Detection of Ciguatoxins. J. AOAC Int. 2014, 97 (2), 373–

379.



Project #13

Title: Development of aptasensor platforms for the determination of histamine in real samples.




Description:

Biogenic amines (BAs) are low-molecular organic compounds produced by microbial decarboxylation of aminoacids or by amination and transamination of aldehydes and ketones. They are found in a variety of foods including fish, meat, cheese, fermented products and beverages as well as in biological fluids such as blood, urine or saliva. Among all the Bas, histamine plays an important role in gastric secretion, cellular differentiation and cell grow, neurotransmission and neuromodulation. Histamine also has been involved in allergenic response.

High levels of BAs can induce adverse health effects and histamine is the most relevant one because of its biological toxicity. The analytical determination of histamine is a complicated task due to the complexity of the matrices, the structural similitude with the other biogenic amines and the low concentration in biological fluids. Current methods of detection are based on chromatography (liquid, gas, thin layer) as well as spectroscopic and electrophoretic techniques. These techniques are inherently laboratory based, requiring expensive instrumentation as well as skilled personnel. In the recent years the use of aptamers for the bio-recognition of small molecules has garnered increasing interest due to the stability, low cost and specificity of the aptamers. In this way, different histamine aptamers had been selected in the group using SELEX methodology and one of the most promising aptamers developed was exploited in a magnetic bead-based competitive assay for the ultrasensitive detection of histamine in synthetic urine. The same aptamers was used for the development of a colorimetric aptasensor for determination of histamine in fish samples based on gold nanoparticles.

The main objective of the project is to exploit the selected histamine aptamers for the development of molecular recognition elements in various platforms (ELISA plate and paper-based) and facilitate their specific and sensitive detection in different real samples.





Project #14

Title: Detection of pork adulteration in food products by a nucleic acid lateral flow assay.




Description:

Proper labelling of food products is a major concern for consumers to ensure their authenticity, especially with regards to meat, for health, economic, cultural and religious reasons. Undeclared meat sources can be found in food products because of accidental cross-contamination or as a result of adulteration. Meat adulteration is a practice aiming at replacing high commercial valued meat species with cheaper ones for economic gains and pork is one of the most commonly found adulterant species. Moreover, because of religious restrictions regarding the consumption of pork, its accurate detection especially in processed food products is essential.

The origin of animal species in meat products can be detected using two main approaches: protein-based and DNA-based methods [1,2]. The protein-based methods rely mostly on immunoassays (e.g. ELISA), whereas chromatographic (HPLC, LC-MS/MS, MALDI-TOF) and electrophoretic (SDS-PAGE, IEF, CE) methods have also been reported. The thermal instability of proteins though can prevent correct speciation. On the other hand, DNA-based analysis is more advantageous because of the high discriminating power of amplification techniques like PCR and the high level of sensitivity obtained. Also, the high thermal stability of DNA allows for genomic DNA detection even after thermal processing. DNA-based methods are also easier to perform and more cost effective compared to the protein-based strategies.

The current project focuses on the development of a quick and easy DNA-based assay for the detection of undeclared pork in food products. A simple DNA extraction protocol will be initially developed and the crude extracts obtained from commercial food products will be used for direct analysis. Species-specific tailed primers will be then used for the amplification of the pork target [3]. Universal animal mitochondrial primers will also be included to guarantee proper DNA extraction and successful amplification. Biotinylated nucleotides will be employed during amplification to introduce multiple biotin labels into the amplicon. A lateral flow assay will finally be designed for the detection of the biotin-labelled tailed amplicons [4]. Signal will be generated on the strips using a neutravidin-carbon nanoparticles conjugate via the affinity interaction of neutravidin with the biotin labels. Signal amplification strategies will also be evaluated in order to increase the sensitivity of the assay and detect low traces of pork.

References

1. Rahmati S., Julkapli N.M., Yehye W.A. Food Control 2016, 68: 379–390.

2. Iammarino M., Marino R. Albenzio M. Int J Food Sc Technol 2017, 52: 851–863.

3. Skouridou V., Tomaso H., Rau R., Bashammakh A.S., El-Shahawi M.S., Alyoubi A.O., O’Sullivan C.K Food Chem 2019, 287:354–362 .

4. Jauset-Rubio M., Svobodová M., Mairal T., McNeil C., Keegan N., Saeed A., Abbas M.N., El-Shahawi M.S., Bashammakh A.S., Alyoubi A.O., O’Sullivan C.K Sci Rep 2016, 6:37732.





Project #15

Title: Emulsion-based microencapsulation with novel food-grade ingredients from insects

Supervisors: Dr. C. Güell; Dr. M. Ferrando; Dr. S. de Lamo

Contact: [email protected]; [email protected]; [email protected]

Research Group: Food Innovation & Engineering (FoodIE, http://www.etseq.urv.es/foodie/)

Description:

World population is likely to grow up to 9 billion in 2050, inducing a dramatic increase of food demand from an already pressured agricultural land. Exploring alternate nutrient sources (e.g. protein and fat) can help alleviate pressures on overused sources such as livestock. Insects are an excellent alternative since they are widely integrated in food cultures around the globe, possess high nutritional quality and are relatively sustainable compared with traditional protein and fat sources. While the consumption of whole or low processed insects in western markets is still scarce, efficient processes for the recovery of functional insect-derived fractions, such as protein or fat, are being developed to promote consumer acceptance and to explore potential food and non-food applications. For that, knowledge of their techno-functional properties is of paramount relevance. The use of insect-based fractions such as proteins and fats to encapsulate bioactive compounds or vitamins could be an excellent strategy to incorporate these new ingredients in food and feed production. Moreover, food industry is highly invested in implementing eco-efficient technologies, such as membrane emulsification. Considering all the above, the project plans to apply micro-structured systems to assess the potential of protein and lipid fractions from edible insects to encapsulate bioactive compounds in oil-in-water (O/W) and water-in-oil-in water (W1/O/W2) emulsions. More specifically, the proposed project will link emulsion stability, in terms of coalescence frequency of oil droplets, to the adsorption capacity of enriched protein fractions from insects. Protein extracts from commercial powders of meal worm (Tenebrio molitor), black soldier fly (Hermetia illucens) or Buffalo worm (Alphitobius diaperinus) will be used to stabilize food-grade emulsions. The surface-active properties of enriched protein fractions from insects will be assessed from on-line measurements in which the protein adsorption to the droplet occurs under controlled conditions, mainly regarding adsorption time. A set of microfluidic devices [1], having different length of the adsorption channel, from Micronit Microfluidics (The Netherlands) will be used for coalescence measurements. In the device, the emulsion droplets are made at a T-shaped junction and then travel through an adsorption channel of various lengths, before entering a coalescence channel. Images will be recorded at the inlet of the coalescence channel, and at the outlet. A high-speed camera will record images of the droplets that will be further processed using Image J and Matlab. From the image analysis, quantitative values of number of coalescence events and frequency will be obtained.

[1] Muijlwijk K., Colijn I., Harsono H., Krebs T., Berton-Carabin C., Schröen K. (2017) Food Hydrocolloids, 70, 96-104





http://www.etseq.urv.es/foodie/


Project #16

Title: Controlled hydrogen peroxide dosing in packed ceramic membrane reactors for Fenton-like processes

Supervisors: Dr. Josep Font


Research Group: Process Intensification, Simulation and Environmental Technology (PISET)

(https://www.researchgate.net/profile/Josep_Font)

Description:

Efficient treatment of wastewater for the subsequent reuse or safe discharge to the environment is becoming an emergent issue in the last decades. Although conventional treatment methods may be quite effective when dealing with typical organic and inorganic contaminants, they have shown very limited capability of removing so called Emergent and Recalcitrant Contaminants. This group of contaminants includes some persistent dyes, antibiotics, hormones, pesticides and other synthetic compounds characterized by high eco-toxicity and low natural degradation rates.

Most of ERCs present in water exhibit undesirable effects very low concentrations. Thus, their selective degradation would imply destruction of very stable organic molecules in the presence of prevailing (by many orders of magnitude) and less degradable natural organic matter, which is extremely difficult to achieve. For this reason, most of depollution methods for ERCs up to date are based on nonselective degradation, commonly, – oxidation, of all the present organic impurities, both toxic and non-toxic. This approach leads to enormous consumptions of oxidants and energy and, hence, to extremely high costs of treatment and severe environmental impact. However, there is at least one promising approach to increase the selectivity of advanced oxidation, which have not been extensively elaborated till now. It is based on combination of advanced oxidation and sorption phenomena.

The main aim of this project is to develop a new technology for treating waste water containing low concentration emergent compounds and azo dyes. The basis is a ceramic membrane, covered by a carbonaceous catalytic material. This carbon layer should process both sorption capacity and catalytic activity towards Fenton-based oxidation. Two general approaches are planned: the use of core-shelled Fe0 (zero-valent iron, ZVI) as a catalyst for reducing ferrous iron and as a source of Fe2+ itself, as well as stabilization of Fe2+ by nitrogen-containing ligands. In both approaches, thiazine-based polymeric carbonization precursors will be used to achieve nitrogen-rich, hydrophilic and mesoporous carbonaceous networks. Stabilization of ZVI or Fe2+ would be designed by altering the coordination ligands loading of the precursor polymers and, hence, the resulting carbons. However, other formulations are also designed based on the general concept of process intensification.

References

• Messele, S.A.; Soares, O.S.G.P.; Órfão, J.J.M.; Bengoa, C.; Font, J. “Zero-valent iron supported on Nitrogen-doped carbon xerogel as catalysts for the oxidation of phenol by Fenton-like system” Environmental Technology 2018, 39, 2951-2958.

• Giménez-Pérez, A.; Bikkarolla, S.K.; Benson, J.; Bengoa, C.; Stüber, F.; Fortuny, A.; Fabregat, A.; Font, J.; Papakonstantinou, P. “Synthesis of N‐doped and non‐doped partially oxidised graphene membranes supported over ceramic materials” Journal of Materials Science 2016, 51, 8346-8360.

• Mezohegyi, G.; van der Zee, F.P.; Font, J.; Fortuny, A.; Fabregat, A. “Towards advanced aqueous dye removal processes: A short review on the versatile role of activated carbon” Journal of Environmental Management 2012, 102, 148-164.

Positions: 1


https://www.researchgate.net/profile/Josep_Font


Project #17

Title: Amine-Free Materials for CO2 Capture

Supervisors: Dr. Alberto Puga


Research Group: MEMTEC (http://www.urv.cat/html/grupsrecerca/reconeguts/general-003404.php)

Description:

The capture and utilisation of CO2 is a crucial—yet challenging—approach to, on one side, help mitigate climate change, and on the other, tackle global recycling of carbon resources. Capture (see Figure) comprises a series of technologies which can be applied to either minimising emissions from large polluting point sources (power plants, cement kilns or steel blast furnaces), or directly capturing atmospheric CO2 (negative emissions scheme). Implementation of such processes entails obvious monetary and energetic costs, a fact often claimed by opponents to CO2 capture; in contrast to this stance, neglecting to capture carbon emissions are generally believed to lead to disastrous and irreversible consequences on health and the environment. Technological advances on CO2 capture technologies are thus required to enhance their efficiency and make them more appealing for widespread deployment. The most common CO2 capture technologies utilise amine sorbent systems. The field of amine-impregnated solids is rapidly growing, especially in the context of direct air capture. Nonetheless, several issues of amine technologies still remain, such as high toxicity, oxidative degradability, high regeneration temperatures and volatility losses.

Non-volatile alternatives, such as those based in ionic sorbents are interesting and deserve further investigation. Chemical

reactivity of ionic liquids for CO2 capture can be tuned by carefully selecting their

basicity. The case of amine-free

carboxylate-based ionic liquids is particularly relevant, since a fine adjustment of their reactivity can be accomplished simply by structural design and control of the moisture content (see Figure on the right), with the added advantage of avoiding the noxious emissions of amine processes.i This project advocates the development of new CO2 sorbents based on phosphonium carboxylate ionic liquids in combination with solid supports.

The student embarking on this project will be involved in experimental tasks on the preparation and testing of these low-emissions solid-supported ionic liquid systems for CO2 capture, in close collaboration with MSc and PhD students, and Professors in MEMTEC. Looking forward to hosting talented and motivated young researchers from NEU!

References


http://www.urv.cat/html/grupsrecerca/reconeguts/general-003404.php


· Sanz-Pérez, E. S., Murdock, C. R., Didas, S. A. & Jones, C. W. Direct Capture of CO2 from Ambient Air. Chem.

Rev. 116, 11840–11876 (2016); Boot-Handford, M. E. et al. Carbon capture and storage update. Energy Environ.

Sci. 7, 130–189 (2014).

· Kramer, D. Can carbon capture from air shift the climate change equation? Phys. Today 71, 26–29 (2018).

· Zeng, S. et al. Ionic-Liquid-Based CO2 Capture Systems: Structure, Interaction and Process. Chem. Rev. 117,

9625–9673 (2017); Cui, G., Wang, J. & Zhang, S. Active chemisorption sites in functionalized ionic liquids for

carbon capture. Chem. Soc. Rev. 45, 4307–4339 (2016).

· Anderson, K. et al. Carbon dioxide uptake from natural gas by binary ionic liquid-water mixtures. Green Chem.

17, 4340–4354 (2015).



Project #18

Title: Cyclodextrin‐derived polymeric materials for the extraction and purification of marine toxins

Supervisors: Dr. Alex Fragoso, Dr. Mabel Torréns

Contact: e-mail: [email protected] Telephone: + 34 977 55 8579

Research Group: INTERFIBIO

Description:

Objective: The main goal of this project is the preparation and characterization cyclodextrin polymers able to capture rare toxins from marine environments.

Introduction: Ciguatera fish poisoning (CFP) is the most commonly reported natural marine toxin‐related illness in the world contracted by humans by consuming fish that contains ciguatoxins (CTXs). Recently, poisoning cases have been reported in Spain (Canary Islands) and Portugal (Madeira) and microalgae of the toxic genera Gambierdiscus and/or Fukuyoa have been detected in several places of the Atlantic Ocean and the Mediterranean basin. A remaining challenge consists in their early detection due to their occurrence in very small amounts

Cyclodextrins (CDs) are cyclic oligosaccharides featuring a truncated cone structure with a central hydrophobic cavity. Although they have been extensively used for pharmaceutical, cosmetic and environmental applications, very little is known about the interaction of marine toxins with CDs. Since CTXs are polycyclic ethers having a worm‐like structure of 2‐3 nm long and 0.5 nm width, their shape and their essentially hydrophobic character makes them perfect guests for CD complexation and thus for the development of extraction/purification/pre‐concentration protocols based on CD‐derived materials.

Structure of CDs and marine toxins

Our group has a vast experience in the preparation of CD derivatives for biomedical, biotechnological and environmental applications. The project thus aims at expanding the current knowledge on these interesting materials with focus in novel applications.

Methodology: CDs will be attached to the side chains of activated polymers or crosslinked with bifunctional agents to prepare insoluble materials. The materials will be characterized by common spectroscopic and microscopy techniques. The toxin extraction capacity of these materials will be assessed using okadaic acid as a model. The extracted toxin will be quantified using a colorimetric ELISA-based assay and the results will be compared with a standard Diaion® resin.



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