Epigenetic editing:Towards sustained gene expression reprogramming in diseases

Veel ziekten zijn geassocieerd met verstoringen in genexpressie door epigenetische mutaties. In tegenstelling tot genetische mutaties, zijn epigenetische mutaties omkeerbaar. Hierdoor is het mogelijk gebleken om abnormale patronen van genexpressie om te draaien. Dit proefschrift heeft betrekking op de ontwikkeling van epigenetische editing-instrumenten om een verstoorde genexpressiepatronen in ziekten te herprogrammeren. Epigenetische editing bestaat uit een DNA-bindend domein (zink vinger, TALEs of CRISPR-Cas) gekoppeld aan het katalytische domein van epigenetische enzymen. Bovendien biedt gen-specifieke epigenetische bewerking de mogelijkheid om de regulatie van genexpessie te bestuderen. In dit proefschrift hebben we de mogelijkheid onderzocht om duurzame genexpressie modulatie te bereiken, door meerdere epigenetische markeringen te combineren en de rol van chromatin micro-omgeving in deze context te evalueren. Door gebruik te maken van verschillende DNA-bindingsplatforms gefuseerd met genexpressie activerende epigenetische enzymen, konden we in verschillende chromatine omgevingen, genexpressieherprogrammering behalen. Met behulp van deze techniek, waren we ook in staat om zowel de functie alsmede de mogelijkheid als nieuw therapeutisch doeleiwit van verschillende genen die betrokken zijn bij ziekten zoals kanker en COPD te onderzoeken. We gebruikten epigenetische editing om aan te tonen dat de overexpressie van TCTN2 een nieuw therapeutisch doeleiwit in kanker is. We hebben de eerste stappen gezet om de dubbele rol van RASSF1-isovormen bij de ontwikkeling van kanker te analyseren. Verder waren we in staat om aan te tonen dat remming van de activiteit van het slijmproductie-gen in COPD cellen inderdaad resulteert in minder slijmproductie. Onze aanpak heeft dus de belofte om verstoorde genexpressie patronen bij vele ziekte te kunnen herstellen.The reversible nature of epigenetic (mis)regulation, in contrast to genetic mutations, allows for the possibility of reverting abnormal patterns of gene expression at a molecular level. This thesis deals with the development of epigenetic editing tools in order to reprogram aberrantly expressed gene patterns in diseases. Epigenetic Editing consists of a DNA binding domain (Zinc finger, TALEs or CRISPR-Cas) linked to the catalytic domain of epigenetic enzymes. Furthermore, site-specific epigenetic editing provides the opportunity to study the regulation of gene expression. In this thesis, we addressed the possibility of achieving sustained gene expression modulation, by combining several epigenetic marks, and evaluating the role of chromatin microenvironment in this context. By using several DNA binding platforms fused to transcriptional activating epigenetic enzymes, we were able to show in different chromatin microenvironments, the possibility of achieving sustained gene expression reprogramming. Using this technique, we were also able to address the function and possible therapeutic use of several genes that are involved in diseases such as cancer and COPD. We used epigenetic editing to show that TCTN2 overexpression is a therapeutic target in cancer, and also to analyze the dual role of RASSF1 isoforms in the development of cancer. Furthermore, we were able to show that downregulation of a mucus production gene in cells might have therapeutic relevance for COPD. In conclusion, our approach has the potency to achieve gene expression reprogramming in diseases

Kinetic study of methanol dehydration over Zro2 supported-activated carbons

The growing concerns about climate change and energy consumption have been the driving force in seek of alternative fuels such as DME, mainly produced via methanol dehydration over a solid acid catalyst. The use of activated carbons for this aim has been little studied up to date. Only a few studies can be found in the literature, reporting all of them materials with a low thermal stability of the acid surface groups, which results into a fast deactivation of the catalyst. In this work, the preparation of activated carbons via chemical activation with phosphoric acid, their modification with different ZrO2 loads, and their application as methanol dehydration catalysts have been studied. The catalytic results showed that the best methanol conversion and selectivity towards DME were achieved with the activated carbon prepared with an impregnation mass ratio value (H3PO4 /precursor) of 2 and an activation temperature of 800 ºC, loaded with a 7 % (wt) of ZrO2 . This catalyst exhibits high steady state methanol conversion values even at temperatures as high as 400 ºC (XCH3OH= 80%, 0.1 g·s/μmol, PCH3OH= 0.08 atm in helium), keeping a selectivity to DME higher than 96%. The effect of oxygen in the reaction atmosphere was also analysed. In this sense, an increase of 15 % in the DME yield was obtained when using air instead of helium as reaction atmosphere (350 ºC, 0.1 g·s/μmol, PCH3OH= 0.04 atm). A kinetic study has been carried out on this catalyst in which two mechanisms (Eley Rideal and Langmuir Hinshelwood) for methanol dehydration have been analysed. The models proposed also consider the presence of oxygen in the reaction media.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. MINECO (CTQ2015-68654-R). MECD (FPU13/02413)

Socioeconomic Factors and the Consumption of Wine in Tenerife

In this paper we measure the impact of an individual's socioeconomic conditions on the decision to consume wine in a traditionally wine-producing area. Based on the data obtained in an exhaustive survey on wine consumption and through discrete choice models, we assess the changes which come about in the decisions to consume the different types of wine under consideration, and we obtain the most relevant distinctive and differentiated characteristics for each one of them.wine consumption, socioeconomic characteristics, discrete choice models, Food Consumption/Nutrition/Food Safety,

Epigenetic Editing:On the Verge of Reprogramming Gene Expression at Will

Genome targeting has quickly developed as one of the most promising fields in science. By using programmable DNA-binding platforms and nucleases, scientists are now able to accurately edit the genome. These DNA-binding tools have recently also been applied to engineer the epigenome for gene expression modulation. Such epigenetic editing constructs have firmly demonstrated the causal role of epigenetics in instructing gene expression. Another focus of epigenome engineering is to understand the order of events of chromatin remodeling in gene expression regulation. Groundbreaking approaches in this field are beginning to yield novel insights into the function of individual chromatin marks in the context of maintaining cellular phenotype and regulating transient gene expression changes. This review focuses on recent advances in the field of epigenetic editing and highlights its promise for sustained gene expression reprogramming

The forensic analysis of soil by FTIR with multivariate analysis

Over the past few years more and more studies have been carried out in an attempt to utilize chemical profiles of soil using a wide variety of analytical methods. The value of soil as evidence rests with its prevalence at crime scenes and its transferability between the scene and the criminal. This can be of value for comparison if the scene of crime is known, but could also be so in the identification of a scene. The main basis for the comparison of sites to determine provenance is that soils vary from one place to another. The aim of this work is to find simple methods to identify soil provenance based on FTIR and multivariate analysi

Strategic situation, design and simulation of a biorefinery in Andalusia

In a lignocellulosic feedstock based biorefinery, biomass can be transformed in several products, with different chemical and/or agro-industrial applications, and energy (biofuels). The use of biomass waste is strongly advocated under European Union (EU) legislation in order to help achieve the climate and energy targets of the EU for 2020 and beyond. In this context, this study was focussed on the design and simulation of a biorefinery to mainly obtain ethanol and DME. These biofuels were obtained from waste forestry and agricultural waste biomass collected near the area where the biorefinery plant was proposed to be installed, to minimize the transportation costs and to promote the valorization of the biomass waste generated in this region. Moreover, the industrial applications of the possible obtained by-products were evaluated to minimize the environmental impacts and to make the biorefinery more sustainable. The central area of Andalusia was selected as the most adequate area to develop the installation of the biorefinery plant. Two commercial simulation software, such as Aspen HYSYS® and UniSim®, were used to design and size the equipments and to simulate both production lines. One of the most important achievements of this biorefinery is the possibility of obtaining 42,700 T y-1 of ethanol with a purity of 96%, which supposes a 16.5% of the Spanish national production in 2016, and 137,850 T y-1 of DME, with a purity of 99.99%. from these biomass waste Both compounds can be used as alternative fuels or energy sources. A techno-economic analysis was performed, obtaining a minimum selling price of 0.58 $/L for bioethanol, 1.15$/kg for DME and 0.65 $/kg for HMF and LA, respectively. These prices are comparable to those found in the literature. Furthermore, the implementation of the biorefinery in this strategic area promotes its economic and social development, improving the use of the natural resources to obtain competitive products to fossil fuels.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. This work was supported by the Spanish MINECO under CTQ2015-68654-R project. MCGM gratefully acknowledges the assistance of Spanish Ministry of Economy and Competitiveness through a “Juan de la Cierva – Formación” fellowship (FJCI-2015-25788)

Dielectric Characterization of Materials at 5G mm-Wave Frequencies

The development of the next-generation 5G wireless networks depends critically on the engineering of optimized high-frequency devices, employing dielectric materials. This work presents a comprehensive broadband dielectric characterization of polymers, ceramics and glasses from 5 GHz until 115 GHz.Various measurement techniques including split-post, split cavity, open resonator and free-space transmission are utilized to obtain wideband spectra. The frequency-dependent permittivity and losstangent are analyzed to identify suitable candidate materials exhibiting minimal dispersion and loss in the 5G millimeter-wave bands. The characterization reveals almost constant permittivity and a loss tangent that increases linearly with the frequency