Small RNAs in Tomato: from defence to development

RNA silencing is a major regulator of gene expression in plants, controlling from development to transposable element silencing and stress responses. As part of the silencing machinery, micro (mi)RNAs orchestrate silencing of their targets, either directly or through cascades of secondary small interfering (si)RNAs. To investigate the role of RNA silencing in plant immunity, I chose to focus on the miR482/2118 family, because of its diversity and presence in many plant species since the appearance of seed plants, with most genomes containing several copies, and because its members target sequences conserved in a family of disease resistance genes known Nucleotide biding site leucine-rich repeat (NLR) genes. In this dissertation, I wanted to address the extent to which the miRNA family and its derived phasiRNAs regulate expression of defence genes as well as contribute to quantitative resistance in crops. I explore the structural differences of miR482/2118 members in Solanum lycopersicum and show that they are functionally significant and affect their target preferences. My approach was based on small RNA sequencing and degradome data to characterize targets of these miRNAs, including the recently discovered tomato TAS5 locus. I also generated transgenic tomatoes constitutively expressing target mimic RNAs that sequester different miR482/2118 members. These tomato mimic RNA lines were less susceptible than their non-transgenic precursors to pathogens Phytophthora infestans and Pseudomonas syringae. Additionally, I investigated the role of small RNAs and their effector proteins during vegetative and reproductive development in tomato. I employed transcript and small RNA sequencing and CRISPR-Cas9 techniques of gene editing to investigate the impact of these factors in gamete viability and transposable element silencing in vegetative meristems. The results presented here provide new evidence about the extent that RNA silencing contributes to the regulation of vital processes in plants. My study primarily explores the extent to which structural differences between the members of the miR482/2118 family affect their range of action, and the use of target mimics against these miRNAs as biotechnological approach for enhancing disease resistance in highly bred cultivars.My work on this thesis was supported by the European Research Council Advanced Investigator Grant ERC-2013-AdG 340642 awarded to Professor David Baulcomb

A novel DCL2-dependent miRNA pathway in tomato affects susceptibility to RNA viruses.

Tomato Dicer-like2 (slDCL2) is a key component of resistance pathways against potato virus X (PVX) and tobacco mosaic virus (TMV). It is also required for production of endogenous small RNAs, including miR6026 and other noncanonical microRNAs (miRNAs). The slDCL2 mRNAs are targets of these slDCL2-dependent RNAs in a feedback loop that was disrupted by target mimic RNAs of miR6026. In lines expressing these RNAs, there was correspondingly enhanced resistance against PVX and TMV. These findings illustrate a novel miRNA pathway in plants and a crop protection strategy in which miRNA target mimicry elevates expression of defense-related mRNAs

Luminescent CdSe Quantum Dot Arrays for Rapid Sensing of Explosive Taggants

Chemical sensors based on fluorescent quantum dots have attracted intense interest because of their excellent optical and electronic properties compared to the routinely employed fluorescent organic dyes. This study reports a CdSe QD-polymer-based luminescent chemosensor, which is based on an array containing either green-emitting or red-emitting CdSe QDs embedded in polycaprolactone as a polymer host matrix. We evaluate the sensing capability of the nanocomposites by exposing both sensors to vapors of explosive taggants, explosive-like molecules, and some common solvents. Both nanocomposites exhibit a very fast response time of <30 s. The limit of detection of the sensors for 3-nitrotoluene, 4-nitrotoluene 2,3-dimethyl-2,3-dinitrobutane, and picric acid was found to be 0.055, 2.7, 0.7 and 916.4 ng, respectively. The sensor array constitutes a powerful tool to discriminate between explosive taggants (3-nitrotoluene, 4-nitrotoluene, and 2,3-dimethyl-2,3-dinitrobutane) and shows specific molecular recognition toward picric acid. This type of miniaturized luminescent QD-based nanocomposites might form the basis of a sensing platform technology to perform effective chemical detection and identification of explosive taggants preblast and postblast

El Puzle i l'aprenentatge de la programació

Es descriu una experiència formalitzada d’aprenentatge cooperatiu mitjançant la utilització de la metodologia del puzle. Concretament, s’utilitza el puzle com una eina de suport en una assignatura organitzada d’acord amb l’estratègia de l’aprenentatge basat en projectes. La metodologia del puzle s’ha aplicat a tots els grups de les assignatures següents des del curs 2004-2005: 1. Enginyeria Tècnica de Telecomunicació, Introducció als Computadors (Q1 de primer curs, 4,5 crèdits) i Laboratori de Programació (Q2 de primer curs, 4,5 crèdits), i 2. Graus en Enginyeria de Sistemes de Telecomunicació i en Enginyeria Telemàtica, Introducció als Ordinadors (Q1 de primer curs, 6 crèdits ECTS) i Projecte de Programació (Q2 de primer curs, 6 crèdits ECTS). Com a part d’assignatures basades en l’aprenentatge basat en projectes es va començar a experimentar en dues assignatures a l’Escola d'Enginyeria de Telecomunicació i Aeroespacial de Castelldefels (EETAC), en el marc de les proves pilot d’adaptació de titulacions a l'espai europeu d'educació superior (EEES), impulsades per l’antic Departament d'Universitats, Recerca i Societat de la Informació (DURSI) de la Generalitat de Catalunya. Posteriorment es va seguir aplicant aquest model en els nous graus. Aquests anys d’experimentació ens han proporcionat una bona quantitat de resultats i conclusions.Peer Reviewe

Incidence of epidural haematoma and neurological injury in cardiovascular patients with epidural analgesia/anaesthesia: systematic review and meta-analysis

BACKGROUND: Epidural anaesthesia is used extensively for cardiothoracic and vascular surgery in some centres, but not in others, with argument over the safety of the technique in patients who are usually extensively anticoagulated before, during, and after surgery. The principle concern is bleeding in the epidural space, leading to transient or persistent neurological problems. METHODS: We performed an extensive systematic review to find published cohorts of use of epidural catheters during vascular, cardiac, and thoracic surgery, using electronic searching, hand searching, and reference lists of retrieved articles. RESULTS: Twelve studies included 14,105 patients, of whom 5,026 (36%) had vascular surgery, 4,971 (35%) cardiac surgery, and 4,108 (29%) thoracic surgery. There were no cases of epidural haematoma, giving maximum risks following epidural anaesthesia in cardiac, thoracic, and vascular surgery of 1 in 1,700, 1 in 1,400 and 1 in 1,700 respectively. In all these surgery types combined the maximum expected rate would be 1 in 4,700. In all these patients combined there were eight cases of transient neurological injury, a rate of 1 in 1,700 (95% confidence interval 1 in 3,300 to 1 in 850). There were no cases of persistent neurological injury (maximum expected rate 1 in 4,600). CONCLUSION: These estimates for cardiothoracic epidural anaesthesia should be the worst case. Limitations are inadequate denominators for different types of surgery in anticoagulated cardiothoracic or vascular patients more at risk of bleeding

Patient preferences and treatment safety for uncomplicated vulvovaginal candidiasis in primary health care

<p>Abstract</p> <p>Background</p> <p>Vaginitis is a common complaint in primary care. In uncomplicated candidal vaginitis, there are no differences in effectiveness between oral or vaginal treatment. Some studies describe that the preferred treatment is the oral one, but a Cochrane's review points out inconsistencies associated with the report of the preferred way that limit the use of such data. Risk factors associated with recurrent vulvovaginal candidiasis still remain controversial.</p> <p>Methods/Design</p> <p>This work describes a protocol of a multicentric prospective observational study with one year follow up, to describe the women's reasons and preferences to choose the way of administration (oral vs topical) in the treatment of not complicated candidal vaginitis. The number of women required is 765, they are chosen by consecutive sampling. All of whom are aged 16 and over with vaginal discharge and/or vaginal pruritus, diagnosed with not complicated vulvovaginitis in Primary Care in Madrid.</p> <p>The main outcome variable is the preferences of the patients in treatment choice; secondary outcome variables are time to symptoms relief and adverse reactions and the frequency of recurrent vulvovaginitis and the risk factors. In the statistical analysis, for the main objective will be descriptive for each of the variables, bivariant analysis and multivariate analysis (logistic regression).. The dependent variable being the type of treatment chosen (oral or topical) and the independent, the variables that after bivariant analysis, have been associated to the treatment preference.</p> <p>Discussion</p> <p>Clinical decisions, recommendations, and practice guidelines must not only attend to the best available evidence, but also to the values and preferences of the informed patient.</p

The microRNA response associated with methyl jasmonate-induced resistance in Norway spruce bark

Methyl jasmonate (MeJA) treatment elicits induced resistance (IR) against pests and diseases in Norway spruce (Picea abies). We recently demonstrated using mRNA-seq that this MeJA-IR is associated with both a prolonged upregulation of inducible defenses and defense priming. Gene expression can be regulated at both a transcriptional and post-transcriptional level by small RNAs, including microRNAs (miRNAs). Here we explore the effects of MeJA treatment and subsequent challenge by wounding on the Norway spruce miRNA transcriptome. We found clusters of prolonged down- or upregulated miRNAs as well as miRNAs whose expression was primed after MeJA treatment and subsequent wounding challenge. Differentially expressed miRNAs included miR160, miR167, miR172, miR319, and the miR482/2118 superfamily. The most prominent mRNA targets predicted to be differentially expressed by miRNA activity belonged to the nucleotide-binding site leucine-rich repeat (NBS-LRR) family. Among other predicted miRNA targets were genes regulating jasmonic acid biosynthesis. Our results indicate that miRNAs have an important role in the regulation of MeJA-IR in Norway spruce

Measurement of the Top Pair Production Cross Section in the Dilepton Decay Channel in ppbar Collisions at sqrt s = 1.96 TeV

Submitted to Phys. Rev. DA measurement of the \ttbar production cross section in \ppbar collisions at $\sqrt{{\rm s}}$ = 1.96 TeV using events with two leptons, missing transverse energy, and jets is reported. The data were collected with the CDF II Detector. The result in a data sample corresponding to an integrated luminosity 2.8 fb$^{-1}$ is: \sigma_{\ttbar} = 6.27 $\pm$ 0.73(stat) $\pm$ 0.63(syst) $\pm$ 0.39(lum) pb. for an assumed top mass of 175 GeV/$c^{2}$.A measurement of the tt̅ production cross section in pp̅ collisions at √s=1.96  TeV using events with two leptons, missing transverse energy, and jets is reported. The data were collected with the CDF II detector. The result in a data sample corresponding to an integrated luminosity 2.8  fb-1 is σtt̅ =6.27±0.73(stat)±0.63(syst)±0.39(lum)  pb. for an assumed top mass of 175  GeV/c2.Peer reviewe