Carvalho, Teresa, Faculdade de Ciências da Universidade do Porto, Portugal Veiga, Marlene, Faculdade de Ciências da Universidade do Porto, Portugal
Fernandez-Gonzalez, Maria, Faculdade de Ciências da Universidade do Porto, Portugal Ribeiro, Helena, Faculdade de Ciências da Universidade do Porto, Portugal
Abreu, Ilda, Faculdade de Ciências da Universidade do Porto, Portugal
Pollen grains are bioaerosols produced by the higher plants during the pollination process. While airborne, pollen can interact with atmospheric pollutants leading to changes in its function. The aim of this study was to evaluate the effects of ozone in Platanus x acerifolia pollen fertility, ROS production and in the NAD(P)H oxidase activity.
Platanus x acerifolia pollen was collected from different branches of trees in the FCUP surrounding area during its flowering. Pollen samples were exposed in vitro for 6 h to O3 concentrations of half, equal and twice the standard limit value for human health protection according to the European Union Directive 2008/50/EC on ambient air quality and cleaner air for Europe. Pollen viability was determined by FDA test and germination rate was determined using a solid medium containing H3BO3, CaCl2 and sucrose. The ROS detection was evaluated through a fluorescent ROS indicator dye (DCFH2-DA) and the NAD(P)H oxidase activity was measured using the nitroblue tetrazolium (NBT) assay by spectrophotometry.
Pollen viability and germination rate significantly decreased after exposure to O3. Also an increase in ROS production was observed although not statistically significant compared with the control sample. Changes in the ROS-generating enzyme NAD(P)H oxidase activity were quantified. Our results suggest that pollen can be modified by the increasing atmospheric pollution, which can lead to a decrease in its ability to perform its vital task of reproduction with severe consequences in ecosystems balance.
Acknowledgments: National Funds through FCT - Foundation for Science and Technology in the project PTDC/ATP-EAM/0817/2014. Institute of Earth Sciences (ICT) funds, under contract with FCT and FEDER through the Operational Program Competitiveness Factors - COMPETE (UID/GEO/04683/2013 (POCI-01-0145-FEDER-007690)).
15334 | Heart rate variability in postural changes
Pereira, Margarida, Faculdade de Ciências da Universidade do Porto, Universidade do Porto, Portugal
Amado, Celestino, Faculdade de Ciências da Universidade do Porto, Universidade do Porto, Portugal
Dias, Celeste, Faculdade de Medicina da Universidade do Porto, Centro Hospitalar São João, Universidade do Porto, Portugal
Rocha, Ana Paula, Faculdade de Ciências da Universidade do Porto, CMUP, Universidade do Porto, Portugal
The autonomic nervous system (ANS) exerts rigorous control over essential functions of our body. Patients with severe acute brain injury (ABI) may have ANS dysfunction which may aggravate critical illness. Use of tilt-table in early rehabilitation of these patients contributes to neuronal recovery although introducing risks, e.g. orthostatic intolerance.
The aim of this work is to study the modelling of heart rate variability and arterial blood pressure to analyse the relation between the ANS and the interplay between cardiovascular variables and postural changes, using recent methodologies such as described in [1-2, and ref. therein]. In particular, to describe the dynamic complexity of the processes, we extended a parametric framework which exploits the theory of state space models to provide the multiscale representation of ARFIMA processes. This enables a reliable evaluation of multiscale entropy across multiple time scales, incorporating the effects of short-term dynamics and long-range correlations [3].
Physiological time series measured during resting state and postural stress in healthy volunteers and in patients after ABI were analysed. We were able to confirm the relationship between cardiovascular variables and the postural changes induced.
Acknowledgments: Work supported by CMUP, UID/MAT/00144/2019, funded by FCT (Portugal), (MEC, FEDER), under PT2020
References:
[1] Almeida R et al. ARFIMA-GARCH Modeling of HRV: Clinical Application in Acute Brain Injury. In Barbieri R. et al. (eds) Complexity and Nonlinearity in Cardiovascular Signals, Springer International Publishing, pg 451-468, 2017
[2] Kamath M et al. (eds) Heart Rate Variability (HRV) Signal Analysis: Clinical Applications. CRC Press, 2013
[3] Faes L, Pereira M, Silva ME, Pernice R, Busacca A, Javorka M, Rocha AP. Multiscale complexity of cardiovascular variability signals assessed by fractionally-integrated autoregressive models. ESCGO 2018 Abstract Book, pg 33. ISBN 978-3-9503613-1-5
15338 | Ozone effects on Betula pendula pollen allergenicity
Veiga, Marlene, Faculdade de Ciências da Universidade do Porto, Portugal Carvalho, Teresa, Faculdade de Ciências da Universidade do Porto, Portugal
Fernandez-Gonzalez, Maria, Faculdade de Ciências da Universidade do Porto, Portugal Ribeiro, Helena, Faculdade de Ciências da Universidade do Porto, Portugal
Abreu, Ilda, Faculdade de Ciências da Universidade do Porto, Portugal
Pollen related respiratory allergies affects the life quality of earth inhabitants being a general worldwide health problem. They are associated to an allergic response of susceptible individuals to the airborne pollen of several anemophilous trees. Birch is one of the major pollen-allergen- producer in north, central, and eastern Europe and is starting to be a significant problem in the Mediterranean area due to its popularity as ornamental plant among landscape architects. In this study we aimed to test the effects of ozone in Betula pendula pollen allergenicity.
Male catkins of Betula pendula was collected during its flowering and the anthers were dried, gently crushed and the pollen thus released was passed through different grades of sieves. An environmental chamber was used to expose the pollen samples to O3. This chamber is equipped with temperature and relative humidity sensors and a sunlight simulator. The pollen was exposed for 6 h to three concentration levels, chosen based on the European Union Directive 2008/50/EC of 21 May 2008 on ambient air quality and cleaner air for Europe: half, equal and twice the standard limit value. Pollen soluble proteins were extracted and the polypeptide profiles were determined by SDS-PAGE. The allergenicity was assessed by Western blot and ELISA using sera from patients allergic to birch pollen.
It was observed a qualitative and quantitative differences in IgE reactivity to the protein extracts of the pollen exposed to ozone compared with the non-exposed one. These changes suggest that ozone can induce alterations in the antigenic characteristics of the pollen leading to a allergenicity potency modification.
Acknowledgments: National Funds through FCT - Foundation for Science and Technology in the project PTDC/ATP-EAM/0817/2014. Institute of Earth Sciences (ICT) funds, under contract with FCT and FEDER through the Operational Program Competitiveness Factors - COMPETE (UID/GEO/04683/2013 (POCI-01-0145-FEDER-007690).
15343 | Intronic cis-regulatory elements regulate CDH1 gene expression and
function
Matos, Rita, i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, Portugal
Mesquita, Bárbara, i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, Portugal
Oliveira, Patrícia, i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, Portugal
Valente, Sofia, i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, Portugal
Pinheiro, Hugo, i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, Portugal
Carvalho, Joana, i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, Portugal
Huntsman, David, Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada; Centre for Translational and Applied Genomics (CTAG), BC Cancer Agency, Vancouver, British Columbia, Canada; Genetic Pathology Evaluation Centre, University of British Columbia and Vancouver General Hospital, Vancouver, British Columbia, Canada
Ferro, Anabela, i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, Portugal
Oliveira, Carla, i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal; IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, Portugal
Cis-regulatory elements (CREs) are non-coding DNA regions, capable of integrating different protein signals important for the expression of specific genes in a controlled temporal and spatial manner. Genomic structural variations in CREs may affect gene regulatory networks, explaining certain disease etiologies. In this work, we are studying CDH1, a tumor suppressor gene widely disrupted in epithelial cancers. We aim at unveiling the role of potential intronic CREs (iCREs) in CDH1 expression regulation, since its exonic alterations do not explain all the abnormal CDH1- associated phenotypes.
Bioinformatics analyses on CDH1 mining ENCODE data for chromatin accessibility, epigenetic marks, transcription factor binding sites and other regulatory elements, allowed to identify putative CDH1 iCREs. To ascertain their functional relevance, we edited each iCRE separately in a gastric cancer cell line by using CRISPR-Cas9, as well as CDH1 exon2 as positive control. All engineered cell clones were purified by single-cell sorting and gene editing was confirmed by sequencing. CDH1 expression was assessed by qRT-PCR and a single base primer-extension assay, while E-cadherin expression level and pattern were determined by western blotting and immunocytochemistry, respectively.
We identified two putative iCREs (iCRE1 and iCRE8) of CDH1 and successfully generated homogeneous clonal cell lines with fine-mapped deletions and inversions in clones´ DNA sequences. Our results indicate that intronic rearrangements at iCRE1 region impair CDH1/E- cadherin expression, possibly leading to allelic imbalance through the differential binding of transcription factors. Moreover, iCRE1-edited cells can have a similar phenotype to those harboring exon2 deletions. No obvious CDH1 loss of function was detected for iCRE8-edited clones. This study highlights iCRE1 as a CRE of CDH1/E-cadherin expression, supporting a potential