Search results for: Helena M. Wilhelm

Authors: Marina Bubalo, Davor Romić, Stjepan Husnjak, Helena Bakić

Abstract:

Even though European Council Directive 91/676/EEC known as Nitrates Directive was adopted in 1991, the issue of water quality preservation in areas of intensive agricultural production still persist all over Europe. High nitrate nitrogen concentrations in surface and groundwater originating from diffuse sources are one of the most important environmental problems in modern intensive agriculture. The fate of nitrogen in soil, surface and groundwater in agricultural area is mostly affected by anthropogenic activity (i.e. agricultural practice) and hydrological and climatological conditions. The aim of this study was to identify impact of land use, soil type, soil vulnerability to pollutant percolation, and natural aquifer vulnerability to nitrate occurrence in surface and groundwater within an intensive agricultural area. The study was set in Varaždin County (northern Croatia), which is under significant influence of the large rivers Drava and Mura and due to that entire area is dominated by alluvial soil with shallow active profile mainly on gravel base. Negative agricultural impact on water quality in this area is evident therefore the half of selected county is a part of delineated nitrate vulnerable zones (NVZ). Data on water quality were collected from 7 surface and 8 groundwater monitoring stations in the County. Also, recent study of the area implied detailed inventory of agricultural production and fertilizers use with the aim to produce new agricultural land use database as one of dominant parameters. The analysis of this database done using ArcGIS 10.1 showed that 52,7% of total County area is agricultural land and 59,2% of agricultural land is used for intensive agricultural production. On the other hand, 56% of soil within the county is classified as soil vulnerable to pollutant percolation. The situation is similar with natural aquifer vulnerability; northern part of the county ranges from high to very high aquifer vulnerability. Statistical analysis of water quality data is done using SPSS 13.0. Cluster analysis group both surface and groundwater stations in two groups according to nitrate nitrogen concentrations. Mean nitrate nitrogen concentration in surface water – group 1 ranges from 4,2 to 5,5 mg/l and in surface water – group 2 from 24 to 42 mg/l. The results are similar, but evidently higher, in groundwater samples; mean nitrate nitrogen concentration in group 1 ranges from 3,9 to 17 mg/l and in group 2 from 36 to 96 mg/l. ANOVA analysis confirmed statistical significance between stations that are classified in the same group. The previously listed parameters (land use, soil type, etc.) were used in factorial correspondence analysis (FCA) to detect importance of each stated parameter in local water quality. Since stated parameters mostly cannot be altered, there is obvious necessity for more precise and more adapted land management in such conditions.

Keywords: agricultural area, nitrate, factorial correspondence analysis, water quality

Procedia PDF Downloads 237

Authors: Laura Helena Caicedo Lopez, Luis Miguel Contreras Medina, Ramon Gerardo Guevara Gonzales, Juan E. Andrade

Abstract:

In this study, we evaluated the effect of three different hydric stress conditions: Low (LHS), medium (MHS), and high (HHS) on capsaicinoid content and enzyme regulation of C. annuum plants. Five main peaks were detected using a 2 Hz resolution vibrometer laser (Polytec-B&K). These peaks or “characteristic frequencies” were used as acoustic emissions (AEs) treatment, transforming these signals into audible sound with the frequency (Hz) content of each hydric stress. Capsaicinoids (CAPs) are the main, secondary metabolites of chili pepper plants and are known to increase during hydric stress conditions or short drought-periods. The AEs treatments were applied in two plant stages: the first one was in the pre-anthesis stage to evaluate the genes that encode the transcription of enzymes responsible for diverse metabolic activities of C. annuum plants. For example, the antioxidant responses such as peroxidase (POD), superoxide dismutase (Mn-SOD). Also, phenyl-alanine ammonia-lyase (PAL) involved in the biosynthesis of the phenylpropanoid compounds. The chalcone synthase (CHS) related to the natural defense mechanisms and species-specific aquaporin (CAPIP-1) that regulate the flow of water into and out of cells. The second stage was at 40 days after flowering (DAF) to evaluate the biochemical effect of AEs related to hydric stress on capsaicinoids production. These two experiments were conducted to identify the molecular responses of C. annuum plants to AE. Moreover, to define AEs could elicit any increase in the capsaicinoids content after a one-week exposition to AEs treatments. The results show that all AEs treatment signals (LHS, MHS, and HHS) were significantly different compared to the non-acoustic emission control (NAE). Also, the AEs induced the up-regulation of POD (~2.8, 2.9, and 3.6, respectively). The gene expression of another antioxidant response was particularly treatment-dependent. The HHS induced and overexpression of Mn-SOD (~0.23) and PAL (~0.33). As well, the MHS only induced an up-regulation of the CHs gene (~0.63). On the other hand, CAPIP-1 gene gas down-regulated by all AEs treatments LHS, MHS, and HHS ~ (-2.4, -0.43 and -6.4, respectively). Likewise, the down-regulation showed particularities depending on the treatment. LHS and MHS induced downregulation of the SOD gene ~ (-1.26 and -1.20 respectively) and PAL (-4.36 and 2.05, respectively). Correspondingly, the LHS and HHS showed the same tendency in the CHs gene, respectively ~ (-1.12 and -1.02, respectively). Regarding the elicitation effect of AE on the capsaicinoids content, additional treatment controls were included. A white noise treatment (WN) to prove the frequency-selectiveness of signals and a hydric stressed group (HS) to compare the CAPs content. Our findings suggest that WN and NAE did not present differences statically. Conversely, HS and all AEs treatments induced a significant increase of capsaicin (Cap) and dihydrocapsaicin (Dcap) after one-week of a treatment. Specifically, the HS plants showed an increase of 8.33 times compared to the NAE and WN treatments and 1.4 times higher than the MHS, which was the AEs treatment with a larger induction of Capsaicinoids among treatments (5.88) and compared to the controls.

Keywords: acoustic emission, capsaicinoids, elicitors, hydric stress, plant signaling

Procedia PDF Downloads 136

Authors: Catelyn Coyle, Helena Kwakwa

Abstract:

Objective: As novel and better tolerated therapies become available, effective HCV testing and care models become increasingly necessary to not only identify individuals with active infection but also link them to HCV providers for medical evaluation and treatment. Our aim is to describe an effective HCV testing and linkage to care model piloted in a network of five community health centers located in Philadelphia, PA. Methods: In October 2012, National Nursing Centers Consortium piloted a routine opt-out HCV testing model in a network of community health centers, one of which treats HCV, HIV, and co-infected patients. Key aspects of the model were medical assistant initiated testing, the use of laboratory-based reflex test technology, and electronic medical record modifications to prompt, track, report and facilitate payment of test costs. Universal testing on all adult patients was implemented at health centers serving patients at high-risk for HCV. The other sites integrated high-risk based testing, where patients meeting one or more of the CDC testing recommendation risk factors or had a history of homelessness were eligible for HCV testing. Mid-course adjustments included the integration of dual HIV testing, development of a linkage to care coordinator position to facilitate the transition of HIV and/or HCV-positive patients from primary to specialist care, and the transition to universal HCV testing across all testing sites. Results: From October 2012 to June 2015, the health centers performed 7,730 HCV tests and identified 886 (11.5%) patients with a positive HCV-antibody test. Of those with positive HCV-antibody tests, 838 (94.6%) had an HCV-RNA confirmatory test and 590 (70.4%) progressed to current HCV infection (overall prevalence=7.6%); 524 (88.8%) received their RNA-positive test result; 429 (72.7%) were referred to an HCV care specialist and 271 (45.9%) were seen by the HCV care specialist. The best linkage to care results were seen at the test and treat the site, where of the 333 patients were current HCV infection, 175 (52.6%) were seen by an HCV care specialist. Of the patients with active HCV infection, 349 (59.2%) were unaware of their HCV-positive status at the time of diagnosis. Since the integration of dual HCV/HIV testing in September 2013, 9,506 HIV tests were performed, 85 (0.9%) patients had positive HIV tests, 81 (95.3%) received their confirmed HIV test result and 77 (90.6%) were linked to HIV care. Dual HCV/HIV testing increased the number of HCV tests performed by 362 between the 9 months preceding dual testing and first 9 months after dual testing integration, representing a 23.7% increment. Conclusion: Our HCV testing model shows that integrated routine testing and linkage to care is feasible and improved detection and linkage to care in a primary care setting. We found that prevalence of current HCV infection was higher than that seen in locally in Philadelphia and nationwide. Intensive linkage services can increase the number of patients who successfully navigate the HCV treatment cascade. The linkage to care coordinator position is an important position that acts as a trusted intermediary for patients being linked to care.

Keywords: HCV, routine testing, linkage to care, community health centers

Procedia PDF Downloads 332

Authors: Patrícia Branco, Catarina Prista, Helena Albergaria

Abstract:

Industrial fuel-ethanol fermentations are carried out under non-sterile conditions, which favors the development of microbial contaminants, leading to huge economic losses. Wild yeasts such as Brettanomyces bruxellensis and lactic acid bacteria are the main contaminants of industrial bioethanol fermentation, affecting Saccharomyces cerevisiae performance and decreasing ethanol yields and productivity. In order to control microbial contaminations, the fuel-ethanol industry uses different treatments, including acid washing and antibiotics. However, these control measures carry environmental risks such as acid toxicity and the rise of antibiotic-resistant bacteria. Therefore, it is crucial to develop and apply less toxic and more environmentally friendly biocontrol methods. In the present study, an industrial fuel-ethanol starter, S. cerevisiae Ethanol-Red, was genetically modified to over-express AMPs with activity against fuel-ethanol microbial contaminants and evaluated regarding its biocontrol effect during mixed-culture alcoholic fermentations artificially contaminated with B. bruxellensis. To achieve this goal, S. cerevisiae Ethanol-Red strain was transformed with a plasmid containing the AMPs-codifying genes, i.e., partial sequences of TDH1 (925-963 bp) and TDH2/3 (925-963 bp) and a geneticin resistance marker. The biocontrol effect of those genetically modified strains was evaluated against B. bruxellensis and compared with the antagonistic effect exerted by the modified strain with an empty plasmid (without the AMPs-codifying genes) and the non-modified strain S. cerevisiae Ethanol-Red. For that purpose, mixed-culture alcoholic fermentations were performed in a synthetic must use the modified S. cerevisiae Ethanol-Red strains together with B. bruxellensis. Single-culture fermentations of B. bruxellensis strains were also performed as a negative control of the antagonistic effect exerted by S. cerevisiae strains. Results clearly showed an improved biocontrol effect of the genetically-modified strains against B. bruxellensis when compared with the modified Ethanol-Red strain with the empty plasmid (without the AMPs-codifying genes) and with the non-modified Ethanol-Red strain. In mixed-culture fermentation with the modified S. cerevisiae strain, B. bruxellensis culturability decreased from 5×104 CFU/mL on day-0 to less than 1 CFU/mL on day-10, while in single-culture B. bruxellensis increased its culturability from 6×104 to 1×106 CFU/mL in the first 6 days and kept this value until day-10. Besides, the modified Ethanol-Red strain exhibited an enhanced antagonistic effect against B. bruxellensis when compared with that induced by the non-modified Ethanol-Red strain. Indeed, culturability loss of B. bruxellensis after 10 days of fermentation with the modified Ethanol-Red strain was 98.7 and 100% higher than that occurred in fermentations performed with the non-modified Ethanol-Red and the empty-plasmid modified strain, respectively. Therefore, one can conclude that the S. cerevisiae genetically modified strain obtained in the present work may be a valuable solution for the mitigation of microbial contamination in fuel-ethanol fermentations, representing a much safer and environmentally friendly preservation strategy than the antimicrobial treatments (acid washing and antibiotics) currently applied in fuel-ethanol industry.

Keywords: antimicrobial peptides, fuel-ethanol microbial contaminations, fuel-ethanol fermentation, biocontrol agents, genetically-modified yeasts

Procedia PDF Downloads 74

Authors: Katarína Koňariková, Georgios A. Perdikaris, Lucia Andrezálová, Zdeňka Ďuračková, Lucia Laubertová, Helena Gbelcová, Ingrid Žitňanová

Abstract:

Introduction: The continuous demand for new anti-cancer drugs has stimulated chemotherapeutic research based on the use of essential metalloelements with the aim to develop potential drugs with lower toxicity and higher antiproliferative activity against tumors. Copper(II) and its complexes play an important role as suitable species for antiproliferative tests. Objectives: The central objective of the current study was to investigate the potential in vitro anti-proliferative effects of N-salicylidene-L-glutamato copper (II) complexes and molecular mechanism of apoptosis induced by tested complexes. In our project we tested N-salicylidene-L-glutamato copper (II) complexes ZK1 - [Cu(N-salicylidene-L-glutamato)(H2O)2].H2O; MK0 - ([Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O); MK1 - [Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O; MK3 - transbis(ethanol)tetrakis(imidazol)Cu(II)(2+)bis(N-salicylidene-D,L-glutamato-N,O)-KO:KO´-(imidazol); MK5 - [Cu(N-salicylidene-D,L- glutamato)(2-methylimidazol] at concentration range 0.001-100 µmol/L against human colon carcinoma cell line HT-29 and human breast carcinoma cell line MCF-7. Methods: Viability was assessed by direct counting of 0.4% trypan blue dye-excluding cells after 24, 48 and 72 hour cultivations with or without copper complex and by MTT assay. To analyze the type of cell death and its mechanism induced by our copper complex we used different methods. To distinguish apoptosis from necrosis we used electrophoretic analysis, to study the activity of caspases 8 and 9 – luminometric analysis and caspase activity 3 colorimetric assay. Results: The observed anti-proliferative effect of the copper complexes appeared to be dose-, time- and cell line- dependent. Human colon carcinoma cells HT-29 appeared to be more sensitive to the complex MK0 ([Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O) than to ZK1 ([Cu(N-salicylidene-L-glutamato)(H2O)2].H2O) and MK1 ([Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O)). Human colon carcinoma cells HT-29 appeared to be more sensitive to the complex than human breast carcinoma cells MCF-7. IC50 decreased with time of incubation (24, 48 and 72h) for HT-29, but increased for MCF-7. By electrophoresis we found apoptotic cell death induced by our copper complexes in HT-29 at concentrations 1, 10, 50 and 100 µmol/L after 48h (ZK1) and 72h (MK0, MK1) and in MCF-7 we did not find apoptosis. We also studied molecular mechanism of apoptosis in HT-29 induced by copper complexes. We found active caspase 9 in HT-29 after ZK1 ([Cu(N-salicylidene-L-glutamato)(H2O)2].H2O) and MK1 ([Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O)) influence and active caspase 8 after MK0 ([Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O) influence. Conclusion: Our copper complexes showed cytotoxic activities against human colon carcinoma cells HT-29 and breast cancer cell line MCF-7 in vitro. Apoptosis was activated by mitochondrial pathway (intrinsic pathway) in case of ZK1 [Cu(N-salicylidene-L-glutamato)(H2O)2].H2O; MK1 [Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O; MK3 - transbis(ethanol)tetrakis(imidazol)Cu(II)(2+)bis(N-salicylidene-D,L-glutamato-N,O)-KO:KO´-(imidazol) and MK5 - [Cu(N-salicylidene-D,L- glutamato)(2-methylimidazol] copper complexes and by death receptors (extrinsic pathway) in case of MK0 [Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O copper complex in HT-29.

Keywords: apoptosis, copper complex, cancer, carcinoma cell line

Procedia PDF Downloads 265

Authors: Fabio N. Moreno, Letícia B. Marinho, Beatriz D. Ruiz, Maria Helena R. B. Martins

Abstract:

Brazil is a top consumer of pesticides worldwide, and the São Paulo State is one of the highest consumers among the Brazilian federative states. However, representative data about the occurrence of pesticides in surface waters of the São Paulo State is scarce. This paper aims to present the results of pesticides monitoring executed within the Water Quality Monitoring Network of CETESB (The Environmental Agency of the São Paulo State) between the 2018-2022 period. Surface water sampling points (21 to 25) were selected within basins of predominantly agricultural land-use (5 to 85% of cultivated areas). The samples were collected throughout the year, including high-flow and low-flow conditions. The frequency of sampling varied between 6 to 4 times per year. Selection of pesticide molecules for monitoring followed a prioritizing process from EMBRAPA (Brazilian Agricultural Research Corporation) databases of pesticide use. Pesticides extractions in aqueous samples were performed according to USEPA 3510C and 3546 methods following quality assurance and quality control procedures. Determination of pesticides in water (ng L-1) extracts were performed by high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS) and by gas chromatography with nitrogen phosphorus (GC-NPD) and electron capture detectors (GC-ECD). The results showed higher frequencies (20- 65%) in surface water samples for Carbendazim (fungicide), Diuron/Tebuthiuron (herbicides) and Fipronil/Imidaclopride (insecticides). The frequency of observations for these pesticides were generally higher in monitoring points located in sugarcane cultivated areas. The following pesticides were most frequently quantified above the Aquatic life benchmarks for freshwater (USEPA Office of Pesticide Programs, 2023) or Brazilian Federal Regulatory Standards (CONAMA Resolution no. 357/2005): Atrazine, Imidaclopride, Carbendazim, 2,4D, Fipronil, and Chlorpiryfos. Higher median concentrations for Diuron and Tebuthiuron in the rainy months (october to march) indicated pesticide transport through surface runoff. However, measurable concentrations in the dry season (april to september) for Fipronil and Imidaclopride also indicates pathways related to subsurface or base flow discharge after pesticide soil infiltration and leaching or dry deposition following pesticide air spraying. With exception to Diuron, no temporal trends related to median concentrations of the most frequently quantified pesticides were observed. These results are important to assist policymakers in the development of strategies aiming at reducing pesticides migration to surface waters from agricultural areas. Further studies will be carried out in selected points to investigate potential risks as a result of pesticides exposure on aquatic biota.

Keywords: pesticides monitoring, são paulo state, water quality, surface waters

Procedia PDF Downloads 31

Authors: Isabel N. Sierra-Garcia, Maria J. Ferreira, Sandro Figuereido, Newton Gomes, Helena Silva, Angela Cunha

Abstract:

Plant-associated microbial communities are considered crucial in the adaptation of halophytes to coastal environments. The plant microbiota can be horizontally acquired from the environment or vertically transmitted from generation to generation via seeds. Recruiting of the microbial communities by the plant is affected by geographical location, soil source, host genotype, and cultivation practice. There is limited knowledge reported on the microbial communities in halophytes the influence of biotic and abiotic factors. In this work, the microbiota associated with the halophyte Salicornia ramosissima was investigated to determine whether the structure of bacterial communities is influenced by host genotype or soil source. For this purpose, two contrasting sites where S. ramosissima is established in the estuarine system of the Ria de Aveiro were investigated. One site corresponds to a natural salt marsh where S. ramosissima plants are present (wild plants), and the other site is a former salt pan that nowadays are subjected to intensive crop production of S. ramosissima (crop plants). Bacterial communities from the rhizosphere, seeds and root endosphere of S. ramossisima from both sites were investigated by sequencing bacterial 16S rRNA gene using the Illumina MiSeq platform. The analysis of the sequences showed that the three plant-associated compartments, rhizosphere, root endosphere, and seed endosphere, harbor distinct microbiomes. However, bacterial richness and diversity were higher in seeds of wild plants, followed by rhizosphere in both sites, while seeds in the crop site had the lowest diversity. Beta diversity measures indicated that bacterial communities in root endosphere and seeds were more similar in both wild and crop plants in contrast to rhizospheres that differed by local, indicating that the recruitment of the similar bacterial communities by the plant genotype is active in regard to the site. Moreover, bacterial communities from the root endosphere and rhizosphere were phylogenetically more similar in both sites, but the phylogenetic composition of seeds in wild and crop sites was distinct. These results indicate that cultivation practices affect the seed microbiome. However, minimal vertical transmission of bacteria from seeds to adult plants is expected. Seeds from the crop site showed higher abundances of Kushneria and Zunongwangia genera. Bacterial members of the classes Alphaprotebacteria and Bacteroidia were the most ubiquitous across sites and compartments and might encompass members of the core microbiome. These findings indicate that bacterial communities associated with S. ramosissima are more influenced by host genotype rather than local abiotic factors or cultivation practices. This study provides a better understanding of the composition of the plant microbiota in S. ramosissima , which is essential to predict the interactions between plant and associated microbial communities and their effects on plant health. This knowledge is useful to the manipulations of these microbial communities to enhance the health and productivity of this commercially important plant.

Keywords: halophytes, plant microbiome, Salicornia ramosissima, agriculture

Procedia PDF Downloads 128

Authors: Adriana Anselmo Oliveira

Abstract:

The present project showcases a tile work from the Franco-Portuguese artist Maria Helena Vieira da Silva (1908-1992). It is a set of 8 panels composed of figurative and geometric tiles, with extra tiles framing nearby doors and windows in a study room in the (UFRRJ, Universidade Federal Rural do Rio de Janeiro). The aforementioned work was created between 1942 and 1943, during the artist's 6 year exile in the Brazilian city. This one-of-a-kind tileset was designed and made by Vieira da Silva between 1942 and 1943. Over the years, several units were lost, which led to their replacement in the nineties. However, these replacements don't do justice to the original work of art. In 2007, a project was initiated to fully repair and maintain the set. Three panels are removed and restored, but the project is halted. To this day, the three fully restored panels remain in boxes. In 2016 a new restoration project is submitted by the (Faculdade de Belas Artes da Universidade de Lisboa) in collaboration with de (Fundacão Árpád Szenes-Vieira da Silva). There are many varied opinions on restoring and conserving older pieces of art, however, we have the moral duty to safeguard the original materials used by the artist along with the artists original vision and also to care for the future generations of students who will use the space in which the tile-work was inserted. Many tiles have been replaced by white tiles, tiles with a divergent colour pallet and technique, and in a few cases, the incorrect place or way around. These many factors make it increasingly difficult to maintain the artists original vision and destroy and chance of coherence within the artwork itself. The conservative technician cannot make new images to fill the empty spaces or mark the remaining images with their own creative input. with reliable photographic documentation that can provide us with the necessary vision to allow us to proceed with an accurate reconstruction, we have the obligation to proceed and return the piece of art to its true form, as in its current state, it is impossible to maintain its original glory. Using the information we have, we must find a way to differentiate the original tiles from the reconstructions in order to recreate and reclaim the original message from the artist. The objective of this project is to understand the significance of tiles in Vieira da Silva's art as well as the influence they had on the artist's pictorial language since the colour definition on tile work is vastly different from the painting process as the materials change during their merger. Another primary goal is to understand what the previous interventions achieved besides increasing the artworks durability. The main objective is to submit a proposal that can salvage the artist's visual intention and supports it for posteriority. In summary, this proposal goes further than the usual conservative interventions as it intends to recreate the original artistic worth, prioritising the aesthetics and keeping its soul alive.

Keywords: Vieira da Silva, tiles, conservation, restoration

Procedia PDF Downloads 123

Authors: Ana Paula Soares, Alexandrina Lages, Helena Oliveira, Francisco-Javier Gutiérrez-Domínguez, Marisa Lousada

Abstract:

From an early age, exposure to a spoken language allows us to implicitly capture the structure underlying the succession of the speech sounds in that language and to segment it into meaningful units (words). Statistical learning (SL), i.e., the ability to pick up patterns in the sensory environment even without intention or consciousness of doing it, is thus assumed to play a central role in the acquisition of the rule-governed aspects of language and possibly to lie behind the language difficulties exhibited by children with development language disorder (DLD). The research conducted so far has, however, led to inconsistent results, which might stem from the behavioral tasks used to test SL. In a classic SL experiment, participants are first exposed to a continuous stream (e.g., syllables) in which, unbeknownst to the participants, stimuli are grouped into triplets that always appear together in the stream (e.g., ‘tokibu’, ‘tipolu’), with no pauses between each other (e.g., ‘tokibutipolugopilatokibu’) and without any information regarding the task or the stimuli. Following exposure, SL is assessed by asking participants to discriminate between triplets previously presented (‘tokibu’) from new sequences never presented together during exposure (‘kipopi’), i.e., to perform a two-alternative-forced-choice (2-AFC) task. Despite the widespread use of the 2-AFC to test SL, it has come under increasing criticism as it is an offline post-learning task that only assesses the result of the learning that had occurred during the previous exposure phase and that might be affected by other factors beyond the computation of regularities embedded in the input, typically the likelihood two syllables occurring together, a statistic known as transitional probability (TP). One solution to overcome these limitations is to assess SL as exposure to the stream unfolds using online techniques such as event-related potentials (ERP) that is highly sensitive to the time-course of the learning in the brain. Here we collected ERPs to examine the neurofunctional correlates of SL in preschool children with DLD, and chronological-age typical language development (TLD) controls who were exposed to an auditory stream in which eight three-syllable nonsense words, four of which presenting high-TPs and the other four low-TPs, to further analyze whether the ability of DLD and TLD children to extract-word-like units from the steam was modulated by words’ predictability. Moreover, to ascertain if the previous knowledge of the to-be-learned-regularities affected the neural responses to high- and low-TP words, children performed the auditory SL task, firstly, under implicit, and, subsequently, under explicit conditions. Although behavioral evidence of SL was not obtained in either group, the neural responses elicited during the exposure phases of the SL tasks differentiated children with DLD from children with TLD. Specifically, the results indicated that only children from the TDL group showed neural evidence of SL, particularly in the SL task performed under explicit conditions, firstly, for the low-TP, and, subsequently, for the high-TP ‘words’. Taken together, these findings support the view that children with DLD showed deficits in the extraction of the regularities embedded in the auditory input which might underlie the language difficulties.

Keywords: development language disorder, statistical learning, transitional probabilities, word segmentation

Procedia PDF Downloads 159

Authors: Elena Filova, Ondrej Kaplan, Marie Markova, Helena Dragounova, Roman Matejka, Eduard Brynda, Lucie Bacakova

Abstract:

Decellularized vessels have been evaluated as small-diameter vascular prostheses. Reseeding autologous cells onto decellularized tissue prior implantation should prolong prostheses function and make them living tissues. Suitable cell types for reseeding are both endothelial cells and bone marrow-derived stem cells, with a capacity for differentiation into smooth muscle cells upon mechanical loading. Endothelial cells assure antithrombogenicity of the vessels and MSCs produce growth factors and, after their differentiation into smooth muscle cells, they are contractile and produce extracellular matrix proteins as well. Fibrin is a natural scaffold, which allows direct cell adhesion based on integrin receptors. It can be prepared autologous. Fibrin can be modified with bound growth factors, such as basic fibroblast growth factor (FGF-2) and vascular endothelial growth factor (VEGF). These modifications in turn make the scaffold more attractive for cells ingrowth into the biological scaffold. The aim of the study was to prepare thin surface-attached fibrin assemblies with bound FGF-2 and VEGF, and to evaluate growth and differentiation of bone marrow-derived mesenchymal stem cells on the fibrin (Fb) assemblies. Following thin surface-attached fibrin assemblies were prepared: Fb, Fb+VEGF, Fb+FGF2, Fb+heparin, Fb+heparin+VEGF, Fb+heparin+FGF2, Fb+heparin+FGF2+VEGF. Cell culture poly-styrene and glass coverslips were used as controls. Human MSCs (passage 3) were seeded at the density of 8800 cells/1.5 mL alpha-MEM medium with 2.5% FS and 200 U/mL aprotinin per well of a 24-well cell culture. The cells have been cultured on the samples for 6 days. Cell densities on day 1, 3, and 6 were analyzed after staining with LIVE/DEAD cytotoxicity/viability assay kit. The differentiation of MSCs is being analyzed using qPCR. On day 1, the highest density of MSCs was observed on Fb+VEGF and Fb+FGF2. On days 3 and 6, there were similar densities on all samples. On day 1, cell morphology was polygonal and spread on all sample. On day 3 and 6, MSCs growing on Fb assemblies with FGF2 became apparently elongated. The evaluation of expression of genes for von Willebrand factor and CD31 (endothelial cells), for alpha-actin (smooth muscle cells), and for alkaline phosphatase (osteoblasts) is in progress. We prepared fibrin assemblies with bound VEGF and FGF-2 that supported attachment and growth of mesenchymal stem cells. The layers are promising for improving the ingrowth of MSCs into the biological scaffold. Supported by the Technology Agency of the Czech Republic TA04011345, and Ministry of Health NT11270-4/2010, and BIOCEV – Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University” project (CZ.1.05/1.1.00/02.0109), funded by the European Regional Development Fund for their financial supports.

Keywords: fibrin assemblies, FGF-2, mesenchymal stem cells, VEGF

Procedia PDF Downloads 293

Authors: Shosh Shahrabani, Sharon Teitler-Regev, Helena Desivilya Syna, Fotini Voulgaris, Evangelos Tsoukatos, Vitor Ambrosio, Sandra M. Correia Loureiro

Abstract:

International crises that affect tourism, such as terror attacks, political unrest, and economic crises have become more frequent, and their influence has become broader. The influence of such extreme events depends on their salience in the tourists' awareness. Hence, it is important to understand the mechanisms underlying tourists' selection of travel destinations, especially their perceptions of crisis-related events and the impact of the sociopolitical and economic context in their countries of origin. The current study examined how the socio-political and economic context in the home countries of potential young tourists affected their selection of travel destinations. The objective was to elucidate how the salience of various crises (economic and political) in the tourists' perceptions, due to their experiences at home, color their construal of destinations affected by similar hazards and influence their travel intentions. The study focused on student tourists from Israel, Greece, and Portugal. Today about a fifth of international tourism is based on young people, especially students. These countries were chosen since Greece and Portugal are in the midst of economic crises. In addition, Greece and Portugal have experienced political instability, while Israel has security-related problems (including terrorist incidents). In 2013, a total of 648 students, responded to a questionnaire that included questions concerning attitudes and risk perceptions regarding travel to destinations with various risk hazards as well as socio-demographic details. The results indicate that over half of the Israelis intend to visit Greece or Portugal. The majority of the Portuguese intend to visit Greece, while less than a third of them intend to visit Israel. About half of the Greeks intend to visit Portugal, and most of them do not intend to visit Israel. The results indicate that greater perceived importance of economic crises mitigates the intention to travel to destinations with economic crises for tourists from origin countries that are also marked by economic crises, such as Greece and Portugal. However, for tourists from Israel, a country with a relatively stable economy, issues related to the economy barely affect their intention to travel to the other two countries. The findings also suggest that Greeks and Portuguese who are highly concerned about political unrest are unlikely to select Israel as a tourist destination. In addition, strong apprehension regarding terrorism impedes the intention to travel to destinations marked by terrorist incidents, such as Israel. The current research contributes to the existing literature by highlighting the impact of travelers' personal previous experience with crisis on their risk perceptions and in turn on their intentions to travel to countries with similar risks. Therefore, in a world where such incidents are on the rise, understanding tourists' risk perceptions and behavior and the factors influencing their destination-related decisions are crucial for countries that wish to increase the numbers of incoming tourists.

Keywords: economic crises, political instability, risk perception, young tourists

Procedia PDF Downloads 439

Authors: Bonnie R. Brammall, Rhonda M. Garad, Helena J. Teede, Cheryce L. Harrison

Abstract:

Introduction: Reproductive aged women are at high-risk for accelerated weight gain and obesity development, with pregnancy recognised as a critical contributory life phase. Healthy lifestyle interventions during the preconception and antenatal period improve maternal and infant health outcomes. Yet, interventions from preconception through to postpartum and translation and implementation into real-world healthcare settings remain limited. OptimalMe is a randomised, hybrid implementation effectiveness study of evidence-based healthy lifestyle intervention. Here, we report engagement, acceptability of the intervention during preconception, and self-reported behaviour change outcomes as a result of the preconception phase of the intervention. Methods: Reproductive aged women who upgraded their private health insurance to include pregnancy and birth cover, signalling a pregnancy intention, were invited to participate. Women received access to an online portal with preconception health and lifestyle modules, goal-setting and behaviour change tools, monthly SMS messages, and two coaching sessions (randomised to video or phone) prior to pregnancy. Results: Overall n=527 expressed interest in participating. Of these, n=33 did not meet inclusion criteria, n=8 were not contactable for eligibility screening, and n=177 failed to engage after the screening, leaving n=309 who were enrolled in OptimalMe and randomised to intervention delivery method. Engagement with coaching sessions dropped by 25% for session two, with no difference between intervention groups. Women had a mean (SD) age of 31.7 (4.3) years and, at baseline, a self-reported mean BMI of 25.7 (6.1) kg/m², with 55.8% (n=172) of a healthy BMI. Behaviour was sub-optimal with infrequent self-weighing (38.1%), alcohol consumption prevalent (57.1%), sub-optimal pre-pregnancy supplementation (61.5%), and incomplete medical screening. Post-intervention 73.2% of women reported engagement with a GP for preconception care and improved lifestyle behaviour (85.5%), since starting OptimalMe. Direct pre-and-post comparison of individual participant data showed that of 322 points of potential change (up-to-date cervical screening, elimination of high-risk behaviours [alcohol, drugs, smoking], uptake of preconception supplements and improved weighing habits) 158 (49.1%) points of change were achieved. Health coaching sessions were found to improve accountability and confidence, yet further personalisation and support were desired. Engagement with video and phone sessions was comparable, having similar impacts on behaviour change, and both methods were well accepted and increased women's accountability. Conclusion: A low-intensity digital health and lifestyle program with embedded health coaching can improve the uptake of preconception care and lead to self-reported behaviour change. This is the first program of its kind to reach an otherwise healthy population of women planning a pregnancy. Women who were otherwise healthy showed divergence from preconception health and lifestyle objectives and benefited from the intervention. OptimalMe shows promising results for population-based behaviour change interventions that can improve preconception lifestyle habits and increase engagement with clinical health care for pregnancy preparation.

Keywords: preconception, pregnancy, preventative health, weight gain prevention, self-management, behaviour change, digital health, telehealth, intervention, women's health

Procedia PDF Downloads 66

Authors: Thiago E. Goto, Carla C. Lopes, Helena B. Nader, Anielle C. A. Silva, Noelio O. Dantas, José R. Siqueira Jr., Luciano Caseli

Abstract:

Quantum dots (QD) are semiconductor nanocrystals that can be employed in biological research as a tool for fluorescence imagings, having the potential to expand in vivo and in vitro analysis as cancerous cell biomarkers. Particularly, cadmium selenide (CdSe) magic-sized quantum dots (MSQDs) exhibit stable luminescence that is feasible for biological applications, especially for imaging of tumor cells. For these facts, it is interesting to know the mechanisms of action of how such QDs mark biological cells. For that, simplified models are a suitable strategy. Among these models, Langmuir films of lipids formed at the air-water interface seem to be adequate since they can mimic half a membrane. They are monomolecular films formed at liquid-gas interfaces that can spontaneously form when organic solutions of amphiphilic compounds are spread on the liquid-gas interface. After solvent evaporation, the monomolecular film is formed, and a variety of techniques, including tensiometric, spectroscopic and optic can be applied. When the monolayer is formed by membrane lipids at the air-water interface, a model for half a membrane can be inferred where the aqueous subphase serve as a model for external or internal compartment of the cell. These films can be transferred to solid supports forming the so-called Langmuir-Blodgett (LB) films, and an ampler variety of techniques can be additionally used to characterize the film, allowing for the formation of devices and sensors. With these ideas in mind, the objective of this work was to investigate the specific interactions of CdSe MSQDs with tumorigenic and non-tumorigenic cells using Langmuir monolayers and LB films of lipids and specific cell extracts as membrane models for diagnosis of cancerous cells. Surface pressure-area isotherms and polarization modulation reflection-absorption spectroscopy (PM-IRRAS) showed an intrinsic interaction between the quantum dots, inserted in the aqueous subphase, and Langmuir monolayers, constructed either of selected lipids or of non-tumorigenic and tumorigenic cells extracts. The quantum dots expanded the monolayers and changed the PM-IRRAS spectra for the lipid monolayers. The mixed films were then compressed to high surface pressures and transferred from the floating monolayer to solid supports by using the LB technique. Images of the films were then obtained with atomic force microscopy (AFM) and confocal microscopy, which provided information about the morphology of the films. Similarities and differences between films with different composition representing cell membranes, with or without CdSe MSQDs, was analyzed. The results indicated that the interaction of quantum dots with the bioinspired films is modulated by the lipid composition. The properties of the normal cell monolayer were not significantly altered, whereas for the tumorigenic cell monolayer models, the films presented significant alteration. The images therefore exhibited a stronger effect of CdSe MSQDs on the models representing cancerous cells. As important implication of these findings, one may envisage for new bioinspired surfaces based on molecular recognition for biomedical applications.

Keywords: biomembrane, langmuir monolayers, quantum dots, surfaces

Procedia PDF Downloads 164

Authors: Marie-Helena Docherty, Sion Edryd Williams

Abstract:

The prevalence of diabetes mellitus is increasing worldwide. In Sub-Saharan Africa, type 2 diabetes represents over 90% of all types of diabetes with the number of diabetic patients expected to rise. This represents a huge economic burden in an area already contending with high rates of other significant diseases, including the highest worldwide prevalence of HIV. Diabetic complications considerably impact on morbidity and mortality. The epidemiological data for the region quotes high rates of retinopathy (7-63%), neuropathy (27-66%) and microalbuminuria (10-83%). It is therefore imperative that diabetic screening programmes are established. It is recognised that in many parts of the developing world the implementation and management of such programmes is limited by a lack of available resources. The International Diabetes Federation produced guidelines in 2012 taking these limitations into account suggesting that all diabetic patients should have access to basic screening. These guidelines are consistent with the national diabetic guidelines produced by the Lesotho Medical Council. However, diabetic care in Lesotho is delivered at the local level, with variable levels of quality. A cross sectional study was performed in the outpatient department of Maluti Hospital in Mapoteng, Lesotho, a busy rural hospital in the Berea district. Demographic data on gender, age and modality of treatment were collected over a six-week time period. Information regarding 3 basic screening parameters was obtained. These parameters included eye screening (defined as a documented ophthalmology review within the last 12 months), foot screening (defined as a documented foot health assessment by any health care professional within the last 12 months) and secondary prevention (defined as a documented blood pressure and lipid profile reading within the last 12 months). These parameters were selected on the basis of the absolute minimum level of resources in Maluti Hospital. Renal screening was excluded, as the hospital does not have access to reliable renal profile checks or urinalysis. There is however a fully functioning on-site ophthalmology department run by a senior ophthalmologist with the ability to provide retinal photography, retinal surgery and photocoagulation therapy. Data was collected on 183 type 2 diabetics. 112 patients were male and 71 were female. The average age was 43 years. 4 patients were diet controlled, 140 patients were on oral hypoglycaemic agents (metformin and/or glibenclamide), and 39 patients were on a combination of insulin and oral hypoglycaemics. In the preceding 12 months, 5 patients had undergone eye screening (3%), 24 patients had undergone foot screening (13%), and 31 patients had lipid profile testing (17%). All patients had a documented blood pressure reading (100%). Our results show that screening is poorly performed in the basic indicators suggested by the IDF and the Lesotho Medical Council. On the basis of these results, a screening programme was developed using the mnemonic SaFE; secondary prevention, foot and eye care. This is simple, memorable and transferable between healthcare professionals. In the future, the expectation would be to expand upon this current programme to include renal screening, and to further develop screening pertaining to secondary prevention.

Keywords: Africa, complications, rural, screening

Procedia PDF Downloads 259

Authors: Helena Pera, Arcadi Sanmartin, Albert Falques, Rafael Rebolo, Xavier Ametller, Heiko Zillgen, Cecile Prum, Boris Even, Eric Kapornyai

Abstract:

Sheet piles system can be an interesting solution when dealing with harbors or quays designs. However, current design methods lead to conservative approaches due to the lack of specific basis of design. For instance, some design features still deal with pseudo-static approaches, although being a dynamic problem. Under this concern, the study particularly focuses on hydrodynamic water pressure definition and stability analysis of sheet pile system under seismic loads. During a seismic event, seawater produces hydrodynamic pressures on structures. Currently, design methods introduce hydrodynamic forces by means of Westergaard formulation and Eurocodes recommendations. They apply constant hydrodynamic pressure on the front sheet pile during the entire earthquake. As a result, the hydrodynamic load may represent 20% of the total forces produced on the sheet pile. Nonetheless, some studies question that approach. Hence, this study assesses the soil-structure-fluid interaction of sheet piles under seismic action in order to evaluate if current design strategies overestimate hydrodynamic pressures. For that purpose, this study performs various simulations by Plaxis 2D, a well-known geotechnical software, and CFD models, which treat fluid dynamic behaviours. Knowing that neither Plaxis nor CFD can resolve a soil-fluid coupled problem, the investigation imposes sheet pile displacements from Plaxis as input data for the CFD model. Then, it provides hydrodynamic pressures under seismic action, which fit theoretical Westergaard pressures if calculated using the acceleration at each moment of the earthquake. Thus, hydrodynamic pressures fluctuate during seismic action instead of remaining constant, as design recommendations propose. Additionally, these findings detect that hydrodynamic pressure contributes a 5% to the total load applied on sheet pile due to its instantaneous nature. These results are in line with other studies that use added masses methods for hydrodynamic pressures. Another important feature in sheet pile design is the assessment of the geotechnical overall stability. It uses pseudo-static analysis since the dynamic analysis cannot provide a safety calculation. Consequently, it estimates the seismic action. One of its relevant factors is the selection of the seismic reduction factor. A huge amount of studies discusses the importance of it but also about all its uncertainties. Moreover, current European standards do not propose a clear statement on that, and they recommend using a reduction factor equal to 1. This leads to conservative requirements when compared with more advanced methods. Under this situation, the study calibrates seismic reduction factor by fitting results from pseudo-static to dynamic analysis. The investigation concludes that pseudo-static analyses could reduce seismic action by 40-50%. These results are in line with some studies from Japanese and European working groups. In addition, it seems suitable to account for the flexibility of the sheet pile-soil system. Nevertheless, the calibrated reduction factor is subjected to particular conditions of each design case. Further research would contribute to specifying recommendations for selecting reduction factor values in the early stages of the design. In conclusion, sheet pile design still has chances for improving its design methodologies and approaches. Consequently, design could propose better seismic solutions thanks to advanced methods such as findings of this study.

Keywords: computational fluid dynamics, hydrodynamic pressures, pseudo-static analysis, quays, seismic design, steel sheet pile

Procedia PDF Downloads 117

Authors: Elsie M. Nolte, Annie M. Joubert, Roy Lakier, Maryke Etsebeth, Jolene M. Helena, Marcel Verwey, Laurence Lafanechere, Anne E. Theron

Abstract:

Treatment regimens for breast- and lung cancers may include both radiation- and chemotherapy. Ideally, a pharmaceutical agent which selectively sensitizes cancer cells to gamma (γ)-radiation would allow administration of lower doses of each modality, yielding synergistic anti-cancer benefits and lower metastasis occurrence, in addition to decreasing the side-effect profiles. A range of 2-methoxyestradiol (2-ME) analogues, namely 2-ethyl-3-O-sulphamoyl-estra-1,3,5 (10) 15-tetraene-3-ol-17one (ESE-15-one), 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10),15-tetraen-17-ol (ESE-15-ol) and 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16) were in silico-designed by our laboratory, with the aim of improving the parent compound’s bioavailability in vivo. The main effect of these compounds is the disruption of microtubule dynamics with a resultant mitotic accumulation and induction of programmed cell death in various cancer cell lines. This in vitro study aimed to determine the cellular responses involved in the radiation sensitization effects of these analogues at low doses in breast- and lung cancer cell lines. The oestrogen receptor positive MCF-7-, oestrogen receptor negative MDA-MB-231- and triple negative BT-20 breast cancer cell lines as well as the A549 lung cancer cell line were used. The minimal compound- and radiation doses able to induce apoptosis were determined using annexin-V and cell cycle progression markers. These doses (cell line dependent) were used to pre-sensitize the cancer cells 24 hours prior to 6 gray (Gy) radiation. Experiments were conducted on samples exposed to the individual- as well as the combination treatment conditions in order to determine whether the combination treatment yielded an additive cell death response. Morphological studies included light-, fluorescence- and transmission electron microscopy. Apoptosis induction was determined by flow cytometry employing annexin V, cell cycle analysis, B-cell lymphoma 2 (Bcl-2) signalling, as well as reactive oxygen species (ROS) production. Clonogenic studies were performed by allowing colony formation for 10 days post radiation. Deoxyribonucleic acid (DNA) damage was quantified via γ-H2AX foci and micronuclei quantification. Amplification of the p53 signalling pathway was determined by western blot. Results indicated that exposing breast- and lung cancer cells to nanomolar concentrations of these analogues 24 hours prior to γ-radiation induced more cell death than the compound- and radiation treatments alone. Hypercondensed chromatin, decreased cell density, a damaged cytoskeleton and an increase in apoptotic body formation were observed in cells exposed to the combination treatment condition. An increased number of cells present in the sub-G1 phase as well as increased annexin-V staining, elevation of ROS formation and decreased Bcl-2 signalling confirmed the additive effect of the combination treatment. In addition, colony formation decreased significantly. p53 signalling pathways were significantly amplified in cells exposed to the analogues 24 hours prior to radiation, as was the amount of DNA damage. In conclusion, our results indicated that pre-treatment of breast- and lung cancer cells with low doses of 2-ME analogues sensitized breast- and lung cancer cells to γ-radiation and induced apoptosis more so than the individual treatments alone. Future studies will focus on the effect of the combination treatment on non-malignant cellular counterparts.

Keywords: cancer, microtubule dynamics, radiation therapy, radiosensitization

Procedia PDF Downloads 178

Authors: Jolene M. Helena, Annie M. Joubert, Peace Mabeta, Magdalena Coetzee, Roy Lakier, Anne E. Mercier

Abstract:

Targeting the distant tumour and its microenvironment whilst preserving bone density is important in improving the outcomes of patients with bone metastases. 2-Ethyl-3-O-sulphamoyl-estra1,3,5(10)16-tetraene (ESE-16) is an in-silico-designed 2- methoxyestradiol analogue which aimed at enhancing the parent compound’s cytotoxicity and providing a more favourable pharmacokinetic profile. In this study, the potential radiosensitization effects of ESE-16 were investigated in an in vitro bone metastasis model consisting of murine pre-osteoblastic (MC3T3-E1) and pre-osteoclastic (RAW 264.7) bone cells, metastatic prostate (DU 145) and breast (MDA-MB-231) cancer cells, as well as human umbilical vein endothelial cells (HUVECs). Cytotoxicity studies were conducted on all cell lines via spectrophotometric quantification of 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide. The experimental set-up consisted of flow cytometric analysis of cell cycle progression and apoptosis detection (Annexin V-fluorescein isothiocyanate) to determine the lowest ESE-16 and radiation doses to induce apoptosis and significantly reduce cell viability. Subsequent experiments entailed a 24-hour low-dose ESE-16-exposure followed by a single dose of radiation. Termination proceeded 2, 24 or 48 hours thereafter. The effect of the combination treatment was investigated on osteoclasts via tartrate-resistant acid phosphatase (TRAP) activity- and actin ring formation assays. Tumour cell experiments included investigation of mitotic indices via haematoxylin and eosin staining; pro-apoptotic signalling via spectrophotometric quantification of caspase 3; deoxyribonucleic acid (DNA) damage via micronuclei analysis and histone H2A.X phosphorylation (γ-H2A.X); and Western blot analyses of bone morphogenetic protein-7 and matrix metalloproteinase-9. HUVEC experiments included flow cytometric quantification of cell cycle progression and free radical production; fluorescent examination of cytoskeletal morphology; invasion and migration studies on an xCELLigence platform; and Western blot analyses of hypoxia-inducible factor 1-alpha and vascular endothelial growth factor receptor 1 and 2. Tumour cells yielded half-maximal growth inhibitory concentration (GI50) values in the nanomolar range. ESE-16 concentrations of 235 nM (DU 145) and 176 nM (MDA-MB-231) and a radiation dose of 4 Gy were found to be significant in cell cycle and apoptosis experiments. Bone and endothelial cells were exposed to the same doses as DU 145 cells. Cytotoxicity studies on bone cells reported that RAW 264.7 cells were more sensitive to the combination treatment than MC3T3-E1 cells. Mature osteoclasts were more sensitive than pre-osteoclasts with respect to TRAP activity. However, actin ring morphology was retained. The mitotic arrest was evident in tumour and endothelial cells in the mitotic index and cell cycle experiments. Increased caspase 3 activity and superoxide production indicated pro-apoptotic signalling in tumour and endothelial cells. Increased micronuclei numbers and γ-H2A.X foci indicated increased DNA damage in tumour cells. Compromised actin and tubulin morphologies and decreased invasion and migration were observed in endothelial cells. Western blot analyses revealed reduced metastatic and angiogenic signalling. ESE-16-induced radiosensitization inhibits metastatic signalling and tumour cell survival whilst preferentially preserving bone cells. This low-dose combination treatment strategy may promote the quality of life of patients with metastatic bone disease. Future studies will include 3-dimensional in-vitro and murine in-vivo models.

Keywords: angiogenesis, apoptosis, bone metastasis, cancer, cell migration, cytoskeleton, DNA damage, ESE-16, radiosensitization.

Procedia PDF Downloads 131

Authors: Maria João Ferreira, Natalia Sierra-Garcia, Javier Cremades, Carla António, Ana M. Rodrigues, Helena Silva, Ângela Cunha

Abstract:

Salicornia ramosissima, a halophyte that grows naturally in coastal areas of the northern hemisphere, is often considered the most promising halophyte candidate for extensive crop cultivation and saline agriculture practices. The expanding interest in this plant surpasses its use as gourmet food and includes their potential application as a source of bioactive compounds for the pharmaceutical industry. Despite growing well in saline soils, sustainable and ecologically friendly techniques to enhance crop production and the nutritional value of this plant are still needed. The root microbiome of S. ramosissima proved to be a source of taxonomically diverse plant growth-promoting bacteria (PGPB). Halotolerant strains of Bacillus, Salinicola, Pseudomonas, and Brevibacterium, among other genera, exhibit a broad spectrum of plant-growth promotion traits [e.g., 3-indole acetic acid (IAA), 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, siderophores, phosphate solubilization, Nitrogen fixation] and express a wide range of extracellular enzyme activities. In this work, three plant growth-promoting bacteria strains (Brevibacterium casei EB3, Pseudomonas oryzihabitans RL18, and Bacillus aryabhattai SP20) isolated from the rhizosphere and the endosphere of S. ramosissima roots from different saltmarshes along the Portuguese coast were inoculated in S. ramosissima seeds. Plants germinated from inoculated seeds were grown for three months in pots filled with a mixture of perlite and estuarine sediment (1:1) in greenhouse conditions and later transferred to a growth chamber, where they were maintained two months with controlled photoperiod, temperature, and humidity. Pots were placed on trays containing the irrigation solution (Hoagland’s solution 20% added with 10‰ marine salt). Before reaching the flowering stage, plants were collected, and the fresh and dry weight of aerial parts was determined. Non-inoculated seeds were used as a negative control. Selected dried stems from the most promising treatments were later analyzed by GC-TOF-MS for primary metabolite composition. The efficiency of inoculation and persistence of the inoculum was assessed by Next Generation Sequencing. Inoculations with single strain EB3 and co-inoculations with EB3+RL18 and EB3+RL18+SP20 (All treatment) resulted in significantly higher biomass production (fresh and dry weight) compared to non-inoculated plants. Considering fresh weight alone, inoculation with isolates SP20 and RL18 also caused a significant positive effect. Combined inoculation with the consortia SP20+EB3 or SP20+RL18 did not significantly improve biomass production. The analysis of the profile of primary metabolites will provide clues on the mechanisms by which the growth-enhancement effect of the inoculants operates in the plants. These results sustain promising prospects for the use of rhizospheric and endophytic PGPB as biofertilizers, reducing environmental impacts and operational costs of agrochemicals and contributing to the sustainability and cost-effectiveness of saline agriculture. Acknowledgments: This work was supported by project Rhizomis PTDC/BIA-MIC/29736/2017 financed by Fundação para a Ciência e Tecnologia (FCT) through the Regional Operational Program of the Center (02/SAICT/2017) with FEDER funds (European Regional Development Fund, FNR, and OE) and by FCT through CESAM (UIDP/50017/2020 + UIDB/50017/2020), LAQV-REQUIMTE (UIDB/50006/2020). We also acknowledge FCT/FSE for the financial support to Maria João Ferreira through a PhD grant (PD/BD/150363/2019). We are grateful to Horta dos Peixinhos for their help and support during sampling and seed collection. We also thank Glória Pinto for her collaboration providing us the use of the growth chambers during the final months of the experiment and Enrique Mateos-Naranjo and Jennifer Mesa-Marín of the Departamento de Biología Vegetal y Ecología, the University of Sevilla for their advice regarding the growth of salicornia plants in greenhouse conditions.

Keywords: halophytes, PGPB, rhizosphere engineering, biofertilizers, primary metabolite profiling, plant inoculation, Salicornia ramosissima

Procedia PDF Downloads 124