| 1. |
- Beal, Jacob, et al.
(författare)
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Robust estimation of bacterial cell count from optical density
- 2020
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Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 3:1
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Tidskriftsartikel (refereegranskat)abstract
- Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data.
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| 2. |
- Carraminana, Albert, et al.
(författare)
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Rationale and Study Design for an Individualized Perioperative Open Lung Ventilatory Strategy in Patients on One-Lung Ventilation (iPROVE-OLV)
- 2019
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Ingår i: Journal of Cardiothoracic and Vascular Anesthesia. - : W B SAUNDERS CO-ELSEVIER INC. - 1053-0770 .- 1532-8422. ; 33:9, s. 2492-2502
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Tidskriftsartikel (refereegranskat)abstract
- Objective: The aim of this clinical trial is to examine whether it is possible to reduce postoperative complications using an individualized perioperative ventilatory strategy versus using a standard lung-protective ventilation strategy in patients scheduled for thoracic surgery requiring one-lung ventilation. Design: International, multicenter, prospective, randomized controlled clinical trial. Setting: A network of university hospitals. Participants: The study comprises 1,380 patients scheduled for thoracic surgery. Interventions: The individualized group will receive intraoperative recruitment maneuvers followed by individualized positive end-expiratory pressure (open lung approach) during the intraoperative period plus postoperative ventilatory support with high-flow nasal cannula, whereas the control group will be managed with conventional lung-protective ventilation. Measurements and Main Results: Individual and total number of postoperative complications, including atelectasis, pneumothorax, pleural effusion, pneumonia, acute lung injury; unplanned readmission and reintubation; length of stay and death in the critical care unit and in the hospital will be analyzed for both groups. The authors hypothesize that the intraoperative application of an open lung approach followed by an individual indication of high-flow nasal cannula in the postoperative period will reduce pulmonary complications and length of hospital stay in high-risk surgical patients. (C) 2019 Published by Elsevier Inc.
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| 3. |
- Batista-Santos, Paula, et al.
(författare)
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Is salt stress tolerance in Casuarina glauca Sieb. ex Spreng. associated with its nitrogen-fixing root-nodule symbiosis? An analysis at the photosynthetic level
- 2015
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Ingår i: Plant physiology and biochemistry (Paris). - : Elsevier BV. - 0981-9428 .- 1873-2690. ; 96, s. 97-109
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Tidskriftsartikel (refereegranskat)abstract
- Casuarina glauca is an actinorhizal tree which establishes root-nodule symbiosis with N-2-fixing Frankia bacteria. This plant is commonly found in saline zones and is widely used to remediate marginal soils and prevent desertification. The nature of its ability to survive in extreme environments and the extent of Frankia contribution to stress tolerance remain unknown. Thus, we evaluated the ability of C. glauca to cope with salt stress and the influence of the symbiosis on this trait. To this end, we analysed the impact of salt on plant growth, mineral contents, water relations, photosynthetic-related parameters and nonstructural sugars in nodulated vs. non-nodulated plants. Although the effects on photosynthesis and stomatal conductance started to become measurable in the presence of 200 mM NaCl, photochemical (e.g., photosynthetic electron flow) and biochemical (e.g., activity of photosynthetic enzymes) parameters were only strongly impaired when NaCl levels reached 600 mM. These results indicate the maintenance of high tissue hydration under salt stress, probably associated with enhanced osmotic potential. Furthermore, the maintenance of photosynthetic assimilation potential (A(max)), together with the increase in the quantum yield of down-regulated energy dissipation of PSII (Y-NPQ), suggested a down-regulation of photosynthesis instead of photo-damaging effects. A comparison of the impact of increasing NaCl levels on the activities of photosynthetic (RubisCO and ribulose-5 phosphate kinase) and respiratory (pyruvate kinase and NADH-dependent malate dehydrogenase) enzymes vs. photosynthetic electron flow and fluorescence parameters, revealed that biochemical impairments are more limiting than photochemical damage. Altogether, these results indicate that, under controlled conditions, C glauca tolerates high NaCl levels and that this capacity is linked to photosynthetic adjustments.
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| 4. |
- Dias, Nicole Geovana, et al.
(författare)
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Intimate Partner Violence and Use of Primary and Emergency Care : The Role of Informal Social Support
- 2020
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Ingår i: Health & Social Work. - : Oxford academic. - 0360-7283 .- 1545-6854. ; 45:2, s. 91-100
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Tidskriftsartikel (refereegranskat)abstract
- Social support may encourage victims to disclose their experiences of intimate partner violence (IPV), but also to seek the appropriate help and care in the social and health services. Using data from a multicenter European project, DOVE (Domestic Violence Against women/men in Europe-prevalence, determinants, effects, and policies/practices), the present study aimed at measuring the frequency of primary care and emergency use according to IPV types of victimization, and to investigate whether victims receiving different levels of informal social support are using health care differently. Results suggested a significant association between IPV types and use of emergency services, and no association was found regarding primary care services. Victims of physical abuse and sexual coercion went to the emergency department (ED) more frequently (more than once a year). Also, victims of physical abuse receiving low social support visited an ED more frequently than those with high social support, whereas victims of sexual coercion with high informal social support went more often to the ED compared with victims of sexual coercion with low social support, even after controlling for other covariates. These results seem to suggest that social support has a significant role in the decision to use health care among victims of IPV.
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| 5. |
- Fernandes, Isabel, et al.
(författare)
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Salt Stress Tolerance in Casuarina glauca : Insights from the Branchlets Transcriptome
- 2022
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Ingår i: PLANTS. - : MDPI AG. - 2223-7747. ; 11:21
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Tidskriftsartikel (refereegranskat)abstract
- Climate change and the accelerated rate of population growth are imposing a progressive degradation of natural ecosystems worldwide. In this context, the use of pioneer trees represents a powerful approach to reverse the situation. Among others, N2-fixing actinorhizal trees constitute important elements of plant communities and have been successfully used in land reclamation at a global scale. In this study, we have analyzed the transcriptome of the photosynthetic organs of Casuarina glauca (branchlets) to unravel the molecular mechanisms underlying salt stress tolerance. For that, C. glauca plants supplied either with chemical nitrogen (KNO3+) or nodulated by Frankia (NOD+) were exposed to a gradient of salt concentrations (200, 400, and 600 mM NaCl) and RNA-Seq was performed. An average of ca. 25 million clean reads was obtained for each group of plants, corresponding to 86,202 unigenes. The patterns of differentially expressed genes (DEGs) clearly separate two groups: (i) control- and 200 mM NaCl-treated plants, and (ii) 400 and 600 mM NaCl-treated plants. Additionally, although the number of total transcripts was relatively high in both plant groups, the percentage of significant DEGs was very low, ranging from 6 (200 mM NaCl/NOD+) to 314 (600 mM NaCl/KNO3+), mostly involving down-regulation. The vast majority of up-regulated genes was related to regulatory processes, reinforcing the hypothesis that some ecotypes of C. glauca have a strong stress-responsive system with an extensive set of constitutive defense mechanisms, complemented by a tight mechanism of transcriptional and post-transcriptional regulation. The results suggest that the robustness of the stress response system in C. glauca is regulated by a limited number of genes that tightly regulate detoxification and protein/enzyme stability, highlighting the complexity of the molecular interactions leading to salinity tolerance in this species.
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| 6. |
- Fortunato, Ana, et al.
(författare)
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Isolation and characterization of cgchi3, a nodule-specific gene from Casuarina glauca encoding a class III chitinase
- 2007
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Ingår i: Physiologia Plantarum. - 0031-9317. ; 130:3, s. 418-426
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Tidskriftsartikel (refereegranskat)abstract
- Chitinases (EC 3.2.1.14) catalyse the hydrolysis of chitin, a homopolymer of beta-1,4-linked N-acetyl-D-glucosamine residues. Plant chitinases are involved in a wide variety of processes; in particular, their expression has been found to be enhanced in symbiotic and pathogenic plant-microbe interactions. During this work we have cloned and characterized a gene encoding a class III chitinase from actinorhizal nodules of Casuarina glauca (cgchi3). CGCHI3 was found to be encoded by a single gene that was specifically activated in nodules as compared with uninoculated control roots and leaves. The expression of this gene was further enhanced in nodules after salicylic acid treatment and completely repressed after wounding. In situ hybridisation analysis revealed that cgchi3 is an early nodulin gene, being expressed in the meristem and in the uninfected cortical cells of young nodules. Based on the obtained results we suggest that this gene is involved in nodule development. This is the first report on a class III chitinase coding gene that is specifically activated during actinorhizal symbiosis.
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| 7. |
- Pocas, Juliana, et al.
(författare)
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Syndecan-4 is a maestro of gastric cancer cell invasion and communication that underscores poor survival
- 2023
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences (PNAS). - 0027-8424 .- 1091-6490. ; 120:20
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Tidskriftsartikel (refereegranskat)abstract
- Gastric cancer is a dominating cause of cancer-associated mortality with limited therapeutic options. Here, we show that syndecan-4 (SDC4), a transmembrane pro-teoglycan, is highly expressed in intestinal subtype gastric tumors and that this sig -nature associates with patient poor survival. Further, we mechanistically demonstrate that SDC4 is a master regulator of gastric cancer cell motility and invasion. We also find that SDC4 decorated with heparan sulfate is efficiently sorted in extracellular vesicles (EVs). Interestingly, SDC4 in EVs regulates gastric cancer cell-derived EV organ distribution, uptake, and functional effects in recipient cells. Specifically, we show that SDC4 knockout disrupts the tropism of EVs for the common gastric cancer metastatic sites. Our findings set the basis for the molecular implications of SDC4 expression in gastric cancer cells and provide broader perspectives on the development of therapeutic strategies targeting the glycan-EV axis to limit tumor progression.
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| 8. |
- Scotti-Campos, Paula, et al.
(författare)
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Antioxidative ability and membrane integrity in salt-induced responses of Casuarina glauca Sieber ex Spreng. in symbiosis with N-2-fixing Frankia Thr or supplemented with mineral nitrogen
- 2016
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Ingår i: Journal of plant physiology (Print). - : Elsevier BV. - 0176-1617 .- 1618-1328. ; 196-197, s. 60-69
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Tidskriftsartikel (refereegranskat)abstract
- The actinorhizal tree Casuarina glauca tolerates extreme environmental conditions, such as high salinity. This species is also able to establish a root-nodule symbiosis with N-2-fixing bacteria of the genus Frankia. Recent studies have shown that C. glauca tolerance to high salt concentrations is innate and linked to photosynthetic adjustments. In this study we have examined the impact of increasing NaCl concentrations (200, 400 and 600 mM) on membrane integrity as well as on the control of oxidative stress in branchlets of symbiotic (NOD+) and non-symbiotic (KNO3+) C. glauca. Membrane selectivity was maintained in both plant groups at 200 mM NaCl, accompanied by an increase in the activity of antioxidative enzymes (superoxide dismutase, ascorbate peroxidase, glutathione reductase and catalase). Regarding cellular membrane lipid composition, linolenic acid (C18:3) showed a significant decline at 200 mM NaCl in both NOD+ and KNO3+ plants. In addition, total fatty acids (TFA) and C18:2 also decreased in NOD+ plants at this salt concentration, resulting in malondialdehyde (MDA) production. Such initial impact at 200 mM NaCl is probably due to the fact that NOD+ plants are subjected to a double stress, i.e., salinity and low nitrogen availability. At 400 mM NaCl a strong reduction of TFA and C18:3 levels was observed in both plant groups. This was accompanied by a decrease in the unsaturation degree of membrane lipids in NOD+. However, in both NOD+ and KNO3+ lipid modifications were not reflected by membrane leakage at 200 or 400 mM, suggesting acclimation mechanisms at the membrane level. The fact that membrane selectivity was impaired only at 600 mM NaCl in both groups of plants points to a high tolerance of C. glauca to salt stress independently of the symbiotic relation with Frankia.
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| 9. |
- Dardouri, Maïssa, et al.
(författare)
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Assuring the Biofunctionalization of Silicone Covalently Bonded to Rhamnolipids : Antibiofilm Activity and Biocompatibility
- 2022
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Ingår i: Pharmaceutics. - : MDPI. - 1999-4923. ; 14:9
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Tidskriftsartikel (refereegranskat)abstract
- Silicone-based medical devices composed of polydimethylsiloxane (PDMS) are widely used all over the human body (e.g., urinary stents and catheters, central venous catheters stents) with extreme clinical success. Nevertheless, their abiotic surfaces, being prone to microorganism colonization, are often involved in infection occurrence. Improving PDMS antimicrobial properties by surface functionalization with biosurfactants to prevent related infections has been the goal of different works, but studies that mimic the clinical use of these novel surfaces are missing. This work aims at the biofunctional assessment of PDMS functionalized with rhamnolipids (RLs), using translational tests that more closely mimic the clinical microenvironment. Rhamnolipids were covalently bonded to PDMS, and the obtained surfaces were characterized by contact angle modification assessment, ATR-FTIR analysis and atomic force microscopy imaging. Moreover, a parallel flow chamber was used to assess the Staphylococcus aureus antibiofilm activity of the obtained surfaces under dynamic conditions, and an in vitro characterization with human dermal fibroblast cells in both direct and indirect characterization assays, along with an in vivo subcutaneous implantation assay in the translational rabbit model, was performed. A 1.2 log reduction in S. aureus biofilm was observed after 24 h under flow dynamic conditions. Additionally, functionalized PDMS lessened cell adhesion upon direct contact, while supporting a cytocompatible profile, within an indirect assay. The adequacy of the biological response was further validated upon in vivo subcutaneous tissue implantation. An important step was taken towards biofunctional assessment of RLs-functionalized PDMS, reinforcing their suitability for medical device usage and infection prevention.
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| 10. |
- Duro, Nuno, et al.
(författare)
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The impact of salinity on the symbiosis between Casuarina glauca Sieb. ex Spreng. and N-2-fixing Frankia bacteria based on the analysis of Nitrogen and Carbon metabolism
- 2016
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Ingår i: Plant and Soil. - : Springer Science and Business Media LLC. - 0032-079X .- 1573-5036. ; 398:1-2, s. 327-337
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Tidskriftsartikel (refereegranskat)abstract
- Casuarina glauca is an actinorhizal plant that establishes root-nodule symbiosis with N-2-fixing bacteria of the genus Frankia. This plant is highly recalcitrant to extreme environmental conditions such as salinity and drought. The aim of this study was to evaluate the impact of salt stress on the symbiotic relationship between C. glauca and Frankia Thr, focusing on N and C metabolism. Symbiotic and non-symbiotic plants were exposed to 0, 200, 400 and 600 mM NaCl. The following analyses were performed: stable carbon (delta C-13) and nitrogen (delta N-15) isotope signature; nitrogenase activity in nodules (acetylene reduction assay); and gene expression of a set of genes involved in nodule infection and N/C metabolism (qRT-PCR). Data were analysed using two-way ANOVA. Salt stress induced an enrichment in delta C-13 and delta N-15, reflecting a negative impact of salt in the relative water content and N-2 fixation, respectively. Furthermore, nitrogenase activity in nodules was insignificant already at 200 mM NaCl, consistent with the expression patterns of nifH as well as of plant genes involved in nodule induction and metabolism. The ability of C. glauca to thrive under highly saline environments is not dependent on the symbiosis with Frankia.
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