Response to fungal exudates of the rhizosphere isolate Pseudomonas sp. UMAF110 involves a GGDEF/EAL domain-containing protein

Pseudomonas sp. UMAF110, isolated from rhizosphere soil in Spain, display in vitro antagonism towards the pythopathogenic fungus Rosellinia necatrix and is able grow in fungal exudates (BM-RE medium). A transposon mutant library of this strain was constructed and several mutants were selected by their reduced competitiveness in BM-RE medium. Pseudomonas sp. UMAF110-G3, which contains the transposon into a gene encoding a putative REC/PAS/GGDEF/EAL protein, was selected for further characterization. Blastn searches using the sequence of the gene interrupted by the transposon in UMAF110-G3, here called cmpA (c-di-GMP Metabolizing Protein), yielded a single positive hit (98% cover, 78% identity) with a gene from a terpene-degrading Pseudomonas sp. strain isolated from soil. Context analysis of the cmpA gene in Pseudomonas sp. UMAF110 showed that this gene is located downstream from several genes involved in flagellar motility/chemotaxis. RT-PCR experiments further confirmed that cmpA form a transcriptional unit with the che gene cluster. Expression analysis of cmpA by qRT-PCR clearly showed upregulation of this gene after transfer of Pseudomonas sp. UMAF110 cells to BM-RE medium, suggesting a role for this operon in response to fungal exudates. Deletion of cmpA in Pseudomonas sp. UMAF110 did not affect the ability of the strain to form biofilms under the conditions tested. However, overexpression of wild type CmpA in Pseudomonas putida KT2440 negatively regulated biofilm formation in this strain. Together, these results suggest that CmpA could be involved in signal transduction pathways regulating flagellar motility/chemotaxis in response to fungal exudates.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Sequential extraction of almond hull biomass with pulsed electric fields (PEF) and supercritical CO2 for the recovery of lipids, carbohydrates and antioxidants

This work reports the first example of combined sequential extraction by pulsed electric fields (PEF) (3 kV/cm, 100 kJ/kg, 2 Hz, 100 ms) and supercritical (SC) fluid extraction (SFE) (15 MPa, 25 mL/min, 50ºC, 60 min) with CO2 (SC-CO2) for the valorisation of almond hull (AH) biomass. PEF+SFE boosted the efficiency of the protocol up to 77% for total antioxidant capacity and 20% in terms of polyphenols recovery compared to the traditional soaking. Triple-TOF-LC-MS-MS analysis provided the phenolic profiles for the PEF and SC-CO2 extracts, observing significant differences in the polyphenol profile according to the technology applied. Additionally, NMR analysis detected the presence of the carbohydrate soluble (mainly glucose, fructose and sucrose) and lipidic fractions, both selectively extracted by PEF or SC-CO2, respectively. Finally, the post-extraction residual solid biomass was characterized by several techniques such as TGA, FT-IR and SEM. For the latter, the formation of surface pores after PEF and a high fibre compaction after SFE was observed. On the other hand, DTG curves allowed to firmly propose concurrent valorisation routes for this solid, in agreement with a zero-waste approach

The Holocene Cedrus pollen record from Sierra Nevada (S Spain), a proxy for climate change in N Africa

Comprehending the effects of climate variability and disturbance on forested ecosystems is paramount to successfully managing forest environments under future climate scenarios (e.g., global warming, aridi-fication increase). Changes in fossil pollen abundance in sedimentary archives record past vegetation dynamics at regional scales, mainly related to climate changes and, in the last few millennia, to human impact. Pollen records can thus provide long databases with information on how the environment reacted to climate change before the historical record. In this study, we synthesized fossil pollen data from seven sites from the Sierra Nevada in southern Spain to investigate the response of forests in the western Mediterranean area to millennial-scale climate changes and to human impact during the Holocene. In particular, here we focused on Cedrus pollen abundances, which most-likely originated from Northern Africa and were carried to Sierra Nevada by wind. Cedrus pollen has received little attention in the Iberian Peninsula palynological records, for it occurs in low concentrations and has an African source, and thus this article explores the potential to reconstruct its past history and climate. Although Cedrus abundances are generally lower than 1% in the studied pollen samples, a comparison with North African (Moroccan) Cedrus pollen records shows similar trends at long- and short-term time-scales. Therefore, this record could be used as a proxy for changes in this forest species in North Africa. As observed in the Sierra Nevada synthetic record, the increasing trend of Cedrus pollen during the Middle and Late Holocene closely correlates with decreasing summer insolation. This would have produced overall cooler annual temperatures in Northern Africa (Middle Atlas and Rif Mountains) as well as lower summer evaporation, benefiting the growth of this cool-adapted montane tree species while increasing available moisture during the summer, which is critical for this water-demanding species. Millennial-scale variability also characterizes the Sierra Nevada Cedrus synthetic pollen record. Cedrus abundance oscillations co-vary with well-known millennial-scale climatic variability that controlled cedar abundance and altitudinal distribution in montane areas of N Africa. (C) 2020 Elsevier Ltd. All rights reserved.Peer reviewe

Valorization of wastewater from table olives: NMR identification of antioxidant phenolic fraction and microwave single-phase reaction of sugary fraction

The table olive industry is producing a huge amount of wastewater, which is a post-processing cost and an environmental concern. The present study aims to valorize this processing by-product to obtain a value-added product, thereby enhancing resource efficiency and contributing to achieving sustainable development goals (SDGs). In this sense, a chemical reaction-based platform was developed to obtain valuable components, such as levulinic acid (LA) and 5-hydromethylfurfural (HMF). The products were then analyzed using NMR identification of the antioxidant phenolic fraction and microwave single-phase reaction of the sugary fraction. According to the results, the highest concentration of phenolic compounds does not correspond to the sample directly obtained from NaOH treatment (S1), indicating that water washing steps (S2-S5) are fundamental to recover phenolic substances. Moreover, glucose was presented in the sugary fraction that can be transformed into levulinic acid by a single-phase reaction under microwave irradiation. The information provided in this manuscript suggests that the wastewater from the olive processing industry can be valorized to obtain valuable products

Table olive wastewater as a potential source of biophenols for valorization : a mini review

The table olive industry generates high amounts of wastewater annually during the alkaline treatment, fermentation, and washing steps of olives. High conductivity and salt content, as well as the high organic and biophenol contents of these waters, is a worldwide problem, especially in the Mediterranean region, which is the major table olive producing area. There is a wide variety of bioactives found in wastewater derived from table olive processing. The main compounds of table olive wastewater, such as those derived from phenolic, hydrocarbon, and sugar fractions, can be recovered and reused. In this review, the table olive manufacturing processes and the volumes and composition of wastewater generated from the different methods of table olive processing are discussed. In addition, biophenols of table olive water and their biological activities are also introduced. The high concentrations of valuable biophenols, such as tyrosol and hydroxytyrosol, show promising potential for valorizing table olive wastewater; however, more research is needed in this area

A Hyaluronic Acid Demilune Scaffold and Polypyrrole-Coated Fibers Carrying Embedded Human Neural Precursor Cells and Curcumin for Surface Capping of Spinal Cord Injuries

[EN] Tissue engineering, including cell transplantation and the application of biomaterials and bioactive molecules, represents a promising approach for regeneration following spinal cord injury (SCI). We designed a combinatorial tissue-engineered approach for the minimally invasive treatment of SCI¿a hyaluronic acid (HA)-based scaffold containing polypyrrole-coated fibers (PPY) combined with the RAD16-I self-assembling peptide hydrogel (Corning® PuraMatrix¿peptide hydrogel (PM)), human induced neural progenitor cells (iNPCs), and a nanoconjugated form of curcumin (CURC). In vitro cultures demonstrated that PM preserves iNPC viability and the addition of CURC reduces apoptosis and enhances the outgrowth of Nestin-positive neurites from iNPCs, compared to nonembedded iNPCs. The treatment of spinal cord organotypic cultures also demonstrated that CURC enhances cell migration and prompts a neuron-like morphology of embedded iNPCs implanted over the tissue slices. Following sub-acute SCI by traumatic contusion in rats, the implantation of PMembedded iNPCs and CURC with PPY fibers supported a significant increase in neuro-preservation (as measured by greater III-tubulin staining of neuronal fibers) and decrease in the injured area (as measured by the lack of GFAP staining). This combination therapy also restricted platelet-derived growth factor expression, indicating a reduction in fibrotic pericyte invasion. Overall, these findings support PM-embedded iNPCs with CURC placed within an HA demilune scaffold containing PPY fibers as a minimally invasive combination-based alternative to cell transplantation alone.This research was funded by the Science by Women program, Women for Africa Foundation to H.E. and the grants FEDER/Ministerio de Ciencia e Innovacion-Agencia Estatal de Investigacion [RTI2018-095872-B-C21 and -C22/ERDF]; Part of the equipment employed in this work was funded by Generalitat Valenciana and cofinanced with ERDF funds (OP ERDF of Comunitat Valenciana 2014-2020). RISEUP project FetOpen in H2020 Program: H2020-FETOPEN-2018-2019-2020-01.Elkhenany, H.; Bonilla, P.; Giraldo-Reboloso, E.; Alastrue Agudo, A.; Edel, MJ.; Vicent, MJ.; Gisbert-Roca, F.... (2021). A Hyaluronic Acid Demilune Scaffold and Polypyrrole-Coated Fibers Carrying Embedded Human Neural Precursor Cells and Curcumin for Surface Capping of Spinal Cord Injuries. Biomedicines. 9(12):1-19. https://doi.org/10.3390/biomedicines9121928S11991

Long-term coexistence between the macroalga Caulerpa prolifera and the seagrass Cymodocea nodosa in a Mediterranean lagoon

En prensa1,51

Paleoenvironmental evolution of Laguna Seca lake (Sierra Nevada, southern Iberia) since the Late Glacial

Comunicación oral en XXI INQUA Congress. Rome (Italy). 14-20th july 2023Laguna Seca lake at 2259 masl has provided the longest alpine sedimentary record in southern Iberia, registering the last ~18 kyr in a ~14-meter-long sediment core. The oldest part of the sedimentary record represents a phase of subaerial debris flows and a small glacier/nivation hollow. The sediment characteristics abruptly changed at ~15.7 cal kyr BP, when a lake environment was established. A multi-proxy approach (magnetic susceptibility, organic geochemical analyses in bulk sediment, XRF core scanner data, and algae identification) has allowed the characterization of three different environmental phases in this lake. Deep lake conditions are identified from ~15.7 to ~10.6 cal kyr BP, agreeing with overall increasing precipitation in southern Iberia coinciding with augmenting summer insolation. This part of the record is characterized by grey lutites with high total organic carbon (TOC) content, high algae productivity, high vascular plant inputs (high C/N ratio) from the catchment and low Fe/S ratio, suggesting low oxygen conditions in the water-sediment interphase. Between ~10.6 and 8.2-8.0 cal kyr BP higher TOC and low Fe/S ratio are also recorded as well as higher algae content and low C/N ratio suggesting high aquatic production and more algae contribution to the local organic matter pool. This period registered the highest lake levels agreeing with summer insolation maximum and highest precipitation in southern Iberia. An abrupt lowering of the lake level is recorded after 8.0 cal kyr BP in the area. This is deduced by the decrease in TOC and algae in the sediments and more siliciclastic contribution from the catchment basin, evidenced by a high increase in siliciclastic elements (Si, Al, K, Ti, among others), with increased oxic conditions in the water-sediment interphase pointed out by the high increase in Fe/S ratio. Additionally, a potential increase in north African aeolian inputs (rich in Fe cations and Fe compounds) can be interpreted for the Middle and Late Holocene, evidenced by high Fe/Al values. This environmental change agrees with an aridification trend previously observed in the southern Iberian Peninsula

Breast cancer PAM50 signature: correlation and concordance between RNA-Seq and digital multiplexed gene expression technologies in a triple negative breast cancer series

[Background]: Full RNA-Seq is a fundamental research tool for whole transcriptome analysis. However, it is too costly and time consuming to be used in routine clinical practice. We evaluated the transcript quantification agreement between RNA-Seq and a digital multiplexed gene expression platform, and the subtype call after running the PAM50 assay in a series of breast cancer patients classified as triple negative by IHC/FISH. The goal of this study is to analyze the concordance between both expression platforms overall, and for calling PAM50 triple negative breast cancer intrinsic subtypes in particular.[Results]: The analyses were performed in paraffin-embedded tissues from 96 patients recruited in a multicenter, prospective, non-randomized neoadjuvant triple negative breast cancer trial (NCT01560663). Pre-treatment core biopsies were obtained following clinical practice guidelines and conserved as FFPE for further RNA extraction. PAM50 was performed on both digital multiplexed gene expression and RNA-Seq platforms. Subtype assignment was based on the nearest centroid classification following this procedure for both platforms and it was concordant on 96% of the cases (N = 96). In four cases, digital multiplexed gene expression analysis and RNA-Seq were discordant. The Spearman correlation to each of the centroids and the risk of recurrence were above 0.89 in both platforms while the agreement on Proliferation Score reached up to 0.97. In addition, 82% of the individual PAM50 genes showed a correlation coefficient > 0.80.[Conclusions]: In our analysis, the subtype calling in most of the samples was concordant in both platforms and the potential discordances had reduced clinical implications in terms of prognosis. If speed and cost are the main driving forces then the preferred technique is the digital multiplexed platform, while if whole genome patterns and subtype are the driving forces, then RNA-Seq is the preferred method.M.M was supported by two research grants from Ministry of Economy and Competitiveness ISCIII-FIS grants (PI 12/02684): “Predictores genómicos de respuesta a la quimioterapia neoadyuvante con docetaxel-carboplatino en pacientes con cáncer de mama triple negativo”/“Genomic predictors of response to neoadjuvant chemotherapy with docetaxel-carboplatin in patients with triple negative breast cancer”; and (PI 15/00117): “Cáncer de mama triple negative: Predicción de respuesta a docetaxel-carboplatino neoadyuvante mediante caracterización de TILs y firmas inmunes basadas en secuenciación masiva de RNA”/” Triple negative breast cancer: Prediction of response to neoadjuvant docetaxel-carboplatin by characterization of TILs and immune signatures based on massive RNA sequencing”. C.M.P was supported by funds from the NCI Breast SPORE program (P50-CA58223).Peer reviewe