Molecular regulation of apple and grape ripening: exploring common and distinct transcriptional aspects of representative climacteric and non-climacteric fruits

Fleshy fruits of angiosperms are organs specialized for promoting seed dispersal by attracting herbivores and enticing them to consume the organ and the seeds it contains. Ripening can be broadly defined as the processes serving as a plant strategy to make the fleshy fruit appealing to animals, consisting of a coordinated series of changes in color, texture, aroma, and flavor that result from an intricate interplay of genetically and epigenetically programmed events. The ripening of fruits can be categorized into two types: climacteric, which is characterized by a rapid increase in respiration rate typically accompanied by a burst of ethylene production, and non-climacteric, in which this pronounced peak in respiration is absent. Here we review current knowledge of transcriptomic changes taking place in apple (Malus x domestica, climacteric) and grapevine (Vitis vinifera, non-climacteric) fruit during ripening, with the aim of highlighting specific and common hormonal and molecular events governing the process in the two species. With this perspective, we found that specific NAC transcription factor members participate in ripening initiation in grape and are involved in restoring normal physiological ripening progression in impaired fruit ripening in apple. These elements suggest the existence of a common regulatory mechanism operated by NAC transcription factors and auxin in the two species.This review summarizes the main transcriptional events coordinating the ripening processes in both climacteric (apple) and non-climacteric (grape) models, focusing on transcription factors and hormonal regulation

Exosome-delivered microRNAs promote IFN-α secretion by human plasmacytoid DCs via TLR7

The excessive production of type I IFNs is a hallmark and a main pathogenic mechanism of many autoimmune diseases, including systemic lupus erythematosus (SLE). In these pathologies, the sustained secretion of type I IFNs is dependent on the improper activation of plasmacytoid DCs (pDCs) by self-nucleic acids. However, the nature and origin of pDC-activating self-nucleic acids is still incompletely characterized. Here, we report that exosomes isolated from the plasma of SLE patients can activate the secretion of IFN-α by human blood pDCs in vitro. This activation requires endosomal acidification and is recapitulated by microRNAs isolated from exosomes, suggesting that exosome-delivered microRNAs act as self-ligands of innate single-stranded endosomal RNA sensors. By using synthetic microRNAs, we identified an IFN induction motif that is responsible for the TLR7-dependent activation, maturation, and survival of human pDCs. These findings identify exosome-delivered microRNAs as potentially novel TLR7 endogenous ligands able to induce pDC activation in SLE patients. Therefore, microRNAs may represent novel pathogenic mediators in the onset of autoimmune reactions and potential therapeutic targets in the treatment of type I IFN-mediated diseases

Tangential Flow Filtration for Highly Efficient Concentration of Extracellular Vesicles from Large Volumes of Fluid

Concentration of extracellular vesicles (EVs) from biological fluids in a scalable and reproducible manner represents a major challenge. This study reports the use of tangential flow filtration (TFF) for the highly efficient isolation of EVs from large volumes of samples. When compared to ultracentrifugation (UC), which is the most widely used method to concentrate EVs, TFF is a more efficient, scalable, and gentler method. Comparative assessment of TFF and UC of conditioned cell culture media revealed that the former concentrates EVs of comparable physicochemical characteristics, but with higher yield, less single macromolecules and aggregates (<15 nm in size), and improved batch-to-batch consistency in half the processing time (1 h). The TFF protocol was then successfully implemented on fluids derived from patient lipoaspirate. EVs from adipose tissue are of high clinical relevance, as they are expected to mirror the regenerative properties of the parent cells

Residual matrix from different separation techniques impacts exosome biological activity

Exosomes are gaining a prominent role in research due to their intriguing biology and several therapeutic opportunities. However, their accurate purification from body fluids and detailed physicochemical characterization remain open issues. We isolated exosomes from serum of patients with Multiple Myeloma by four of the most popular purification methods and assessed the presence of residual contaminants in the preparations through an ad hoc combination of biochemical and biophysical techniques - including Western Blot, colloidal nanoplasmonics, atomic force microscopy (AFM) and scanning helium ion microscopy (HIM). The preparations obtained by iodixanol and sucrose gradients were highly pure. To the contrary, those achieved with limited processing (serial centrifugation or one step precipitation kit) resulted contaminated by a residual matrix, embedding the exosomes. The contaminated preparations showed lower ability to induce NfkB nuclear translocation in endothelial cells with respect to the pure ones, probably because the matrix prevents the interaction and fusion of the exosomes with the cell membrane. These findings suggest that exosome preparation purity must be carefully assessed since it may interfere with exosome biological activity. Contaminants can be reliably probed only by an integrated characterization approach aimed at both the molecular and the colloidal length scales

Comparative analysis of antioxidant activity and capacity in apple varieties: Insights from stopped flow DPPH• kinetics, mass spectrometry and electrochemistry

The aim of this research was to investigate the antioxidant kinetics of different apple varieties, a red-flesh variety (‘R201’), a non-browning (‘Majda’), and a ‘Golden Delicious’. Kinetic approaches for antioxidant activity provide more detailed information than conventional assays by examining both the quantity and velocity of active molecules in their reaction with radicals. In this study, DPPH• stopped flow method was applied to study the antioxidant activity and capacity of the three apples varieties, allowing the determination of the reaction rates. The results show that the antioxidant activity of ‘R201’ was not significantly different from ‘Golden Delicious’. Instead, the activity of the non-browning variety was 20 times higher than the others according to the DPPH• kinetic approach, despite having the lowest phenolic content. To further understand their reactivity, the antioxidant molecules were identified using HPLC-HRMS/MS coupled with a coulometric array detector, which validated the DPPH• kinetics. This analysis also found differences in the phenolic profile of the three varieties, attributed to the enhanced antioxidant activity of ’Majda’ to its high content of ascorbic acid. Overall, the research highlights that the antioxidant behavior of apples primarily depends on the velocity of the antioxidants rather than the amount of phenolic compound

A Comparison of the Laser Powder Interaction in Laser Powder Bed Fusion and Direct Energy Deposition Processes

Metal additive manufacturing (AM) is a class of innovative production technologies that allows the production of metal components layer by layer directly from a Computer Aided Design (CAD) model. The AM process of aluminium and aluminium alloys gained much interest in past years especially thanks to the high geometrical freedom, the peculiar microstructures and enhanced mechanical properties it is possible to achieve. However, the quality of final metal components strongly depends on the stability and the quality of each single scan track. Therefore, the understanding of the laser-powder interaction and of the phenomena that arise in the melt pool is a key aspect for the development of these technologies. In this work, AlSi10Mg single scan tracks (SSTs), produced by laser powder bed fusion (LPBF) and direct energy deposition (DED), were analysed and compared in order to select the most suitable range of parameters for each building process

Nanoanalytical analysis of bisphosphonate-driven alterations of microcalcifications using a 3D hydrogel system and in vivo mouse model

Vascular calcification predicts atherosclerotic plaque rupture and cardiovascular events. Retrospective studies of women taking bisphosphonates (BiPs), a proposed therapy for vascular calcification, showed that BiPs paradoxically increased morbidity in patients with prior acute cardiovascular events but decreased mortality in event-free patients. Calcifying extracellular vesicles (EVs), released by cells within atherosclerotic plaques, aggregate and nucleate calcification. We hypothesized that BiPs block EV aggregation and modify existing mineral growth, potentially altering microcalcification morphology and the risk of plaque rupture. Three-dimensional (3D) collagen hydrogels incubated with calcifying EVs were used to mimic fibrous cap calcification in vitro, while an ApoE−/− mouse was used as a model of atherosclerosis in vivo. EV aggregation and formation of stress-inducing microcalcifications was imaged via scanning electron microscopy (SEM) and atomic force microscopy (AFM). In both models, BiP (ibandronate) treatment resulted in time-dependent changes in microcalcification size and mineral morphology, dependent on whether BiP treatment was initiated before or after the expected onset of microcalcification formation. Following BiP treatment at any time, microcalcifications formed in vitro were predicted to have an associated threefold decrease in fibrous cap tensile stress compared to untreated controls, estimated using finite element analysis (FEA). These findings support our hypothesis that BiPs alter EV-driven calcification. The study also confirmed that our 3D hydrogel is a viable platform to study EVmediated mineral nucleation and evaluate potential therapies for cardiovascular calcification

Country-level gender inequality is associated with structural differences in the brains of women and men

Gender inequality across the world has been associated with a higher risk to mental health problems and lower academic achievement in women compared to men. We also know that the brain is shaped by nurturing and adverse socio-environmental experiences. Therefore, unequal exposure to harsher conditions for women compared to men in gender-unequal countries might be reflected in differences in their brain structure, and this could be the neural mechanism partly explaining women´s worse outcomes in gender-unequal countries. We examined this through a random-effects meta-analysis on cortical thickness and surface area differences between adult healthy men and women, including a meta-regression in which country-level gender inequality acted as an explanatory variable for the observed differences. A total of 139 samples from 29 different countries, totaling 7,876 MRI scans, were included. Thickness of the right hemisphere, and particularly the right caudal anterior cingulate, right medial orbitofrontal, and left lateral occipital cortex, presented no differences or even thicker regional cortices in women compared to men in gender-equal countries, reversing to thinner cortices in countries with greater gender inequality. These results point to the potentially hazardous effect of gender inequality on women´s brains and provide initial evidence for neuroscience-informed policies for gender equality.Fil: Zugman, André. National Institutes of Health; Estados UnidosFil: Alliende, Luz María. Pontificia Universidad Católica de Chile; Chile. Universidad Católica de Chile; Chile. Northwestern University; Estados UnidosFil: Medel, Vicente. Universidad Adolfo Ibañez; ChileFil: Bethlehem, Richard A.I.. University of Cambridge; Estados UnidosFil: Seidlitz, Jakob. University of Pennsylvania; Estados UnidosFil: Ringlein, Grace. National Institutes of Health; Estados UnidosFil: Arango, Celso. Universidad Complutense de Madrid; EspañaFil: Arnatkevičiūtė, Aurina. Monash University; AustraliaFil: Asmal, Laila. Stellenbosch University; SudáfricaFil: Bellgrove, Mark. Monash University; AustraliaFil: Benegal, Vivek. National Institute Of Mental Health And Neuro Sciences; IndiaFil: Bernardo, Miquel. Universidad de Barcelona; EspañaFil: Billeke, Pablo. Universidad del Desarrollo; ChileFil: Bosch Bayard, Jorge. McGill University. Montreal Neurological Institute and Hospital; Canadá. Université Mcgill; CanadáFil: Bressan, Rodrigo. Universidade Federal de Sao Paulo; BrasilFil: Busatto, Geraldo F.. Universidade de Sao Paulo; BrasilFil: Castro, Mariana Nair. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires; Argentina. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; ArgentinaFil: Chaim Avancini, Tiffany. Universidade de Sao Paulo; BrasilFil: Compte, Albert. Institut d’Investigacions Biomèdiques August Pi i Sunyer; EspañaFil: Costanzi, Monise. Hospital de Clinicas de Porto Alegre; BrasilFil: Czepielewski, Leticia. Hospital de Clinicas de Porto Alegre; Brasil. Universidade Federal do Rio Grande do Sul; BrasilFil: Dazzan, Paola. Kings College London (kcl);Fil: de la Fuente-Sandoval, Camilo. Instituto Nacional de Neurología y Neurocirugía; MéxicoFil: Gonzalez Campo, Cecilia. Universidad de San Andrés; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Zamorano, Francisco. Universidad del Desarrollo; Chile. Universidad San Sebastián; ChileFil: Zanetti, Marcus V.. Universidade de Sao Paulo; BrasilFil: Winkler, Anderson M.. University of Texas; Estados UnidosFil: Pine, Daniel S.. National Institutes of Health; Estados UnidosFil: Evans Lacko, Sara. School of Economics and Political Science; Reino UnidoFil: Crossley, Nicolas A.. Pontificia Universidad Católica de Chile; Chile. Universidad Católica de Chile; Chile. University of Oxford; Reino Unid

Altered structural brain asymmetry in autism spectrum disorder in a study of 54 datasets

Altered structural brain asymmetry in autism spectrum disorder (ASD) has been reported. However, findings have been inconsistent, likely due to limited sample sizes. Here we investigated 1,774 individuals with ASD and 1,809 controls, from 54 independent data sets of the ENIGMA consortium. ASD was significantly associated with alterations of cortical thickness asymmetry in mostly medial frontal, orbitofrontal, cingulate and inferior temporal areas, and also with asymmetry of orbitofrontal surface area. These differences generally involved reduced asymmetry in individuals with ASD compared to controls. Furthermore, putamen volume asymmetry was significantly increased in ASD. The largest case-control effect size was Cohen's d = -0.13, for asymmetry of superior frontal cortical thickness. Most effects did not depend on age, sex, IQ, severity or medication use. Altered lateralized neurodevelopment may therefore be a feature of ASD, affecting widespread brain regions with diverse functions. Large-scale analysis was necessary to quantify subtle alterations of brain structural asymmetry in ASD