Short-Term Gaseous Treatments Improve Rachis Browning in Red and White Table Grapes Stored at Low Temperature: Molecular Mechanisms Underlying Its Beneficial Effect.

Short-term gaseous treatments improve rachis quality during table grape postharvest, but little is known about the mechanisms involved. In this work, we observed that the application of a 3-day CO2 treatment at 0 °C improved rachis browning of Superior Seedless and Red Globe bunches, affecting the non-enzymatic antioxidant system by reducing the total phenolic content, the antioxidant activity and the expression of different stilbene synthase genes. Lipid peroxidation levels revealed lower oxidative stress in CO2-treated rachis of both cultivars linked to the activation of the enzymatic antioxidant system. Furthermore, whereas a positive correlation was denoted between rachis browning and the accumulation of key ABA regulatory genes in Red Globe bunches, this effect was restricted to ACS1, a key synthetic ethylene gene, in Superior Seedless clusters. This work also corroborated the important role of ethylene-responsive factors in the beneficial effect of the gaseous treatment, not only in the berries but also in the rachis. Finally, the application of the gaseous treatment avoided the induction of cell wall-degrading enzyme-related genes in both cultivars, which could favor the maintenance of rachis quality. This work provides new insight into specific responses modulated by the gaseous treatment focused on mitigating rachis browning independently of the cultivar.

The Effect of Ethanol Treatment on the Quality of a New Table Grape Cultivar It 681-30 Stored at Low Temperature and after a 7-Day Shelf-Life Period at 20 °C: A Molecular Approach.

Despite the fact that many studies have examined the effectiveness of different gaseous postharvest treatments applied at low temperature to maintain table grape quality, the use of ethanol vapor has hardly been investigated. Thus, this work has studied the effectiveness of ethanol vapor-generating sachets in the maintenance of It 681-30 table grape quality, a new cultivar, during storage at low temperature and after the shelf-life period at 20 °C. To this end, various quality assessments have been carried out and the effect of the ethanol treatment on the expression of different genes (phenylpropanoids, transcription factors, PRs, and aquaporins) was determined. The results indicated that the application of ethanol vapor reduced the total decay incidence, weight loss, and the rachis browning index in It 681-30 grapes stored at 0 °C and after the shelf-life period at 20 °C, as compared to non-treated samples. Moreover, the modulation of STS7 and the different PR genes analyzed seems to play a part in the molecular mechanisms activated to cope with fungal attacks during the postharvest of It 681-30 grapes, and particularly during the shelf-life period at 20 °C. Furthermore, the expression of aquaporin transcripts was activated in samples showing higher weight loss. Although further work is needed to elucidate the role of ethanol in table grape quality, the results obtained in this work provide new insight into the transcriptional regulation triggered by ethanol treatment.

Partial contributions and temporal trends of leading causes of death during the last four decades in Spain.

OBJECTIVES: The study was conducted to assess time trend shifts of leading causes of death and their partial contributions over the years 1975-2016 in Spain. STUDY DESIGN: A longitudinal ecological epidemiologic design was conducted to analyse linear trend period shifts using joinpoint regression as the annual percentage of change (APC) in the period 1975-2016. The partial contributions were illustrated as the rate ratio of a singular-cause to their major-cause shift periods. RESULTS: HIV/AIDS shaped the increasing trend period of infectious diseases in 1989-1995 (APC = 25.3, P < 0.05) and the decreasing trend in 1995-1999 and 1999-2016. Lung cancer fell gradually from 1994 in men (-0.4, P < 0.05); however, in women, the condition continued increasing from 1990 (P < 0.05). Dementia types influenced mental and neurological disease drifts. The recent trend for circulatory periods (1980-2016) was mainly modulated by cardiac ischaemia, with increased partial contributions (25%, 32% and 30%). Traffic accidents defined the descending tendency of external causes. CONCLUSIONS: Spain showed a Western pattern in descended rates, including non-decreasing trends in mental and neurological diseases, pancreatic cancer, drug abuse and suicide. Trend shifts and partial contributions illustrated targets for further mortality reduction.

Multiwalled carbon nanotubes enter broccoli cells enhancing growth and water uptake of plants exposed to salinity.

BACKGROUND: Carbon nanotubes have been shown to improve the germination and growth of some plant species, extending the applicability of the emerging nano-biotechnology field to crop science. RESULTS: In this work, exploitation of commercial multiwalled carbon nanotubes (MWCNTs) in control and 100 mM NaCl-treated broccoli was performed. Transmission electron microscopy demonstrated that MWCNTs can enter the cells in adult plants with higher accumulation under salt stress. Positive effect of MWCNTs on growth in NaCl-treated plants was consequence of increased water uptake, promoted by more-favourable energetic forces driving this process, and enhanced net assimilation of CO2. MWCNTs induced changes in the lipid composition, rigidity and permeability of the root plasma membranes relative to salt-stressed plants. Also, enhanced aquaporin transduction occurred, which improved water uptake and transport, alleviating the negative effects of salt stress. CONCLUSION: Our work provides new evidences about the effect of MWCNTs on plasma membrane properties of the plant cell. The positive response to MWCNTs in broccoli plants opens novel perspectives for their technological uses in new agricultural practices, especially when 1plants are exposed to saline environments.

Non-traumatic spontaneous abdominal haemorrhage.

Spontaneous abdominal hemorrhage is defined as intra-abdominal hemorrhage from a non-traumatic cause. It is a challenging clinical situation and in most cases the diagnosis is made on the basis of imaging findings. CT is the technique of choice for the detection, localization and extension of bleeding. Objective is to review the main imaging findings expected in spontaneous abdominal hemorrhage as well as its main etiologies.

Are the Blueberries We Buy Good Quality? Comparative Study of Berries Purchased from Different Outlets.

Blueberries (Vaccinium corymbosum L.) are becoming increasingly popular for their nutritional and health benefits, and their economic value is therefore increasing. The loss of quality that can occur due to softening and fungal attack is an important consideration when marketing blueberries. Despite the added value of blueberries, no studies have been carried out on how the fruit arrives at the outlets just before purchase by the consumer in terms of firmness, physico-chemical parameters, phenolic compounds, and fungal growth. The aim of this work has been, therefore, to investigate possible differences in quality parameters between blueberries purchased from ten different outlets, regardless of the supplier. The results showed that all the samples were of acceptable quality, although they all had a low maturity index at the point of sale. None of the samples studied showed clear signs of fungal decay at the time of purchase, although we were able to grow and identify some pathogen specimens after cultivation. In terms of total phenolic and anthocyanin content, as well as antioxidant activity, all the samples showed low values, possibly due to their postharvest storage, but they were within the expected range for this fruit. On the other hand, differences in the measured parameters were observed between samples of the same cultivar while no differences were found between conventionally and organically grown blueberries. This suggests that preharvest (such as edaphoclimatic conditions, agricultural practices, and cultivars) and postharvest factors (such as treatments used, storage, and transport temperatures) could influence the berry quality when they reach the consumer.

The Adolescent Problem Gambling Prevalence Associated with Leisure-Time Activities and Risky Behaviors in Southern Spain.

Gambling addiction is increasing and is becoming a public health concern due to the rise of gambling-related harms affecting the youth. Previous studies suggest a strong link between problem gambling (PG) and substance use and psychosocial and familial factors. Our main objective was to analyze the association between PG and factors like sport, leisure-time activities, and risk-taking behaviors in adolescents. A survey on substance use and addictive behaviors was performed in 2019 on a representative sample of 2240 subjects (14-18 years) from the Southern Spain Region. Data variables like socio-demographic characteristics, sport, leisure and free time activities, family environment, PG (Lie-Bet Scale), compulsive internet use (CIUS Scale), and consumption of alcohol and cannabis were collected using a standardized questionnaire. Weighted PG prevalence was estimated in either sex, as well as the differences between various levels of PG using chi-square tests. Crude and adjusted weighted logistic regression models were used to identify predictors associated with PG. The prevalence of PG was associated with shopping frequency, compulsive internet use, cannabis use in the previous month, higher family economic status, and having a homemaker father which increased the likelihood of PG. On the contrary, cultural hobbies such as playing an instrument, painting, singing, and writing, and having a working mother were inversely associated with PG. Our results suggest that encouraging participation in creative activities along with supervised shopping and monitoring compulsive internet use and cannabis consumption may contribute some protection against adolescent PG. Supplementary Information: The online version contains supplementary material available at 10.1007/s11469-022-00950-7.

Postharvest High-CO2 Treatments on the Quality of Soft Fruit Berries: An Integrated Transcriptomic, Proteomic, and Metabolomic Approach.

Soft fruits are appreciated for their taste qualities and for being a source of health-promoting compounds. However, their postharvest is affected by their high respiratory rates and susceptibility to fungal decay. Our aim here is to provide a perspective on the application of short-term high-CO2 treatments at a low temperature to maintain the postharvest quality of soft fruits. This work also suggests using a multi-omics approach to better understand the role of the cell wall and phenolic compounds in maintaining quality. Finally, the contribution of high-throughput transcriptomic technologies to understand the mechanisms modulated by the short-term gaseous treatments is also highlighted.

Analysis of root plasma membrane aquaporins from Brassica oleracea: post-translational modifications, de novo sequencing and detection of isoforms by high resolution mass spectrometry.

Plasma membrane Intrinsic Proteins (PIPs), a subfamily of aquaporins, are ubiquitous membrane channel proteins that play a crucial role in water uptake in plants. The use of high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) analysis of peptides has previously shown to be a valuable tool to differentiate among PIP homologues sharing a high sequence homology and also to characterize their post-translational modifications (PTMs). The recent introduction of mass spectrometers able to measure peptide mass with high mass accuracy, together with new alternative ways of peptide fragmentation allows the identification and characterization of proteins from nonsequenced organisms, such as broccoli. In this study, we combined three endoproteases (trypsin, Glu-C and Lys-C) with HPLC-MS/MS analysis and two types of peptide fragmentations, CID (collision induced dissociation) and HCD (higher-energy C-trap dissociation), to identify PIP isoforms and PTMs from broccoli roots. After de novo sequencing analysis, eight peptides showing homology to Arabidopsis thaliana PIPs were identified. Although Arabidopsis nomenclature of PIP isoforms has not been defined for broccoli, our results agree with the occurrence of seven AtPIP isoforms (PIP 1;1, PIP 1;2, PIP 1;3 and PIP2;2, PIP 2;3, PIP2;1 and PIP2;7) in broccoli roots, as compared to the plant model A. thaliana. To our knowledge, these results represent the deepest characterization of the PIPs isolated from the roots of broccoli, a crop with increasing agronomical interest.

Effect of high CO2 levels and low temperature on stilbene biosynthesis pathway gene expression and stilbenes production in white, red and black table grape cultivars during postharvest storage.

Phenolic compounds, such as phytoalexin resveratrol, can be induced in grapes in response to biotic and abiotic stresses and have been related in many healthy effects. Stilbene synthases (STSs) are the key enzyme responsible for resveratrol biosynthesis. They have been already isolated and characterized from several plant species, however, VviSTS is a multigene family and little is known about their modulation in response to the application of gaseous treatments that maintain table grapes quality during postharvest. In this work, we have analyzed the effect of a 3-day CO2 treatment on the modulation of 4 STSs (VviSTS6, VviSTS7, VviSTS16 and VviSTS46) and on the accumulation of different stilbene compounds (resveratrol, resveratrol-glucoside, trans-piceatannol, z-miyabenol and pallidol) during the postharvest storage at 0 °C of white (Superior Seedless, Dominga), red (Red Globe) and black (Autumn Royal) table grapes. Results indicated that the accumulation of the stilbene compounds by the application of CO2 and low temperature storage were cultivar dependent. In white Dominga fruit, accumulation of stilbene compounds increased in CO2-treated samples what seems to be modulated by VviSTS6, VviSTS7 and VviSTS46. However, in Red Globe the accumulation of compounds was mainly due to the cold storage in air and seems to be also mediated by the induction of the same VviSTSs. By contrast, in Superior Seedless and Autumn Royal table grapes the modulation of VviSTSs genes and the stilbene accumulation was independent of the atmosphere storage. Further studies would be needed to elucidate the possible role of transcription factors involved on VviSTSs modulation.

Discerning the mechanism of the multiwalled carbon nanotubes effect on root cell water and nutrient transport.

Multiwalled carbon nanotubes (MWCNTs) are tubular carbon structures that are able to enter cells through holes in the plasma membrane and produce changes in gene expression. In this work, we compared the functionality of carbon nanotubes with the electroporation that perforates membranes, in Brassica oleracea var. Italica (broccoli) root protoplasts. For this, we combined those treatments with control conditions and abiotic stress (salinity) in order to elucidate if the response is related to conditions optimal for the plant. The measurement of the osmotic water permeability (Pf), mineral concentrations and expression levels of aquaporins (PIP1s and PIP2s) revealed that the physiological action of the nanotubes was similar to that achieved with electroporation for both Pf and the concentrations of nutrients in the protoplasts. On the other hand, PIP1s and PIP2s expression was increased in the protoplasts receiving the control plus MWCNTs treatment but not in those treated with electroporation. This opens new and interesting lines, as it shows that nanotubes are able to modulate the expression of aquaporins.

Regulation of flavonoid biosynthesis pathway by a single or dual short-term CO2 treatment in black table grapes stored at low temperature.

The application of one or two short-term treatments with high levels of CO2 during 3 days at 0 °C maintained the quality of Autumn Royal table grapes (Vitis vinifera) during storage at 0 °C. We have analyzed how the application of a 3-day gaseous treatment, for one or two times at 0 °C, influences on common (VviPAL, VviCHS, VviCHI, VviF3'H, VviF3'5'H, VviF3H and VviLDOX) and branch-specific (VviFLS1, VviLAR1, VviLAR2, VviANR and VviUFGT) flavonoid gene expression in the skin of Autumn Royal table grapes. Likewise, the content of flavonols, flavan-3-ols and anthocyanins were identified with Q-TOF equipment and quantified by HPLC-quadrupole together with the total phenolic content and the antioxidant capacity by the ABTS and FRAP methods. Moreover, we have also used a solid-state voltammetry methodology to compare the effect of the application of one or two gaseous treatments in the skin of table grapes stored at 0 °C. Results revealed that the application of one or two gaseous treatments modulated the expression of flavonoid gene expression and the levels of catechin, in the case of one application, or quercetin-3-glucoside and five anthocyanins in fruit treated twice, maintaining their levels after 28 days of storage at 0 °C similar to those recorded in freshly harvested fruit. Satisfactorily, the electrochemical approach was useful to distinguish between treated and non-treated samples not only in the first stage of storage but also after 16 days.

Water relations of the tos1 tomato mutant at contrasting evaporative demand.

The tos1 (tomato osmotically sensitive) mutant, isolated from an in vitro screen of root growth during osmotic stress, was less sensitive to exogenous ABA, but accumulated more ABA under osmotic stress than WT plants. We assessed growth and water relations characteristics of hydroponically grown tos1 seedlings (in the absence of osmotic stress) at low and high evaporative demands. Growth of tos1 was severely inhibited at both high and low evaporative demands. Twenty DAS, WT and tos1 genotypes had a similar leaf water and turgor potential, but mature tos1 plants (45 day old) showed a significant diurnal loss of leaf turgor, with recovery overnight. Increased evaporative demand increased turgor loss of tos1 plants. High evaporative demand at the beginning of the day decreased stomatal conductance of tos1, without diurnal recovery, thus whole plant transpiration was decreased. De-topped tos1 seedlings showed decreased root hydraulic conductance and had a 1.4-fold increase in root ABA concentration. Impaired root function of tos1 plants failed to meet transpirational water demand and resulted in shoot turgor loss, stomatal closure and growth inhibition.

Anthocyanin, antioxidant activity and stress-induced gene expression in high CO2-treated table grapes stored at low temperature.

A pretreatment with 20kPa CO2+20 kPa O2+60 kPa N2 for 3 days proved effective in maintaining the fruit quality and controlling decay in table grapes (Vitis vinifera cv. Cardinal) stored at 0 degrees C. In the present work, we analyzed whether total anthocyanin content, the molecular mechanism implicated in their biosynthesis and antioxidant activity is related to the beneficial effect of this gaseous treatment. We isolated partial cDNAs that codified for enzymes implicated in the anthocyanin biosynthesis such as l-phenylalanine ammonia-lyase (PAL) and chalcone synthase (CHS), and an antioxidant enzyme such as ascorbate peroxidase (APX). Low temperatures induced an accumulation of total anthocyanin content in the skin of both treated and non-treated grapes, although levels were lower in CO2-treated fruit. By contrast, antioxidant activity decreased during storage at 0 degrees C in non-treated grapes but did not change in CO2-treated grapes. The up-regulation of anthocyanin biosynthesis gene expression and VcAPX mRNA observed in non-treated grape is not enhanced in CO2-treated grapes, which presented low total decay. These results point out the ability of CO2-treated grapes to prevent the generation of reactive oxygen species rather than their inactivation by means of induction of studied defense systems.

Taking stock of Roma health policies in Spain: Lessons for health governance.

Roma health inequities are a wicked problem. Despite concerted efforts to reduce them under the Decade of Roma Inclusion initiative, the health gap between Roma and non-Roma populations in Europe persists. To address this problem, the European Commission devised the National Roma Integration Strategies (NRIS). This paper provides a critical assessment of the implementation of the NRIS' health strand (NRIS-H) in Spain and proposes an evaluation tool to monitor Roma health policies - the Roma Health Integration Policy Index (RHIPEX). It also makes recommendations to promote Roma health governance. To achieve these goals, four community forums, 33 stakeholder interviews and a scoping review were conducted. Results show that the NRIS-H implementation is hindered by lack of political commitment and poor resource allocation. This has a negative impact on Roma's entitlement to healthcare and on their participation in decision-making processes, jeopardising the elimination of the barriers that undermine their access to healthcare and potentially contributing to reproduce inequalities. These unintended effects point out the need to rethink Roma health governance by strengthening intersectional and intersectoral policies, enabling transformative Roma participation in policymaking and guaranteeing shared socio-political responsibility and accountability.

The Expanding Role of Vesicles Containing Aquaporins.

In animals and plants, membrane vesicles containing proteins have been defined as key for biological systems involving different processes such as trafficking or intercellular communication. Docking and fusion of vesicles to the plasma membrane occur in living cells in response to different stimuli, such as environmental changes or hormones, and therefore play an important role in cell homeostasis as vehicles for certain proteins or other substances. Because aquaporins enhance the water permeability of membranes, their role as proteins immersed in vesicles formed of natural membranes is a recent topic of study. They regulate numerous physiological processes and could hence serve new biotechnological purposes. Thus, in this review, we have explored the physiological implications of the trafficking of aquaporins, the mechanisms that control their transit, and the proteins that coregulate the migration. In addition, the importance of exosomes containing aquaporins in the cell-to-cell communication processes in animals and plants have been analyzed, together with their potential uses in biomedicine or biotechnology. The properties of aquaporins make them suitable for use as biomarkers of different aquaporin-related diseases when they are included in exosomes. Finally, the fact that these proteins could be immersed in biomimetic membranes opens future perspectives for new biotechnological applications.

Deciphering the Role of CBF/DREB Transcription Factors and Dehydrins in Maintaining the Quality of Table Grapes cv. Autumn Royal Treated with High CO2 Levels and Stored at 0°C.

C-repeat/dehydration-responsive element binding factors (CBF/DREB) are transcription factors which play a role in improving plant cold stress resistance and recognize the DRE/CRT element in the promoter of a set of cold regulated genes. Dehydrins (DHNs) are proteins that accumulate in plants in response to cold stress, which present, in some cases, CBF/DREB recognition sequences in their promoters and are activated by members of this transcription factor family. The application of a 3-day gaseous treatment with 20 kPa CO2 at 0°C to table grapes cv. Autumn Royal maintained the quality of the bunches during postharvest storage at 0°C, reducing weight loss and rachis browning. In order to determine the role of CBF/DREB genes in the beneficial effect of the gaseous treatment by regulating DHNs, we have analyzed the gene expression pattern of three VviDREBA1s (VviDREBA1-1, VviDREBA1-6, and VviDREBA1-7) as well as three VviDHNs (VviDHN1a, VviDHN2, and VviDHN4), in both alternative splicing forms. Results showed that the differences in VviDREBA1s expression were tissue and atmosphere composition dependent, although the application of high levels of CO2 caused a greater increase of VviDREBA1-1 in the skin, VviDREBA1-6 in the pulp and VviDREBA1-7 in the skin and pulp. Likewise, the application of high levels of CO2 regulated the retention of introns in the transcripts of the dehydrins studied in the different tissues analyzed. The DHNs promoter analysis showed that VviDHN2 presented the cis-acting DRE and CRT elements, whereas VviDHN1a presented only the DRE motif. Our electrophoretic mobility shift assays (EMSA) showed that VviDREBA1-1 was the only transcription factor that had in vitro binding capacity to the CRT element of the VviDHN2 promoter region, indicating that the transcriptional regulation of VviDHN1a and VviDHN4 would be carried out by activating other independent routes of these transcription factors. Our results suggest that the application of high CO2 levels to maintain table grape quality during storage at 0°C, leads to an activation of CBF/DREBs transcription factors. Among these factors, VviDREBA1-1 seems to participate in the transcriptional activation of VviDHN2 via CRT binding, with the unspliced form of this DHN being activated by high CO2 levels in all the tissues analyzed.

Plant aquaporins: new perspectives on water and nutrient uptake in saline environment.

The mechanisms of salt stress and tolerance have been targets for genetic engineering, focusing on ion transport and compartmentation, synthesis of compatible solutes (osmolytes and osmoprotectants) and oxidative protection. In this review, we consider the integrated response to salinity with respect to water uptake, involving aquaporin functionality. Therefore, we have concentrated on how salinity can be alleviated, in part, if a perfect knowledge of water uptake and transport for each particular crop and set of conditions is available.

Expression Profiles and DNA-Binding Affinity of Five ERF Genes in Bunches of Vitis vinifera cv. Cardinal Treated with High Levels of CO2 at Low Temperature.

Ethylene response factors (ERFs) play an important role in plants by regulating defense response through interaction with various stress pathways. After harvest, table grapes (Vitis vinifera L.) are subject to a range of problems associated with postharvest storage at 0°C, such as fungal attack, water loss and rachis browning. The application of a 3-day high CO2 treatment maintained fruit quality and activated the induction of transcription factors belonging to different families such as ERF. In this paper, we have isolated five VviERFs from table grapes cv. Cardinal, whose deduced amino acid sequence contained the conserved apetalous (AP2)/ERF domain. The phylogeny and putative conserved motifs in VviERFs were analyzed and compared with those previously reported in Vitis. VviERFs-c gene expression was studied by quantitative real-time RT-PCR in the different tissues of bunches stored at low temperature and treated with high levels of CO2. The results showed that in most of the tissues analyzed, VviERFs-c gene expression was induced by the storage under normal atmosphere although the application of high levels of CO2 caused a greater increase in the VviERFs-c transcript accumulation. The promoter regions of two PRs (pathogenesis related proteins), Vcchit1b and Vcgns1, were obtained and the in silico analysis revealed the presence of a cis-acting ethylene response element (GCC box). In addition, expression of these two PR genes was analyzed in the pulp and rachis of CO2-treated and non-treated table grapes stored at 0°C and results showed significant correlations with VviERF2-c and VviERF6L7-c gene expression in rachis, and between VviERF11-c and Vcchit1b in pulp. Finally by using electro mobility shift assays, we denoted differences in binding of VviERFs to the GCC sequences present in the promoters of both PRs, with VviERF6L7-c being the only member which did not bind to any tested probe. Overall, our results suggest that the beneficial effect of high CO2 treatment maintaining table grape quality seems to be mediated by the regulation of ERFs and in particular VviERF2-c might play an important role by modulating the expression of PR genes.