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.

Short-Term Exposure to High CO2 and O2 Atmospheres to Inhibit Postharvest Gray Mold of Pomegranate Fruit.

The effect of short-term exposure to high CO2 or O2 atmospheres, alone or in combination with heat (35°C), for the control of postharvest gray mold was evaluated on 'Mollar de Elche' pomegranate fruit artificially inoculated with Botrytis cinerea and stored at 20 or 5°C. Exposure to high CO2 for 48 h at 20°C effectively reduced gray mold on pomegranate fruit incubated at 20°C for 5 days in a concentration-based manner. Furthermore, gaseous treatments with partial pressures of 95 kPa CO2 or 30 kPa O2 + 70 kPa CO2 for 48 h significantly reduced gray mold incidence and severity on fruit regardless of storage temperature. Moreover, for fruit receiving gaseous treatments at 20°C then cold stored for 12 weeks, there were no apparent negative effects of the treatments on their quality (weight loss, skin color, maturity index, pH, and sensory quality). In general, compared with those applied at 20°C, treatments at 35°C did not improve gray mold inhibition and adversely affected some quality parameters. In conclusion, 48-h exposures to specific atmospheres at 20°C may be a suitable treatment to extend pomegranate storage life and could be part of integrated control programs to control postharvest decay.

The Associations of Mental Health Disturbances, Self-Esteem, and Partner Violence Victimization with Condom Use in Spanish Adolescents.

Risky sexual behaviors, such as not using a condom for preventing sexually transmittable diseases and unwanted pregnancies, are associated with emotional and psychological problems in adolescence, including higher levels of depression, anxiety, stress, and low self-esteem. Adolescents with a history of violence victimization are also at increased risk of engaging in sexual risk behaviors. In this study, we examined the associations between mental health disturbances and partner violence victimization and the non-use of condoms among young people in Spain. We also examined self-esteem as a mediator of mental health problems and unprotected sexual relationships. A cross-sectional study was conducted on a sample of 831 adolescents randomly selected from 25 high schools in Spain. From the initial sample, 285 students (144 men and 141 women) from high schools in Alicante (Spain) who reported having experienced sexual activity with at least one partner were approached. The results showed that one in three adolescents between 14 and 19 years old did not use condoms during sexual intercourse. The factors associated with the non-use of condoms in the estimated models of Poisson robust variance were having a history of physical, psychological, or sexual violence; having been expelled from school because of behavioral problems; and having higher levels of depression, anxiety, and stress and lower levels of self-esteem. Self-esteem was independently associated with the non-use of condoms. Having lower levels of self-esteem increased the risk of not using a condom during participants' last sexual intercourse. These findings suggest the importance of developing public health strategies for mental health promotion to increase condom use among adolescents.

Effects of a School-Based Intervention for Preventing Substance Use among Adolescents at Risk of Academic Failure: A Pilot Study of the Reasoning and Rehabilitation V2 Program.

Tobacco, alcohol and cannabis use are important health problems that contribute greatly to causes of death in worldwide. Early onset of substance use increases rapidly during adolescence and it has been associated with a wide range of adverse events. Because substance use is associated with dramatic consequences, delaying the initiation of substance use among adolescents remains a major public priority. This study examined the effectiveness of a school-based intervention program based on the application of the Reasoning and Rehabilitation V2 (R&R2) program for preventing substance use among adolescents at risk of academic failure. A sample of 142 participants (aged 13-17 years old) who were studying alternative education provision in Spain were randomly allocated to two conditions (68 experimental group, 74 control group). A pre-test survey assessing past and current substance use was conducted prior the implementation of the program, while a post-test survey was conducted about 12 months after the pre-test. The results showed a significant effect of the R&R program in the reduction of cigarette smoking, episodes of drunkenness, alcohol consumption and cannabis use. However, for daily smoking and episodes of drunkenness such benefits showed a reduction over time. These findings offer additional evidence of the effectiveness of the Reasoning and Rehabilitation V2 program in Spanish adolescent students who are exposed to substance use and suggest areas of future research.

Effectiveness of the Reasoning and Rehabilitation V2 Programme for Improving Personal and Social Skills in Spanish Adolescent Students.

Significant progress has been made in developing intervention programmes for adolescents at high risk of delinquency, school failure and emotional problems. The most effective programmes incorporate behavioural and skills training aimed at changing attitudes and promoting psychosocial and emotional skills in adolescents. This study examined the effectiveness of a school-based intervention programme based on the Reasoning and Rehabilitation V2 (R&R2). R&R2 is a cognitive behavioural programme developed using psychological theories about the aetiology of delinquency, as well as the cognitive, behavioural and socioemotional deficits in high-risk youth populations. A sample of 142 students (aged 13-17 years old) who were attending alternative education provision in Spain were randomly assigned to two experimental conditions (68 experimental group, 74 control group). The results showed that the R&R2 improved participants' self-esteem, social skills, empathy and rational problem-solving with a medium-large effect size (η2 = 0.08 to 0.26). The effects of the programme were significant after controlling for age and the pre-test scores in baseline. These findings confirm the effectiveness of the Reasoning and Rehabilitation V2 programme in Spanish adolescent students and offer additional evidence regarding the implementation of the R&R2 programme in both alternative educational and mainstream school settings.

Relationships between Problematic Cannabis Use and Risky Behaviors in Spanish Adolescents.

This study examined the relations between problematic cannabis use, physical assault, and getting involved in a motor vehicle accident under the influence of cannabis in a sample of adolescents randomly selected from 25 public and semiprivate high schools in Alicante (Spain). Participants (n = 648) completed The Spanish National Standardized Survey about drug use in high school adolescents (ESTUDES, 2017), which includes the cannabis abuse screening test (CAST). Prevalence of cannabis use across the life-span and within the past 30 days was 37.5% and 17.4%, respectively. CAST scores were associated with an increased risk of driving under the effects of cannabis, riding shotgun, and physical assault, but not with an increased risk of having a motor vehicle accident. There were no differences between boys and girls in the association of problematic cannabis use with risky behaviors. This result highlights the importance of comprehensive prevention and education strategies for adolescents at high risk of cannabis use.

Crop losses due to olive moth mediated by ethylene.

BACKGROUND: Studies for nine years in southern Spain on the olive moth, Prays oleae Bern., have tested the possibility of altering oviposition behaviour on newly formed olive fruits by means of a single ethylene application. RESULTS: A single spraying of ethylene on the olive trees at the beginning of fruiting significantly decreased the olive moth egg populations and consequent the olive fruit fall. At the same time, no negative effect was found in the activity of natural oophagous predators of the olive moth. CONCLUSIONS: The ethylene treatments against P. oleae yielded substantial economical benefits each year (a mean of euro526 ha(-1)), benefits that fluctuated depending on the olive crop and on the annual fruit fall caused by this major pest.

Low Temperature and Short-Term High-CO2 Treatment in Postharvest Storage of Table Grapes at Two Maturity Stages: Effects on Transcriptome Profiling.

Table grapes (Vitis vinifera cv. Cardinal) are highly perishable and their quality deteriorates during postharvest storage at low temperature mainly because of sensitivity to fungal decay and senescence of rachis. The application of a 3-day CO2 treatment (20 kPa CO2 + 20 kPa O2 + 60 kPa N2) at 0°C reduced total decay and retained fruit quality in early and late-harvested table grapes during postharvest storage. In order to study the transcriptional responsiveness of table grapes to low temperature and high CO2 levels in the first stage of storage and how the maturity stage affect these changes, we have performed a comparative large-scale transcriptional analysis using the custom-made GrapeGen GeneChip®. In the first stage of storage, low temperature led to a significantly intense change in grape skin transcriptome irrespective of fruit maturity, although there were different changes within each stage. In the case of CO2 treated samples, in comparison to fruit at time zero, only slight differences were observed. Functional enrichment analysis revealed that major modifications in the transcriptome profile of early- and late-harvested grapes stored at 0°C are linked to biotic and abiotic stress-responsive terms. However, in both cases there is a specific reprogramming of the transcriptome during the first stage of storage at 0°C in order to withstand the cold stress. Thus, genes involved in gluconeogenesis, photosynthesis, mRNA translation and lipid transport were up-regulated in the case of early-harvested grapes, and genes related to protein folding stability and intracellular membrane trafficking in late-harvested grapes. The beneficial effect of high CO2 treatment maintaining table grape quality seems to be an active process requiring the induction of several transcription factors and kinases in early-harvested grapes, and the activation of processes associated to the maintenance of energy in late-harvested grapes.

Differential regulation of dehydrin expression and trehalose levels in Cardinal table grape skin by low temperature and high CO2.

Dehydrins and trehalose are multifunctional protective biomolecules that play a role in counteracting cellular damage during dehydrative stresses. In this paper, we studied dehydrin isoform patterns, dehydrin gene expression and trehalose levels in the skin of Cardinal (Vitis vinifera L.) table grapes, along with their regulation by different cold postharvest storage conditions. Immunoanalysis with K-segment antibody recognizes four constitutive dehydrins (from 17 to 44 kDa) that are tightly regulated by low temperature and high CO2. Phosphatase treatment showed that DHN44 and DHN22 isoforms are phosphorylated polypeptides, while MALDI-TOF MS and MS/MS analysis suggested that 44 kDa polypeptide may be a dehydrin homodimer. At the transcriptional level, dehydrins are also regulated by low temperature and high CO2, showing a fairly good correlation with their mRNA levels. Trehalose was quantified by high performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD), revealing a progressive increase of this metabolite throughout storage at 0 °C and the sudden transitory increases in short-term high CO2-treated fruit. We propose that the constitutive presence and up-regulation of dehydrins and trehalose during low temperature postharvest storage could be positively correlated with the relative chilling tolerance of table grapes and the adaptive responses activated by high CO2 levels to preserve cell water status and to counteract the disruption of physiological processes during cold storage.

Effects of high CO2 levels on fermentation, peroxidation, and cellular water stress in Fragaria vesca stored at low temperature in conditions of unlimited O2.

To better understand the tolerance of strawberries (Fragaria vesca L.) to high CO2 in storage atmospheres, fermentation and cellular damage were investigated. Fruits were stored for 3 and 6 days at 0 °C in the presence of different CO2 levels (0, 20, or 40%) with 20% O2. Changes in pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) gene expression and in fermentative metabolites, as well as in bound water and malondialdehyde (MDA) concentrations, were analyzed. In strawberries stored without added CO2, up-regulation of PDC and ADH was not associated with an increase in fermentative metabolites. By contrast, moderate ethanol fermentation in fruits exposed to 20% CO2 seems to be essential to maintain fruit metabolism, reducing both lipid peroxidation and cellular water stress. However, if the CO2 concentration increases (40%), the excess acetaldehyde and ethanol produced were closely correlated with a decrease in bound water and production of MDA.

The crucial role of Φ- and K-segments in the in vitro functionality of Vitis vinifera dehydrin DHN1a.

Dehydrins (DHNs), group II LEA (Late Embryogenesis Abundant) proteins, are among the most commonly observed proteins which accumulate in plants in response to cold and any other environmental factors, causing the dehydration of cells. In previous studies, we isolated a YSK2-type VvcDHN1a gene from table grapes (Vitis vinifera cv. Cardinal) which presented two spliced variants (the spliced, DHN1a_s and the unspliced, DHN1a_u). Their expression was induced by low temperature storage and CO2, although with different accumulation patterns. DHN1a_u codifies for a truncated YS protein lacking Ф- and K-segments, which might affect its functionality. In this work, we expressed both DHN1a_s and DHN1a_u recombinant proteins in Escherichia coli. We carried out a number of in vitro assays to analyze the implications that Ф- and K-segments have in the protective role of VvcDHN1 against different abiotic stresses and their antifungal activity against the fungal pathogen Botrytis cinerea. Our results showed that unlike DHN1a_u, DHN1a_s has a potent cryoprotective effect on lactate dehydrogenase activity, protects malate dehydrogenase against dehydration and partially inhibits B. cinerea growth. Moreover, the DHN1a promoter presented cis-regulatory elements related to cold and drought, as well as biotic stress-related elements. We also observed that both spliced variants interact weakly with DNA, suggesting that K-segments are not involved in DNA binding. Overall, this work highlights the crucial role of Ф- and K-segments in DHNs function in plant response to abiotic stress showing for the first time, the potential role of the V. vinifera DHN1a_s in the protection against freezing and dehydration as well as inhibiting B. cinerea growth.

Unraveling the roles of CBF1, CBF4 and dehydrin 1 genes in the response of table grapes to high CO2 levels and low temperature.

CBFs (C-repeat binding factors) are transcription factors that are rapidly induced by low temperature and that recognize the CRT/DRE element in the promoter of a set of cold regulated genes, the CBF regulon. Dehydrins are proteins that accumulate in plants under stress conditions, such as low temperature, and some form part of the CBF regulon. To investigate their role in the response of table grape clusters to 0°C long storage as well as to 3-day high CO2 postharvest treatment, we isolated two partial CBF genes (VvcCBF1 and VvcCBF4) and a full-length dehydrin (VvcDHN1a) from Vitis vinifera cv. Cardinal. Hydrophobic cluster analysis (HCA) identified differences in the secondary and tertiary structure between Vitis CBF4s and CBF1s. Overall, our results showed that, in table grapes, the expression of CBF genes is induced mainly in response to CO2 treatment, suggesting that the response of DHN1a in this fruit could be attributed to a cold-inducible CBF-independent pathway.