Spanish Melon Landraces: Revealing Useful Diversity by Genomic, Morphological, and Metabolomic Analysis.

Spain is a secondary centre of the diversification of the melon (Cucumis melo L.), with high diversity represented in highly appreciated landraces belonging to the Flexuosus and Ibericus groups. A collection of 47 accessions of Flexuosus, Chate, Piel de Sapo, Tendral, Amarillo, Blanco, and Rochet was analysed using a genotyping-by-sequencing (GBS) approach. A total of 66,971 quality SNPs were identified. Genetic analysis differentiated Ibericus accessions and exotic materials (Ameri, Momordica, Kachri, and Agrestis), while Flexuous accessions shared ancestry between them. Within the Ibericus group, no clear genomic distinction could be identified for the different landraces evaluated, with accessions of different landraces showing high genetic similarity. The morphological characterization confirmed that the external colour and fruit shape had been used as recognition patterns for Spanish melon landraces, but variability within a landrace exists. Differences were found in the sugars and acid and volatile profiles of the materials. Flexuosus and Chate melons at the immature commercial stage accumulated malic acid and low levels of hexoses, while Ibericus melons accumulated high contents of sucrose and citric acid. Specific trends could be identified in the Ibericus landraces. Tendral accumulated low levels of sugars and citric acid and high of malic acid, maintaining higher firmness, Rochet reached higher levels of sugars, and Amarillo tended to lower malic acid contents. Interestingly, high variability was found within landraces for the acidic profile, offering possibilities to alter taste tinges. The main volatile organic compounds (VOCs) in Flexuosus and Chate were aldehydes and alcohols, with clear differences between both groups. In the Ibericus landraces, general trends for VOC accumulation could be identified, but, again, a high level of variation exists. This situation highlights the necessity to develop depuration programs to promote on-farm in situ conservation and, at the same time, offers opportunities to establish new breeding program targets and to take advantage of these sources of variation.

A Patient Self-Made Point-of-Care Fecal Test Improves Diagnostic Accuracy Compared with Fecal Calprotectin Alone in Inflammatory Bowel Disease Patients.

BACKGROUND: Monitoring inflammatory bowel disease patients may be challenging. Fecal calprotectin is one of the most performed tests. Other fecal biomarkers are less used in clinical practice. Rapid fecal tests that could be performed by patients may be a useful strategy to closely monitor disease activity. METHODS: We performed a prospective observational study including consecutive inflammatory bowel disease patients referred for colonoscopy in a single center. Certest FOB + Transferrin + Calprotectin + Lactoferrin® (Certest Biotec S.L, Zaragoza, Spain), a one-step point-of-care test which simultaneously detects these four biomarkers was performed. Endoscopic inflammatory activity was defined using the Mayo score (≥1) in ulcerative colitis, SES-CD (>3) and Rutgeerts scores (≥1) for Crohn's disease. RESULTS: Out of a total of 106 patients (56.5% female, mean age 51 years), 54 (50.9%) were diagnosed with ulcerative colitis and 52 (49.1%) with Crohn's disease. Endoscopic activity was detected in 42 patients (39.0%). Fecal calprotectin provided the best sensitivity (97.6%), with limited specificity (34.4%). Compared to calprotectin, the other 3 fecal biomarkers showed better specificity (87.5-92.1%) and lower sensitivity (45.2-59.5%). Patients with a negative result in all biomarkers (19/106-17.9%) had 100% (CI 95% 97.4-100) negative predictive value, while patients with the 4 biomarkers positive (13/106-12.3%) had 100% (CI 95% 96.1-100) positive predictive value of endoscopic inflammatory activity. AUROC of this 4 biomarker point-of-care test was 0.845 (95% CI 0.771-0.920), significantly higher than the AUROCs of any of the 4 biomarkers. CONCLUSIONS: This test may be a useful strategy to monitor inflammatory activity in clinical practice by excluding or prioritizing patients in need of a colonoscopy.

Optimization of electronic nose drift correction applied to tomato volatile profiling.

E-noses can be routinely used to evaluate the volatile profile of tomato samples once the sensor drift and standardization issues are adequately solved. Short-term drift can be corrected using a strategy based on a multiplicative drift correction procedure coupled with a PLS adaptation of the component correction. It must be performed specifically for each sequence, using all sequence signals data. With this procedure, a drastic reduction of sensor signal %RSD can be obtained, ranging between 91.5 and 99.7% for long sequences and between 75.7 and 98.8% for short sequences. On the other hand, long-term drift can be fixed up using a synthetic reference standard mix (with a representation of main aroma volatiles of the species) to be included in each sequence that would enable sequence standardization. With this integral strategy, a high number of samples can be analyzed in different sequences, with a 94.4% success in the aggrupation of the same materials in PLS-DA two-dimensional graphical representations. Using this graphical interface, e-noses can be used to developed expandable maps of volatile profile similitudes, which will be useful to select the materials that most resemble breeding objectives or to analyze which preharvest and postharvest procedures have a lower impact on the volatile profile, avoiding the costs and sample limitations of gas chromatography.

Grafting Snake Melon [Cucumis melo L. subsp. melo Var. flexuosus (L.) Naudin] in Organic Farming: Effects on Agronomic Performance; Resistance to Pathogens; Sugar, Acid, and VOC Profiles; and Consumer Acceptance.

The performance of snake melon [Cucumis melo var. flexuosus (L.)] in organic farming was studied under high biotic and salt stress conditions. Soilborne diseases (mainly caused by Macrophomina phaseolina and Neocosmospora falciformis), combined with virus incidence [Watermelon mosaic virus (WMV), Zucchini yellow mosaic virus (ZYMV), and Tomato leaf curl New Delhi virus (ToLCNDV)] and Podosphaera xanthii attacks, reduced yield by more than 50%. Snake melon susceptibility to M. phaseolina and Monosporascus cannonballus was proved in pathogenicity tests, while it showed some degree of resistance to Neocosmospora keratoplastica and N. falciformis. On the contrary, salt stress had a minor impact, although a synergic effect was detected: yield losses caused by biotic stress increased dramatically when combined with salt stress. Under biotic stress, grafting onto the melon F1Pat81 and wild Cucumis rootstocks consistently reduced plant mortality in different agroecological conditions, with a better performance compared to classic Cucurbita commercial hybrids. Yield was even improved under saline conditions in grafted plants. A negative effect was detected, though, on consumer acceptability, especially with the use of Cucurbita rootstocks. Cucumis F1Pat81 rootstock minimized this side effect, which was probably related to changes in the profile of sugars, acids, and volatiles. Grafting affected sugars and organic acid contents, with this effect being more accentuated with the use of Cucurbita rootstocks than with Cucumis. In fact, the latter had a higher impact on the volatile organic compound profile than on sugar and acid profile, which may have resulted in a lower effect on consumer perception. The use of Cucumis rootstocks seems to be a strategy to enable organic farming production of snake melon targeted to high-quality markets in order to promote the cultivation of this neglected crop.

The targeted overexpression of SlCDF4 in the fruit enhances tomato size and yield involving gibberellin signalling.

Tomato is one of the most widely cultivated vegetable crops and a model for studying fruit biology. Although several genes involved in the traits of fruit quality, development and size have been identified, little is known about the regulatory genes controlling its growth. In this study, we characterized the role of the tomato SlCDF4 gene in fruit development, a cycling DOF-type transcription factor highly expressed in fruits. The targeted overexpression of SlCDF4 gene in the fruit induced an increased yield based on a higher amount of both water and dry matter accumulated in the fruits. Accordingly, transcript levels of genes involved in water transport and cell division and expansion during the fruit enlargement phase also increased. Furthermore, the larger amount of biomass partitioned to the fruit relied on the greater sink strength of the fruits induced by the increased activity of sucrose-metabolising enzymes. Additionally, our results suggest a positive role of SlCDF4 in the gibberellin-signalling pathway through the modulation of GA4 biosynthesis. Finally, the overexpression of SlCDF4 also promoted changes in the profile of carbon and nitrogen compounds related to fruit quality. Overall, our results unveil SlCDF4 as a new key factor controlling tomato size and composition.

High throughput FT-MIR indirect analysis of sugars and acids in watermelon.

Indirect measurements of taste-related compounds are required when a high number of samples has to be analyzed in a short period of time, with a minimum cost. For this purpose, FT-MIR partial least square (PLS) regression models for the prediction of total soluble solids, sugars and organic acids have been developed using three sample sets including breeding lines and commercial varieties of watermelon. Specific models with excellent performance were obtained only for sugars. Nevertheless, a general model supposed a compromise between the best and worse models and offered %RMSEP values of 11.3%, 11.1% and 11.7% for fructose, glucose and sucrose respectively. The model was applied to the selection of high content samples (selection pressure 20% and 30%) obtaining good sensitivity levels and mean percentile of selected samples close to the expected values (100% sensitivity). The robustness of FT-MIR models was assessed with predictions of external assays, obtaining reasonable performances.

Influence of controlled deficit irrigation on tomato functional value.

The effect of controlled deficit irrigation (CDI) on the accumulation of carotenoids, polyphenols and l-ascorbic acid was studied in conventional and high lycopene tomato cultivars. Plants were initially irrigated to cover 100%ETc and after the fruit set phase, the dose was reduced to 75% or 50% of ETc. CDI had no significant effect on the accumulation of carotenoids, while it increased the levels of the hydroxycinnamic acids chlorogenic and ferulic acids, the flavonoid rutin and l-ascorbic acid. Nevertheless, there were important interactions and this effect was highly dependent on the year and site of cultivation. Certain growing areas would be more favorable to supply high quality markets, and, fortunately, CDI would maximize polyphenol (100-75%ETc) and l-ascorbic acid (100-50%ETc) in these areas. A combination of the best genotype and growing area with CDI would offer high quality products, preserving a scarce resource: water.

Polyphenol and l-ascorbic acid content in tomato as influenced by high lycopene genotypes and organic farming at different environments.

The accumulation of polyphenols and l-ascorbic acid was evaluated under conventional (integrated pest management, IPM) and organic farming, as means to increase the accumulation of chemoprotective compounds. The effect of genotype was considerably higher than the growing system, in fact it is determining. 'Kalvert', a high-lycopene cultivar, outstood for the accumulation of most polyphenols, though low-carotenoid cultivars with high accumulation were also detected. Organic farming significantly increased the levels of caffeic acid by 20%, but reduced those of ferulic acid and naringenin by 13% and 15% respectively. A strong interaction with the environment was detected: in Navarra the differences were limited, while in Extremadura lower contents of ferulic acid and higher contents of chlorogenic acid and rutin were found in organic farming for certain cultivars. The effect of organic farming on l-ascorbic acid was dependent on cultivar and environment and it only led to an increase in Extremadura by 58%.

Simultaneous determination of main phenolic acids and flavonoids in tomato by micellar electrokinetic capillary electrophoresis.

A methodology for the determination of tomato phenolic acids and flavonoids has been developed combining MEKC and DAD detection. The influence on polyphenol separation of pH and background electrolyte, BGE (borax, acetonitrile, methanol and SDS concentrations), was studied and optimized using response surface methodology and weighted desirability function. Separation of polyphenols was achieved within 20min at 15°C using 11.3mM borax and 11.2mM SDS adjusted to pH 8.5 as BGE. Validation was performed using standards and tomato extracts. Recoveries ranged from 77 to 106%. Acceptable repeatabilities were obtained for peak area (%RSD <3.1% and <3.7%) and migration times (%RSD <0.2% and <1.4%) for intra- and inter-day respectively. Detection limits ranged between 0.8 and 3.8mgkg-1. Five and seven of these polyphenols were determined in samples of tomato and related species. This methodology will be valuable tool in breeding programs, analyzing a large number of samples.

Tomato as a Source of Carotenoids and Polyphenols Targeted to Cancer Prevention.

A diet rich in vegetables has been associated with a reduced risk of many diseases related to aging and modern lifestyle. Over the past several decades, many researches have pointed out the direct relation between the intake of bioactive compounds present in tomato and a reduced risk of suffering different types of cancer. These bioactive constituents comprise phytochemicals such as carotenoids and polyphenols. The direct intake of these chemoprotective molecules seems to show higher efficiencies when they are ingested in its natural biological matrix than when they are ingested isolated or in dietary supplements. Consequently, there is a growing trend for improvement of the contents of these bioactive compounds in foods. The control of growing environment and processing conditions can ensure the maximum potential accumulation or moderate the loss of bioactive compounds, but the best results are obtained developing new varieties via plant breeding. The modification of single steps of metabolic pathways or their regulation via conventional breeding or genetic engineering has offered excellent results in crops such as tomato. In this review, we analyse the potential of tomato as source of the bioactive constituents with cancer-preventive properties and the result of modern breeding programs as a strategy to increase the levels of these compounds in the diet.

Fast simultaneous determination of prominent polyphenols in vegetables and fruits by reversed phase liquid chromatography using a fused-core column.

A reversed-phase high-performance liquid chromatography method with photodiode array detection has been developed enabling the joint determination of 17 prominent flavonoids and phenolic acids in vegetables and fruits. A multi-segmented gradient program using a fused-core column for the separation of several phenolic classes (phenolic acids and flavonoids) has been optimised. The influence of extraction conditions (sample freeze-drying, ultrasound extraction, solvent composition and extraction time) has been also optimised using response surface methodology with tomato samples as a model. Complete recoveries (76-108%) were obtained for the phenolic compounds present in tomato. The developed method provided satisfactory repeatability in terms of peak area (RSD<2.9%) and retention time (RSD<0.2%) both for standards and real samples. Detection limits ranged between 3 and 44µgkg(-1) for the detected polyphenols. This method is recommended for routine analysis of large number of samples typical of production quality systems or plant breeding programs.

Clinical features of patients inappropriately undiagnosed of pulmonary embolism.

PURPOSES: The objective of this study was to identify clinical factors associated with delayed diagnosis of acute pulmonary embolism (PE) in the emergency department (ED). BASIC PROCEDURES: A retrospective observational study was performed at three University affiliated Hospitals; 436 consecutive patients who presented to the ED with an acute PE confirmed by chest computed tomography from 2008 to 2011 were included. Patients were divided into 3 groups: group 1, PE was diagnosed while the patient was still in the ED; group 2, PE was diagnosed during hospitalization; group 3, patients who were sent home with a wrong alternative diagnosis and returned to the ED and were diagnosed of PE. MAIN FINDINGS: One hundred forty-six patients (33.5%) had a delayed diagnosis of PE--21.5% belong to group 2 and 11.9% to Group 3. Chronic coexisting medical conditions like asthma or chronic obstructive pulmonary disease were independent predictors of a delayed diagnosis in patients who were admitted to hospital whereas non-specific and less severe symptoms like the presence of pleuro-mechanic thoracic pain, fever, hemoptysis, or the presence of a pulmonary infiltrate in chest x-ray were independent predictors of a delayed diagnosis in patients who were sent home. PRINCIPAL CONCLUSIONS: Delay in diagnosis of acute PE is frequent despite current diagnostic strategies. Patients are sent home or admitted to hospital with a wrong diagnosis depending on clinical presentation or coexisting medical conditions.