Strawberry phenotypic plasticity in flowering time is driven by interaction between genetic loci and temperature.

The flowering time (FT), which determines when fruits or seeds can be harvested, is subject to phenotypic plasticity, i.e. the ability of a genotype to display different phenotypes in response to environmental variations. Here, we investigated how the environment affects the genetic architecture of FT in cultivated strawberry (Fragaria ×ananassa) and modifies its QTL effects. To this end, we used a bi-parental segregating population grown for two years at widely divergent latitudes (5 European countries) and combined climatic variables with genomic data (Affymetrix® SNP array). Examination, using different phenological models, of the response of FT to photoperiod, temperature and global radiation, indicated that temperature is the main driver of FT in strawberry. We next characterized in the segregating population the phenotypic plasticity of FT by using three statistical approaches that generated plasticity parameters including reaction norm parameters. We detected 25 FT QTL summarized into 10 unique QTL. Mean values and plasticity parameters QTL were co-localized in three of them, including the major 6D_M QTL whose effect is strongly modulated by temperature. The design and validation of a genetic marker for the 6D_M QTL offers great potential for breeding programs, for example for selecting early-flowering strawberry varieties well adapted to different environmental conditions.

From by-products to new application opportunities: the enhancement of the leaves deriving from the fruit plants for new potential healthy products.

In the last decades, the world population and demand for any kind of product have grown exponentially. The rhythm of production to satisfy the request of the population has become unsustainable and the concept of the linear economy, introduced after the Industrial Revolution, has been replaced by a new economic approach, the circular economy. In this new economic model, the concept of "the end of life" is substituted by the concept of restoration, providing a new life to many industrial wastes. Leaves are a by-product of several agricultural cultivations. In recent years, the scientific interest regarding leaf biochemical composition grew, recording that plant leaves may be considered an alternative source of bioactive substances. Plant leaves' main bioactive compounds are similar to those in fruits, i.e., phenolic acids and esters, flavonols, anthocyanins, and procyanidins. Bioactive compounds can positively influence human health; in fact, it is no coincidence that the leaves were used by our ancestors as a natural remedy for various pathological conditions. Therefore, leaves can be exploited to manufacture many products in food (e.g., being incorporated in food formulations as natural antioxidants, or used to create edible coatings or films for food packaging), cosmetic and pharmaceutical industries (e.g., promising ingredients in anti-aging cosmetics such as oils, serums, dermatological creams, bath gels, and other products). This review focuses on the leaves' main bioactive compounds and their beneficial health effects, indicating their applications until today to enhance them as a harvesting by-product and highlight their possible reuse for new potential healthy products.

Special Issue "Dietary Bioactive Components in Inflammatory Bowel Disease".

Inflammatory bowel diseases (IBD) comprise chronic debilitating inflammatory disorders that can affect different parts of the gastrointestinal tract and are commonly correlated to two main diseases: Crohn's disease (CD) and ulcerative colitis (UC) [...].

Application of Near Infrared Spectroscopy for the Rapid Assessment of Nutritional Quality of Different Strawberry Cultivars.

Strawberry is the most cultivated berry fruit globally and it is really appreciated by consumers because of its characteristics, mainly bioactive compounds with antioxidant properties. During the breeding process, it is important to assess the quality characteristics of the fruits for a better selection of the material, but the conventional approaches involve long and destructive lab techniques. Near infrared spectroscopy (NIR) could be considered a valid alternative for speeding up the breeding process and is not destructive. In this study, a total of 216 strawberry fruits belonging to four different cultivars have been collected and analyzed with conventional lab analysis and NIR spectroscopy. In detail, soluble solid content, acidity, vitamin C, anthocyanin, and phenolic acid have been determined. Partial least squares discriminant analysis (PLS-DA) models have been developed to classify strawberry fruits belonging to the four genotypes according to their quality and nutritional properties. NIR spectroscopy could be considered a valid non-destructive phenotyping method for monitoring the nutritional parameters of the fruit and ensuring the fruit quality, speeding up the breeding program.

QTL analysis for ascorbic acid content in strawberry fruit reveals a complex genetic architecture and association with GDP-L-galactose phosphorylase.

Strawberry (Fragaria × ananassa) fruits are an excellent source of L-ascorbic acid (AsA), a powerful antioxidant for plants and humans. Identifying the genetic components underlying AsA accumulation is crucial for enhancing strawberry nutritional quality. Here, we unravel the genetic architecture of AsA accumulation using an F1 population derived from parental lines 'Candonga' and 'Senga Sengana', adapted to distinct Southern and Northern European areas. To account for environmental effects, the F1 and parental lines were grown and phenotyped in five locations across Europe (France, Germany, Italy, Poland and Spain). Fruit AsA content displayed normal distribution typical of quantitative traits and ranged five-fold, with significant differences among genotypes and environments. AsA content in each country and the average in all of them was used in combination with 6,974 markers for quantitative trait locus (QTL) analysis. Environmentally stable QTLs for AsA content were detected in linkage group (LG) 3A, LG 5A, LG 5B, LG 6B and LG 7C. Candidate genes were identified within stable QTL intervals and expression analysis in lines with contrasting AsA content suggested that GDP-L-Galactose Phosphorylase FaGGP(3A), and the chloroplast-located AsA transporter gene FaPHT4;4(7C) might be the underlying genetic factors for QTLs on LG 3A and 7C, respectively. We show that recessive alleles of FaGGP(3A) inherited from both parental lines increase fruit AsA content. Furthermore, expression of FaGGP(3A) was two-fold higher in lines with high AsA. Markers here identified represent a useful resource for efficient selection of new strawberry cultivars with increased AsA content.

An Expectation-Maximization Algorithm for Including Oncological COVID-19 Deaths in Survival Analysis.

We address the problem of how COVID-19 deaths observed in an oncology clinical trial can be consistently taken into account in typical survival estimates. We refer to oncological patients since there is empirical evidence of strong correlation between COVID-19 and cancer deaths, which implies that COVID-19 deaths cannot be treated simply as non-informative censoring, a property usually required by the classical survival estimators. We consider the problem in the framework of the widely used Kaplan-Meier (KM) estimator. Through a counterfactual approach, an algorithmic method is developed allowing to include COVID-19 deaths in the observed data by mean-imputation. The procedure can be seen in the class of the Expectation-Maximization (EM) algorithms and will be referred to as Covid-Death Mean-Imputation (CoDMI) algorithm. We discuss the CoDMI underlying assumptions and the convergence issue. The algorithm provides a completed lifetime data set, where each Covid-death time is replaced by a point estimate of the corresponding virtual lifetime. This complete data set is naturally equipped with the corresponding KM survival function estimate and all available statistical tools can be applied to these data. However, mean-imputation requires an increased variance of the estimates. We then propose a natural extension of the classical Greenwood's formula, thus obtaining expanded confidence intervals for the survival function estimate. To illustrate how the algorithm works, CoDMI is applied to real medical data extended by the addition of artificial Covid-death observations. The results are compared with the estimates provided by the two naïve approaches which count COVID-19 deaths as censoring or as deaths by the disease under study. In order to evaluate the predictive performances of CoDMI an extensive simulation study is carried out. The results indicate that in the simulated scenarios CoDMI is roughly unbiased and outperforms the estimates obtained by the naïve approaches. A user-friendly version of CoDMI programmed in R is freely available.

Survival Analysis Using the Covid-death Mean-imputation (CoDMI) Algorithm: A First Clinical Application in Radiation Oncology.

BACKGROUND/AIM: To report long-term survival results after trimodal approach for locally advanced rectal cancer (LARC) in the Covid-19 era. We herein illustrate a clinical application of Covid-death mean-imputation (CoDMI) algorithm in LARC patients with Covid-19 infection. PATIENTS AND METHODS: We analyzed 94 patients treated for primary LARC. Overall survival was calculated in months from diagnosis to first event (last follow-up/death). Because Covid-19 death events potentially bias survival estimation, to eliminate skewed data due to Covid-19 death events, the observed lifetime of Covid-19 cases was replaced by its corresponding expected lifetime in absence of the Covid-19 event using the CoDMI algorithm. Patients who died of Covid-19 (DoC) are mean-imputed by the Kaplan-Meier estimator. Under this approach, the observed lifetime of each DoC patient is considered as an "incomplete data" and is extended by an additional expected lifetime computed using the classical Kaplan-Meier model. RESULTS: Sixteen patients were dead of disease (DoD), 1 patient was DoC and 77 cases were censored (Cen). The DoC patient died of Covid-19 52 months after diagnosis. The CoDMI algorithm computed the expected future lifetime provided by the Kaplan-Meier estimator applied to the no-DoC observations as well as to the DoC data itself. Given the DoC event at 52 months, the CoDMI algorithm estimated that this patient would have died after 79.5 months of follow-up. CONCLUSION: The CoDMI algorithm leads to "unbiased" probability of overall survival in LARC patients with Covid-19 infection, compared to that provided by a naïve application of Kaplan-Meier approach. This allows for a proper interpretation/use of Covid-19 events in survival analysis. A user-friendly version of CoDMI is freely available at https://github.com/alef-innovation/codmi.

Branch Numbers and Crop Load Combination Effects on Production and Fruit Quality of Flat Peach Cultivars (Prunus persica (L.) Batsch) Trained as Catalonian Vase.

Thinning and pruning are expensive cultural practices in peach cultivation, but essential to obtain adequate production. This study evaluated the effects of combining two pruning (four and six scaffold branches) and three thinning (low, medium, and high crop load) levels on yield and fruit quality of four different flat peach cultivars, trained as Catalonian vase in 2017-2018 in Italy. Productive (average fruit weight, plant total production, and fruit circumference), qualitative (fruit firmness and overcolor, Soluble Solids Content, and Titratable Acidity), and nutritional (Total Antioxidant Capacity, and Total Phenol Content) parameters were evaluated. For productive parameters, a high crop load level led to a decrease in fruit weight and circumference, while a high crop load resulted in higher plant yield. Regarding the qualitative parameters, fruit SSC significantly increased with the diminution of the crop load level in both years of study, while TA was not influenced by crop load and number of branches. Both the total antioxidant capacity and the polyphenol content decreased with an increase in branches number. The findings derived from this study will help growers to select the most suitable combination among genotypes and plant management, to obtain the desired productive or qualitative goals.

Organic vs conventional plant-based foods: A review.

Organic farming is characterized by the prohibition of the use of chemical synthetic fertilizers, pesticides, feed additives and genetically modified organisms and by the application of sustainable agricultural technologies based on ecological principles and natural rules. Organic products are believed to be more nutritious and safer foods compared to the conventional alternatives by consumers, with the consequent increase of demand and price of these foodstuffs. However, in academic circles there is much debate on these issues, since there is not a clear scientific evidence of the difference on the environmental impact and on the nutritional quality, safety and health effects between conventional and organic foods. Therefore, this work aims to describe and update the most relevant data on organic foods, by describing the impact of this practice on environment, producers, consumers and society, as well as by comparing the physicochemical, nutritional and phytochemical quality of conventional and organic plant foods.

Yield and nutritional quality of highbush blueberry genotypes trialled in a Mediterranean hot summer climate.

BACKGROUND: Cultivation of highbush blueberry (Vaccinium corymbosum L.) has been increasing in Europe in recent years, in particular due to the availability of new genotypes suitable for cultivation in many different environmental conditions. The aim of this study was to evaluate the resilience and nutritional quality of 11 highbush blueberry cultivars and two new selections (from the New Zealand Institute for Plant and Food Research Ltd breeding program) in Mediterranean hot summer climate conditions by measuring plant yield, seasonality, fruit sensorial traits, and phytochemical content in the fruits. RESULTS: The new blueberry genotype PFR005 showed a high level of adaptability to these environmental conditions, with the highest total plant yield, whereas PFR075 was the best genotype for nutritional characteristics. Among the cultivars, 'Cosmopolitan' showed the maximum average fruit weight, 'Blueray' and 'Hortblue Poppins' demonstrated a good sensorial profile, while the best cultivars from the nutritional point of view were 'Hortblue Poppins', 'Hortblue Petite', and 'Early Blue'. CONCLUSION: New cultivars and new genotypes able to satisfy the needs of different users, have been identified. The integration of germplasm evaluation with a tailored breeding program will help to create new cultivars that will be useful for the expansion of blueberry cultivation in hot Mediterranean summer climate conditions, which up to now have limited this crop. © 2020 Society of Chemical Industry.

Application of the Non-Destructive NIR Technique for the Evaluation of Strawberry Fruits Quality Parameters.

The determination of strawberry fruit quality through the traditional destructive lab techniques has some limitations related to the amplitude of the samples, the timing and the applicability along all phases of the supply chain. The aim of this study was to determine the main qualitative characteristics through traditional lab destructive techniques and Near Infrared Spectroscopy (NIR) in fruits of five strawberry genotypes. Principal Component Analysis (PCA) was applied to search for spectral differences among all the collected samples. A Partial Least Squares regression (PLS) technique was computed in order to predict the quality parameters of interest. The PLS model for the soluble solids content prediction was the best performing-in fact, it is a robust and reliable model and the validation values suggested possibilities for its use in quality applications. A suitable PLS model is also obtained for the firmness prediction-the validation values tend to worsen slightly but can still be accepted in screening applications. NIR spectroscopy represents an important alternative to destructive techniques, using the infrared region of the electromagnetic spectrum to investigate in a non-destructive way the chemical-physical properties of the samples, finding remarkable applications in the agro-food market.

Isolation of strawberry anthocyanin-rich fractions and their mechanisms of action against murine breast cancer cell lines.

The aim of this study was the evaluation of the effects of strawberry anthocyanin extract treatment on two in vitro models of murine breast cancer cell lines, in an attempt to detect a specific pathway (AMP-activated protein kinase or AMPK) through which strawberries exert their anticancer activity. The anticancer activity of purified anthocyanin extracts from an Alba cultivar on two murine cancer cell lines, N202/1A (with high levels of the HER2/neu oncogene) and N202/1E (with low levels of the HER2/neu oncogene), was evaluated after 48 and 72 h of treatment. The cell viability and apoptosis, intracellular ROS rates, and cell oxidative damage were assessed. Western blot assays were performed to analyze the expression of several proteins related to apoptosis, autophagy, metastasis, the oxidative status, mitochondrial functionality, and the AMPK pathway. This study demonstrated that the anthocyanin extract of Alba strawberry shows an antiproliferative effect on cancer cells, through the induction of apoptosis and oxidative stress, by stimulating different molecular pathways. This study is one of the first studies that have tried to deepen the understanding of a candidate pathway for the explanation of the effects of strawberry on cancer cells. A relationship between the AMPK pathway and the anticancer effects of strawberries was demonstrated.

Romina: A powerful strawberry with in vitro efficacy against uterine leiomyoma cells.

Uterine leiom yomas are benign tumors highly prevalent in reproductive women. In thecurrent study, initially, we aimed to screen five different strawberry cultivars (Alba, Clery, Portola, Tecla, and Romina) to identify efficient cultivars in terms of phytochemical characterization and biological properties by measuring phenolic and anthocyanin content as well as antioxidant capacity, and by measuring apoptotic rate and reactive oxygen species (ROS) production in uterine leiomyoma cells. Next, we focused on the most efficient ones, cultivar Alba (A) and Romina (R) as well as Romina anthocyanin (RA) fraction for their ability to regulate oxidative phosphorylation (oxygen consumption rate [OCR]) glycolysis (extracellular acidification rate [ECAR]), and also fibrosis. Leiomyoma and myometrial cells were treated with a methanolic extract of A and R (250 µg/ml) or with RA (50 µg/ml) for 48 hr to measure OCR and ECAR, as well as gene expression associated with fibrosis. In the leiomyoma cells, RA was more effective in inducing apoptosis and increasing intracellular ROS levels, followed by R and A. In myometrial cells, all strawberry treatments increased the cellular viability and decreased ROS concentrations. Leiomyoma cells showed also a significant decrease in ECAR, especially after RA treatment, while OCR was slightly increased in both myometrial and leiomyoma cells. R and RA treatment significantly decreased collagen 1A1, fibronectin, versican, and activin A messenger RNA expression in leiomyoma cells. In conclusion, this study suggests that Romina, or its anthocyanin fraction, can be developed as a therapeutic and/or preventive agent for uterine leiomyomas, confirming the healthy effects exerted by these fruits and their bioactive compounds.

Phytochemical Composition and Cytotoxic Effects on Liver Hepatocellular Carcinoma Cells of Different Berries Following a Simulated In Vitro Gastrointestinal Digestion.

Berry fruits are rich in nutrients and polyphenols, providing potential health benefits. Understanding the factors that affect their bioavailability is becoming of utmost importance for evaluating their biological significance and efficacy as functional food. In this study, the phytochemical composition and the total antioxidant capacity of different varieties of five berries (blackberry, blackcurrant, blueberry, raspberry, and strawberry) were evaluated after an in vitro gastrointestinal digestion process. The cultivar of each berry that showed the higher content of total phenols and flavonoids was selected to study its cytotoxic effect on human hepatoma cells. Digestion resulted in a high reduction (p ˂ 0.05) of total phenolic, flavonoid and anthocyanin contents and total antioxidant capacity, in the "IN" samples compared to the "OUT" extracts, which represent the "serum-available" and the "colon-available" fractions, respectively. Incubation of the digested fraction for 24 h didn't exert any effect on cellular viability, while a dose- and time-dependent cytotoxicity was observed after 48 h and 72 h of incubation for all the berries analyzed. Our results suggest that the approach proposed in this work may represent a rapid tool for evaluating and identifying new berries with increased phytochemical bioavailability, highlighting their antiproliferative agents after an in vitro digestion.

Comparison of the Antimicrobial Activities of Four Honeys From Three Countries (New Zealand, Cuba, and Kenya).

Skin and chronic wound infections are an increasing and urgent health problem worldwide. Their management is difficult and the development of antibiotic resistance by both planktonic and biofilm-associated bacteria necessitates the use of alternative treatments. The purpose of this study was to compare the antimicrobial activity of four honeys from different floral and geographical origins: Melipona beecheii honey (Cuba) and three Apis mellifera honeys [Manuka honey (New Zealand), A. mellifera honey (Cuba), and African honey (Kenya)]. The physicochemical parameters were within the ranges reported for these honeys and M. beecheii honey stood out due to its acidic character. An agar incorporation technique was used to determine the minimum active dilution of each honey against 52 clinical isolates (34 Gram-positive, 17 Gram-negative, and 1 Candida albicans). The antibiofilm activity of honeys was tested by assessing their ability to inhibit biofilm formation and to disrupt preformed biofilms. Overall, M. beecheii honey had the highest antimicrobial and antibiofilm activity, although a marked disruption in preformed biofilms was shared by all tested honeys. Structural changes induced by M. beecheii honey on Staphylococcus aureus and Pseudomonas aeruginosa cells were observed by transmission electron microscopy suggesting that this honey has a potent antimicrobial action and may be an excellent candidate for the development of topical preparations for the treatment of infected wounds.

Manuka honey attenuates oxidative damage induced by H2O2 in human whole blood in vitro.

Manuka honey has been widely researched regarding its biological properties, in particular its antimicrobial and antioxidant capacities. We tested the genotoxic and genoprotective properties of Manuka honey, ranging from 25-1000 µg/mL, by performing an in vitro comet assay after exposure to human whole blood. No genotoxic effect on whole blood cells was observed within the tested concentration range (p = 0.154). Then, the antigenotoxic potency of Manuka honey against oxidative DNA damage to whole blood cells was assessed. Prior to Manuka honey treatment a modest decrease of H2O2-induced DNA damage was detected in cells, with no statistical significance (p = 0.087). Post-treatment, Manuka honey displayed a stronger potential to attenuate damaged cells at all tested concentrations, with a statistical significant difference (p < 0.001), where concentrations of 25 and 100 µg/mL were most efficient. Manuka honey exhibited a marked potential to protect DNA of whole blood cells from oxidative damage induced by hydrogen peroxide in vitro.

Late sodium current inhibitors to treat exercise-induced obstruction in hypertrophic cardiomyopathy: an in vitro study in human myocardium.

BACKGROUND AND PURPOSE: In 30-40% of hypertrophic cardiomyopathy (HCM) patients, symptomatic left ventricular (LV) outflow gradients develop only during exercise due to catecholamine-induced LV hypercontractility (inducible obstruction). Negative inotropic pharmacological options are limited to ß-blockers or disopyramide, with low efficacy and tolerability. We assessed the potential of late sodium current (INaL )-inhibitors to treat inducible obstruction in HCM. EXPERIMENTAL APPROACH: The electrophysiological and mechanical responses to ß-adrenoceptor stimulation were studied in human myocardium from HCM and control patients. Effects of INaL -inhibitors (ranolazine and GS-967) in HCM samples were investigated under conditions simulating rest and exercise. KEY RESULTS: In cardiomyocytes and trabeculae from 18 surgical septal samples of patients with obstruction, the selective INaL -inhibitor GS-967 (0.5 µM) hastened twitch kinetics, decreased diastolic [Ca2+ ] and shortened action potentials, matching the effects of ranolazine (10µM). Mechanical responses to isoprenaline (inotropic and lusitropic) were comparable in HCM and control myocardium. However, isoprenaline prolonged action potentials in HCM myocardium, while it shortened them in controls. Unlike disopyramide, neither GS-967 nor ranolazine reduced force at rest. However, in the presence of isoprenaline, they reduced Ca2+ -transient amplitude and twitch tension, while the acceleration of relaxation was maintained. INaL -inhibitors were more effective than disopyramide in reducing contractility during exercise. Finally, INaL -inhibitors abolished arrhythmias induced by isoprenaline. CONCLUSIONS AND IMPLICATIONS: Ranolazine and GS-967 reduced septal myocardium tension during simulated exercise in vitro and therefore have the potential to ameliorate symptoms caused by inducible obstruction in HCM patients, with some advantages over disopyramide and ß-blockers.

Guava (Psidium guajava L. cv. Red Suprema) Crude Extract Protect Human Dermal Fibroblasts against Cytotoxic Damage Mediated by Oxidative Stress.

We analyzed guava fruits (Psidium guajava L. cv. Red Suprema) from Cuba to determine their chemical composition, total antioxidant capacity, as well as their protective effect against oxidative damage using an in vitro model of human dermal fibroblasts. The guava fruit is a natural source of bioactive compounds, such as polyphenols, vitamin C, folates and beta carotenes with proven health benefits. Human dermal fibroblasts were pre-incubated with different concentrations of guava crude extract and then subjected to oxidative stress using the AAPH stressor. The number of apoptotic and dead cells, as well as the markers of oxidative damage such as lipid and protein oxidation significantly decreased when cells were pre-incubated with guava crude extract and then exposed to the stressor. The activity of antioxidant enzymes also improved when cells were pre-incubated with guava crude extract in comparison to cells subjected to stress without prior pre-incubation with the guava extract. The results obtained in this study highlight the health benefits of guava regarding oxidative stress, proving it to be an important source of bioactive compounds associated with important biological properties.