Cell-penetrating peptides are widely used as molecular transporters for the internalization inside cells of various cargo, including proteins and nucleic acids. A special role is played by arginine-rich peptides and oligoarginines covalently linked or simply mixed with the cargo. Here we report cell-penetrating agents in which arginine units are clustered on a macrocyclic scaffold. Instead of using long peptides, four single arginine units were covalently attached to either the upper or lower rim of a calix[4]arene, kept in the cone conformation building a ‘parallel’ cyclic array. These new macrocyclic carriers show high efficiency in DNA delivery and transfection in a variety of cell lines.
Here, we report on a mechanistic investigation based on DFT calculations and kinetic measures aimed at determining the energetics related to the cysteine nucleophilic attack on nitrile-carrying compounds. Activation energies were found to correlate well with experimental kinetic measures of reactivity with cysteine in phosphate buffer. The agreement between computations and experiments points to this DFT-based approach as a tool for predicting both nitrile reactivity toward cysteines and the toxicity of nitriles as electrophile agents.
Biostimulants have been found effective in enhancing plant resistance toward stressful conditions. The aim of the present study was to evaluate the efficacy of selected biostimulants to overcome the negative effects of nutrient limitation on the growth performances and on the fruit quality of soilless cultivated strawberry plants. The condition of nutrient limitation was imposed by supplying the plants with only a single fertilization at transplantation and by excluding any further nutrient supply for the entire duration of the experiment (three months, from May to July). Strawberry plants were treated seven times during the period from preflowering up to berry maturation with different classes of biostimulants (humic acids, alfalfa hydrolysate, macroseaweed extract and microalga hydrolysate, amino acids alone or in combination with zinc, B-group vitamins, chitosan, and a commercial product containing silicon) at commercial dosages. The use of alfalfa hydrolysate, vitamins, chitosan, and silicon was able to promote biomass accumulation in roots (four to seven folds) and fruits (+20%) of treated plants, whereas the total leaf area increased by 15%–30%. Nutrient concentrations in leaves and roots showed variations for microelements (e.g., Fe, B, Zn, and Si) in response to biostimulant applications, whereas no significant differences were observed for macronutrient contents among treatments. Final berry yield was found around 20% higher in chitosan- and silicon-treated plants. Chitosan treatment significantly increased pulp firmness (by 20%), while a high nutritional value (e.g., phenolic compounds concentration) was observed in alfalfa- and seaweed-treated fruits (+18%–20% as compared to control). The overall outcomes of the present experiment show that selected biostimulants can be considered as a valid agronomic tool able to contrast the negative consequence of growing crops under insufficient nutritional conditions.
The experiment was conducted during two consecutive seasons (years 2016 and 2017) in an organic apple orchard of the cultivar Jonathan. Several biostimulants were tested (10 in total), including humic acids, macro and micro seaweed extracts, alfalfa protein hydrolysate, amino acids alone or in combination with zinc, B-group vitamins, chitosan and a commercial product containing silicon. Treatments were performed at weekly intervals, starting from the end of May until mid-August. The macroseaweed extract was effective in stimulate tree growth potential in both years, as shown by a significantly larger leaf area (+20% as compared to control) and by an higher chlorophyll content and leaf photosynthetic rate in year 2016. As for the yield performances and apples quality traits at harvest (average fruit weight, soluble solids content, titratable acidity, and flesh firmness), they were generally affected by the different climatic conditions that characterized the two growing seasons (year 2017 being characterized by higher maximal and average temperatures and by limited rainfalls at the beginning of the season). Treatments with macroseaweed extract, B-group vitamins and alfalfa protein hydrolysate were able to significantly improve the intensity and extension of the red coloration of apples at harvest. Correspondingly, the anthocyanin content in the skin of apples treated with the same biostimulants resulted significantly higher than control, highlighting the potential influence of these substances on the synthesis of secondary metabolites in apple. The incidence of physiological disorders was also monitored during apple storage period. Amino acids plus zinc application was effective in reducing (more than 50%) the incidence of the “Jonathan spot,” the main post-harvest disorder for this cultivar.
The present work was designed to compare the vitamin C (ascorbic acid, AsA) content of pulp, peel, and juice of 64 apple cultivars. These cultivars were carefully identified as 'true to type' by molecular genetic tools, grown in the same site under identical conditions and processed by a standardized protocol. Twenty-one of them, accounting for more than 95% of the apple production of South Tyrol, were chosen to represent the current market, 16 were old or local cultivars formerly grown in the area, and 27 were new cultivars, including 15 with scab resistance and 12 with red flesh fruit. For the determination of the AsA content, a new High-Performance Liquid Chromatography with Diode-Array Detection method was developed and validated. While old cultivars stood out for their high AsA content in pulp and peel, the red-fleshed cultivars are the ones maintaining most of their AsA content during processing. Our data thus suggest a potential for old and red-fleshed cultivars for healthy juices or further processed food components.
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