Deep eutectic solvent-based shaking-assisted extraction for determination of bioactive compounds from Norway spruce roots.

Polyphenolic compounds play an essential role in plant growth, reproduction, and defense mechanisms against pathogens and environmental stresses. Extracting these compounds is the initial step in assessing phytochemical changes, where the choice of extraction method significantly influences the extracted analytes. However, due to environmental factors, analyzing numerous samples is necessary for statistically significant results, often leading to the use of harmful organic solvents for extraction. Therefore, in this study, a novel DES-based shaking-assisted extraction procedure for the separation of polyphenolic compounds from plant samples followed by LC-ESI-QTOF-MS analysis was developed. The DES was prepared from choline chloride (ChCl) as the hydrogen bond acceptor (HBA) and fructose (Fru) as the hydrogen bond donor (HBD) at various molar ratios with the addition of 30% water to reduce viscosity. Several experimental variables affecting extraction efficiency were studied and optimized using one-variable-at-a-time (OVAT) and confirmed by response surface design (RS). Nearly the same experimental conditions were obtained using both optimization methods and were set as follows: 30 mg of sample, 300 mg of ChCl:Fru 1:2 DES containing 30% w/w of water, 500 rpm shaking speed, 30 min extraction time, 10°C extraction temperature. The results were compared with those obtained using conventional solvents, such as ethanol, methanol and water, whereby the DES-based shaking-assisted extraction method showed a higher efficiency than the classical procedures. The greenness of the developed method was compared with the greenness of existing procedures for the extraction of polyphenolic substances from solid plant samples using the complementary green analytical procedure index (ComplexGAPI) approach, while the results for the developed method were better or comparable to the existing ones. In addition, the practicability of the developed procedure was evaluated by application of the blue applicability grade index (BAGI) metric. The developed procedure was applied to the determination of spruce root samples with satisfactory results and has the potential for use in the analysis of similar plant samples.

Element content and expression of genes of interest in guard cells are connected to spatiotemporal variations in stomatal conductance.

Element content and expression of genes of interest on single cell types, such as stomata, provide valuable insights into their specific physiology, improving our understanding of leaf gas exchange regulation. We investigated how far differences in stomatal conductance (gs ) can be ascribed to changes in guard cells functioning in amphistomateous leaves. gs was measured during the day on both leaf sides, on well-watered and drought-stressed trees (two Populus euramericana Moench and two Populus nigra L. genotypes). In parallel, guard cells were dissected for element content and gene expressions analyses. Both were strongly arranged according to genotype, and drought had the lowest impact overall. Normalizing the data by genotype highlighted a structure on the basis of leaf sides and time of day both for element content and gene expression. Guard cells magnesium, phosphorus, and chlorine were the most abundant on the abaxial side in the morning, where gs was at the highest. In contrast, genes encoding H+ -ATPase and aquaporins were usually more abundant in the afternoon, whereas genes encoding Ca2+ -vacuolar antiporters, K+ channels, and ABA-related genes were in general more abundant on the adaxial side. Our work highlights the unique physiology of each leaf side and their analogous rhythmicity through the day.

Nutrition habits in 24-hour mountain bike racers.

We investigated seventy-four ultra-mountain bikers (MTBers) competing in the solo category in the first descriptive field study to detail nutrition habits and the most common food before during and after the 24 hour race using questionnaires. During the race, bananas (86.5%), energy bars (50.0%), apples (43.2%) and cheese (43.2%) were the most commonly consumed food, followed by bread (44.6%), rice (33.8%) and bananas (33.8%) after the race. Average fluid intake was 0.5 ± 0.2 l/h. The main beverage was isotonic sports drink (82.4%) during and pure water (66.2%) after the race. The most preferred four supplements in the four weeks before, the day before, during and after the race were vitamin C (35.1%), magnesium (44.6%), magnesium (43.2%) and branched-chain amino acids (24.3%), respectively. Total frequency of food intake (30.6 ± 10.5 times/24 hrs) was associated with fluid intake (r = 0.43, P = 0.04) and both were highest at the beginning of the race and lower during the night hours and the last race segment in a subgroup of twenty-three ultra-MTBers. Supplement intake frequency (6.8 ± 8.4 times/24 hrs) was highest during the night hours and lower at the beginning and end of the race. Elevated food and fluid intake among participants tracked across all race segments (P < 0.001). In conclusion, the nutrition strategy employed by ultra-MTBers was similar to those demonstrated in previous studies of ultra-cyclists with some exceptions among selected individuals.