Acrylamide (AA) is a probable human carcinogen present in several highly consumed starchy foods like potato chips. Due to the lack of important chromophore groups in its chemical structure, it is generally derivatized to be detected by ultraviolet (UV) or fluorescence (FL). The objective of this work was to develop a novel high-throughput high-performance thin-layer chromatography method to determine AA in starchy foods using the fluorescent probe 7-mercapto-4-methylcoumarin (7M4MC). The main derivatization factors for AA and N,N-Dimethylacrylamide (DMAA, internal standard) were optimized using a Face-centered Central Composite Design, establishing the following optimal conditions: temperature (50 °C), time (10 min), and catalyst concentration (0.5 mmol/L of 4-dimethylaminopyridine). Chromatography was carried out on silica gel 60 F254 HPTLC plates using a mobile phase composed of dichloromethane, acetic acid, and acetone (94.5: 3.9: 1.6 v/v/v). The analytical method was validated according to the International Conference on Harmonization (ICH) guidelines. Analytical response in the range from 5 to 400 ng/band showed an R2 > 0.998. Repeatability and intermediate precision showed RSD values lower than 7

Endophyte fungi (EF) are considered a new and valuable reservoir of bioactive molecules of biotechnological interest for pharmacy, agricultural and forestry industries. In this study, thirty EFs, isolated from three Chilean Nothofagus species (N. alpina, N. dombeyi, N. oblicua) were identified and cultured in submerged liquid fermentations aimed at searching for natural active substances. The extracts obtained were evaluated against pathogenic bacteria and fungi. Sixteen extracts (53.3%) presented antibacterial and fourteen (46.6%) presented antifungal activities in different intensities. Extracts from isolates Coryneum sp.-72 and P. cinnamomea-78 exhibited the highest antimicrobial activity. Using bioautography, the compounds responsible for the antimicrobial activity exhibited by Coryneum sp.-72 and P. cinnamomea-78 were detected and characterized. Coryneum sp.-72 showed bactericidal properties at 200 μg/mL and bacteriostatic effects at 50 μg/mL against B. cereus, B. subtilis, L. monocytogenes and S. aureus. MIC values indicated that P. cinnamomea-78 exhibited a strong fungistatic and fungicidal effect against B. cinerea and C. gloesporioides at 10–50 μg/mL. Isolates were grouped in the following order: Botryosphaeriales, Diaporthales, Eurotiales, Helotiales, Hypocreales, Pleosporales, Magnaporthales, Sordariales and Polyporales. EF isolated, identified and evaluated constitute the first report for Chilean Nothofagus genus.

The PLE grape pomace extract successfully inhibits the AA formation in ASN–GLC and bread models showing reductions of ca. 33% and 41% respectively epicatechin, catechin, and gallic acid were the primary inhibitors of AA formation.

Previous studies indicate that the alkali-induced oxidation of quercetin (Que) or kaempferol (Kae) converts them into hydroxylated-benzoyl-benzofuranones (BZFs), a type of metabolite whose antioxidant potency is substantially higher. To explore whether the former conversion can take place under a more biologically relevant oxidizing setting, these flavonols were incubated in a nitrite-containing acidic medium. Under such condition, the concentrations of Que or Kae rapidly decayed, simultaneously generating their corresponding BZFs. When Que and Kae were co-incubated, the depletion of the parent flavonols accelerated without changes in BZF formation. Similar results were observed when either Que or Kae were incubated with some of their corresponding 3-O-glycosides. In turn, when Que was co-incubated with certain flavanols or phenolic acids, the latter compounds nearly doubled not only the oxidative disappearance of Que but also its BZF formation. Interestingly, the concentrations of Que-BZF and Kae-BZF reached under all former incubation scenarios are greater than those previously reported to be needed to protect intestinal epithelial cells against the oxidative and lytic damage induced by ROS. We suggest that, if these conversions also took place in vivo, the gastric fluid could be potentially seen as a chemical milieu where the antioxidant properties of Que and Kae could be enormously amplified.

The pisco industry generates significant environmental waste, particularly grape pomace, which is a rich source of phenolic compounds. Emerging extraction technologies offer promising alternatives for recovering these bioactive components. This study evaluated enzyme-assisted extraction (EAE) and pressurized liquid extraction (PLE) techniques using response surface methodology to optimize phenolic compound yield and antioxidant capacity. Specifically, a D-optimal design was applied for EAE, and a Box–Behnken design was applied for PLE. The optimal extraction conditions for EAE were 0.75 U/mL of tannase, 40 U/mL of cellulase, 20 °C, and 15 min. For PLE, the optimal parameters were 54% ethanol, 113 °C, and three extraction cycles. These conditions yielded 38.49 mg GAE g−1 dw and 50.03 mg GAE g−1 dw of total polyphenols and antioxidant capacities of 342.47 μmol TE g−1 dw and 371.00 μmol TE g−1 dw, respectively. The extracts obtained under optimal conditions were further characterized through chromatographic techniques to determine their phenolic profiles. Seven phenolic compounds were identified: gallic acid, catechin, epicatechin, 4-hydroxybenzoic acid, quercetin-3-rutinoside hydrate, quercetin-3-O-rhamnoside, and kaempferol. PLE extracts exhibited the highest concentration of these compounds. These findings demonstrate that recovering antioxidant-rich phenolic compounds from pisco grape pomace using innovative extraction methods is a viable strategy for obtaining functional ingredients and supporting sustainable industrial practices.

This work reports the development and validation of a fast and selective high-performance thin-layer chromatography (HPTLC) method for determining the most relevant biogenic amines (BAs) present in beers, namely, spermine, spermidine, putrescine, cadaverine, tryptamine, histamine, tyramine, and 2-phenylethylamine. Dansyl-BAs were separated on dipotassium hydrogen phosphate-treated HPTLC silica gel 60 plates, using a mobile phase composed of tert-butyl methyl ether‒toluene‒triethylamine (2:2.5:6, V/V). Densitometric quantification was carried out by fluorescence at 366/> 400 nm. Validation was established according to the International Conference on Harmonisation (ICH) guidelines. Each calibration data (in matrix) fitted a polynomial regression model with R2 > 0.996. Repeatability (n = 6) and intermediate precision (n = 6) in matrix showed relative standard deviation (RSD) values < 3.57 and < 2.58

Black Chiloe’s giant garlic is a functional food produced by a mild Maillard reaction that contains relevant bioactive molecules like organosulfur compounds (OSCs) and (poly)phenols (PPs). Compared with raw garlic, black garlic has a higher content of PPs and S-allyl cysteine (SAC), a key OSC due to its bioactivities. The objective of the present work was to optimize by chemometric tools a green microwave-assisted extraction (MAE) of SAC and PPs present in black Chiloe’s giant garlic to detect and identify novel bioactive molecules with antioxidant and/or inhibitory activities over cyclooxygenase, α-glucosidase, and acetylcholinesterase enzymes. The MAE factors were optimized using a central composite design, establishing optimal PP and SAC yields at 67 °C, 0% ethanol, 12 min and 30 °C, 40% ethanol, 3 min, respectively. PP and SAC values were 9.19 ± 0.18 mg GAE/g DW and 2.55 ± 0.10 mg SAC/g DW. Applying effect-directed analysis using high-performance thin layer chromatography-bioassay and mass spectrometry, the bioactive molecules present in the MAE extract with antioxidant and inhibitory activities over cyclooxygenase, α-glucosidase, and acetylcholinesterase enzymes were identified as N-fructosyl-glutamyl-S-(1-propenyl)cysteine, N-fructosyl-glutamylphenylalanine, and Harmane.

The present work reports the chemical and functional profile of Chiloe's giant garlic. Applying AOAC's official methods the proximal content of giant garlic was determined finding a moisture content of 62.34 +/- 0.35%, carbohydrates 20.64 +/- 0.01%, protein 2.80 +/- 0.10%, fat (crude) 0.09 +/- 0.00%, ash 0.74 +/- 0.02%, and fiber (crude) 13.04 +/- 0.01%. Three saccharides [sucrose (5.92 +/- 0.02 mg g(-1)), glucose (0.11 +/- 0.00 mg g(-1)) and fructose (0.46 +/- 0.01 mg g(-1))], four fatty acids [linoleic acid (57.67 +/- 0.00%), palmitic acid (23.46 +/- 0.00%), oleic acid (7.20 +/- 0.00%), and alpha-linolenic acid (5.06 +/- 0.00%)], and three sulfoxide compounds [alliin (2.66 +/- 0.90 mg g1), methiin (9.61 +/- 0.33 mg g(-1)) and isoalliin (5.02 +/- 1.24 mg g(-1))] were determined by high-performance thin-layer chromatography (HPTLC), gas chromatography 0.250 +/- 0.001 mmol TE per 100 g(-1) and a total (poly)phenols content (TPC) of 40.66 +/- 2.08 mg EAG 100 g(-1). (Poly)phenols profile analyzed by liquid chromatography (LC)/MS showed only the presence of caffeic acid (0.57 +/- 0.05 mu g g(-1)) and rutin (at traces level). Bioactive molecules with antioxidant (DPPH) and COX-2 inhibition activities were identified through HPTLC (bio)autography and MS analysis, finding the presence of tryptophan (antioxidant) and gamma-glutamyl-S-allyl-L- cysteine (GSAC), gamma-glutamyl-S-(trans-1-propenyl)-L- cysteine (GSPC), alliin and isoalliin with antioxidant and COX-2 inhibitory activity.

Priming effect is the enhanced microbial degradation of refractory organic carbon in the presence of labile organic matter. It has been proposed to explain the disappearance of supposedly recalcitrant terrestrial organic matter that reaches the coastal ocean. Here, we experimentally evaluated whether labile organics boosts the degradation of persistent pollutants, phenanthrene and pyrene (polycyclic aromatic hydrocarbons, PAHs). Microcosm incubations were conducted during 11 and 50 days using marine surface sediments from Concepción Bay, a PAH-impacted upwelling ecosystem in central Chile, and the Almirante Montt Gulf, a rather pristine Patagonian Fjord. Addition of yeast extract enhanced the degradation of phenanthrene and pyrene by 14-170 %, while microalgal extract increased the decay rate constant of phenanthrene by 67 %. PAH degradation was accompanied by a pronounced enrichment of hydrocarbonoclastic bacteria, which increased approximately 840-fold (average Log₂FC = 9.7) with rather pristine Patagonian sediments but only 12-fold increase (average Log₂FC = 3.6) in PAH-impacted Concepción Bay sediments. The broader shift in microbial community structure detected in Patagonian fjords coincides with low ambient PAH contents, as expected in unpopulated areas. Considering the long record and intensity of industrial activity off Concepción Bay and the surrounding area, we suggest that those sediments contain a core microbiome with a toolkit of potential enzymes involved in PAH-degradation pathways capable of processing high inputs of PAHs. This finding may imply that coastal marine sediments store microbial capacity for degradation of contaminants induced by a long history of PAH emissions, and this ability can be further enhanced by exposure to primers.

Quercetin oxidation leads to the formation of a metabolite, 2-(3,4-dihydroxybenzoyl)-2,4,6-trihydroxy-3(2H)-benzofuranone, whose antioxidant potency was recently reported to be a 1000-fold higher than that of its precursor. The formation of similar metabolites (BZF) is limited to certain flavonols (FL), among which are myricetin, fisetin, and morin. Here we addressed the consequences of inducing the auto-oxidation of these flavonols in terms of their antioxidant properties (assessed in ROS-exposed Caco-2 cells). The mixtures that result from their oxidation (FLox) exhibited antioxidant activities 10-to-50-fold higher than those of their precursors. Such amplification was fully attributable to the presence of BZF in each FLox (established by HPLC-ESI-MS/MS and chemical subtraction techniques). An identical amplification was also found when the antioxidant activities of BZF, isolated from each FLox, and FL were compared. These findings warrant the search of these BZF in edible plants and their subsequent evaluation as a new type of functional food ingredients.

The search and identification of inhibitory molecules from novel natural sources, such as pisco grape pomace extract obtained by green techniques, may help to develop agents with therapeutic potential that are beneficial to health with fewer adverse effects than drugs. Many drugs act as enzyme inhibitors, decreasing their activity and thus correcting a metabolic imbalance. This study aims to identify bioactive molecules with antioxidant and inhibitory activity over acetylcholinesterase and cyclooxygenase enzymes present in pisco grape pomace green extracts. Bioactive molecules were detected and identified applying directed effect analysis on planar chromatography coupled to mass spectrometry. For the first time, the presence of antioxidant molecules (quercetin-3-O-glucuronide, quercetin-3-O-glucoside, and gallic acid) and inhibitors of acetylcholinesterase (kaempferol-3-O-glucoside) and cyclooxygenase (gallic acid) enzymes are reported in pisco grape pomace. According to the results, grape pomace could be an alternative to develop novel functional foods and nutraceuticals that provide health benefits and, at the same time, generate a circular economy in the industry.

Polycyclic aromatic hydrocarbons (PAH) are semi-volatile, lipophilic, and harmful compounds that can persist for decades in a range of marine environments. There are several marine and soil microorganisms that possess enzymes involved in arene degradation. Here, we analyzed the structure (16S rRNA amplicons) and metabolic potential (inferred using phylogenetic placement) of the bacterial community in surface marine sediments from coastal waters off Concepci & oacute;n, Chile, and describe how microbial community patterns are shaped and altered by PAH contamination. Two depositional zones were identified, a "High PAH" area containing a mix of high and low molecular weight PAH of up to 10,350 ng & sum;PAH gdw-1 and with high organic matter content; and a "Low PAH" zone mostly characterized by low molecular weight PAH of up to 1810 ng & sum;PAH gdw-1 and lower levels of organic matter. We identified 53 hydrocarbonoclastic bacteria genera, with eight showing relatively high abundances at High PAH sites, although known PAH degrader clades were also present at Low PAH sites. With potential enzymes inferred in almost all samples, we suggest that breakdown of PAH is widespread in this area, likely resulting from the long history of local PAH emissions that may have promoted a stored microbial capacity for these degradation processes.

The pisco produced in Chile is a distillate from pisco grape varieties, which generates a large amount of grape pomace. However, with the growing interest in reducing the environmental impact and converting these residues into novel by-products, the research and their evaluation in each stage can contribute significantly to ensuring their recovery. This study evaluated the kinetics of vacuum and convective drying from grape pomace at different temperatures and the influence on the retention of polyphenolic compounds and antioxidant activity as scalable alternatives for agro-industry. The pisco grape pomace was dried by two drying methods at different temperatures: Vacuum drying (50–100 °C; 100 mbar) and convective drying (40–80 °C). Liquid chromatography with diode-array detection and spectrophotometry were used to characterize the polyphenol profile and evaluate the antioxidant activity. The best drying grape pomace process was vacuum drying at 60 °C with a short drying time (210 min), and diffusivity values were higher (6.64 × 10−10 m2s−1) than those obtained by other drying conditions. The model that best fits the experimental data was the Midilli Kucuk model. Vacuum drying at 60 °C obtained the highest content of polyphenols and flavonoids, and significant antioxidant capacity was obtained. The gallic acid, 4-hydroxybenzoic acid, catechin, epicatechin, and rutin were found in grape pomace. Vacuum drying at 60 °C is a viable alternative to stabilize pisco grape pomace, achieving good drying times and generating an environmentally friendly solution.

Ingredients rich in phenolic compounds and antioxidants of winemaking wastes, which play an important role in the prevention of various diseases and the control of viruses, are being explored. Currently, there is a concern about honeybee population loss, with deformed wing virus (DWV) being the most common virus infecting apiaries and one of the main causes of honeybee decline. Hence, the effect of grape pomace powder (GPP) as a dietary supplement to enhance the immune system of honeybees affected by DWV was evaluated. The characteristics of the ingredient GPP, obtained by spray-drying, revealed a high anthocyanin content (1102.45 mg 100 g−1), and it was applied at doses of 0.5, 1, 2.5 and 5% as a dietary supplement for bees infected by DWV. The results showed that the GPP treatments strengthened the immune response of honeybees against DWV. Moreover, the expression of the Relish gene was significantly higher in bees fed with GPP compared to the infected control. This study, which is framed in the search of food waste valorization for environmental sustainability, proves the feasibility of using grape wastes as dietary supplements for pollinators, and provides knowledge of the influence of polyphenols on the expression profiles of immune-related genes in honeybees.

This research aimed to evaluate the possible runoff and leaching of atrazine in corn crops during high-intensity rainfall and the influence of summer and winter seasons on this herbicide's environmental distribution. Leaching and suction lysimeters were used, with atrazine mobility evaluated during simulated precipitations 24 and 48 h after the atrazine spraying during corn development during the summer and winter seasons for two years. Adsorption studies at the laboratory were conducted to understand soil adsorption better. The K-d and K-OC at 25 degrees C of 2.59 L Kg(-1) and 281.64 L Kg(-1) indicate moderate atrazine retention, with soil adsorption being exothermic, reversible, and spontaneous. Multilayer adsorption of atrazine is enhanced by soil organic matter by about 20.3%. During winter, atrazine retention in soil increases by 2.2x, while its loss by runoff and leaching decreases by 6.75 and 3.0x. Losses by runoff during rainfall simulations are 14.5x greater at 24 h than 48 h after spray, while atrazine loss by leaching is 1.7x greater at 48 h than 24 h after spray. Runoff or leached water contained much higher levels of atrazine than the acceptable national or international standards for drinking water, natural waters, or effluent discharge, indicating that extreme weather could increase environmental contamination.

Endothelial progenitor cells (EPCs) are stem cells mainly derived from bone marrow; from where they migrate to repair and regenerate damaged tissues. eEPCs have been classified into two sub-populations, early (eEPC) and late EPCs (lEPC), depending on maturation stages in vitro. In addition, eEPC release endocrine mediators, including small extracellular vesicles (sEVs), which in turn may enhance the eEPC-mediated wound healing properties. Nevertheless, adenosine contributes to angiogenesis by recruiting eEPC at the injury site. However, whether ARs may enhance the secretome of eEPC, including sEVs, is unknown. Therefore, we aimed to inves-tigate whether AR activation increase the release of sEVs in eEPC, which in turn has paracrine effects on recipient endothelial cells. Results shown that 5 '-N-ethylcarboxamidoadenosine (NECA), a non-selective agonist, increase both the protein levels of the vascular endothelial growth factor (VEGF), and the number of sEVs released to the conditioned medium (CM) in primary culture of eEPC. Importantly, CM and EVs harvested from NECA-stimulated eEPC promote in vitro angiogenesis, without changes in cell proliferation, in recipient ECV-304 endothelial cells. This constitutes the first evidence showing that adenosine enhances sEVs release from eEPC, which has pro-angiogenic capacity on recipient endothelial cells.

BACKGROUND:The exceptional capacities of aquaporins in terms of water permeation and selectivity have made them an interesting system for membrane applications. Despite the multiple attempts for immobilizing the aquaporins over a porous substrate, there is a lack of studies related to the purification and reconstitution steps, principally associated with the use of detergents in solubilization and destabilization steps. This study analyzed the effect of detergents in Aquaporin Z solubilization, considering the purity and structural homogeneity of the protein.METHODS:The extraction process was optimized by the addition of detergent at the sonication step, which enabled the omission of the ultracentrifugation and resuspension steps. Two detergents, Triton X-100, and octyl-glucoside were also evaluated. Destabilization mediated by detergents was used as reconstitution method. Saturation and solubilization points were defined by detergent concentration and both, liposomes and proteoliposomes, were analyzed by size distribution and permeability assays. Detergent removal with Bio-beads was also analyzed.RESULTS:Octyl glucoside ensures structural stability and homogeneity of Aquaporin Z. However, high concentrations of detergents induce the presence of defects in proteoliposomes. While saturated liposomes create homogeneous and functional structures, solubilized liposomes get affected by a reassembly process, creating vesicle defects with anomalous permeability profiles.CONCLUSIONS:Detergent concentration affects the structural conformation of proteoliposomes in the reconstitution process.GENERAL SIGNIFICANCE:Since the destabilization process is dependent on vesicle, detergent, and buffer composition, optimization of this process should be mandatory for further studies. All these considerations will allow achieving the potential of Aquaporins and any other integral membrane protein in their applications for industrial purposes.

Caseins are the principal milk proteins and serve as an important source of bioactive peptides with diverse beneficial effects for human health, such as antihypertensive, immunomodulating, anti-inflammatory, and antithrombotic. The objective of this study was to optimize, using the design of the experiment, the production of bioactive peptides from alpha-casein applying microwave -assisted enzyme digestion (MAED). The optimal MAED conditions (time, temperature, and enzyme/protein ratio) were established for pepsin (digestion time 4 min, temperature 41 degrees C, and E:P ratio 1:40) and trypsin (digestion time 10 min, temperature 37 degrees C and E:P ratio 1:200.) enzymes. Digestion yields and the intensity of different bioactivities, i.e., antimicrobial, antioxidant, and acetylcholinesterase and alpha glucosidase inhibitory activities, evaluated by the HPTLC-bioassay technique were used as response variables. Trypsin-formed alpha-casein peptides showed antimicrobial, antioxidant, and acetylcholinesterase inhibition activities, while pepsin -formed showed antimicrobial, antioxidant, and alpha-glucosidase inhibition activities. These results demonstrate that MAED is a fast and effective technique for bioactive peptide production from casein proteins.

This work aimed at studying the effect of molecular weight (MW) and deacetylation degree (DD) of chitosan on the quercetin bioaccessibility encapsulated in alginate/chitosan-coated zein nanoparticles (alg/chiZN). The chitosan coating layer produced nanoparticulate systems with good stability parameters, high encapsulation efficiency (EE) and a higher bioaccessibilty of quercetin after in-vitro digestion. By increasing the DD of chitosan, the ζ-potential of the colloidal system significantly increased (≥27.1 mV), while low and very low MW chitosans generated systems with smaller particle sizes (≤ 277.8 nm) and polydispersity index [PDI (0.189)]. The best results, in terms of EE (≥84.44) and bioaccessibility (≥76.70), were obtained when the systems were prepared with low MW chitosan and high DD. Thus, the alg/chiZN nanocapsules may be a promising delivery system for improving the quercetin bioaccessibility or other compounds with a similar chemical nature, especially when higher DD and lower MWs are used.