In contrast to the well-studied canonical regulatory mechanisms, the way by which the recently discovered Src N-terminal regulatory element (SNRE) modulates Src activity is not yet well understood. ...Phosphorylation of serine and threonine residues modulates the charge distribution along the disordered region of the SNRE and may affect a fuzzy complex with the SH3 domain that is believed to act as an information transduction element. The pre-existing positively charged sites can interact with the newly introduced phosphate groups by modulating their acidity, introducing local conformational restrictions, or by coupling various phosphosites into a functional unit. In this paper, we use pH-dependent NMR measurements combined with single point mutations to identify the interactions of basic residues with physiologically important phosphorylated residues and to characterize the effect of these interactions in neighbor residues, thus providing insight into the electrostatic network in the isolated disordered regions and in the entire SNRE. From a methodological point of view, the linear relationships observed between the mutation-induced pKa changes of the phosphate groups of phosphoserine and phosphothreonine and the pH-induced chemical shifts of the NH groups of these residues provide a very convenient alternative to identify interacting phosphate groups without the need to introduce point mutations on specific basic residues.
Knowledge of composition of beverages volatile fraction is essential for understanding their sensory attributes. Analysis of volatile compounds predominantly resorts to gas chromatography coupled ...with mass spectrometry (GC-MS). Often a previous concentration step is required to quantify compounds found at low concentrations. This work presents a liquid-liquid microextraction method combined with GC-MS (LLME/GC-MS) for the analysis of compounds in fermented beverages and spirits. The method was validated for a set of compounds typically found in fermented beverages comprising alcohols, esters, volatile phenols, and monoterpenic alcohols. The key requirements for validity were observed, namely linearity, sensitivity in the studied range, accuracy, and precision within the required parameters. Robustness of the method was also evaluated with satisfactory results. Thus, the proposed LLME/GC-MS method may be a useful tool for the analysis of several fermented beverages, which is easily implementable in a laboratory equipped with a GC-MS.
Plant proteins have been proposed as an alternative to animal-origin proteins in the wine industry because they are allergen-free and vegan-friendly. The aim of this study was to evaluate the ...effectiveness of plant proteins as fining agents on red wines with different phenolic composition. Two formulations for commercially available vegetal proteins (potato and pea origin) were assessed at two doses to modulate the fining treatment to the wine phenolic profile. The results evidenced that fining agents derived from plants have different levels of effectiveness on the removal of phenolic compounds depending on the origin, the formulation used, dose applied, and also wine characteristics. On Nebbiolo wine, the study was particularly significant due to its phenolic composition. One pea-based fining agent had an effect comparable to gelatin (animal origin) on the removal of polymeric flavanols with a minor loss of anthocyanins and therefore better preserving the wine color in terms of intensity and hue. For Primitivo, Montepulciano, and Syrah wines, even though there was a formulation-dependent effect, vegetal proteins gave more balanced reductions in terms of target phenolic compounds contributing to astringency and color perception.
Ozone, a powerful oxidative stressor, has been recently used in wine industry as sanitizing agent to reduce spoilage microflora on grapes. In this study, we evaluated ozone-induced metabolic and ...molecular responses during postharvest grape dehydration. Ozone increased the contents of total volatile organic compounds (VOCs), which have a great impact on the organoleptic properties of grapes and wines. Among terpenes, responsible for floral and fruity aroma, linalool, geraniol and nerol were the major aromatic markers of Moscato bianco grapes. They were significantly affected by the long-term ozone treatment, increasing their concentration in the last phases of dehydration (>20% weight loss). At molecular level, our results demonstrated that both postharvest dehydration and ozone exposure induce the biosynthesis of monoterpenes via methylerythritol phosphate (MEP) pathway and of aldehydes from lipoxygenase-hydroperoxide lyase (LOX-HPL) pathway. Therefore, transcriptional changes occurred and promoted the over-production of many important volatile compounds for the quality of white grapes.
Changes in volatile composition (mean and standard deviation) for Betanzos cultivars in two ripening dates: 23.08.2007 (A) and 30.08.2007 (B). Display omitted
► Changes in volatile composition at ...last ripening stages for Vitis vinifera cv. ► Significant volatile increase between the ripening stages. ► Free C6-compounds and bound alcohols showed the highest concentration. ► Strong influence by the cultivar and the ripening stage on wine volatiles. ► Significant interaction cultivar*ripening date for 93.8% of volatile compounds.
The volatile composition of white Agudelo, Blancolexitimo, Godello and red Serradelo cultivars (NW Spain) harvested at two different stages of ripening have been evaluated. C6-compounds, alcohols, volatile fatty acids, monoterpenes, C13-norisoprenoids, volatile phenols and carbonyl compounds were identified and quantified in free and glycosidically bound forms by gas chromatography–mass spectrometry (GC–MS). The total volatile concentration showed a significant increase between the two ripening stages studied for all cultivars. The free volatile composition increased during maturity for Godello and Serradelo cultivars; however the glycosidically bound concentration increases for all cultivars with exception of B. lexitimo. Free C6-compounds ((E)-2-hexanal, 1-hexanol and (E)-2-hexen-1-ol) and bound alcohols (benzyl alcohol and 2-phenylethanol) showed the highest concentrations of volatile compounds for all grape cultivars in the two dates studied. Godello cultivar showed the highest change of volatile concentration between two ripening dates because of the high value of free C6-compounds. B. lexitimo was the most terpene-rich cultivar at the last ripening stage due to linalool; however C13-norisoprenoids in free form were detected in low concentrations for all cultivars but not in Godello and B. lexitimo cultivars at the last ripening stage. Free hexanoic acid increased during ripening in all cultivars. The evolution of volatiles during ripening of grape juice from the cultivars studied was not proportional to the changes in sugar content, which shows that the technological and aromatic maturities did not occur at the same time in these cultivars. The results also showed the cultivar*ripening date interaction for all, free and bound, groups of compounds.
► Vitis vinifera Albariño was supplemented with diammonium phosphate (DAP) in the range 250–450mgN/L. ► DAP has a significant effect on the chemical and volatile composition of Albariño wines. ► ...Moderate nitrogen supplementation contributed to the fruity/floral aroma.
Vitis vinifera cv. Albariño is an aromatic variety characterised by terpenes present largely as glycosidic flavour precursor compounds, which depends on fermentation to reveal its aromatic varietal potential. Clarified Albariño must containing 250mgN/L (control) was supplemented with diammonium phosphate to 350 and 450mgN/L before fermentation with a high-nitrogen-demand wine strain of Saccharomyces cerevisiae (M05). Ammonium supplementation had a significant effect on the chemical composition (titratable acidity and ethanol) and volatile profile of Albariño wines. Varietal compounds in the free fraction (limonene, linalool, α-terpineol, α-ionone and β-damascenone) and bound (limonene, linalool and α-terpineol), as well as most yeast-derived fermentation products (including esters, higher alcohols and volatile acids) were associated with moderate nitrogen concentrations. Free β-ionone, bound geraniol, bound α-ionone, bound β-damascenone and 1-hexanol were associated with high nitrogen concentrations. OAV wines made with moderate nitrogen exhibited a higher total odorant activity, driven by ethyl esters, terpenes and C13-norisoprenoids, resulting in fruity and floral aroma attributes.
Cassava plays a key role in the food production and economies of several countries worldwide. Due to its starch content, alcoholic fermentation is a promising transformation process for adding value ...to cassava. However, most of the existing cassava beverages are from traditional origin, with the yields and quality often poorly known or controlled due to the use of artisanal production processes. This work aims at the application of easily implementable biotechnological tools for the production of cassava spirits, in order to add value to this raw material. Cassava flour was liquefied and saccharified using enzymatic cocktails, generating a fermentable broth with ~184 g L
of fermentable sugars. This was then fermented into an alcoholic product with ~10% ethanol by volume and distilled for spirit production. Cassava spirits with 40% ethanol by volume, with or without application of oak wood, were produced. For further valorization, volatile fractions of cassava spirits were characterized by gas chromatography-flame ionization detection (GC-FID) and GC-MS. These showed a predominance of yeast fermentation metabolites, complemented by wood extractives where oak chips were applied. Both produced spirits showed desirable sensory traits, receiving good acceptance by experienced tasters, demonstrating the feasibility of the proposed process to add value to cassava surplus.
Premature birth, perinatal inflammation, and life-saving therapies such as postnatal oxygen and mechanical ventilation are strongly associated with the development of bronchopulmonary dysplasia ...(BPD); these risk factors, alone or combined, cause lung inflammation and alter programmed molecular patterns of normal lung development. The current knowledge on the molecular regulation of lung development mainly derives from mechanistic studies conducted in newborn rodents exposed to postnatal hyperoxia, which have been proven useful but have some limitations.
Here, we used the rabbit model of BPD as a cost-effective alternative model that mirrors human lung development and, in addition, enables investigating the impact of premature birth per se on the pathophysiology of BPD without further perinatal insults (e.g., hyperoxia, LPS-induced inflammation). First, we characterized the rabbit's normal lung development along the distinct stages (i.e., pseudoglandular, canalicular, saccular, and alveolar phases) using histological, transcriptomic and proteomic analyses. Then, the impact of premature birth was investigated, comparing the sequential transcriptomic profiles of preterm rabbits obtained at different time intervals during their first week of postnatal life with those from age-matched term pups.
Histological findings showed stage-specific morphological features of the developing rabbit's lung and validated the selected time intervals for the transcriptomic profiling. Cell cycle and embryo development, oxidative phosphorylation, and WNT signaling, among others, showed high gene expression in the pseudoglandular phase. Autophagy, epithelial morphogenesis, response to transforming growth factor β, angiogenesis, epithelium/endothelial cells development, and epithelium/endothelial cells migration pathways appeared upregulated from the 28th day of gestation (early saccular phase), which represents the starting point of the premature rabbit model. Premature birth caused a significant dysregulation of the inflammatory response. TNF-responsive, NF-κB regulated genes were significantly upregulated at premature delivery and triggered downstream inflammatory pathways such as leukocyte activation and cytokine signaling, which persisted upregulated during the first week of life. Preterm birth also dysregulated relevant pathways for normal lung development, such as blood vessel morphogenesis and epithelial-mesenchymal transition.
These findings establish the 28-day gestation premature rabbit as a suitable model for mechanistic and pharmacological studies in the context of BPD.
The aim of this work was to study the influence of early basal leaf removal on Aglianico wines produced in the Apulia region (Italy). Three treatments were carried out, where 100% of fruit-zone ...leaves on the north (DN), south (DS) and on both sides of the canopy (DNS) were removed. A control (CT), where all basal leaves were retained, was also performed. Instrumental (HPLC-DAD-MS and GC-MS) and sensory analysis (QDA) were used to evaluate the treatment effect on the phenolic and volatile compositions and on the sensory descriptors of wines. DNS reached the highest amounts of phenolic compounds, showing a change in the phenolic pattern from flavonols and anthocyanins. Moreover, leaf removal influenced the levels of 37.8% of volatile compounds, quantified by increasing the concentration when early leaf removal was applied on the north side of the canopy (DN), with respect to the south (DS) and both sides (DNS). In the sensory analysis, Aglianico wines were defined by 16 sensory attributes with GM > 30%, where the highest values were reached for defoliation treatments vs. control. In conclusion, early leaf removal treatments allowed us to modulate the phenolic and volatile concentrations of Aglianico wines.
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