The Management of Irrigation and Potassium Fertilization to Mitigate the Effect of Light Frosts on the Phenolic and Volatile Compounds in Virgin Olive Oils.

The frequency of early frosts has increased in recent years, which are injurious to olive growing, causing losses in the yield and quality of virgin olive oil. In this research, it was studied how the management of agronomic factors mitigates frost damage in Arbequina olives, minimizing the loss of phenols and volatiles in virgin olive oil, at different fruit ripening stages. A Box-Behnken design and multivariate analysis were performed, with three levels of irrigation, potassium fertilization, and foliar copper application (15 treatments). Virgin olive oil was extracted from fresh and frozen olives. Light frost caused a significant decrease in the total phenols and secoiridoid compounds in and the antioxidant capacity of the frost-affected oils, which were perceived as more pungent and had the slight defect of "frostbitten olives". According to the Box-Behnken design, an 86% reference evapotranspiration (ET0) or higher with 100 potassium oxide units (UK2O) and a 100% ET0 or higher with 250 UK2O would be required to minimize the effect of light frost on phenols and volatiles. Partial Least Squares Regression-Discriminant Analysis (PLS-DA) differentiated the virgin olive oils according to their ripening stage and fresh and frost conditions. Moreover, PLS-DA positively correlated a 75-100% ET0 and 0 Uk2O with the dialdehydic form of the decarboxymethyl ligstroside aglycone (p-HPEA-EDA), the dialdehydic form of the decarboxymethyl oleuropein aglycone (3,4-DHPEA-EDA), the dialdehydic form of the ligstroside aglycone (p-HPEA-EDA-DLA), and with fruity, pungent, and bitter attributes. Precision agronomic management based on the needs of the crop itself would avoid unnecessary stress on olive trees and oil damage.

Gradual Changes of the Protective Effect of Phenols in Virgin Olive Oils Subjected to Storage and Controlled Stress by Mesh Cell Incubation.

The oxidation reactions that take place in virgin olive oil under moderate conditions involved the combined effect of antioxidant and prooxidant compounds. Given the complexity of oxidation processes of multicomponent matrices, there is still a need to develop new methods with a dynamic approach to study the persistence of the compounds with healthy properties. This work studied the joint evolution of them, including phenols and pheophytin a, modeling their tendency during a real storage. The regression equations performed with the total phenol concentration showed that around 2% of the concentration was lost every month. Simultaneously, the progress of oxidation was evaluated by mesh cell incubation and Fourier transform infrared analysis. This method pointed out that, in the presence of light, the prooxidant effect of pigments was able to mask the protective effect of phenols, until the pheophytin a concentration was lower than 1 mg/kg. The antioxidant effect of phenols was less remarkable when the concentration loss was 35% or more.

Comparing the potential of IR-spectroscopic techniques to gas chromatography coupled to ion mobility spectrometry for classifying virgin olive oil categories.

Virgin olive oil (OO) can be classified into three different categories: extra virgin, virgin and lampante. The official method for this classification, based on physicochemical analysis and sensory tasting, is considered useful and effective, although it is a costly and time-consuming process. The aim of this study was to assess the potential of some analytical techniques for classifying and predicting different OO categories to support official methods and to provide olive oil companies with a rapid tool to assess product quality. Thus, mid and near infrared spectroscopies (MIR and NIR) have been compared by using different instruments and with head-space gas chromatography coupled to an ion mobility spectrometer (HS-GC-IMS). High classification success rates in validation models were obtained using IR spectrometers (>70% and > 80% in average for ternary and binary classifications, respectively), although HS-GC-IMS showed greater classification potential (>85% and > 90%).

Extracellular matrix protein anosmin-1 overexpression alters dopaminergic phenotype in the CNS and the PNS with no pathogenic consequences in a MPTP model of Parkinson's disease.

The development and survival of dopaminergic neurons are influenced by the fibroblast growth factor (FGF) pathway. Anosmin-1 (A1) is an extracellular matrix protein that acts as a major regulator of this signaling pathway, controlling FGF diffusion, and receptor interaction and shuttling. In particular, previous work showed that A1 overexpression results in more dopaminergic neurons in the olfactory bulb. Prompted by those intriguing results, in this study, we investigated the effects of A1 overexpression on different populations of catecholaminergic neurons in the central (CNS) and the peripheral nervous systems (PNS). We found that A1 overexpression increases the number of dopaminergic substantia nigra pars compacta (SNpc) neurons and alters the striosome/matrix organization of the striatum. Interestingly, these numerical and morphological changes in the nigrostriatal pathway of A1-mice did not confer an altered susceptibility to experimental MPTP-parkinsonism with respect to wild-type controls. Moreover, the study of the effects of A1 overexpression was extended to different dopaminergic tissues associated with the PNS, detecting a significant reduction in the number of dopaminergic chemosensitive carotid body glomus cells in A1-mice. Overall, our work shows that A1 regulates the development and survival of dopaminergic neurons in different nuclei of the mammalian nervous system.

miRNAs as biomarkers of autism spectrum disorder: a systematic review and meta-analysis.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder with complex clinical manifestations that arise between 18 and 36 months of age. Social interaction deficiencies, a restricted range of interests, and repetitive stereotyped behaviors are characteristics which are sometimes difficult to detect early. Several studies show that microRNAs (miRs/miRNAs) are strongly implicated in the development of the disorder and affect the expression of genes related to different neurological pathways involved in ASD. The present systematic review and meta-analysis addresses the current status of miRNA studies in different body fluids and the most frequently dysregulated miRNAs in patients with ASD. We used a combined approach to summarize miRNA fold changes in different studies using the mean values. In addition, we summarized p values for differential miRNA expression using the Fisher method. Our literature search yielded a total of 133 relevant articles, 27 of which were selected for qualitative analysis based on the inclusion and exclusion criteria, and 16 studies evaluating miRNAs whose data were completely reported were ultimately included in the meta-analysis. The most frequently dysregulated miRNAs across the analyzed studies were miR-451a, miR-144-3p, miR-23b, miR-106b, miR150-5p, miR320a, miR92a-2-5p, and miR486-3p. Among the most dysregulated miRNAs in individuals with ASD, miR-451a is the most relevant to clinical practice and is associated with impaired social interaction. Other miRNAs, including miR19a-3p, miR-494, miR-142-3p, miR-3687, and miR-27a-3p, are differentially expressed in various tissues and body fluids of patients with ASD. Therefore, all these miRNAs can be considered candidates for ASD biomarkers. Saliva may be the optimal biological fluid for miRNA measurements, because it is easy to collect from children compared to other biological fluids.

Effect of Controlled Oxygen Supply during Crushing on Volatile and Phenol Compounds and Sensory Characteristics in Coratina and Ogliarola Virgin Olive Oils.

In virgin olive oil industries, the technological choices of the production plant affect the biochemical activities that take place in the olives being processed throughout the entire process, thereby affecting the quality of the final product. The lipoxygenase pool enzymes that operated their activity during the first phases of the process need the best conditions to work, especially concerning temperature and oxygen availability. In this study, a system was equipped to supply oxygen in the crusher at a controllable concentration in an industrial olive oil mill at pilot plant scale, and four oxygen concentrations and two cultivars, Coratina and Ogliarola, were tested. The best concentration for oxygen supply was 0.2 L/min at the working capacity of 0.64 Ton/h. Further, using this addition of oxygen, it was possible to increase the compound's concentration, which is responsible for the green, fruity aroma. The effect on volatile compounds was also confirmed by the sensory analyses. However, at the same time, it was possible to maintain the concentration of phenols in a good quality olive oil while also preserving all the antioxidant properties of the product due to the presence of phenols. This study corroborates the importance of controlling oxygen supply in the first step of the process for process management and quality improvement in virgin olive oil production.

Method for the analysis of volatile compounds in virgin olive oil by SPME-GC-MS or SPME-GC-FID.

During the course of the EU H2020 OLEUM project, a harmonized method was developed to quantify volatile markers of the aroma of virgin olive oil with the aim to support the work of sensory panel test to assess the quality grade. A peer validation of this method has been carried out, with good results in terms of analytical quality parameters. The method allows the quantification of volatile compounds by SPME-GC with two possible detectors, flame ionization detector and mass spectrometry, depending on the technical facilities of the labs applying this method. The method was optimized for the quantification of 18 volatile compounds that were selected as being markers responsible for positive attributes (e.g. fruity) and sensory defects (e.g. rancid and winey-vinegary). The quantification is carried out with calibration curves corrected by the internal standards. Additionally, a protocol is provided to prepare the calibration samples. This procedure enhances reproducibility between labs since one of the main sources of errors is the application of different procedures in calibration.

Contribution of specific volatile markers to green and ripe fruity attributes in extra virgin olive oils studied with three analytical methods.

An objective sensory evaluation of extra virgin olive oil (EVOO), involving the chemical characterization of positive attributes, is of interest. These attributes are objectively divided, according to fruitiness, into "green" and "ripe" fruity. This work studied the differentiation in the volatile profile of EVOOs into these two classes, obtained by three analytical methods, including different extraction techniques and detectors and two data processing strategies, and their relation with sensory results. According to the results, each method allowed the characterization of the two classes, providing information on different volatile compounds, which increased in number through PARADISe software (14 more than the conventional processing). Moreover, some volatile compounds showed significant differences between the two classes, 16 highlighted by the variables with importance in projection (VIP) for green fruity (e.g. (Z)-3-hexen-1-ol, methyl ether) and 23 for ripe fruity EVOOs (e.g. (Z)-2-hexen-1-ol), which could be considered as useful markers to complement quality assessment.

Upper cortical layer-driven network impairment in schizophrenia.

Schizophrenia is one of the most widespread and complex mental disorders. To characterize the impact of schizophrenia, we performed single-nucleus RNA sequencing (snRNA-seq) of >220,000 neurons from the dorsolateral prefrontal cortex of patients with schizophrenia and matched controls. In addition, >115,000 neurons were analyzed topographically by immunohistochemistry. Compositional analysis of snRNA-seq data revealed a reduction in abundance of GABAergic neurons and a concomitant increase in principal neurons, most pronounced for upper cortical layer subtypes, which was substantiated by histological analysis. Many neuronal subtypes showed extensive transcriptomic changes, the most marked in upper-layer GABAergic neurons, including down-regulation in energy metabolism and up-regulation in neurotransmission. Transcription factor network analysis demonstrated a developmental origin of transcriptomic changes. Last, Visium spatial transcriptomics further corroborated upper-layer neuron vulnerability in schizophrenia. Overall, our results point toward general network impairment within upper cortical layers as a core substrate associated with schizophrenia symptomatology.

The Impact of Mild Frost Occurring at Different Harvesting Times on the Volatile and Phenolic Composition of Virgin Olive Oil.

The organoleptic characteristics of virgin olive oil (VOO), together with its nutritional and health properties, have led the olive tree to be cultivated beyond the Mediterranean basin, reaching latitudes with colder climates, with minimum temperatures below 0 °C and with a higher probability of early frosts. The freezing of olives generates destruction within the tissues and promotes degradation reactions. In this study, the impact of mild frost occurring at different harvesting times on the composition of volatiles and phenolic compounds in VOO were investigated. Arbequina variety olives were harvested at different stages of ripening. Half of the olives were subjected to oil extraction and the other half were frozen at -3 ± 1 °C for 12 h prior to oil extraction. A significant decrease of phenolic compounds with harvesting time was observed in both types of oils (fresh and frozen olives). Oils from frozen olives presented a slightly higher content of total phenols, except in the advanced ripening stage (September), and a slightly lower content of volatile compounds at all harvesting times. In addition, a higher content of 3,4-DHPEA-EDA was observed in oils from frozen olives, which is attributed to an early action of the endogenous ß-glucosidase enzyme on oleuropein in freeze-damaged olive fruits. Principal component analysis and Discriminant Partial Least Square Regression allowed the oils to be classified according to the type of fruit (fresh and frozen) and the month of harvest. This study would indicate that mild frost would have a low impact on the chemical composition of virgin olive oil, although, this depends on the ripening stage.

An International Survey on Olive Oils Quality and Traceability: Opinions from the Involved Actors.

A survey was launched to understand the current problems and sensitivities of the olive oil market through a series of questions clustered around topics related to quality, traceability, regulation, standard methods and other issues. The questions were selected after a series of interviews with different actors to identify those aspects where some disagreement or different points of view may exist. These questions were grouped in topics such as geographical traceability, consumer perception and quality management. The survey was addressed to eight different olive oil actors independently: producers, retailers, importers, exporters, analysts, workers at regulatory bodies, and consumers. Approximately half of the respondents (67.0% for consumers and 56.0% for the rest of olive oil actors) claimed to understand the importance of the protected designation of origin. In fact, the traceability objectives that were selected as the most relevant were those related with geographical traceability (19.3%) followed by the detection of adulteration (15.6%). Most of the respondents (80%) would agree to share data for a common database; however, some concerns exist about the use of these data and the issue of paying to have access to this database. The respondents mostly expressed an affirmative answer concerning the efficiency of panel test (74%) and a negative answer (90%) concerning the proposal of removing from regulation, although 42% agree with their revision for improvement. The opinions on "best before" date and their relationship with quality and the willingness to apply non-targeted methods were also surveyed.

Neurogenesis of medium spiny neurons in the nucleus accumbens continues into adulthood and is enhanced by pathological pain.

In mammals, most adult neural stem cells (NSCs) are located in the ventricular-subventricular zone (V-SVZ) along the wall of the lateral ventricles and they are the source of olfactory bulb interneurons. Adult NSCs exhibit an apico-basal polarity; they harbor a short apical process and a long basal process, reminiscent of radial glia morphology. In the adult mouse brain, we detected extremely long radial glia-like fibers that originate from the anterior-ventral V-SVZ and that are directed to the ventral striatum. Interestingly, a fraction of adult V-SVZ-derived neuroblasts dispersed in close association with the radial glia-like fibers in the nucleus accumbens (NAc). Using several in vivo mouse models, we show that newborn neurons integrate into preexisting circuits in the NAc where they mature as medium spiny neurons (MSNs), i.e., a type of projection neurons formerly believed to be generated only during embryonic development. Moreover, we found that the number of newborn neurons in the NAc is dynamically regulated by persistent pain, suggesting that adult neurogenesis of MSNs is an experience-modulated process.

Monitoring Virgin Olive Oil Shelf-Life by Fluorescence Spectroscopy and Sensory Characteristics: A Multidimensional Study Carried Out under Simulated Market Conditions.

The control of virgin olive oil (VOO) freshness requires new tools that reflect the diverse chemical changes that take place during the market period. Fluorescence spectroscopy is one of the techniques that has been suggested for controlling virgin olive oil (VOO) freshness during its shelf-life. However, a complete interpretation of fluorescence spectra requires analyzing multiple parameters (chemical, physical-chemical, and sensory) to evaluate the pace of fluorescence spectral changes under moderate conditions with respect to other changes impacting on VOO quality. In this work, four VOOs were analyzed every month with excitation-emission fluorescence spectra. The same samples were characterized with the concentration of fluorophores (phenols, tocopherols, chlorophyll pigments), physical-chemical parameters (peroxide value, K232, K270, free acidity), and sensory attributes (medians of defects and of the fruity attribute). From the six components extracted with parallel factor analysis (PARAFAC), two components were assigned to chlorophyll pigments and those assigned to tocopherols, phenols, and oxidation products were selected for their ability to discriminate between fresh and aged oils. Thus, the component assigned to oxidation products correlated with K270 in the range 0.80-0.93, while the component assigned to tocopherols-phenols correlated with the fruity attribute in the range 0.52-0.90. The sensory analysis of the samples revealed that the changes of these PARAFAC components occurred at the same time as, or even before, the changes of the sensory characteristics.

Formulations of Rancid and Winey-Vinegary Artificial Olfactory Reference Materials (AORMs) for Virgin Olive Oil Sensory Evaluation.

Sensory assessment of virgin olive oil ("panel test") is the only sensory method included in international regulations of edible oils and its application is compulsory. Even if its application has been a success in quality control, improving the quality of virgin olive oils over the last 30 years, at present, there is no reference material (RM), in the strict sense of the term, to be used as a validated standard for sensory defects of virgin olive oil with which tasters can be trained. Usually, real samples of virgin olive oils assessed by many panels for the International Olive Council (IOC) ring tests are used as materials of reference in panel training and control. The latter are highly representative of the main perceived defects, but availability is limited, samples are not homogeneous year after year, and other secondary defects can be present. Thus, in order to provide solutions, this work describes an analytical procedure for implementing olfactory formulations that emulate rancid and winey-vinegary defects found in virgin olive oils with the aim of providing reproducible RMs that can be prepared on demand. A strategy for designing RMs for aroma is presented and the optimization process to obtain the best formulation is described. Under the criteria of representativeness, verified with the advice of the IOC, aroma persistence, and simplicity in formulation, two RMs for winey-vinegary and rancid were obtained by diluting acetic acid and ethanol (winey-vinegary defect) and hexanal (rancid defect) together with other compounds that are used to modify aroma and avoid non-natural sensory notes.

Supporting the Sensory Panel to Grade Virgin Olive Oils: An In-House-Validated Screening Tool by Volatile Fingerprinting and Chemometrics.

The commercial category of virgin olive oil is currently assigned on the basis of chemical-physical and sensory parameters following official methods. Considering the limited number of samples that can be analysed daily by a sensory panel, an instrumental screening tool could be supportive by reducing the assessors' workload and improving their performance. The present work aims to in-house validate a screening strategy consisting of two sequential binary partial least squares-discriminant analysis (PLS-DA) models that was suggested to be successful in a proof-of-concept study. This approach is based on the volatile fraction fingerprint obtained by HS-SPME-GC-MS from more than 300 virgin olive oils from two crop seasons graded by six different sensory panels into extra virgin, virgin or lampante categories. Uncertainty ranges were set for the binary classification models according to sensitivity and specificity by means of receiver operating characteristics (ROC) curves, aiming to identify boundary samples. Thereby, performing the screening approach, only the virgin olive oils classified as uncertain (23.3%) would be assessed by a sensory panel, while the rest would be directly classified into a given commercial category (78.9% of correct classification). The sensory panel's workload would be reduced to less than one-third of the samples. A highly reliable classification of samples would be achieved (84.0%) by combining the proposed screening tool with the reference method (panel test) for the assessment of uncertain samples.

Flash Gas Chromatography in Tandem with Chemometrics: A Rapid Screening Tool for Quality Grades of Virgin Olive Oils.

This research aims to develop a classification model based on untargeted elaboration of volatile fraction fingerprints of virgin olive oils (n = 331) analyzed by flash gas chromatography to predict the commercial category of samples (extra virgin olive oil, EVOO; virgin olive oil, VOO; lampante olive oil, LOO). The raw data related to volatile profiles were considered as independent variables, while the quality grades provided by sensory assessment were defined as a reference parameter. This data matrix was elaborated using the linear technique partial least squares-discriminant analysis (PLS-DA), applying, in sequence, two sequential classification models with two categories (EVOO vs. no-EVOO followed by VOO vs. LOO and LOO vs. no-LOO followed by VOO vs. EVOO). The results from this large set of samples provide satisfactory percentages of correctly classified samples, ranging from 72% to 85%, in external validation. This confirms the reliability of this approach in rapid screening of quality grades and that it represents a valid solution for supporting sensory panels, increasing the efficiency of the controls, and also applicable to the industrial sector.

An HS-GC-IMS Method for the Quality Classification of Virgin Olive Oils as Screening Support for the Panel Test.

Sensory evaluation, carried out by panel tests, is essential for quality classification of virgin olive oils (VOOs), but is time consuming and costly when many samples need to be assessed; sensory evaluation could be assisted by the application of screening methods. Rapid instrumental methods based on the analysis of volatile molecules might be considered interesting to assist the panel test through fast pre-classification of samples with a known level of probability, thus increasing the efficiency of quality control. With this objective, a headspace gas chromatography-ion mobility spectrometer (HS-GC-IMS) was used to analyze 198 commercial VOOs (extra virgin, virgin and lampante) by a semi-targeted approach. Different partial least squares-discriminant analysis (PLS-DA) chemometric models were then built by data matrices composed of 15 volatile compounds, which were previously selected as markers: a first approach was proposed to classify samples according to their quality grade and a second based on the presence of sensory defects. The performance (intra-day and inter-day repeatability, linearity) of the method was evaluated. The average percentages of correctly classified samples obtained from the two models were satisfactory, namely 77% (prediction of the quality grades) and 64% (prediction of the presence of three defects) in external validation, thus demonstrating that this easy-to-use screening instrumental approach is promising to support the work carried out by panel tests.

Tracking Sensory Characteristics of Virgin Olive Oils During Storage: Interpretation of Their Changes from a Multiparametric Perspective.

Virgin olive oil is inevitably subject to an oxidation process during storage that can affect its stability and quality due to off-flavors that develop before the oil surpasses its 'best before' date. Many parameters are involved in the oxidation process at moderate conditions. Therefore, a multiparametric study is necessary to establish a link between physico-chemical changes and sensory quality degradation in a real storage experiment. In this context, a storage experiment of 27 months was performed for four monovarietal virgin olive oils, bottled in transparent 500-mL PET bottles and subjected to conditions close to a supermarket scenario. Volatile composition, quality parameters and phenolic compounds were determined monthly. Simultaneously, an accredited sensory panel assessed their sensory characteristics. The stability of the fresh samples was also studied with the oxidative stability index (OSI) and mesh cell-FTIR. (E)-2-hexenal, (Z)-3-hexen-1-ol and (E)-2-hexen-1-ol were identified as markers of the fruity attribute. Hexanal and nonanal were also identified as compounds that were associated with the rise of median of defect during storage. Some disagreements were observed between the sensory assessment and the OSI analyzed by Rancimat. However, the increase of concentration of rancid markers agreed with the increase of aldehyde band measured with mesh cell-FTIR.

Alignment and Proficiency of Virgin Olive Oil Sensory Panels: The OLEUM Approach.

A set of 334 commercial virgin olive oil (VOO) samples were evaluated by six sensory panels during the H2020 OLEUM project. Sensory data were elaborated with two main objectives: (i) to classify and characterize samples in order to use them for possible correlations with physical-chemical data and (ii) to monitor and improve the performance of panels. After revision of the IOC guidelines in 2018, this work represents the first published attempt to verify some of the recommended quality control tools to increase harmonization among panels. Specifically, a new "decision tree" scheme was developed, and some IOC quality control procedures were applied. The adoption of these tools allowed for reliable classification of 289 of 334 VOOs; for the remaining 45, misalignments between panels of first (on the category, 21 cases) or second type (on the main perceived defect, 24 cases) occurred. In these cases, a "formative reassessment" was necessary. At the end, 329 of 334 VOOs (98.5%) were classified, thus confirming the effectiveness of this approach to achieve a better proficiency. The panels showed good performance, but the need to adopt new reference materials that are stable and reproducible to improve the panel's skills and agreement also emerged.