In-House Validation of Four Duplex Droplet Digital PCR Assays to Quantify GM Soybean Events.

Due to the increasing number of authorized events in the European Union, it is crucial for the official laboratories to enforce market control to detect and quantify genetically modified organisms. In this study, an in-house validation of quantitative duplex ddPCR methods was performed involving MON87701, MON87769, MON89788 and CV-127-9 assays with respect to the lectin reference gene. Since the ddPCR methods provide accurate quantification, show less sensitivity to PCR inhibitors and are more suitable for multiplexing compared to the real-time PCR, the optimization of the existing assays was performed with the exception of MON87701, according to the JRC Guidance documents and technical reports.

Comparative assessment of three commercial kits and in house optimized PCR assays for GMO screening in food and feed.

Screening strategies for GMO detection in food and feed are a crucial aspect in GMO testing laboratories for streamlining the analytical workflow and reducing turnaround time and costs. These strategies can be more or less complex or even be targeted according to the ingredients in the product, but whatever the choice, a good basic approach is generally based on the search for 35S promoter (P35S), nos-terminator (T-nos) and FMV promoter (P-FMV). In this study, we compare the singleplex real time PCR method for P35S, T-nos and P-FMV detection currently adopted by the Italian National Reference Laboratory for GM food and feed (NRL) with three commercial kits available on the market for giving a greater choice to consider the best approach suitable to the official control laboratories that are different from each other.

Tick salivary proteome and lipidome with low glycan content correlate with allergic type reactions in the zebrafish model.

Ticks, as hematophagous ectoparasites, can manipulate host immune and metabolic processes, causing tick-borne allergies such as α-Gal syndrome (AGS). Glycolipids with bound galactose-alpha-1-3-galactose (α-Gal) are potential allergenic molecules associated with AGS. Nevertheless, proteins and lipids lacking α-Gal modifications may contribute to tick salivary allergies and be linked to AGS.

Protein hydrolysates enhance recovery from drought stress in tomato plants: phenomic and metabolomic insights.

Introduction: High-throughput phenotyping technologies together with metabolomics analysis can speed up the development of highly efficient and effective biostimulants for enhancing crop tolerance to drought stress. The aim of this study was to examine the morphophysiological and metabolic changes in tomato plants foliarly treated with two protein hydrolysates obtained by enzymatic hydrolysis of vegetal proteins from Malvaceae (PH1) or Fabaceae (PH2) in comparison with a control treatment, as well as to investigate the mechanisms involved in the enhancement of plant resistance to repeated drought stress cycles.

Methods: A phenotyping device was used for daily monitoring morphophysiological traits while untargeted metabolomics analysis was carried out in leaves of the best performing treatment based on phenotypic results.

Plant biostimulants as natural alternatives to synthetic auxins in strawberry production: physiological and metabolic insights.

The demand for high-quality strawberries continues to grow, emphasizing the need for innovative agricultural practices to enhance both yield and fruit quality. In this context, the utilization of natural products, such as biostimulants, has emerged as a promising avenue for improving strawberry production while aligning with sustainable and eco-friendly agricultural approaches. This study explores the influence of a bacterial filtrate (BF), a vegetal-derived protein hydrolysate (PH), and a standard synthetic auxin (SA) on strawberry, investigating their effects on yield, fruit quality, mineral composition and metabolomics of leaves and fruits.

Arthroscopic Treatment of a Subchondral Bone Cyst via Stem Cells Application: A Case Study in Equine Model and Outcomes.

Subchondral bone cysts in horses represent one of the main causes of lameness that can occur in different anatomical locations. The study describes the treatment in regenerative therapy of the intracystic implantation of adipose tissue mesenchymal stromal cells (AMSCs) included in platelet-rich plasma (PRP). The ability of AMSCs to differentiate in osteogenic cells was tested in vitro and in vivo.

Shedding light on human olfaction: Electrophysiological recordings from sensory neurons in acute slices of olfactory epithelium.

The COVID-19 pandemic brought attention to our limited understanding of human olfactory physiology. While the cellular composition of the human olfactory epithelium is similar to that of other vertebrates, its functional properties are largely unknown. We prepared acute slices of human olfactory epithelium from nasal biopsies and used the whole-cell patch-clamp technique to record electrical properties of cells.

Recovery from olfactory and gustatory dysfunction following COVID-19 acquired during Omicron BA.1 wave in Italy.

Background: Despite alterations in the sense of smell and taste have dominated the symptoms of SARS-CoV-2 infection, the prevalence and the severity of self-reporting COVID-19 associated olfactory and gustatory dysfunction has dropped significantly with the advent of the Omicron BA.1 subvariant. However, data on the evolution of Omicron-related chemosensory impairment are still lacking.

Optimization of the growth conditions through response surface methodology and metabolomics for maximizing the auxin production by C1.

Introduction: The fermentative production of auxin/indole 3-acetate (IAA) using selected strains can be a promising approach to developing novel plant biostimulants for agriculture use.

Methods: By integrating metabolomics and fermentation technologies, this study aimed to define the optimal culture conditions to obtain auxin/IAA-enriched plant postbiotics using strain C1. Metabolomics analysis allowed us to demonstrate that the production of a selected.

The Functional Profile and Antioxidant Capacity of Tomato Fruits Are Modulated by the Interaction between Microbial Biostimulants, Soil Properties, and Soil Nitrogen Status.

The application of microbial biostimulants to plants has revealed positive effects related to nutrients uptake, stress tolerance, root development and phenological growth. However, little information is available exploiting the potential synergistic biostimulant action of microbes on the functional quality of the yields. The current research elucidated the effect of single or coupled action of biostimulants, associated with either optimal or reduced nitrogen application, on the functional quality of tomato fruits.

A response surface methodology approach to improve nitrogen use efficiency in maize by an optimal mycorrhiza-to- co-inoculation rate.

Co-inoculation of arbuscular mycorrhizal fungi (AMF) and bacteria can synergically and potentially increase nitrogen use efficiency (NUE) in plants, thus, reducing nitrogen (N) fertilizers use and their environmental impact. However, limited research is available on AMF-bacteria interaction, and the definition of synergisms or antagonistic effects is unexplored. In this study, we adopted a response surface methodology (RSM) to assess the optimal combination of AMF () and (a PGPR-plant growth promoting rhizobacteria) formulations to maximize agronomical and chemical parameters linked to N utilization in maize ( L.

The Hierarchical Contribution of Organic vs. Conventional Farming, Cultivar, and Terroir on Untargeted Metabolomics Phytochemical Profile and Functional Traits of Tomato Fruits.

In this work, the impact of terroir, cultivar, seasonality, and farming systems on functional traits of tomato was hierarchically investigated. Untargeted metabolomics, antioxidant capacity, colorimetric assays, and enzyme inhibition were determined. The total phenolic and carotenoid contents significantly varied between growing years, whereas an interaction between the farming system and growing year ( < 0.

Nitrogen use efficiency, rhizosphere bacterial community, and root metabolome reprogramming due to maize seed treatment with microbial biostimulants.

Seed inoculation with beneficial microorganisms has gained importance as it has been proven to show biostimulant activity in plants, especially in terms of abiotic/biotic stress tolerance and plant growth promotion, representing a sustainable way to ensure yield stability under low input sustainable agriculture. Nevertheless, limited knowledge is available concerning the molecular and physiological processes underlying the root-inoculant symbiosis or plant response at the root system level. Our work aimed to integrate the interrelationship between agronomic traits, rhizosphere microbial population and metabolic processes in roots, following seed treatment with either arbuscular mycorrhizal fungi (AMF) or Plant Growth-Promoting Rhizobacteria (PGPR).

Changes of Milk Metabolomic Profiles Resulting from a Mycotoxins-Contaminated Corn Silage Intake by Dairy Cows.

In this study, an untargeted metabolomics approach based on ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS) was used for investigating changes in chemical profiles of cow milk considering diets based on mycotoxins-contaminated corn silages. For this purpose, 45 milk samples were classified into five clusters according to the corn silage contamination profile, namely (1) low levels of - and -mycotoxins; (2) low levels of fumonisins and other -mycotoxins; (3) high levels of -mycotoxins; (4) high levels of non-regulated -mycotoxins; (5) high levels of fumonisins and their metabolites, and subsequently analyzed by UHPLC-HRMS followed by a multivariate statistical analysis (both unsupervised and supervised statistical approaches). Overall, the milk metabolomic profile highlighted potential correlations between the quality of contaminated corn silages (as part of the total mixed ration) and milk composition.

An Endophytic Fungi-Based Biostimulant Modulates Volatile and Non-Volatile Secondary Metabolites and Yield of Greenhouse Basil ( L.) through Variable Mechanisms Dependent on Salinity Stress Level.

Salinity in water and soil is one of the major environmental factors limiting the productivity of agronomic and horticultural crops. In basil ( L., Lamiaceae) and other species, information on the plant response to mild salinity levels, often induced by the irrigation or fertigation systems, is scarce.

Plant-Based Biostimulant as Sustainable Alternative to Synthetic Growth Regulators in Two Sweet Cherry Cultivars.

Sweet cherry is a high value crop and the economic success of its cultivation depends not only on yield but also on fruit visual and nutritional quality attributes that influence consumer acceptability, as well as on fruit post-harvest performance and resistance to cracking. During the last few decades, cherry growers have tried to achieve these goals through exogenous applications of synthetic plant hormones and/or nutrients, but there is growing concern about the sustainability of the extensive use of these compounds in agriculture. For this reason, there is increasing interest in the possible adoption of different classes of biostimulants as sustainable alternatives to plant growth regulators.

A Microbial-Based Biostimulant Enhances Sweet Pepper Performance by Metabolic Reprogramming of Phytohormone Profile and Secondary Metabolism.

Microbial-based biostimulants can improve crop productivity by modulating cell metabolic pathways including hormonal balance. However, little is known about the microbial-mediated molecular changes causing yield increase. The present study elucidates the metabolomic modulation occurring in pepper ( L.

A Genetic and Metabolomic Perspective on the Production of Indole-3-Acetic Acid by and Use of Their Metabolites as Biostimulants in Plant Nurseries.

The species includes strains that are agronomically relevant for their growth-promoting or biocontrol traits. Molecular analysis demonstrated that the IPDC pathway involved in the conversion of tryptophan (Trp) to indole-3-acetic acid (IAA) is highly conserved among strains at both gene and protein levels. Results also indicated that the promoter region controlling the inducible expression of gene differs from the model system , which is in accordance with the observation that accumulates higher levels of IAA when cells are collected in the exponential phase of growth.

A lipidome atlas in MS-DIAL 4.

We present Mass Spectrometry-Data Independent Analysis software version 4 (MS-DIAL 4), a comprehensive lipidome atlas with retention time, collision cross-section and tandem mass spectrometry information. We formulated mass spectral fragmentations of lipids across 117 lipid subclasses and included ion mobility tandem mass spectrometry. Using human, murine, algal and plant biological samples, we annotated and semiquantified 8,051 lipids using MS-DIAL 4 with a 1-2% estimated false discovery rate.

Metabolomic Responses of Maize Shoots and Roots Elicited by Combinatorial Seed Treatments With Microbial and Non-microbial Biostimulants.

Microbial and non-microbial plant biostimulants have been successfully used to improve agriculture productivity in a more sustainable manner. Since the mode of action of biostimulants is still largely unknown, the present work aimed at elucidating the morpho-physiological and metabolomic changes occurring in maize ( L.) leaves and roots following seed treatment with (i) a consortium of two beneficial fungi [arbuscular mycorrhizal fungi (AMF) and TK7] and rhizobacteria, (ii) a protein hydrolyzate-based biostimulant (PH) alone, or (iii) in combination with a consortium of TK7 and rhizobacteria.

Retip: Retention Time Prediction for Compound Annotation in Untargeted Metabolomics.

Unidentified peaks remain a major problem in untargeted metabolomics by LC-MS/MS. Confidence in peak annotations increases by combining MS/MS matching and retention time. We here show how retention times can be predicted from molecular structures.

Development and comparative study of a pat/bar real-time PCR assay for integrating the screening strategy of a GMO testing laboratory.

Background: The number and variety of genetically modified organisms (GMOs) used globally for the production of food and feed, and potentially circulating in the European Union (EU), is constantly increasing. This implies an additional effort for the EU enforcement laboratories to optimize available resources, to contain costs and time. A well established approach for streamlining the analytical workflow is the introduction of a screening step, typically based on a smart set of real-time polymerase chain reaction (PCR) screening methods.

A Combined Phenotypic and Metabolomic Approach for Elucidating the Biostimulant Action of a Plant-Derived Protein Hydrolysate on Tomato Grown Under Limited Water Availability.

Plant-derived protein hydrolysates (PHs) are an important category of biostimulants able to increase plant growth and crop yield especially under environmental stress conditions. PHs can be applied as foliar spray or soil drench. Foliar spray is generally applied to achieve a relatively short-term response, whereas soil drench is used when a long-term effect is desired.

Topical Steroids in Rhinosinusitis and Intraoperative Bleeding: More Harm Than Good?

The aim of this study was to assess whether the chronic preoperative administration of intranasal corticosteroids (INCs) in chronic rhinosinusitis with/without nasal polyposis (CRSwNP/CRSsNP, respectively) could significantly influence bleeding during functional endoscopic sinus surgery (FESS). We prospectively enrolled 109 patients (56 CRSwNP and 53 CRSsNP) candidate for FESS who underwent clinical evaluation and anamnestic data collection. They were allocated to 2 groups depending on whether or not they were chronic INC users, as declared at their first medical evaluation: chronic "INC users" represented the treated group, while "INC nonusers" formed the control group.