Cistus incanus: a natural source of antimicrobial metabolites.

The discovery of natural molecules with antimicrobial properties has become an urgent need for the global treatment of bacterium and virus infections. Cistus incanus, a Mediterranean shrub species, represents a valuable source of phytochemicals with an interesting wide-spectrum antimicrobial potential. In this study, we analysed the spectrum of molecules composing a commercial hydroalcoholic extract of C. incanus finding ellagitannins as the most abundant. The effect of the extract and its main constituents (gallic acid, ellagic acid and punicalin) was assessed as co-treatment during viral (HSV-1, HCoV-229E, SARS-CoV-2) and bacterial infection (Staphylococcus aureus and Escherichia coli) of cells and as pre-treatment before virus infections. The results indicated a remarkable antiviral activity of punicalin against SARS-CoV-2 by pre-treating both the viral and the host cells, and a major sensitivity of S. aureus to the C. incanus extract compared to E. coli. The present study highlights broad antimicrobial potential of C. incanus extract.

Exploring Influence of Production Area and Harvest Time on Specialized Metabolite Content of Glycyrrhiza glabra Leaves and Evaluation of Antioxidant and Anti-Aging Properties.

Calabrian Glycyrrhiza glabra is one of the most appreciated licorice varieties worldwide, and its leaves are emerging as a valuable source of bioactive compounds. Nevertheless, this biomass is usually discarded, and its valorization could contribute to boost the economic value of the licorice production chain. In this study, the effects of production area and harvest time on the specialized metabolite content of G. glabra leaves (GGL) and also the antioxidant and anti-aging properties are evaluated to explore the potential of this untapped resource and to select the most optimal harvesting practices. GGL exhibited high levels of specialized metabolites (4-30 g/100 g of dry leaf) and the most abundant ones are pinocembrin, prenylated flavanones (licoflavanone and glabranin), and prenylated dihydrostilbenes. Their levels and antioxidant capacity in extracts are influenced by both production area and harvest time, showing a decisive role on specialized metabolites accumulation. Interestingly, GGL extracts strongly attenuate the toxicity of α-synuclein, the intracellular reactive oxygen species (ROS) content, and cellular senescence on Saccharomyces cerevisiae expressing human α-synuclein model, showing great potential to prevent aging and age-related disorders. These results provide insights into the phytochemical dynamics of GGL, identifying the best harvesting site and period to obtain bioactive-rich sources with potential uses in the food, nutraceutical, and pharmaceutical sectors.

Upadacitinib effectiveness and factors associated with minimal disease activity achievement in patients with psoriatic arthritis: preliminary data of a real-life multicenter study.

BACKGROUND: Upadacitinib (UPA) is a selective JAK inhibitor recently approved for the treatment of psoriatic arthritis (PsA). In this post-approval study, we aimed to evaluate the effectiveness and safety of UPA over 24 weeks and identify clinical predictors of response, in a multicentric cohort of patients affected by PsA. METHODS: One hundred and twenty-six patients with PsA treated with UPA were enrolled in 10 Italian centres. UPA effectiveness outcomes, such as the proportion of patients with MDA status, DAPSA remission, and low disease activity, ASDAS-CRP inactive and low disease activity, and change from baseline in DAPSA and ASDAS-CRP scores, were evaluated every 12 weeks until week 24. The proportion of DAPSA minor, moderate, and major improvement, and ASDAS clinically important improvement (CII) and major improvement (MI) were considered as well. All treatment-related adverse events were collected during the observation period. Clinical predictors of MDA response at week 24 were evaluated through multivariate analysis. RESULTS: At baseline, 124/126 (98%) and 54/126 (43%) patients showed peripheral and axial involvement, respectively; 110 (87%) patients were intolerant or resistant to biologic DMARDs.  At 24 weeks, MDA status, DAPSA remission, and ASDAS-CRP inactive disease were achieved in 47%, 23%, and 48% of patients, respectively. Minor, moderate, and major DAPSA improvement was observed in 67%, 39%, and 23%, respectively; while 65% and 35% achieved ASDAS-CRP CII and MI, respectively. The mean change from baseline was 15.9 ± 13.5 (p < 0.001) for DAPSA and 1.21 ± 0.97 (p < 0.001) for ASDAS-CRP. Thirteen patients (10%) discontinued UPA due to a lack of efficacy or non-serious adverse events. No serious adverse events were observed. Male gender (OR 2.54, 95% CI 1.03-6.25 p = 0.043), being naïve to biological DMARDs (OR 4.13, 95% CI 1.34-12.71, p = 0.013) and elevated baseline CRP (OR 2.49, 95% CI 1.02-6.12, p = 0.046) were associated with MDA response at week 24. CONCLUSIONS: This is one of the first real-life studies supporting the effectiveness of UPA and its safety profile in PsA patients. Furthermore, the study identifies predictors of MDA response to UPA treatment at 6 months.

A multi-analyte screening method for the rapid detection of illicit adulterants in dietary supplements using a portable SERS analyzer.

The popularity and number of dietary supplements on the health market have experienced an unprecedented boost in recent years. Simultaneously, their increased use has been accompanied by an increase in acute intoxication cases linked to the adulteration of these products with illicit and undeclared substances. In this study, a SERS-based screening methodology was developed to rapidly detect illegally added pharmaceutically active substances to dietary supplements. A portable analyzer and silver printed-SERS substrates were used to enhance the signal, requiring less than 20 min of sample preparation prior to the analysis. The method was successful in the qualitative identification of eleven out of twenty-three illicit adulterants in the dietary supplements; it could detect the target compounds at realistic adulteration levels (0.1-5.0% w/w), demonstrating the potential of SERS-based methodologies for forensic rapid screening applications. The developed method is quick, easy to use, requires no skilled technicians and little sample preparation, and allows in-situ analyses. For these reasons, it is suitable for quick screening to be performed by inspectors at customs. Moreover, the low specificity of spectroscopic methods, to which SERS belongs, would benefit the detection of newly synthesized analogues of the target adulterants, which would otherwise be more difficult using common mass spectrometry methods in absence of reference standards.

Phytochemical Extracts of Dittrichia viscosa (L.) Greuter from Agroecological Systems: Seed Antigerminative Properties and Effectiveness in Counteracting Alternaria Leaf Spot Disease on Baby-Leaf Spinach.

Dittrichia viscosa (L.) Greuter subsp. viscosa (Asteraceae) is a perennial species naturally distributed in arid and marginal areas whose agroecological cultivation could be a useful innovation to produce quality biomass to extract phenolic-rich phytochemical blends. Here, biomass-yield trends were profiled at different growth stages under direct cropping, and inflorescences, leaves, and stems were submitted to water extraction and hydrodistillation. Then, four extracts were investigated for their biological activities in invitro and in planta assays. Extracts inhibited cress (Lepidium sativum)- and radish (Raphanus sativus)-seed germination and root elongation. All samples showed dose-dependent antifungal activity in the plate experiments, inhibiting up to 65% of the growth of the fungal pathogen Alternaria alternata, a leaf-spot disease agent of baby spinach (Spinacea oleracea). However, only the extracts from dried green parts and fresh inflorescences at the highest concentration significantly reduced (54%) the extent of Alternaria necrosis on baby spinach. UHPLC-HRMS/MS analysis revealed that the main specialized metabolites of the extracts are caffeoyl quinic acids, methoxylated flavonoids, sesquiterpene compounds such as tomentosin, and dicarboxylic acids, which may explain the observed bioactivity. Plant extracts obtained through sustainable methodology can be effective in biological agricultural applications.

Target screening method for the quantitative determination of 118 pyrrolizidine alkaloids in food supplements, herbal infusions, honey and teas by liquid chromatography coupled to quadrupole orbitrap mass spectrometry.

An analytical procedure for the screening of 118 pyrrolizidine alkaloids (PAs) was successfully validated and applied to their quantitative determination in food supplements, herbal infusions, honey, and teas. It provides the reliable analyte identification by high-resolution tandem mass spectrometry (HRMS/MS), the accurate determination of 21 regulated PAs, and broad contamination profiles. 10% of 281 analyzed samples resulted contaminated at levels above the maximum levels (MLs) of European legislation. The contamination of herbal infusions of mixed plants can represent a possible health concern (23%; mean of PA sum above ML). A high number of PAs not included in the regulation was detected in honey and herbal food supplements, but their contribution was only relevant to the overall level in honey. The results indicate the need to continue collecting contamination data in food supplements and infusions of mixed herbs and to expand the PA-pool to be monitored in honey and related products.

Ultrasonographic non-radiographic erosions could predict the efficacy of belimumab in articular systemic lupus erythematosus.

The aim of this study is to characterise lupus-related arthritis and assess if the presence of ultrasound-detected erosions could be associated with belimumab in the treatment of systemic lupus erythematosus (SLE) articular manifestations. We performed a spontaneous, monocentric, retrospective, and observational study. We enrolled patients affected by SLE with articular involvement treated with belimumab. We excluded patients with positive rheumatoid factor (RF) or anti-citrullinated peptide antibody (ACPA), Jaccoud's arthropathy, and radiographic erosions. Patients were assessed at baseline, 3, and 6 months. We collected laboratory and clinical data from electronic records. Joint disease activity was assessed using disease activity score on 28 joints based on C-reactive protein (DAS28-CRP), swollen and tender joints count. All patients underwent an ultrasound examination of the wrist, metacarpophalangeal, proximal interphalangeal, and metatarsal-phalangeal joints before the initiation of treatment with belimumab. We performed Student's T-test and Mann-Whitney's U-test to assess the difference between means and Fisher's exact test to assess difference in proportions, and linear univariate regression to investigate predictors of disease activity. We enrolled 23 patients (female 82.6%, mean age of 50.65 ± 14.1 years). Seven patients (30.4%) presented bone erosions at baseline. Patients with bone erosions were generally older (61 ± 16.1 vs 46.13 ± 10.7 years, p = 0.016), more frequently male (42.8 vs 6.2%, p = 0.03), with higher baseline CRP levels (10.29 ± 11.6 vs 2.25 ± 3.1 mg/L, p = 0.015) and C4 levels (0.19 ± 0.17 vs 0.1 ± 0.04 g/L, p = 0.05). After 6 months of treatment with belimumab, patients without erosions improved their DAS28-CRP significantly (2.95 ± 0.89 vs 2.26 ± 0.48, p = 0.01), while patients with erosions did not (3.6 ± 0.79 vs 3.2 ± 0.95, p = 0.413). DAS28-CRP did not differ between the two groups at baseline, while it was significantly lower at the other two time points in patients without erosions. The majority of patients achieved remission at 6 months follow-up based on DAS28-CRP criteria (73.9%), with a significant difference between patients with and without erosions (42.8 vs 87.5%, p = 0.045). The presence of articular ultrasound-detected erosions could be predictive of a decreased efficacy of belimumab in the articular manifestations of SLE. A possible explanation is a rheumatoid-like articular phenotype, despite the lack of ACPA-positivity and radiologic erosions. However, due to the small sample population, larger cohorts are needed to assess the possible predictive role of this finding.

Coping strategies, emotional distress and perceived disease severity in a cohort of patients with rheumatoid arthritis: a mediation analysis.

OBJECTIVES: Few data are available on the role of emotional distress as a possible mediator of the relationship between coping strategies and the Patient Global Assessment (PGA). This study aims to investigate, in a large cohort of patients affected by RA, the relationship between specific copying strategies and PGA, and the role of emotional distress as a mediator. METHODS: A total of 490 patients with RA completed a set of standardized assessments including the self-reported PGA, the Coping Orientation to the Problems Experienced (COPE-NVI) and the Hospital Anxiety and Depression Scale (HADS). A mediation analysis was conducted to investigate the role of emotional distress. RESULTS: The effect of coping strategies on the PGA score was significantly mediated by the emotional distress for religious (total effect mediated 42.0%), planning (total effect mediated 17.5%), behavioural disengagement (total effect mediated 10.5%), and focus on and venting of emotions (total effect mediated 9.8%). Seven coping strategies (acceptance, positive reinterpretation and growth, active coping, denial, humour, substance use-mental disengagement) resulted directly associated to PGA total score, but no mediation effect was found. The remaining four coping strategies were not associated to the PGA score. CONCLUSION: This study suggests that coping strategies could be an important factor in the perceived disease severity. Consequently, in order to reduce PGA in patients with RA, a useful tool could be represented by the implementation of psychological interventions aiming to modify the specific coping styles. Moreover, to prevent or treat emotional distress seems to further reduce PGA.

An analytical platform for the screening and identification of pyrrolizidine alkaloids in food matrices with high risk of contamination.

An analytical platform for the detection of pyrrolizidine alkaloids (PAs) in honey, pollen, teas, herbal infusions, and dietary supplements is proposed; it includes a wide-scope suspect screening method, based on a diagnostic product ion filtering strategy for the characterization of PAs, and a target screening and identification method for the high-throughput detection of 118 PAs of a high-resolution mass spectral library. Salting-out assisted liquid-liquid extraction of aqueous extracts combined to ultra-high performance liquid chromatography-high-resolution tandem mass spectrometry was employed. The limit of identification (0.6-30 µg kg-1) of 28 standards were fit-for-purpose in PA-monitoring applications, with a false negative rate <1.3 % at 4 µg L-1. The wide-scope suspect screening method allowed the tentative identification of 88 compounds. The screening of 282 commercial samples revealed a broad contamination of the studied matrices, demonstrating the effectiveness of the platform in detecting and identifying both target and untarget PAs.

Chemical characterization and antioxidant potential of ecuadorian propolis.

The chemical composition and the antioxidant potential of Ecuadorian propolis samples (n = 19) collected in different provinces were investigated. HPLC-DAD-ESI/MSn and GC-EI-MS analysis of the methanol extracts enabled us to define six types of Ecuadorian propolis based on their secondary metabolite composition. 68 compounds were identified, 59 of which are reported for the first time in Ecuadorian propolis. The detected compounds include flavonoids, diterpenes, triterpenes, organic acid derivatives, alkylresorcinol derivatives and nemorosone. Plants belonging to genera Populus, Mangifera and Clusia seemed to be vegetable sources employed by bees to produce Ecuadorian propolis. Total phenolic content and antioxidant activity of propolis extracts were determined by the Folin-Ciocalteu assay and 2,2-diphenyl-1-picrylhydrazyl and ferric reducing/antioxidant potential assays, respectively. As expected, the variable chemical composition affected the differences in terms of antioxidant potential.

Green non-conventional techniques for the extraction of polyphenols from agricultural food by-products: A review.

Food processing industry is accompanied with the generation of a great production of wastes and by-products exceptionally rich in bioactive compounds (especially phenolics), with antioxidant activity. The recovery of these health molecules constitutes a key point for the valorization of by-products, with the possibility of creating new ingredients to be used for the formulation of food and cosmetic products. One of the main limitations to reuse by-products is linked to the high cost to obtain bioactive compounds, consequently in order to exploit these resources commercially valuable it is necessary to develop innovative, economic and environmentally friendly extraction strategies. These extraction methods should be able to reduce petroleum solvents, energy consumption and chemical wastes, protecting both environment and consumers and ensuring safe and high-quality final products. The purpose of this review is to summarize current knowledge and applications of the new extraction techniques such as supercritical fluid extraction, pressurized liquid extraction, ultrasound assisted extraction applied to polyphenols extraction from agricultural food by-products. Particular attention has been paid to theoretical background, highlighting mechanisms and safety precautions. Authors concluded that relevant results of these techniques represent an opportunity to industrial scale-up, improving the extraction yields, minimizing time, costs and environmental impact.

Onion Peel: Turning a Food Waste into a Resource.

Food waste is a serious problem for food processing industries, especially when it represents a loss of a valuable source of nutrients and phytochemicals. Increasing consumer demand for processed food poses the problem of minimizing waste by conversion into useful products. In this regard, onion (Allium cepa) waste consisting mainly of onion skin is rich in bioactive phenolic compounds. Here, we characterized the flavonoid profiles and biological activities of onion skin wastes of two traditional varieties with protected geographical indication (PGI), the red "Rossa di Tropea" and the coppery "Ramata di Montoro", typically cultivated in a niche area in southern Italy. The phytochemical profiles of exhaustive extracts, characterized by ultra-high-performance liquid chromatography coupled with ultraviolet (UV) detection and high-resolution mass spectrometry, revealed that flavonols and anthocyanins were the characteristic metabolite classes of onion skins. Quercetin, quercetin glucosides and their dimer and trimer derivatives, and, among anthocyanins, cyanidin 3-glucoside, were the most abundant bioactive compounds. The potential of onion skins was evaluated by testing several biological activities: ABTS/oxygen radical absorbance capacity (ORAC) and in vitro alpha-glucosidase assays were performed to evaluate the antioxidant and anti-diabetic properties of the extracts and of their main compounds, respectively, and proliferative activity was evaluated by MTT assay on human fibroblasts. In the present study, by observing various biological properties of "Rossa di Tropea" and "Ramata di Montoro" onion-dried skins, we clearly indicated that this agricultural waste can provide bioactive molecules for multiple applications, from industrial to nutraceutical and cosmetical sectors.

Nigerian propolis: chemical composition, antioxidant activity and α-amylase and α-glucosidase inhibition.

Propolis is an attractive natural ingredient to design health products due to its pharmacological effects. Our chemical investigation of a polar extract of Nigerian propolis (NP) led the isolation and identification of five isoflavonoids (1-4, 6), one diarylpropane (5) and one prenylated flavanone (7) by the combination of chromatographic and spectroscopic techniques. Compounds 1, 4 and 7 were found to be the main markers in NP (8.0, 5.0 and 4.0 mg/g of dry extract, respectively). Moreover, NP and its phenolic constituents exhibited in vitro free radical scavenging activity together with a promising antidiabetic effect against α-amylase and α-glucosidase enzymes. Finally, NP showed also a moderate inhibition of Helicobacter pylori growth. These results suggested that NP could be a good candidate in nutraceuticals and food products.

Physiological, Biochemical, and Metabolic Responses to Short and Prolonged Saline Stress in Two Cultivated Cardoon Genotypes.

Cultivated cardoon is a multipurpose crop with adaptability to limiting environments. Two genotypes ("Bianco Avorio" and "Spagnolo") were comparatively characterized in response to short and prolonged 100 mM NaCl stress in hydroponics. Salt induced no growth variations between genotypes or symptoms of NaCl toxicity, but boosted ABA accumulation in roots and leaves. Both genotypes had high constitutive phenol content, whose major components were depleted upon 2 days of stress only in "Bianco Avorio". Prolonged stress stimulated accumulation of proline, phenylpropanoids, and related transcripts, and non-enzymatic antioxidant activity. Decreased antioxidant enzymes activities upon short stress did not occur for APX in "Spagnolo", indicating a stronger impairment of enzymatic defenses in "Bianco Avorio". Nonetheless, H2O2 and lipid peroxidation did not increase under short and prolonged stress in both genotypes. Overall, the two genotypes appear to share similar defense mechanisms but, in the short term, "Bianco Avorio" depends mainly on non-enzymatic antioxidant phenylpropanoids for ROS scavenging, while "Spagnolo" maintains a larger arsenal of defenses. Upon prolonged stress, proline could have contributed to protection of metabolic functions in both genotypes. Our results provide cues that can be exploited for cardoon genetic improvement and highlight genotypic differences for breeding salinity tolerant varieties.

Determination of mycotoxins in beer by multi heart-cutting two-dimensional liquid chromatography tandem mass spectrometry method.

Beer is one the most consumed alcoholic beverage in the world and its contamination with mycotoxins is of public health concern. This study reports a fast and automated analytical procedure based on a multi-heart-cutting two-dimensional liquid chromatography tandem mass spectrometry method using electrospray ionization for the determination of seven mycotoxins (aflatoxins B1, B2, G2 and G1, ochratoxin A, fumonisins B1 and B2) in beers. The developed method was based on the heart-cutting 2D- HPLC technique in which only the specific portions of the first dimension, in the retention time of analytes, were transferred into the second dimension for the further separation and successive determination. The method uses two different chromatographic columns; in the first dimension, 50 µL of sample was injected on first column, and mycotoxins elution regions were collected in a loop and transferred into the second column for the separation of analytes. Each column operated in gradient elution mode in order to eliminate interfering compounds and improve separation and peak shape. After the optimization, the method has been validated according to EU regulation and finally applied for the analysis of forty beer samples collected from Italian supermarkets. Among all mycotoxins studied, fumonisins B1 was the most widely distributed in analysed beers (>21%) in the range from 0.6 to 12.3 ng mL-1. The automated methodology developed was able to determine accurately and simultaneously seven mycotoxins in beer. This provided a significant reduction of sample handle and, consequently of analysis time.

Determination of Selected Pyrrolizidine Alkaloids in Honey by Dispersive Liquid-Liquid Microextraction and Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry.

The contamination of honey with hepatotoxic pyrrolizidine alkaloids (PAs) is an actual concern for food safety. This study reports the first application of dispersive liquid-liquid microextraction (DLLME) in the determination of five relevant PAs, and the relative N-oxide derivatives (PANOs), in honey. The effects of different experimental parameters (pH, ionic strength, type and volume of DLLME solvents) affecting the extraction efficiency were carefully investigated and optimized. PAs were extracted from honey (diluted solution 10% w/v at pH 9.5) by injecting a mixture of chloroform and isopropyl alcohol. A reduction step (zinc powder in acidic aqueous solution) before DLLME was performed to convert PANOs in PAs and to obtain the total PA levels. Both sample preparation protocols (DLLME and Zn-DLLME) showed negligible matrix effects on PA signal intensity in honeys of different botanical origins. The overall recoveries of DLLME and Zn-DLLME ranged from 71 to 102% and from 63 to 103%, respectively, with a good precision (standard deviations in the range from 1 to 12%). The attained method quantification limits stayed between 0.03 and 0.06 µg kg-1, and the linear response range extended to 25 µg kg-1. Additionally, the proposed method provides results comparable to those of the SPE protocol in the analysis of real samples. An analysis of retail honeys revealed PA residues in all analyzed samples, with a maximum level of 17.5 µg kg-1 (total PAs). Globally, the proposed method provides a sensitive and accurate determination of analytes and offers numerous advantages, such as simplicity, low cost, and a high sample throughput, which make it suitable for screening and quality control programs in food chain and occurrence studies.

Plant origin authentication of Sonoran Desert propolis: an antiproliferative propolis from a semi-arid region.

The main chemical composition of Sonoran propolis (SP), as well as its antiproliferative activity on cancer cells through apoptosis induction, has been reported. The chemical constitution of SP remained qualitatively similar throughout the year, whereas the antiproliferative effect on cancer cells exhibited significant differences amongst seasonal samples. The main goal of this study was to provide phytochemical and pharmacological evidence for the botanical source of SP and its antiproliferative constituents. A chemical comparative analysis of SP and plant resins of species found in the surrounding areas of the beehives was carried out by HPLC-UV-DAD, as well as by 1H NMR experiments. The antiproliferative activity on cancerous (M12.C3.F6, HeLa, A549, PC-3) and normal cell lines (L-929; ARPE-19) was assessed through MTT assays. Here, the main polyphenolic profile of SP resulted to be qualitatively similar to Populus fremontii resins (PFR). However, the antiproliferative activity of PFR on cancer cells did not consistently match that exhibited by SP throughout the year. Additionally, SP induced morphological modifications on treated cells characterised by elongation, similar to those induced by colchicine, and different to those observed with PFR treatment. These results suggest that P. fremontii is the main botanical source of SP along the year. Nevertheless, the antiproliferative constituents of SP that induce that characteristic morphological elongation on treated cells are not obtained from PFR. Moreover, the presence of kaempferol-3-methyl-ether in SP could point Ambrosia ambrosioides as a secondary plant source. In conclusion, SP is a bioactive poplar-type propolis from semi-arid zones, in which chemical compounds derived from other semi-arid plant sources than poplar contribute to its antiproliferative activity.

Transcriptome reprogramming, epigenetic modifications and alternative splicing orchestrate the tomato root response to the beneficial fungus Trichoderma harzianum.

Beneficial interactions of rhizosphere microorganisms are widely exploited for plant biofertilization and mitigation of biotic and abiotic constraints. To provide new insights into the onset of the roots-beneficial microorganisms interplay, we characterised the transcriptomes expressed in tomato roots at 24, 48 and 72 h post inoculation with the beneficial fungus Trichoderma harzianum T22 and analysed the epigenetic and post-trascriptional regulation mechanisms. We detected 1243 tomato transcripts that were differentially expressed between Trichoderma-interacting and control roots and 83 T. harzianum transcripts that were differentially expressed between the three experimental time points. Interaction with Trichoderma triggered a transcriptional response mainly ascribable to signal recognition and transduction, stress response, transcriptional regulation and transport. In tomato roots, salicylic acid, and not jasmonate, appears to have a prominent role in orchestrating the interplay with this beneficial strain. Differential regulation of many nutrient transporter genes indicated a strong effect on plant nutrition processes, which, together with the possible modifications in root architecture triggered by ethylene/indole-3-acetic acid signalling at 72 h post inoculation may concur to the well-described growth-promotion ability of this strain. Alongside, T. harzianum-induced defence priming and stress tolerance may be mediated by the induction of reactive oxygen species, detoxification and defence genes. A deeper insight into gene expression and regulation control provided first evidences for the involvement of cytosine methylation and alternative splicing mechanisms in the plant-Trichoderma interaction. A model is proposed that integrates the plant transcriptomic responses in the roots, where interaction between the plant and beneficial rhizosphere microorganisms occurs.

Ultrasound assisted dispersive liquid-liquid microextraction for fast and accurate analysis of chloramphenicol in honey.

Honey is a food produced from honey bee widely used for the sweetening power and for its biological properties. In order to prevent the infection of the hive, different xenobiotics (antibiotics, pesticide) were frequently employed. One of these substances is the chloramphenicol, that given its chemical stability could often found in food. Chloramphenicol have several side effects in humans after their ingestion and for this reason their intake must be avoid. The aim of this study, was developed an ultrasound-assisted dispersive liquid-liquid microextraction method coupled with UHPLC MS/MS determination, for fast and accurate analysis of chloramphenicol in honey. The parameters affecting on extraction efficiency were carefully optimized using an experimental design in order to maximized the recovery reducing matrix effects. After the optimization the method was validated and successfully applied to 66 honey samples.