Tetraploidy was induced using colchicine in the CBD-dominant Cannabis sativa cultivar ‘CBD Zero’ to assess the impact of ploidy change on plant desirable traits. Biometry, stem anatomy productivity, CBD and THC concentrations in the inflorescences, ionome composition of above-ground organs (i.e. stem, leaf and inflorescence) and VOC emission profile from the vegetative to the flowering phases were investigated. Tetraploids exhibited reduced stature and lower inflorescence biomass compared to diploids. These phenotypic shifts are likely a consequence of broad morphological and anatomical reorganization driven by increased genome size, as evidenced by comparative cross-sectional analyses. The tetraploid structural remodelling was characterized by a decreasing trend in stem fiber number and increased tissue thickness in both fan and sugar leaf. These observations were coupled to a distinct nutrient profile of polyploids, which showed higher concentrations of K, Fe and Ni, especially in leaves, and to their enhanced emissions of VOCs. The disparity was most pronounced towards the end of the flowering period, with the tetraploids showing a marked increase in monoterpene (e.g. p-cymene) and sesquiterpene production, compounds of considerable interest for their aromatic properties and potential health benefits. In contrast, the concentrations of the primary cannabinoids CBD and THC in dried buds showed no significant difference between ploidy levels. Some of the novel traits emerging in our induced polyploids, i.e., reduced height and increased emission of specific VOCs following leaf tissue-level changes, offer direct utility for new breeding strategies.

Molecularly imprinted nanoparticles (MIP-NPs) are recognition elements obtained upon the polymerization of functional monomers around the target template, that, once removed, reveal selective binding sites in the polymeric matrix enabling the target binding with high specificity. Due to their small size, the binding sites are mostly located at the surface of the polymer, which definitely improves the binding kinetics. Solid-phase synthesis with an excess of the functional monomer NIPAm (N-isopropylacrylamide) confers a thermoresponsive character to the polymers, thus acquiring a solvated gel-like nanometer-sized feature, while the use of a protein epitope avoids the challenging issues related to the whole protein imprinting. Once produced, epitope-imprinted nanogels (MIP-NGs) can be anchored onto the surface of electrochemical transducers, thus leading to the development of selective electrochemical sensors. In this work, MIP-NGs for the cardiac cell damage biomarker cardiac troponin T (cTnT) were anchored onto screen-printed carbon electrodes, enabling a linear impedimetric response in the concentration range 0.01-0.3 ng/mL with a LOD of 0.004 ng/mL. The sensor showed a negligible response towards interfering proteins and a reliable detection of the target protein in undiluted serum. To the best of our knowledge, for the first time an electrochemical sensor based on molecularly imprinted nanogels produced by solid-phase synthesis in aqueous environment with a protein epitope as template was developed for the detection of cTnT. The sensor featured analytical performances comparable to currently available immunoassays, thus showing a striking potential application as alternative tool for myocardial damage biomarker detection.

Gene therapy represents a promising strategy for the treatment of severe diseases through delivery of genetic material into target cells. Its clinical translation, however, remains limited by the need for safe and efficient vectors. β-Cyclodextrins (β-CDs), owing to their biocompatibility and structural tunability, represent an attractive platform. In this study, three novel cationic β-CD derivatives (CD1, CD2, and CD3), functionalized with amino groups and different lipophilic chains, were evaluated. Preformulation studies were conducted to optimize the composition and preparation of β-CD-based nanovectors (NVs), which were subsequently complexed with a GFP-encoding plasmid (pGFP). Sodium hyaluronate (HA) and polysialic acid (PSA), were incorporated at 10% w/w, to enhance loading capacity and biocompatibility. Physico-chemical characterization included size, polydispersity index (PdI), and zeta potential. Further insights concerning the architecture of NVs were obtained by TEM, Cryo-EM, and synchrotron SAXS, which indicated a hierarchical arrangement in the structure of the complexes, while electrophoresis verified efficient nucleic acid binding. Stability was assessed at 4 °C for one week and in culture medium for 24 h. MTT test was performed in HeLa cells and transfection efficiency was analysed by flow cytometry and fluorescence microscopy. NVs were then complexed with GFP-targeting-siRNA (N/P = 10), and silencing efficiency was assessed in HeLa-GFP cells. All formulations displayed suitable size (<200 nm) and PdI (<0.3). CD1 showed the highest biopharmaceutical performance, achieving GFP expression and silencing efficiency comparable to commercial Lipofectamine. Overall, CD1 emerged as the most promising candidate, supporting its potential as an effective and biocompatible NVs for gene delivery.

The diseases caused by the xylem-inhabiting bacterium Xylella fastidiosa (Xf) are considered a consequence of the drastic reduction in water flow that this endophyte causes in some of its hosts. To date, within the EU, Xf has been found infecting several taxa that are common members of the Mediterranean maquis, a resource-poor environment with very hot and dry summers. To adapt, native plants possess several means to cope with water paucity and derived gas voids, including a high density of small lumen vessels in the wood. We describe for the first time the anatomy of the anisohydric shrub Rhamnus alaternus spontaneously growing within the Xf demarcated area of Monte Argentario (Italy), when naturally infected by X. fastidiosa subsp. multiplex (Xfm) ST87. By comparing infected plants using quantitative PCR and histological analyses, we could not find a clear relationship between the symptoms of scattered leaf chlorosis, premature leaf drop and branch dieback, and the presence and abundance of ST87 in branches, or the fraction of occluded vessels (p > 0.05). In contrast, in leaves, there was a highly significant relationship (p < 0.001) between the presence of ST87 in their midribs and chlorosis of their lamina. We observed here that this host accumulates phenolics-rich mucilage inside vessels in a way that appears to be independent from the physical presence of ST87 in their lumen. Further experiments in a controlled environment are necessary to prove the ability of Xfm ST87 to cause the symptoms of dehydration that are recurrently observed on R. alaternus at Monte Argentario.

Ulvan oligosaccharides, which exhibit greater solubility than high molecular weight ulvan, hold significant potential for the valorization of Ulva biomass. They have been shown to promote the growth of probiotic bacteria in fermentation models and have demonstrated protective and anti-inflammatory effects in animal models. The enzymatic hydrolysis of ulvan using the commercial ulvan lyase PLSV_3875 bypasses the challenge posed by the resistance of ulvanobiuronic acids to conventional acidic hydrolysis. This study aimed to develop a rapid, scalable, and efficient method for a complete ulvan hydrolysis under reduced salt concentrations, yielding oligosaccharides suitable for biological applications. The enzyme's robustness and kinetic parameters were evaluated. Enzyme activity was assessed at pH 6-10.5 with 0-100 mM Tris-HCl, carbonate or phosphate buffer, and 0-200 mM NaCl/KCl. Optimal conditions were identified as 3 mg/mL ulvan, 25 mM KCl, and 100 mM phosphate buffer at pH 7.5. The resulting oligosaccharides were characterized using 2D-NMR and UHPLC-MS/MS.

Fluctuations in emergency department (ED) utilization may be related to pollutant concentration variations that can heighten acute pathologies, exacerbate chronic conditions or indicate increased human activity. This study investigates the association between particulate matter (PM2.5-PM10) concentrations in patients' residential areas and ED visits. Retrospective, single-centre, time-series cohort study on a 100km(2) urban area around a level 1 hospital in Florence, Italy, with 307,279 ED visits of patients of age > 14 years between 01/01/2019 and 31/12/2022. PM2.5 and PM10 daily concentrations, recorded by 14 air-quality stations, were interpolated at patients' residences, in a 250m(2) grid, with inverse distance weighting and averaged on various time windows (lag) up to 30 days before ED visit. The association between ED visits and pollutant concentrations as percent excess of incidence rate (%Er) with 95% confidence intervals were investigated with generalized additive models. Significant association was found with short-term PM2.5 and PM10 fluctuations (PM2.5(Lag1): 1.24%Er, 0.59-1.99 95%CI; PM10(Lag2): 1.79%Er, 1.08-2.72). Such association was stronger for the subgroups of deceased patients (PM2.5(Lag1) 5.71%Er, 1.44-8.99; PM10(Lag2) 6.38%Er, 2.02-10.83), elderly patients presenting with dyspnoea (PM2.5(Lag1) 4.18%Er, 2.36-6.33; PM10(Lag2) 4.99%Er, 2.77-7.32) and younger patients suffering traumatic events (PM2.5(Lag1) 1.29%Er, 0.32-2.51; PM10(Lag2) 2.14%Er, 0.88-3.50). Exposure-response curves showed that particulate matter is associated with ED visits even at concentrations below the WHO guard levels. A short-term rise in PM2.5 and PM10, even below guard levels, was associated with increased ED utilization, particularly related to elderly patients with dyspnoea and the concomitant lurking occurrence of vehicle accidents in the younger population. Trial registration: NCT06491290; 09/07/2024; retrospectively registered.

Oxidative and inflammatory stresses contribute to the development of many intestinal pathologies. This study characterized the polyphenolic profile and biological activity of a hydroalcoholic extract obtained from the fruit pulp of Feijoa sellowiana on HT29 intestinal epithelial cells subjected to oxidative (H2O2) and inflammatory (cytokines) stress. HPLC-DAD-MS analysis revealed an interesting phenolic composition, rich in hydrolyzable tannins (HHDP-glucose, pedunculagin and other ellagic acid derivatives) and condensed tannins (procyanidin dimers), with a total polyphenol content of 8.07 mg/g GAE. The extract was non-cytotoxic up to 160 µg/mL and exerted a protective effect against the cytokine-induced reduction in cell viability. In vitro assays confirmed its strong antioxidant and scavenging capacity. The scratch assay suggested enhanced cell migration. The extract modulated the activity of key metabolic enzymes restoring glucose-6-phosphate dehydrogenase and enolase activity, while supporting glycolytic flux through pyruvate kinase and lactate dehydrogenase. PCA and Pearson correlation analyses confirmed a treatment-dependent modulation of the metabolic and redox profile, suggesting a regulatory role beyond a mere scavenging effect. These findings highlight the nutraceutical potential of feijoa polyphenols, not only as direct antioxidants but also as modulators of cellular metabolism and redox homeostasis, supporting their application in gastrointestinal disorders with oxidative or inflammatory components.

AIMS:A systematic review and meta-analysis of published randomized controlled trials was conducted to collate evidence from studies implementing ancient grains and investigate the impact of ancient grain consumption on health outcomes of patients with Diabetes Mellitus (DM). DATA SYNTHESIS:Twenty-nine randomized controlled trials were included, and 13 were meta-analyzed. Interventions ranged from 1 day to 24 weeks; most samples were affected by DM type 2 (n = 28 studies) and the ancient grains used were oats (n = 10 studies), brown rice (n = 6 studies), buckwheat (n = 4 studies), chia (n = 3 studies), Job's Tears (n = 2 studies), and barley, Khorasan and millet (n = 1 study). Thirteen studies that used oats, brown rice, and chia provided data for a quantitative synthesis. Four studies using oats showed a small to moderate beneficial effect on health outcomes including LDL-c (n = 717, MD: 0.30 mmol/l, 95% CI: 0.42 to -0.17, Z = 4.61, p < 0.05, I2 = 0%), and TC (n = 717, MD: 0.44 mmol/l, 95% CI: 0.63 to -0.24, Z = 4.40, p < 0.05, I2 = 0%). Pooled analyses of studies using chia and millet did not show significant effects on selected outcomes. CONCLUSIONS:For adults affected by DM type 2, the use of oats may improve lipidic profile. Further experimental designs are needed in interventional research to better understand the effects of ancient grains on diabetes health outcomes. PROSPERO REGISTRATION:CRD42023422386.

Pâté is a by-product of olive oil production which represents an abundant source of phenolic compounds and can be used for food formulation, reducing its environmental impact and promoting a circular economy. In this context, the effects of a hydroalcoholic extract of pâté were evaluated for the first time in an AGS human cell line commonly used as model of gastric mucosa. Pâté was obtained from Tuscan olives; the total phenolic content was 16.6 mg/g dried extract, with verbascoside and secoiridoid derivatives as the most abundant phenols. The phenolic pâté extract did not alter viability, distribution of cell cycle phases or proliferation and migration of AGS cells at the tested concentrations. Seven enzymes were chosen to investigate the metabolic effect of the pâté extract in the context of oxidative stress. Pâté produced a statistically significant increase in the activity of key enzymes of some metabolic pathways: Lactate dehydrogenase, Enolase, Pyruvate kinase, Glucose 6-phosphate dehydrogenase, Citrate synthase, 3-Hydroxyacyl-CoA dehydrogenase and Hexokinase. Pre-treatments with the extract of pâté at 100 µg/mL or 200 µg/mL, as observed through PCA analysis, appeared able to counteract the enzymatic activity alterations due to oxidative stress induced by H2O2 1 mM and 2 mM. The results indicate that dried pâté, due to its phenolic components, can be proposed as a new functional food ingredient.

The development of analytical devices that can allow an easy, rapid and cost-effective measurement of multiple markers, such as progesterone and β-hCG, could have a role in decreasing the burden associated with pregnancy-related complications, such as ectopic pregnancies. Indeed, ectopic pregnancies are a significant contributor to maternal morbidity and mortality in both high-income and low-income countries. In this work, an effective and highly performing electrochemical strip for a combo determination of progesterone and β-hCG was developed. Two immunosensing approaches were optimized for the determination of these two hormones on the same strip. The immunosensors were realized using cost-effective disposable electrode arrays and reagent-saving procedures. Each working electrode of the array was modified with both the IgG anti-β-hCG and anti-progesterone, respectively. By adding the specific reagents, progesterone or β-hCG can then be determined. Fast quantitative detection was achieved, with the analysis duration being around 1 h. Sensitivity and selectivity were assessed with a limit of detection of 1.5 × 10–2 ng/mL and 2.45 IU/L for progesterone and β-hCG, respectively. The proposed electrochemical combo-strip offers great promise for rapid, simple, cost-effective, and on-site analysis of these hormones and, thus, for the development of a point-of-care diagnostic tool for early detection of pregnancy-related complications.

The study of azo dyes degradation products and their harmfulness is an important topic because their diffusion in the environment is still a serious problem. We investigated the reduction by pig liver microsomal fraction of four bis-azo dyes, Acid Red 73, Acid Red 150, Direct Red 24, and Direct Red 28, in anaerobic condition. These complex molecules, different for position/nature of substituent groups and for the group interposed between the two azo bonds, allowed to investigate the correlation between substrate molecular structure and action of a subcellular fraction representative of the xenobiotic degradation by a living organism and the consequent possible appearance of harmful reaction products. Dyes were first carefully purified to eliminate interferences in the enzymatic assays. The reduction products were identified by gas chromatography/mass spectrometry. Acid Red 73 and Acid Red 150 were markedly reduced with production of harmful aromatic amines, while Direct Red 24 and Direct Red 28 were neither reduced to their synthesis amines nor to other amines. These results suggest that azo dyes with a benzidine derivative group are not degraded by microsomal fraction and therefore molecules with a similar structure could be classified with a lower risk index not producing any harmful catabolic product.

The effects of the phenolic compounds of extra virgin olive oil (EVOO) on AGS cells have never been studied so far, which is the aim of this study. The profiles of the main phenolic components in EVOOs, mainly secoiridoid compounds derived from the transformation of oleuropein during the olive milling process, were evaluated and compared. Oils of different origins were evaluated aiming at verifying whether chemical differences in the phenolic composition of the dry extracts played a role in the metabolism and in maintaining the cellular redox state of AGS cells. The following key enzymes of some metabolic pathways were studied: lactate dehydrogenase, enolase, pyruvate kinase, glucose 6-phosphate dehydrogenase, citrate synthase, 3-Hydroxyacyl-CoA dehydrogenase and hexokinase. As confirmed through PCA analysis, pretreatments with the dry extracts of EVOOs at different concentrations appeared to be able to counteract the enzymatic activity alterations due to oxidative stress induced by H2O2 1 mM and 2 mM. The studied phytocomplexes showed the ability to protect AGS cells from oxidative damage and the secoiridoid derivatives from both oleuropein and ligstroside contributed to the observed effects. The results suggested that EVOOs with medium to high concentrations of phenols can exert this protection.

Here is presented a setup made of a combination of an injection valve, a hand-made chromatographic microcolumn and an electrochemical detector for the simultaneous separation of Hg2+ and Ag+ ions in water. The microcolumns were packed with exchanger resin and used for the separation of Hg2+ and Ag+ ions, whereas a screen-printed carbon electrode (SPCE) was the amperometric detector. The performances of the SPCE towards ions were firstly studied in a FIA setup. The efficiency of ion separation was then evaluated. The reproducibility, stability, and the regeneration of the obtained microcolumns were also studied and discussed.

In patients with suspected acute aortic syndrome (AAS), standardized clinical risk assessment following the American Heart Association (AHA) and European Society of Cardiology (ESC) recommendations is based on 12 risk markers summarized in the aortic dissection detection (ADD) risk score.1, 2 However, additional risk factors and conditions have been associated with AASs. These include autoimmune diseases, immunosuppressive or glucocorticoid treatment, and use of fluoroquinolones.3-5 The latter association has even led to official prescription warnings from the Food and Drug Administration and other agencies. The underlying evidence, however, has been obtained in retrospective population studies, case series, and anecdotal reports not involving acute patients or the emergency department (ED).3-5 We use the data from the Aortic Dissection Detection Risk Score Plus D-dimer in Suspected Acute Aortic Dissection (ADvISED) prospective multicenter study to evaluate whether these risk factors are associated with diagnoses of AASs in ED patients with an operator-defined clinical suspicion and pertinent symptoms.6 We explored whether routine evaluation of these additional variables may provide any potential diagnostic advantage to physicians when approaching patients with potential AAS symptoms. The characteristics of the ADvISED study methods and cohort have been described elsewhere.6 The present subanalysis was conducted on patients from one of participating EDs (Careggi University Hospital, Florence, Italy). For all patients, ED and hospital charts were retrospectively reviewed for the following variables: (1) use of a fluoroquinolone drug in the past 90 days (at least 5 days of treatment), (2) use of any systemic nonsteroidal immunosuppressive drug or any glucocorticoid drug (defined by an equivalent dose of ≥0.25 mg/kg per day of methylprednisolone) for at least 14 days in the past 90 days, and (3) known systemic autoimmune disease such as connectivities (e.g., rheumatoid arthritis, systemic lupus erythematosus, systemic sclerosis) or vasculitis (e.g., giant cell arteritis, ANCA-associated vasculitis). Sections of medical charts reporting recent and past medical history by the treating physicians were obtained from the ED and hospital database and were evaluated by two independent reviewers, who were blinded to both diagnostic results and final diagnosis. Any discrepancy was adjudicated by discussion between reviewers. Statistical analysis included binomial logistic regression, adjusted with stratification on the main effect of known risk factors. We analyzed data from 657 patients with suspected AASs. The mean ± SD age was 63.4 ± 17 years, and 431 (65.6%) patients were male. Seventy-seven (11.7%) patients had a final diagnosis of AAS: 42 (6.4%) had a type A acute aortic dissection (AD), 17 (2.6%) had a type B AD, 10 (1.5%) had an intramural aortic hematoma, seven (1.1%) had a spontaneous aortic rupture, and one (0.1%) had a penetrating aortic ulcer. In patients with AAS, four had used fluoroquinolone drugs in the past 90 days, five were taking oral glucocorticoids, one was taking oral glucocorticoids plus rituximab, and two had a systemic vasculitis (giant cell arteritis and p-ANCA–associated vasculitis). The prevalence of the analyzed variables and the associated odd ratios (ORs) for diagnosis of AASs are shown in Table 1. None were associated with the diagnosis of AASs, in both univariate and multivariate analysis including the 12 ADD risk markers. Among the subgroup (543 patients) with low likelihood of AAS according with ESC guidelines, those ultimately not diagnosed with AASs (504 patients) were more likely to have a history of fluroquinolone, NSAID, or glucocorticoid use or known autoimmune disease (10.7% vs. 7.7%; p < 0.01). Limits of this study are its retrospective nature, potentially leading to underreporting of study variables, and insufficient power to detect exceedingly small increases in risk. We cannot exclude with certainty some degree of differential suspicion and reporting biases, but both were expected to skew findings toward a positive association between analyzed variables and AASs. In conclusion, in ED patients evaluated for suspected AASs, use of fluoroquinolones, use of immunosuppressive or glucocorticoid drugs, systemic autoimmune diseases, or any of them was not associated with final diagnosis of AASs. Hence, evaluation of these variables, despite potentially relevant in pathophysiologic terms, is unlikely to meaningfully impact on ED diagnostic reasoning.

Bacterial canker disease caused by Pseudomonas syringae pv. actinidiae (Psa) biovar 3 involved all global interest since 2008. We have found that in Psa3 genome, similarly to other P. syringae, there are three putative genes, lscα, lscβ and lscγ, coding for levansucrases. These enzymes, breaking the sucrose moiety and releasing glucose can synthetize the fructose polymer levan, a hexopolysaccharide that is well known to be part of the survival strategies of many different bacteria. Considering lscα non-coding because of a premature stop codon, in the present work we cloned and expressed the two putatively functional levansucrases of Psa3, lscβ and lscγ, in E. coli and characterized their biochemical properties such as optimum of pH, temperature and ionic strength. Interestingly, we found completely different behaviour for both sucrose splitting activity and levan synthesis between the two proteins; lscγ polymerizes levan quickly at pH 5.0 while lscβ has great sucrose hydrolysis activity at pH 7.0. Moreover, we demonstrated that at least in vitro conditions, they are differentially expressed suggesting two distinct roles in the physiology of the bacterium.

Pulmonary embolism (PE) and deep vein thrombosis (DVT) are gathered in venous thromboembolism (VTE) and represent the third cause of cardiovascular diseases. Recent studies suggest that meteorological parameters as atmospheric pressure, temperature, and humidity could affect PE incidence but, nowadays, the relationship between these two phenomena is debated and the evidence is not completely explained. The clinical experience of the Department of Emergency Medicine at AOUC Hospital suggests the possibility that a relationship effectively exists. We have collected data concerning the Emergency Medicine Unit admissions of PE patients to confirm our hypothesis. At the same time, atmospheric parameters are collected from the Lamma Consortium of Tuscany region. We have implemented new IT models and statistic tools by using semi-hourly records of weather time high resolution data to process the dataset. We have carried out tools from econometrics, like mobile means, and we have studied anomalies through the search for peaks and possible patterns. We have created a framework in Python to represent and study time series and to analyze data and plot graphs. The project has been uploaded on GitHub. Our analyses highlighted a strong correlation between the moving averages of atmospheric pressure and those of the hospitalizations number (R= -0.9468, p<0,001) although causality is still unknown. The existence of an increase in the number of hospitalizations in the days following short-to-medium periods of time characterized by a high number of half-hourly pressure changes is also detected. The spectrograms studies obtained by the Fourier transform requires to increase the dataset. The analyzed data (especially hospitalization data) were too few to carry out this kind of analyses.

Scratch assay is an easy and widely used "in vitro" technique to study cell migration and proliferation. In this work we focus on its modelling and on the capability to distinguish between these two phenomena that the simpler and common models are not able to disentangle. We adapted a model based on reaction–diffusion equation for being used with common microscopy instruments/data and therefore taking place in the gap between simpler modelling approaches and complex ones. An optimized image analysis pipeline and numerical least-squares fit provide estimates of the scratch proliferation and diffusion coefficients l and D. This work is intended as a first of a series in which the model is tested and its robustness and reproducibility are evaluated. Test samples were NIH3T3 cells scratch assays with proliferation and migration stimulated by varying the foetal bovine serum amount in the culture medium (10%, 7.5%, 5% and 2.5%). Results demonstrate, notwithstanding an expected l−D anticorrelation, the model capability to disentangle them. The 7.5% serum treatment can be identified as the model sensitivity limit. Treat–control l and D variations showed an intra-experiment reproducibility (∼±0.05∕h and ∼±200μm2∕h respectively) consistent with single fit typical uncertainties (∼±0.02∕h and ∼±300μm2∕h respectively).