Ruxolitinib in patients with polycythemia vera resistant and/or intolerant to hydroxyurea: European observational study.

BACKGROUND: Hydroxyurea (HU) is a commonly used first-line treatment in patients with polycythemia vera (PV). However, approximately 15%-24% of PV patients report intolerance and resistance to HU. METHODS: This phase IV, European, real-world, observational study assessed the efficacy and safety of ruxolitinib in PV patients who were resistant and/or intolerant to HU, with a 24-month follow-up. The primary objective was to describe the profile and disease burden of PV patients. RESULTS: In the 350 enrolled patients, 70% were >60 years old. Most patients (59.4%) had received ≥1 phlebotomy in the 12 months prior to the first dose of ruxolitinib. Overall, 68.2% of patients achieved hematocrit control with 92.3% patients having hematocrit <45% and 35.4% achieved hematologic remission at month 24. 85.1% of patients had no phlebotomies during the study. Treatment-related adverse events were reported in 54.3% of patients and the most common event was anemia (22.6%). Of the 10 reported deaths, two were suspected to be study drug-related. CONCLUSION: This study demonstrates that ruxolitinib treatment in PV maintains durable hematocrit control with a decrease in the number of phlebotomies in the majority of patients and was generally well tolerated.

Carbon Dots in Enantioselective Sensing.

Chirality has a crucial effect on clinical, chemical and biological research since most bioactive compounds are chiral in the natural world. It is thus important to evaluate the enantiomeric ratio (or the enantiopurity) of the selected chiral analytes. To this purpose, fluorescence and electrochemical sensors, in which a chiral modifier is present, are reported in the literature. In this review, fluorescence and electrochemical sensors for enantiorecognition, in which chiral carbon dots (CDs) are used, are reported. Chiral CDs are a novel zero-dimensional carbon-based nanomaterial with a graphitic or amorphous carbon core and a chiral surface. They are nanoparticles with a high surface-to-volume ratio and good conductivity. Moreover, they have the advantages of good biocompatibility, multi-color emission, good conductivity and easy surface functionalization. Their exploitation in enantioselective sensing is the object of this review, in which several examples of fluorescent and electrochemical sensors, containing chiral CDs, are analyzed and discussed. A brief introduction to the most common synthetic procedures of chiral CDs is also reported, evidencing strengths and weaknesses. Finally, consideration concerning the potential challenges and future opportunities for the application of chiral CDs to the enantioselective sensing world are outlined.

Electrochemical Sensing Platform Based on Carbon Dots for the Simultaneous Determination of Theophylline and Caffeine in Tea.

A simple and selective method for the determination of caffeine (CAF) and theophylline (THEO) has been developed for a glassy carbon electrode (GCE) modified with a composite including carbon dots (CDs) and chitosan (CS). To our knowledge, there are no previous studies that analyze a CDs-modified GCE for the presence of CAF and THEO. The electrochemical behavior of a GCE modified with a CDs-CS composite was studied in acidic medium by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Considering the sensor analytical parameters, the same linear concentrations range was found for CAF and THEO ranging from 1 × 10-5 to 5 × 10-3 mol L-1 with the same detection limit (LOD) of 1 × 10-6 mol L-1. The reproducibility and repeatability data were satisfactory in terms of RSD%. Moreover, the storage stability was evaluated, evidencing good results whatever the experimental conditions used. The developed sensor was applied for the simultaneous determination of CAF and THEO in tea and drug, and results were compared with those obtained with HPLC-ESI-MS in SIR mode as an independent method optimized on purpose. The electrochemical sensor presents the undoubled advantages in terms of cheapness, portability, and ease of use, since it does not require skilled personnel.

Not Only Graphene Two-Dimensional Nanomaterials: Recent Trends in Electrochemical (Bio)sensing Area for Biomedical and Healthcare Applications.

Two-dimensional (2D) nanomaterials (e.g., graphene) have attracted growing attention in the (bio)sensing area and, in particular, for biomedical applications because of their unique mechanical and physicochemical properties, such as their high thermal and electrical conductivity, biocompatibility, and large surface area. Graphene (G) and its derivatives represent the most common 2D nanomaterials applied to electrochemical (bio)sensors for healthcare applications. This review will pay particular attention to other 2D nanomaterials, such as transition metal dichalcogenides (TMDs), metal-organic frameworks (MOFs), covalent organic frameworks (COFs), and MXenes, applied to the electrochemical biomedical (bio)sensing area, considering the literature of the last five years (2018-2022). An overview of 2D nanostructures focusing on the synthetic approach, the integration with electrodic materials, including other nanomaterials, and with different biorecognition elements such as antibodies, nucleic acids, enzymes, and aptamers, will be provided. Next, significant examples of applications in the clinical field will be reported and discussed together with the role of nanomaterials, the type of (bio)sensor, and the adopted electrochemical technique. Finally, challenges related to future developments of these nanomaterials to design portable sensing systems will be shortly discussed.

Phytochemical Analysis and In Vitro Antileukemic Activity of Alkaloid-Enriched Extracts from Vinca sardoa (Stearn) Pignatti.

Vinca sardoa (Stearn) Pignatti, known as Sardinian periwinkle, is widely diffused in Sardinia (Italy). This species contains indole alkaloids, which are known to have a great variety of biological activities. This study investigated the antileukemic activity against a B lymphoblast cell line (SUP-B15) of V. sardoa alkaloid-rich extracts obtained from plants grown in Italy, in Iglesias (Sardinia) and Rome (Latium). All the extracts showed a good capacity to induce reductions in cell proliferation of up to 50% at the tested concentrations (1-15 µg/mL). Moreover, none of the extracts showed cytotoxicity on normal cells at all the studied concentrations.

Metabolic Profile of Agropyron repens (L.) P. Beauv. Rhizome Herbal Tea by HPLC-PDA-ESI-MS/MS Analysis.

Agropyron repens (L.) P. Beauv. (couch grass) is a world-wide infesting rhizomatous plant with pharmacological applications. Chemical research is focused on its allelopathic and anti-inflammatory components, which are mainly present in the essential oil. Conversely, the aqueous extracts have been sparingly investigated, although the herbal tea is by far the most used formulation. To fill the gap, the metabolic profile of Agropyron repens rhizome herbal tea was investigated by electrospray ionization (ESI) tandem-mass spectrometry (MS/MS); the phenolic profile was investigated by HPLC-PDA-ESI-MS/MS. ESI-MS fingerprinting was provided, evidencing diagnostic ions for saccharides, organic acids and amino acids. The HPLC-PDA-ESI-MS/MS analysis evidenced at least 20 characteristic phenolic compounds, the most representative being caffeoyl and feruloyl quinic esters, followed by coumaric, caffeic and ferulic acids, and hesperidin among flavonoids. In addition, the essential amino acid tryptophan was identified for the first time. The results suggest new perspectives of applications for Agropyron repens rhizome.

Targeted phenolic profile of radler beers by HPLC-ESI-MS/MS: the added value of hesperidin to beer antioxidants.

The well-known health beneficial properties of beer are mainly due to phenolic antioxidants. Citrus-flavored beers represent a growing side-market in the beer industry, sparingly investigated to date. The phenolic profile of commercial radler beers (R1, R2) was investigated to evaluate the impact of the lemon juice added to beer in the industrial production. Results were compared to those obtained for opportunely chosen commercial beer (B) and lemonade (L). The study was carried out by an HPLC-MS/MS with an electrospray ionization source in selected ion recording mode, analyzing in a single chromatographic run 26 compounds belonging to the different phenolic classes of hydroxybenzoic, hydroxycinnamic and caffeoylquinic acids, flavonoids and prenylflavonoids. Different phenolic profiles were found for R1 and R2, mainly ascribed to different malt/hop/recipe used for the beer. High to very high level of hesperidin were found in the radlers, so that a major impact on phenolic antioxidants of the radlers was due to the lemon. Similarly, a major impact of the lemon aromas was found, D-limonene being the dominant peak resulting from the GC-MS analysis of the volatile fraction of the radlers.

Novel IncFII plasmid harbouring blaNDM-4 in a carbapenem-resistant Escherichia coli of pig origin, Italy.

OBJECTIVES: To the best of our knowledge, we describe the first evidence in Europe of an MDR, blaNDM-4-positive Escherichia coli isolated from a food-producing animal, harboured by a novel IncFII plasmid of which we report the complete sequence. METHODS: One blaNDM-4-positive E. coli isolated in 2019 from the caecal contents of a fattening pig in Italy was in-depth characterized by combined bioinformatic analysis of Oxford Nanopore long reads and Illumina short reads, for in silico typing, determination of the blaNDM-4 genetic context and full reconstruction of the blaNDM-4-carrying plasmid. RESULTS: The isolate belonged to ST641 and to the genoserotype O108:H23 and tested positive for different virulence genes and plasmid replicons. The MDR phenotype of resistance to all ß-lactams, carbapenems, sulfamethoxazole and trimethoprim was mediated by blaTEM-1B, blaNDM-4, sul1/sul3 and dfrA12, respectively. The blaNDM-4 gene was harboured by a novel 53 043 bp IncFII plasmid (pMOL412_FII) composed of four main genetic regions, including an MDR region (MRR-NDM-4) of 16 kb carrying blaNDM-4 and several antimicrobial resistance genes located in a class 1 integron. pMOL412_FII was closely related to another ∼90.3 kb plasmid (pM109_FII) harbouring blaNDM-4 in an E. coli isolated from a human patient in Myanmar. CONCLUSIONS: To the best of our knowledge, we have identified for the first time in Europe an NDM-producing Enterobacterales in livestock and resolved the complete sequence of the novel pMOL412_FII plasmid harbouring blaNDM-4 in an MRR. A global One Health approach, comparing genomic data from different sources and geographical areas, may help to trace back and control possible plasmid-borne carbapenemase gene transmission between animals and humans and along the food chain at international level.

Mechanism of Action of the Tumor Vessel Targeting Agent NGR-hTNF: Role of Both NGR Peptide and hTNF in Cell Binding and Signaling.

NGR-hTNF is a therapeutic agent for a solid tumor that specifically targets angiogenic tumor blood vessels, through the NGR motif. Its activity has been assessed in several clinical studies encompassing tumors of different histological types. The drug's activity is based on an improved permeabilization of newly formed tumor vasculature, which favors intratumor penetration of chemotherapeutic agents and leukocyte trafficking. This work investigated the binding and the signaling properties of the NGR-hTNF, to elucidate its mechanism of action. The crystal structure of NGR-hTNF and modeling of its interaction with TNFR suggested that the NGR region is available for binding to a specific receptor. Using 2D TR-NOESY experiments, this study confirmed that the NGR-peptides binds to a specific CD13 isoform, whose expression is restricted to tumor vasculature cells, and to some tumor cell lines. The interaction between hTNF or NGR-hTNF with immobilized TNFRs showed similar kinetic parameters, whereas the competition experiments performed on the cells expressing both TNFR and CD13 showed that NGR-hTNF had a higher binding affinity than hTNF. The analysis of the NGR-hTNF-triggered signal transduction events showed a specific impairment in the activation of pro-survival pathways (Ras, Erk and Akt), compared to hTNF. Since a signaling pattern identical to NGR-hTNF was obtained with hTNF and NGR-sequence given as distinct molecules, the inhibition observed on the survival pathways was presumably due to a direct effect of the NGR-CD13 engagement on the TNFR signaling pathway. The reduced activation of the pro survival pathways induced by NGR-hTNF correlated with the increased caspases activation and reduced cell survival. This study demonstrates that the binding of the NGR-motif to CD13 determines not only the homing of NGR-hTNF to tumor vessels, but also the increase in its antiangiogenic activity.

A Livestock-Associated, Multidrug-Resistant, Methicillin-Resistant Staphylococcus aureus Clonal Complex 97 Lineage Spreading in Dairy Cattle and Pigs in Italy.

Pandemic methicillin-resistant Staphylococcus aureus (MRSA) clonal complex 97 (CC97) lineages originated from livestock-to-human host jumps. In recent years, CC97 has become one of the major MRSA lineages detected in Italian farmed animals. The aim of this study was to characterize and analyze differences in MRSA and methicillin-susceptible S. aureus (MSSA) mainly of swine and bovine origins. Forty-seven CC97 isolates, 35 MRSA isolates, and 6 MSSA isolates from different Italian pig and cattle holdings; 5 pig MRSA isolates from Germany; and 1 human MSSA isolate from Spain were characterized by macrorestriction pulsed-field gel electrophoresis (PFGE) analysis, multilocus sequence typing (MLST), spa typing, staphylococcal cassette chromosome mec (SCCmec) typing, and antimicrobial resistance pattern analysis. Virulence and resistance genes were investigated by PCR and microarray analysis. Most of the isolates were of SCCmec type V (SCCmec V), except for two German MRSA isolates (SCCmec III). Five main clusters were identified by PFGE, with the German isolates (clusters I and II) showing 60.5% similarity with the Italian isolates, most of which (68.1%) grouped into cluster V. All CC97 isolates were Panton-Valentine leukocidin (PVL) negative, and a few (n = 7) tested positive for sak or scn. All MRSA isolates were multidrug resistant (MDR), and the main features were erm(B)- or erm(C)-mediated (n = 18) macrolide-lincosamide-streptogramin B resistance, vga(A)-mediated (n = 37) pleuromutilin resistance, fluoroquinolone resistance (n = 33), tet(K) in 32/37 tet(M)-positive isolates, and blaZ in almost all MRSA isolates. Few host-associated differences were detected among CC97 MRSA isolates: their extensive MDR nature in both pigs and dairy cattle may be a consequence of a spillback from pigs of a MRSA lineage that originated in cattle as MSSA and needs further investigation. Measures should be implemented at the farm level to prevent spillover to humans in intensive farming areas.

Evaluation of the Antifungal and Antiproliferative Properties of the Lichen Roccella tinctoria DC. Extracts and Main Components.

In this work, phytochemical analysis on different extracts of Roccella tinctoria DC. was reported using different techniques with respect to the past. Twenty volatile and three non-volatile compounds were identified, some of which were found in this species for the first time. The methanolic extracts and their non-volatile components were then evaluated for their antitumor effects in cancerous A549 and Mz-ChA-1 cells and for their tolerability in non-cancerous BEAS-2B and H69 cells, showing IC50 values from 94.6 µg/mL to 416.4 µg/mL, in general. The same extracts and compounds were also tested for their antifungal effects in Candida albicans, with only compound 2 being active, with an MIC50 value of 87 µg/mL. In addition, they were tested for their anti-Candida adhesion activity, anti-Candida biofilm formation, and anti-Candida mature biofilm inhibition, with efficacy percentages generally above 50% but not for all of them. Lastly, the DF3 extract and compounds 1-2 were tested in vivo according to the Galleria mellonella survival assay, showing positive mortality rates above 50% at different concentrations. All these biological assays were conducted on this species for the first time. Comparisons with other lichens and compounds were also presented and discussed.

Phytochemical Characterization of Malt Spent Grain by Tandem Mass Spectrometry also Coupled with Liquid Chromatography: Bioactive Compounds from Brewery By-Products.

BACKGROUND: Brewer's spent grain (BSG) is one of the main by-products of beer industry, little used because of its high moisture making it difficult to transport and store. Mainly used as animal feed and for energy production, the agro-industrial waste have recently attracted attention as source of bioactive compounds, with potential applications in many sectors as food, nutraceutical, pharmaceutical, cosmetic, food packaging. The present work focuses on BSG as potential source of valuable small-size bioactive compounds. METHODS: Laboratory-made BSG was obtained by using four base malts for mashing. After drying, BSG was eco-friendly extracted with water and the extracts analyzed by untargeted ElectroSpray Ionization (ESI)-Mass Spectrometry (MS)/Mass Spectrometry (MS) (ESI-MS/MS) infusion experiments and by targeted High Performance Liquid Chromatography-PhotoDiodeArray-ElectroSpray Ionization-Mass Spectrometry (HPLC-PDA-ESI-MS) in Selected Ion Recording (SIR) mode analysis, to investigate the metabolic profile, the phenolic profile, the individual phenolic content, and tryptophan content. Aqueous extracts of malts and wort samples were also analyzed for a comparison. Data were statistically analyzed by ANOVA test. An explorative analysis based on Principal Component Analysis (PCA) was also carried out on malts, wort and threshes, in order to study correlation among samples and between samples and variables. RESULTS: The untargeted ESI-MS/MS infusion experiments provided the mass spectral fingerprint of BSG, evidencing amino acids (γ-aminobutyric acid, proline, valine, threonine, leucine/isoleucine, lysine, histidine, phenylalanine and arginine) and organic and inorganic acids (pyruvic, lactic, phosphoric, valerianic, malonic, 2-furoic, malic, citric and gluconic acids), besides sugars. γ-Aminobutyric acid and lactic acid resulted predominant among the others. The targeted HPLC-PDA-ESI-MS in SIR mode analysis provided the phenolic profile of the polar fraction of BSG, evidenced tryptophan as the main residual metabolite in BSG (62.33-75.35 µg/g dry BSG), and catechin (1.13-4.24 µg/g dry BSG) as the representative phenolic antioxidant of not pre-treated BSG samples. The chemometric analysis of the individual compounds content in BSG, malt and wort evidenced similarities and differences among the samples. CONCLUSIONS: As main goal, the phytochemical characterization of BSG from base malts highlighted BSG as a potential source of small biomolecules, as tryptophan and catechin, besides γ-aminobutyric acid and lactic acid, opening to new perspectives of application for BSG. Strategies for their recovery are a future challenge. Moreover, ESI-MS/MS analysis was confirmed as a powerful tool for fast characterization of complex matrix. Last, results obtained by chemometric elaboration of data demonstrated the possibility to monitor a small number of molecules to ensure the quality of a final product.

Burden of Illness in Follicular Lymphoma with Multiple Lines of Treatment, Italian RWE Analysis.

This real-world analysis investigated patients with follicular lymphoma in Italy receiving three or more treatment lines (≥3L), focusing on therapeutic pathways with their rebounds on healthcare resource consumptions and costs. Data were retrieved from administrative databases from healthcare entities covering about 13.3 million residents. Adults diagnosed with follicular lymphoma were identified between January 2015 and June 2020, and among them 2434 patients with ≥3L of treatment during the data availability interval (January 2009 to June 2021) were included. Of them, 1318 were in 3L, 494 in 4L and 622 in ≥5L. A relevant proportion of patients (12-32%) switched to a later line within the same calendar year. At 3-year follow-up (median), 34% patients died. Total mean annual expenses were euro 14,508 in the year preceding inclusion and rose to euro 21,081 at 1-year follow-up (on average euro 22,230/patient/year for the whole follow-up), with hospitalization and drug expenses as weightiest cost items. In conclusion, the clinical and economic burden of follicular lymphoma increases along with later treatment lines. The high mortality rates indicate that further efforts are needed to optimize disease management.

Miconazole-like Scaffold is a Promising Lead for Naegleria fowleri-Specific CYP51 Inhibitors.

Developing drugs for brain infection by Naegleria fowleri is an unmet medical need. We used a combination of cheminformatics, target-, and phenotypic-based drug discovery methods to identify inhibitors that target an essential N. fowleri enzyme, sterol 14-demethylase (NfCYP51). A total of 124 compounds preselected in silico were tested against N. fowleri. Nine primary hits with EC50 ≤ 10 µM were phenotypically identified. Cocrystallization with NfCYP51 focused attention on one primary hit, miconazole-like compound 2a. The S-enantiomer of 2a produced a 1.74 Å cocrystal structure. A set of analogues was then synthesized and evaluated to confirm the superiority of the S-configuration over the R-configuration and the advantage of an ether linkage over an ester linkage. The two compounds, S-8b and S-9b, had an improved EC50 and KD compared to 2a. Importantly, both were readily taken up into the brain. The brain-to-plasma distribution coefficient of S-9b was 1.02 ± 0.12, suggesting further evaluation as a lead for primary amoebic meningoencephalitis.

Gold Nanomaterials-Based Electrochemical Sensors and Biosensors for Phenolic Antioxidants Detection: Recent Advances.

Antioxidants play a central role in the development and production of food, cosmetics, and pharmaceuticals, to reduce oxidative processes in the human body. Among them, phenolic antioxidants are considered even more efficient than other antioxidants. They are divided into natural and synthetic. The natural antioxidants are generally found in plants and their synthetic counterparts are generally added as preventing agents of lipid oxidation during the processing and storage of fats, oils, and lipid-containing foods: All of them can exhibit different effects on human health, which are not always beneficial. Because of their relevant bioactivity and importance in several sectors, such as agro-food, pharmaceutical, and cosmetic, it is crucial to have fast and reliable analysis Rmethods available. In this review, different examples of gold nanomaterial-based electrochemical (bio)sensors used for the rapid and selective detection of phenolic compounds are analyzed and discussed, evidencing the important role of gold nanomaterials, and including systems with or without specific recognition elements, such as biomolecules, enzymes, etc. Moreover, a selection of gold nanomaterials involved in the designing of this kind of (bio)sensor is reported and critically analyzed. Finally, advantages, limitations, and potentialities for practical applications of gold nanomaterial-based electrochemical (bio)sensors for detecting phenolic antioxidants are discussed.

Mechanical and Metallurgical Evaluation of 3 Different Nickel-Titanium Rotary Instruments: An In Vitro and In Laboratory Study.

An in-depth evaluation of the mechanical and metallurgical properties of NiTi instruments is fundamental to assess their performance and to compare recently introduced instrument with widespread ones. According to this, since there are no data on this topic, the aim of the study was to mechanically and metallurgically evaluate an instrument recently introduced into the market (ZenFlex (ZF)), by comparing it with two well-known instruments with similar characteristics: Vortex Blue (VB) and EdgeSequel Sapphire (EES). According to this, 195 instruments were selected: 65 ZF, 65 VB and 65 EES. Each group was divided in subgroups according to the mechanical tests (i.e., cyclic fatigue resistance, torsional resistance and bending ability; (n = 20)) and the metallurgical test (differential scanning calorimetry (n = 5)). A scanning electron microscopy was performed to verify the causes of fracture after mechanical tests (cyclic fatigue and torsional tests). According to results, VB showed the highest flexibility and cyclic fatigue resistance in comparison to the other instruments, with a statistically significant difference (p < 0.05). Regarding torsional resistance, EES showed the lowest value of torque at fracture, with a statistically significant difference, whilst the comparison between ZF and VB showed no statistically significant difference (p > 0.05). DSC analysis pointed out that VB had the highest austenite start and finish temperatures, followed by ESS and then ZF. ESS sample showed the highest martensite start and finish temperatures followed by VB and ZF. Considering the results, it can be concluded that VB showed the best mechanical performance during static tests in comparison to ESS and ZF. This is fundamentally due to the interaction of parameters such as instrument design and heat-treatments that are able to enhance its mechanical performance.

Mechanical properties and metallurgical features of two similar endodontic rotary instruments with different heat treatments (FireWire™ and Gold).

The aim of this study was to explain the mechanical differences between EdgeTaper Platinum (ETP) and ProTaper Gold (PTG) determining the transition temperatures and the composition of the alloy of ETP, since no data are present in literature. A total of 130 instruments were selected: 65 ETP F2 and 65 PTG F2. 20 instruments per type were submitted to each mechanical tests. The cyclic fatigue and torsional tests were performed at a pre-set temperature of 35 °C ± 1 °C. During the cyclic fatigue test, Time to fracture, number of cycles to fracture (NCF) and the fragment length (FL) were recorded. During the static torsional, Torque to Fracture (TtF) and FL were recorded. The fracture surface of 5 randomly selected fragments from each test was examined through a Scanning Electron Microscopy (SEM) and an EDX analysis was performed. 20 instruments per type were submitted to a bending test The force generated (grams) to bend each was recorded. Recorded data of mechanical and metallurgical tests were statistically analyzed using a one-way analysis of variance (ANOVA) test with significance set to a 95% confidence level. ETP F2 showed a higher cyclic fatigue resistance and bending ability than PTG F2, with a statistically significant difference (p < 0.05). PTG showed a higher torsional resistance with a statistically significant difference (p < 0.05). DSC analysis of 5 samples for each instrument type pointed out that the austenite finish temperature of PTG was higher than the ETP, respectively of 49.99 ± 3.31 (°C) and 38.92 ± 1.75 (°C). EDX analysis confirmed the near-equiatomic composition of the Ni-Ti alloys, with a presence of a third component recognized as rubidium in the ETP samples. ETP showed higher flexibility in comparison with PTG, despite the latter is characterized by a more martensitic characterization. since its Af temperature is higher than the ETP.

The hazard of carbapenemase (OXA-181)-producing Escherichia coli spreading in pig and veal calf holdings in Italy in the genomics era: Risk of spill over and spill back between humans and animals.

Carbapenemase-producing Enterobacterales (CPE) are considered a major public health issue. In the frame of the EU Harmonized AMR Monitoring program conducted in Italy in 2021, 21 epidemiological units of fattening pigs (6.98%; 95% CI 4.37-10.47%; 21/301) and four epidemiological units of bovines <12 months (1.29%; 95% CI 0.35-3.27%, 4/310) resulted positive to OXA-48-like-producing E. coli (n = 24 OXA-181, n = 1 OXA-48). Whole Genome Sequencing (WGS) for in-depth characterization, genomics and cluster analysis of OXA-181-(and one OXA-48) producing E. coli isolated, was performed. Tracing-back activities at: (a) the fattening holding of origin of one positive slaughter batch, (b) the breeding holding, and (c) one epidemiologically related dairy cattle holding, allowed detection of OXA-48-like-producing E. coli in different units and comparison of further human isolates from fecal samples of farm workers. The OXA-181-producing isolates were multidrug resistant (MDR), belonged to different Sequence Types (STs), harbored the IncX and IncF plasmid replicons and multiple virulence genes. Bioinformatics analysis of combined Oxford Nanopore Technologies (ONT) long reads and Illumina short reads identified bla OXA-181 as part of a transposon in IncX1, IncX3, and IncFII fully resolved plasmids from 16 selected E. coli, mostly belonging to ST5229, isolated during the survey at slaughter and tracing-back activities. Although human source could be the most likely cause for the introduction of the bla OXA-181-carrying IncX1 plasmid in the breeding holding, concerns arise from carbapenemase OXA-48-like-producing E. coli spreading in 2021 in Italian fattening pigs and, to a lesser extent, in veal calf holdings.

Critical role of flanking residues in NGR-to-isoDGR transition and CD13/integrin receptor switching.

Various NGR-containing peptides have been exploited for targeted delivery of drugs to CD13-positive tumor neovasculature. Recent studies have shown that compounds containing this motif can rapidly deamidate and generate isoaspartate-glycine-arginine (isoDGR), a ligand of alphavbeta3-integrin that can be also exploited for drug delivery to tumors. We have investigated the role of NGR and isoDGR peptide scaffolds on their biochemical and biological properties. Peptides containing the cyclic CNGRC sequence could bind CD13-positive endothelial cells more efficiently than those containing linear GNGRG. Peptide degradation studies showed that cyclic peptides mostly undergo NGR-to-isoDGR transition and CD13/integrin switching, whereas linear peptides mainly undergo degradation reactions involving the alpha-amino group, which generate non-functional six/seven-membered ring compounds, unable to bind alphavbeta3, and small amount of isoDGR. Structure-activity studies showed that cyclic isoDGR could bind alphavbeta3 with an affinity >100-fold higher than that of linear isoDGR and inhibited endothelial cell adhesion and tumor growth more efficiently. Cyclic isoDGR could also bind other integrins (alphavbeta5, alphavbeta6, alphavbeta8, and alpha5beta1), although with 10-100-fold lower affinity. Peptide linearization caused loss of affinity for all integrins and loss of specificity, whereas alpha-amino group acetylation increased the affinity for all tested integrins, but caused loss of specificity. These results highlight the critical role of molecular scaffold on the biological properties of NGR/isoDGR peptides. These findings may have important implications for the design and development of anticancer drugs or tumor neovasculature-imaging compounds, and for the potential function of different NGR/isoDGR sites in natural proteins.

Impact of Dealcoholization by Osmotic Distillation on Metabolic Profile, Phenolic Content, and Antioxidant Capacity of Low Alcoholic Craft Beers with Different Malt Compositions.

Beer antioxidants originate mainly from malts, classified as colored, caramel, and roasted, according to the malting process. This study aimed to characterize, in terms of phenolic antioxidants, three types of Pale Ale craft beers brewed using increasing percentage of dark malt (0, 5, and 15% Caraamber malt, called PA100, PA95, PA85, respectively) and to evaluate the impact of dealcoholization by osmotic distillation (OD) on the same antioxidants. All the alcoholic (PA, 6.2-6.8 vol %) and low alcoholic (LA-PA, 1 vol %) beers were analyzed by HPLC-ESI-MS/MS, total phenolic content (TPC), and antioxidant activity (AA): similar phenolic profiles were evidenced and 43 compounds identified or tentatively identified. Some differences were found among PA100, PA95, and PA85: PA85 was richer in free phenolic compounds (10.55 mg/L) and had a higher TPC (463.7 GAE mg/L) and AA (852.1 TE mg/L). LA-PA beers showed the same phenolic profile and similar TPC and AA compared to PA beers; however, there were some differences regarding LA-PA85 (5.91 mg/L). Dealcoholization by OD seemed to weakly affect the phenolic fraction. ESI-MS/MS infusion experiments evidenced oligosaccharides, small organic acids, and amino acids, whose presence was confirmed and quantitated by NMR: besides ethanol and other alcohols, weak to strong loss of low-molecular-weight metabolites was evidenced in LA-PA beers.