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Nel Parco Nazionale dei Monti Sibillini sono stati catturati e muniti di trasmittente tre individui di cinghiale: M1, maschio di circa 20 mesi, del peso di Kg 50, catturato il 14/09/03 insieme ad altri due individui coetanei; F1, femmina di circa 36 mesi, del peso di Kg 55, catturata il 08/04/03 insieme ad un?altra femmina coetanea, un subadulto maschio e sei striati; M2 maschio di circa 18 mesi, del peso di Kg 50, catturato il 05/06/03. Il programma di monitoraggio è in corso, i dati qui esposti si riferiscono ad M1 ed F1, raccolti fino al 30 aprile. Per M1 si sono avuti 665 contatti; per F1 si sono avuti 61 contatti. M1 si è mantenuto entro i confini del Parco nei primi due mesi successivi alla cattura, nel mese seguente, dicembre, il cinghiale si è progressivamente e definitivamente allontanato dal Parco. L?estensione dello home range (hr) è stata calcolata con Animal Movement per ArcView mediante MCP 100% e Kernel 90%. Lo hr interno al Parco è risultato essere di 651 ettari (MCP) e 535 ettari (Kernel); quello esterno all?area protetta è di 5231 ettari (MCP) e 3084 ettari (Kernel). Complessivamente M1 si è mosso all?interno di un?area estesa 18509 ettari (MCP). La distanza massima percorsa, entro un periodo massimo di 5 giorni, è di Km 17 in linea retta. M1 si è tenuto lontano dai confini del Parco per una distanza media di m 4322, con un intervallo di variabilità compreso tra m 10160 e m 394 (DS = 2516; IF 95% = 4322 ± 328). L?abbandono dell?area protetta da parte di M1 è avvenuto in piena stagione di caccia al cinghiale in un?area soggetta a battuta- braccata, con almeno un caso accertato di compresenza cinghiale-cacciatori. Lo hr di F1 è di ha 280 (MCP) interamente e permanentemente compresi nei confini del Parco. L?uso dello spazio da parte di M1 sembra contrastare con la tradizionale attribuzione della funzione di ?rifugio? alle aree protette: un cinghiale catturato all?interno di un Parco sembra ?scegliere?, probabilmente insieme al suo gruppo, l?esterno dell?area protetta nel periodo di maggiore pressione venatoria. È ipotizzabile che tale spostamento sia conseguenza della dispersione giovanile e/o della pressione derivante dal controllo in atto nel Parco, operato con tecniche di prelievo all?aspetto, indirizzato esclusivamente a questa specie. Il monitoraggio di F1 ed M2 consentirà il confronto con individui coetanei e sintopici.

Antimicrobial resistance (AMR) poses a serious threat to global health. The rapid emergence of resistance contrasts with the slow pace of antimicrobial development, emphasizing the urgent need for innovative drug discovery approaches. This study addresses a critical bottleneck in early drug development by introducing integral solvent-induced protein precipitation (iSPP) to rapidly assess the target-engagement of lead compounds in extracts of pathogenic microorganisms under close-to-physiological conditions. iSPP measures the change in protein stability against solvent-induced precipitation in the presence of ligands. The iSPP method for bacteria builds upon established SPP procedures and features optimized denaturation gradients and minimized sample input amounts. The effectiveness of the iSPP workflow was initially demonstrated through a multidrug target-engagement study. Using quantitative mass spectrometry (LC-MS/MS), we successfully identified known drug targets of seven different antibiotics in cell extracts of four AMR-related pathogens: the three Gram-negative bacteria Escherichia coli , Klebsiella pneumoniae , Pseudomonas aeruginosa and the Gram-positive bacterium Staphylococcus aureus . The iSPP method was ultimately applied to demonstrate target-engagement of compounds derived from target-based drug discovery. We employed five small molecules targeting three enzymes in the 2- C -methyl-d-erythritol 4-phosphate (MEP) pathway─a promising focus for anti-infective drug development. The study showcases iSPP adaptability and efficiency in identifying anti-infective drug targets, advancing early-stage drug discovery against AMR.

The limited understanding of the mechanism of action (MoA) of several antimalarials and the rise of drug resistance toward existing malaria therapies emphasizes the need for new strategies to uncover the molecular target of compounds in Plasmodium falciparum . Integral solvent-induced protein precipitation (iSPP) is a quantitative mass spectrometry-based (LC-MS/MS) proteomics technique. The iSPP leverages the change in solvent-induced denaturation of the drug-bound protein relative to its unbound state, allowing identification of the direct drug-protein target without the need to modify the drug. Here, we demonstrate proof-of-concept of iSPP in P. falciparum (Pf) lysate. At first, we profiled the solvent-induced denaturation behavior of the Pf proteome, generating denaturation curves and determining the melting concentration ( C M ) of 2712 proteins. We then assessed the extent of stabilization of three antimalarial target proteins in multiple organic solvent gradients, allowing for a rational selection of an optimal solvent gradient. Subsequently, we validated iSPP by successfully showing target-engagement of several standard antimalarials. The iSPP assay allows the testing of multiple conditions within reasonable LC-MS/MS measurement time. Furthermore, it requires a minimal amount of protein input, reducing culturing time and simplifying protein extraction. We envision that iSPP will be useful as a complementary tool for MoA studies for next-generation antimalarials.

The methyl-d-erythritol phosphate (MEP) pathway has emerged as an interesting target in the fight against antimicrobial resistance. The pathway is essential in many human pathogens, including Plasmodium falciparum ( Pf ), but is absent in human cells. In the present study, we report on the discovery of a new chemical class targeting IspD, the third enzyme in the pathway. Exploration of the structure-activity relationship yielded inhibitors with potency in the low-nanomolar range. Moreover, we investigated the whole-cell activity, mode of inhibition, metabolic, and plasma stability of this compound class, and conducted in vivo pharmacokinetic profiling on selected compounds. Lastly, we disclosed a new mass spectrometry (MS)-based enzymatic assay for direct IspD activity determination, circumventing the need for auxiliary enzymes. In summary, we have identified a readily synthesizable compound class, demonstrating excellent activity and a promising profile, positioning it as a valuable tool compound for advancing research on IspD.

AbstractHummingbird flower mites are assumed to monopolize single host plant species owing to sexual selection for unique mating rendezvous sites. We tested the main assumption of the mating rendezvous hypothesis-extreme host specialization-by reconstructing interactions among tropical hummingbird flower mites and their host plants using DNA barcoding and taxonomic identifications. We collected 10,654 mites from 489 flowers. We extracted DNA from 1,928 mite specimens and amplified the cytochrome c oxidase I (CO1) DNA barcode. We analyzed the network structure to assess the degree of generalization or specialization of mites to their host plants. We recorded 18 species of hummingbird flower mites from three genera ( Proctolaelaps , Rhinoseius , and Tropicoseius ) interacting with 14 species of plants. We found that generalist mites are common, and congeneric mite species often share host plants. Our results challenge the assumption of strict specialization that supports this system as an example of mating rendezvous evolution.

Despite human gastrointestinal exposure to nanoparticles (NPs), data on NPs toxicity in intestinal cells are quite scanty. In this study we evaluated the toxicity induced by zinc oxide (ZnO) and titanium dioxide (TiO₂) NPs on Caco-2 cells. Only ZnO NPs produced significant cytotoxicity, evaluated by two different assays. The presence of foetal calf serum in culture medium significantly reduced ZnO NPs toxicity as well as ion leakage and NP-cell interaction. The two NPs increased the intracellular amount of reactive oxygen species (ROS) after 6 h treatment. However, only ZnO NPs increased ROS and induced IL-8 release both after 6 and 24 h. Experimental data indicate a main role of chemical composition and solubility in ZnO NPs toxicity. Moreover our results suggest a key role of oxidative stress in ZnO NPs cytotoxicity induction related both to ion leakage and to cell interaction with NPs in serum-free medium.

In modern taxonomy, DNA barcoding is particularly useful where biometric parameters are difficult to determine or useless owing to the poor quality of samples. These situations are frequent in parasitology. Here, we present an integrated study, based on both DNA barcoding and morphological analysis, on cestodes belonging to the genus Taenia, for which biodiversity is still largely underestimated. In particular, we characterized cestodes from Italian wildcats (Felis silvestris silvestris), free-ranging domestic cats (Felis silvestris catus) and hybrids populations. Adult taeniids were collected by post-mortem examinations of the hosts and morphologically identified as Taenia taeniaeformis. We produced cox1 barcode sequences for all the analysed specimens, and we compared them with reference sequences of individuals belonging to the genus Taenia retrieved from GenBank. In order to evaluate the performance of a DNA barcoding approach to discriminate these parasites, the strength of correlation between species identification based on classical morphology and the molecular divergence of cox1 sequences was measured. Our study provides clear evidence that DNA barcoding is highly efficient to reveal the presence of cryptic lineages within already-described taeniid species. Indeed, we detected three well-defined molecular lineages within the whole panel of specimens morphologically identified as T. taeniaeformis. Two of these molecular groups were already identified by other authors and should be ranked at species level. The third molecular group encompasses only samples collected in Italy during this study, and it represents a third candidate species, still morphologically undescribed., (© 2012 Blackwell Publishing Ltd.)

Having described the morphological and functional characteristics of phagocytic cells, the paper underlines that, in addition to the etiological responsibility of bacterial plaque, the main role in the onset and evolution of periodontitis is played by the host's response. Phagocytic response plays a fundamental role in the host's defence reaction and represents the first barrier to the penetration of bacteria into periodontal tissue. In addition to their defensive role, phagocytic cells may also be responsible for damage to periodontal tissue as a collateral effect of their phagocytic function, thus worsening the periodontal lesion.