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The first case of CWD in a Norwegian red deer was detected by a routine ELISA test and confirmed by western blotting and immunohistochemistry in the brain stem of the animal. Two different western blotting tests were conducted independently in two different laboratories, showing that the red deer glycoprofile was different from the Norwegian CWD reindeer and CWD moose and from North American CWD. The isolate showed nevertheless features similar to the classical BSE (BSE-C) strain. Furthermore, BSE-C could not be excluded based on the PrPSc immunohistochemistry staining in the brainstem and the absence of detectable PrPSc in the lymphoid tissues. Because of the known ability of BSE-C to cross species barriers as well as its zoonotic potential, the CWD red deer isolate was submitted to the EURL Strain Typing Expert Group (STEG) as a BSE-C suspect for further investigation. In addition, different strain typing in vivo and in vitro strategies aiming at identifying the BSE-C strain in the red deer isolate were performed independently in three research groups and BSE-C was not found in it. These results suggest that the Norwegian CWD red deer case was infected with a previously unknown CWD type and further investigation is needed to determine the characteristics of this potential new CWD strain.

After the detection of bovine spongiform encephalopathy (BSE), and a zoonotic transmissible spongiform encephalopathy (TSE) caused by the pathological prion protein (PrP Sc ) in two goats, the investigation of goat prions became of greater interest. Therefore, a broad collection of European goat TSE isolates, including atypical scrapie, CH1641 and goat BSE as reference prion strains were biochemically characterised and subsequently inoculated into seven rodent models for further analysis (already published results of this comprehensive study are reviewed here for com-parative reasons). We report here the histopathological and immunohistochemical data of this goatTSE panel, obtained after the first passage in Tgshp IX (tg-shARQ) mice, which over express the ovine prion protein. In addition to the clear-cut discrimination of all reference prion strains from the classical scrapie (CS) isolates, we were further able to determine three categories of CS strains.The investigation further indicates the occurrence of sub-strains that slightly resemble distant TSE strains, such as BSE or CH1641, reinforcing the theory that CS is not a single strain but a mixture of sub-strains, existing at varying extents in one isolate. This study further proved that Tgshp IX is apotent and reliable tool for the in-depth characterisation of prion strains.

The data set consists of an excel file containing all the data used for the redaction of the systematic review “Combating viruses with cold plasma: A systematic review of cold atmospheric plasma applications in virology” submitted for publication.

Introduction: Lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called nano-sized hybridosomes or nanohybridosomes, NSHs) were considered. Methods: We formed NSHs by mixing appropriate amounts of lecithin, glycerol and supernatant of isolation of extracellular vesicles from spruce needle homogenate. We visualized NSHs by light microscopy and cryogenic transmission electron microscopy and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy, interferometric light microscopy and liquid chromatography–mass spectrometry. Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well on the average hydrodynamic radius of the particles Rh (between 140 and 180 nm), while the concentrations of the particles were in the range between 1013 and 1014/mL indicating that NSHs present a considerable (more than 25%) of the sample which is much more than the yield of natural extracellular vesicles (EVs) from spruce needle homogenate (estimated less than 1%). Spruce specific lipids and proteins were found in hybridosomes. Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that large-scale production of NSHs from lipids and spruce needle homogenate is feasible. Plain Language Summary: Cells shed into their exterior nanoparticles (here referred to as extracellular vesicles – EVs) that are free to move, reach distant cells and are taken up by them. As they carry bioactive constituents, EVs may have important impact on the recipient cells. The mechanisms of EV formation and mediation can be employed in designing therapeutic, prophylactic and diagnostic methods for various medical issues. EVs can

Vesna Spasovski,1,2,* Anna Romolo,1,3,* UrÅ¡ka Zagorc,4 Vesna Arrigler,4 Matic Kisovec,5 Apolonija Bedina Zavec,5 Matevž Arko,1 Adrienn Molnár,6,7 Gitta Schlosser,7 AleÅ¡ Iglič,3,8 Ksenija Kogej,4 Veronika Kralj-Iglič1 1University of Ljubljana, Faculty of Health Sciences, Laboratory of Clinical Biophysics, Ljubljana, Slovenia; 2Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia; 3University of Ljubljana, Faculty of Electrical Engineering, Laboratory of Physics, Ljubljana, Slovenia; 4University of Ljubljana, Faculty of Chemistry and Chemical Technology, Ljubljana, Slovenia; 5National Institute of Chemistry, Department of Molecular Biology and Nanobiotechnology, Ljubljana, Slovenia; 6Hevesy György PhD School of Chemistry, ELTE Eötvös Loránd University, Budapest, Hungary; 7MTA-ELTE Lendület Ion Mobility Mass Spectrometry Research Group, Faculty of Science, Institute of Chemistry, ELTE Eötvös Loránd University, Budapest, Hungary; 8University of Ljubljana, Faculty of Medicine, Laboratory of Clinical Biophysics, Ljubljana, Slovenia*These authors contributed equally to this workCorrespondence: Veronika Kralj-Iglič, Email veronika.kralj-iglic@zf.uni-lj.siIntroduction: Lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called nano-sized hybridosomes or nanohybridosomes, NSHs) were considered.Methods: We formed NSHs by mixing appropriate amounts of lecithin, glycerol and supernatant of isolation of extracellular vesicles from spruce needle homogenate. We visualized NSHs by light microscopy and cryogenic transmission electron microscopy and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy, interferometric light microscopy and liquid chromatography–mass spectrometry.Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow c

Atypical Scrapie, which is not linked to epidemics, is assumed to be an idiopathic spontaneous prion disease in small ruminants. Therefore, its occurrence is unlikely to be controlled through selective breeding or other strategies as it is done for classical scrapie outbreaks. Its spontaneous nature and its sporadic incidence worldwide is reminiscent of the incidence of idiopathic spontaneous prion diseases in humans, which account for more than 85% of the cases in humans. Hence, developing animal models that consistently reproduce this phenomenon of spontaneous PrP misfolding, is of importance to study the pathobiology of idiopathic spontaneous prion disorders. Transgenic mice overexpressing sheep PrPC with I112 polymorphism (TgShI112, 1-2 × PrP levels compared to sheep brain) manifest clinical signs of a spongiform encephalopathy spontaneously as early as 380 days of age. The brains of these animals show the neuropathological hallmarks of prion disease and biochemical analyses of the misfolded prion protein show a ladder-like PrPres pattern with a predominant 7-10 kDa band. Brain homogenates from spontaneously diseased transgenic mice were inoculated in several models to assess their transmissibility and characterize the prion strain generated: TgShI112 (ovine I112 ARQ PrPC), Tg338 (ovine VRQ PrPC), Tg501 (ovine ARQ PrPC), Tg340 (human M129 PrPC), Tg361 (human V129 PrPC), TgVole (bank vole I109 PrPC), bank vole (I109I PrPC), and sheep (AHQ/ARR and AHQ/AHQ churra-tensina breeds). Our analysis of the results of these bioassays concludes that the strain generated in this model is indistinguishable to that causing atypical scrapie (Nor98). Thus, we present the first faithful model for a bona fide, transmissible, ovine, atypical scrapie prion disease.

Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation of extracellular particles from spruce needle homogenate and glycerol. Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and interferometric microscopy. Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well with the determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the mixture, excluding solvent and other small molecules. Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle homogenate is feasible.

[Italiano]: Questo Manuale di geoarcheologia. Un approccio geomorfologico da remoto intende offrirsi come strumento di formazione e conoscenza a quanti, studenti universitari, ricercatori e cultori della materia, intendano approfondire lo studio dell’ambiente considerato come contenitore delle testimonianze del passato nella sua perenne evoluzione di forme e di risorse. Esso raccoglie l’esperienza acquisita sul campo, nel corso di vari decenni di attività di ricognizione e scavo, da parte dei suoi Autori in Italia e in numerosi paesi del vicino e medio-oriente, del centro Asia e del nord Africa. Ricerche sostenute e condotte sempre con l’ausilio di un punto prospettico privilegiato, quale quello offerto dalle varie immagini telerilevate riprese con sensori differenti da piattaforme aeree e satellitari in periodi temporali successivi, la cui interpretazione è stata sempre confrontata con le osservazioni dirette, ovvero con la cosiddetta ‘verità terreno’. Proprio questa stretta integrazione tra paesaggio, risorse del territorio ed emergenze antropiche più o meno antiche costituisce l’elemento portante del libro, declinato attraverso numerosi ed esaustivi esempi di analisi in vari ambienti dell’area mediterranea, del Vicino Oriente e dell’Asia che vedono applicato un approccio teorico ampiamente illustrato nella prima parte del volume. L’augurio è che il produttivo connubio tra tecnologia avanzata e cultura diventi una comune attitudine tra le giovani generazioni di futuri studiosi e che l’esperienza maturata in questo settore da parte degli scriventi venga raccolta e implementata, raggiungendo nuovi traguardi e più fruttuosi risultati./[English]: This Handbook of geoarchaeology. A remote geomorphological approach intends to offer itself as a training and knowledge tool to those who, university students, researchers and connoisseurs of the subject, intend to deepen the study of the environment as a context of the testimonies of the past in its perennial evolution of fo

Introduction: Being of compatible structure with biomembranes, lipid–based nanoparticles are considered as convenient platforms for drug delivery systems. In the proposed work we considered formation of lipid nanovesicles associated with bioactive phytochemicals from spruce needle homogenate (here called hybridosomes). We formed hybridosomes by mixing appropriate amounts of lecithin, supernatant of isolation of extracellular particles from spruce needle homogenate and glycerol. Methods: We visualized hybridosomes by light microscopy and cryogenic transmission electron microscopy and assessed them by flow cytometry, dynamic light scattering, ultraviolet–visual spectroscopy and interferometric microscopy. Results: We found that the particles consisted of a bilayer membrane and a fluid-like interior. Flow cytometry and interferometric light microscopy measurements showed that the majority of the particles were nano-sized. Dynamic light scattering and interferometric light microscopy measurements agreed well with the determined average hydrodynamic radius of the particles Rh (between 140 and 180 nm) while their number densities were in the range between 10^13 and 10^14/mL indicating that hybridosomes present about 2/3 of the mixture, excluding solvent and other small molecules. Discussion: Simple and low-cost preparation method, non-demanding saving process and efficient formation procedure suggest that large scale production of hybridosomes from lipids and spruce needle homogenate is feasible.

Introduction: Blood plasma is used in regenerative medicine for more than 30 years as it was found to have impact on angiogenesis and proliferation of endothelial cells. This effect was initially ascribed to growth factors and cytokines, however, particularly in the last decade researchers are focusing on platelet-derived extracellular vesicles (PEVs) that are also present in the platelet rich plasma. Methods: We prepared platelet and extracellular vesicles-rich plasma (PVRP) by centrifugation of the human blood, and differential (ultra) centrifugation of PVRP to isolate PEVs. We exposed Human Umbilical Vein Endothelial Cells (HUVEC) to 5% PEVs for 24 hours and assessed inflammation markers (Interleukin(IL)-1beta, IL-6 and Tumor necrosis factor (TNF)-alpha using ELISA tests), oxidative stress markers (Cholinesterase (ChE) and glutathione S-transferase (GST) activity by spectrophotometry, as well reactive oxygen species (ROS) and lipid droplets (LD) by flow cytometry). We observed morphological changes in HUVEC indicating angiogenesis by using optical microscopy. Results: We found that after 24 hours caused a decrease of the concentration of IL-6, IL-1beta and TNF-alpha. Also, ChE and GST activity and ROS count were decreased. LD production, which is triggered in need to protect the cells from free radicals and oxidative stress damage, was however higher. Treatment of the cells had impact on cell morphology with progressed formation of the tubes and cell connecting, which is regarded as the beginning of HUVEC angiogenesis process. Discussion: Beneficial effect of PVRP in healing and regeneration may include suppression of inflammation and oxidative stress by PEVs.

Epithelial ovarian cancer (EOC) is the most common type of gynecological tumor, presenting poor prognosis at diagnosis and with recurrences being frequently observed. Reactive species generated by physical plasma and transferred into liquids have shown promising results in cancer therapy. Recently, Ringer's lactate solution was exposed to plasma showing selective anticancer activity on EOC cells. In this work, we compared the effect of plasma treatment, using the kINPen plasma jet, on Ringer's saline and Ringer's lactate solution. These two plasma-treated liquids were analyzed chemically by quantifying reactive species and the extent of lactate oxidation. The biological efficiency of the plasma-treated liquids was explored in EOC cells. The results show that lactate is affected by plasma treatment, displaying a reduction of cytotoxic potential., This publication is based upon work from COST Action Action CA20114 PlasTHER “Therapeutical Applications of Cold Plasmas,” supported by COST. The authors also acknowledge MINECO for PID2019‐103892RB‐I00/AEI/ 12 of 14 | BUCCI ET AL .16128869, 0, Downloaded from https://onlinelibrary.wiley.com/doi/10.1002/ppap.202300093 by Readcube (Labtiva Inc.), Wiley Online Library on [13/11/2023]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License 10.13039/501100011033 project and for PLEC2022‐ 009277/MCIN/AEI/10.13039/501100011033 funded by UE through NextGenerationEU/PRTR. The authors belong to SGR2017‐1165 (Cristina Canal, Miguel Mateu‐Sanz, and Francesco Tampieri) and acknowledge Generalitat de Catalunya for the ICREA Academia Award for Excellence in Research of Cristina Canal., Peer Reviewed, Postprint (published version)

Chitosan films have attracted increased attention in the field of sensors because of chitosan’s unique chemico-physical properties, including high adsorption capacity, filmability and transparency. A chitosan film sensor was developed through the dispersion of an ammonia specific reagent (Nessler’s reagent) into a chitosan film matrix. The chitosan film sensor was characterized to assess the film’s properties by Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), scanning electron microscopy (SEM) and differential scanning calorimetry (DSC). A gas diffusion device was prepared with the chitosan film sensor, enabling the collection and detection of ammonia vapor from biological samples. The chitosan film sensor color change was correlated with the ammonia concentration in samples of human serum and artificial urine. This method enabled facile ammonia detection and concentration measurement, making the sensor useful not only in clinical laboratories, but also for point-of-care devices and wherever there is limited access to modern laboratory facilities.

The contamination of water used in post-harvest handling and processing operations of fresh andfrozen fruit, vegetables and herbs (ffFVHs) is a global concern. The most relevant microbial hazardsassociated with this water are:Listeria monocytogenes,Salmonellaspp., human pathogenicEscherichia coliand enteric viruses, which have been linked to multiple outbreaks associated withffFVHs in the European Union (EU). Contamination (i.e. the accumulation of microbiological hazards) ofthe process water during post-harvest handling and processing operations is affected by severalfactors including: the type and contamination of the FVHs being processed, duration of the operationand transfer of microorganisms from the product to the water and vice versa, etc. For food businessoperators (FBOp), it is important to maintain the microbiological quality of the process water to assurethe safety of ffFVHs. Good manufacturing practices (GMP) and good hygienic practices (GHP) relatedto a water management plan and the implementation of a water management system are critical tomaintain the microbiological quality of the process water. Identified hygienic practices include technicalmaintenance of infrastructure, training of staff and cooling of post-harvest process water. Interventionstrategies (e.g. use of water disinfection treatments and water replenishment) have been suggested tomaintain the microbiological quality of process water. Chlorine-based disinfectants and peroxyaceticacid have been reported as common water disinfection treatments. However, given current practices inthe EU, evidence of their efficacy under industrial conditions is only available for chlorine-baseddisinfectants. The use of water disinfection treatments must be undertaken following an appropriatewater management strategy including validation, operational monitoring and verification. Duringoperational monitoring, real-time information on process parameters related to the process andproduct, as well as the water an

The preparation of autologous platelet and extracellular vesicle-rich plasma (PVRP) has been explored in many medical fields with the aim to benefit from its healing potential. In parallel, efforts are being invested to understand the function and dynamics of PVRP that is complex in its composition and interactions. Some clinical evidence reveals beneficial effects of PVRP, while some report that there were no effects. To optimize the preparation methods, functions and mechanisms of PVRP, its constituents should be better understood. With the intention to promote further studies of autologous therapeutic PVRP, we performed a review on some topics regarding PVRP composition, harvesting, assessment and preservation, and also on clinical experience following PVRP application in humans and animals. Besides the acknowledged actions of platelets, leukocytes and different molecules, we focus on extracellular vesicles that were found abundant in PVRP.

The Middle Ages represented a crucial period for the evolutionary history of wolves (Canis lupus), marked by both significant ecosystem changes, especially through the degradation of wooded landscapes and heavy persecution, that drove this species to a dramatic demographic decline. In Europe, informative and well-documented wolf remains from the Medieval Ages are exceptionally rare and are mostly represented by teeth and postcranial elements. In this study, we describe a well-preserved wolf cranium dated to ca. 967–1157 AD from the Po Valley (northern Italy). The specimen was analysed through a multidisciplinary approach including CT-based, ancient DNA, and palaeopathological analyses. Morphological and genetic data supported the assignment of this sample to Canis lupus species. CT-based analyses indicated a typical wolf-like morphology falling into the extant variability of the medium-sized subspecies C. lupus italicus, whereas palaeopathological analyses indicated a severe periodontitis. Phylogenetic analyses showed that the Po valley wolf had a unique and never described mtDNA control region haplotype, testifying variability in the ancient Italian wolf, which has now been lost. This study provides the first comprehensive description of a wolf from the Middle Ages, adding useful information for a deeper knowledge about population dynamics, variability, and diseases of this species.

[Italiano]: Questo Manuale di geoarcheologia. Un approccio geomorfologico da remoto intende offrirsi come strumento di formazione e conoscenza a quanti, studenti universitari, ricercatori e cultori della materia, intendano approfondire lo studio dell’ambiente considerato come contenitore delle testimonianze del passato nella sua perenne evoluzione di forme e di risorse. Esso raccoglie l’esperienza acquisita sul campo, nel corso di vari decenni di attività di ricognizione e scavo, da parte dei suoi Autori in Italia e in numerosi paesi del vicino e medio-oriente, del centro Asia e del nord Africa. Ricerche sostenute e condotte sempre con l’ausilio di un punto prospettico privilegiato, quale quello offerto dalle varie immagini telerilevate riprese con sensori differenti da piattaforme aeree e satellitari in periodi temporali successivi, la cui interpretazione è stata sempre confrontata con le osservazioni dirette, ovvero con la cosiddetta ‘verità terreno’. Proprio questa stretta integrazione tra paesaggio, risorse del territorio ed emergenze antropiche più o meno antiche costituisce l’elemento portante del libro, declinato attraverso numerosi ed esaustivi esempi di analisi in vari ambienti dell’area mediterranea, del Vicino Oriente e dell’Asia che vedono applicato un approccio teorico ampiamente illustrato nella prima parte del volume. L’augurio è che il produttivo connubio tra tecnologia avanzata e cultura diventi una comune attitudine tra le giovani generazioni di futuri studiosi e che l’esperienza maturata in questo settore da parte degli scriventi venga raccolta e implementata, raggiungendo nuovi traguardi e più fruttuosi risultati./[English]: This Handbook of geoarchaeology. A remote geomorphological approach intends to offer itself as a training and knowledge tool to those who, university students, researchers and connoisseurs of the subject, intend to deepen the study of the environment as a context of the testimonies of the past in its perennial evolution of fo

Neural networks find widespread use in scientific and technological applications, yet their implementations in conventional computers have encountered bottlenecks due to ever-expanding computational needs. Photonic neuromorphic hardware, which manipulates information and represents data continuously in the optical domain, is one of the promising platforms with potential advantages of massive parallelism, ultralow latency, and reduced energy consumption. While linear photonic neural networks are within reach, photonic computing with large-scale optical nonlinear nodes remains largely unexplored. Here, we demonstrate a large-scale, high-performance nonlinear photonic neural system based on a disordered polycrystalline slab composed of lithium niobate nanocrystals. Mediated by random quasi-phase-matching and multiple scattering, linear and nonlinear optical speckle features are generated as the interplay between the simultaneous linear random scattering and the second-harmonic generation, defining a complex neural network in which the second-order nonlinearity acts as internal nonlinear activation functions. Benchmarked against linear random projection, such nonlinear mapping embedded with rich physical computational operations shows improved performance across a large collection of machine learning tasks in image classification, regression, and graph classification with varying complexity. Demonstrating up to 27,648 input and 3,500 nonlinear output nodes, the combination of optical nonlinearity and random scattering serves as a scalable computing engine for diverse applications.

The diversity of goat scrapie strains in Europe has recently been studied using bioassays in a wide collection of rodent models, resulting in the classification of classical scrapie into four different categories. However, the sole use of the first passage does not lead to isolate adaptation and identification of the strains involved and might therefore lead to misclassification of some scrapie isolates. Therefore, this work reports the complete transmission study of a wide collection of goat transmissible spongiform encephalopathy (TSE) isolates by intracranial inoculation in two transgenic mouse lines overexpressing either small ruminant (TgGoat-ARQ) or bovine (TgBov) PrPC. To compare scrapie strains in sheep and goats, sheep scrapie isolates from different European countries were also included in the study. Once the species barrier phenomenon was overcome, an accurate classification of the isolates was attained. Thus, the use of just two rodent models allowed us to fully differentiate at least four different classical scrapie strains in small ruminants and to identify isolates containing mixtures of strains. This work reinforces the idea that classical scrapie in small ruminants is a prion disease caused by multiple different prion strains and not by a single strain, as is the case for epidemic classical bovine spongiform encephalopathy (BSE-C). In addition, the clear dissimilarity between the different scrapie strains and BSE-C does not support the idea that classical scrapie is the origin of epidemic BSE-C.

Bovine Spongiform Encephalopathy (BSE) is the only animal prion which has been recognized as a zoonotic agent so far. The identification of BSE in two goats raised the need to reliably identify BSE in small ruminants. However, our understanding of scrapie strain diversity in small ruminants remains ill-defined, thus limiting the accuracy of BSE surveillance and spreading fear that BSE might lurk unrecognized in goats. We investigated prion strain diversity in a large panel of European goats by a novel experimental approach that, instead of assessing the neuropathological profile after serial transmissions in a single animal model, was based on the direct interaction of prion isolates with several recipient rodent models expressing small ruminants or heterologous prion proteins. The findings show that the biological properties of scrapie isolates display different patterns of geographical distribution in Europe and suggest that goat BSE could be reliably discriminated from a wide range of biologically and geographically diverse goat prion isolates. Finally, most field prion isolates showed composite strain features, with discrete strain components or sub-strains being present in different proportions in individual goats or tissues. This has important implications for understanding the nature and evolution of scrapie strains and their transmissibility to other species, including humans.

[EN] Intra- and inter-specific gene flow are natural evolutionary processes. However, human-induced hybridization is a global conservation concern across taxa, and the development of discriminant genetic markers to differentiate among gene flow processes is essential. Wolves (Canis lupus) are affected by hybridization, particularly in southern Europe, where ongoing recolonization of historic ranges is augmenting gene flow among divergent populations. Our aim was to provide diagnostic canid markers focused on the long-divergent Iberian, Italian and Dinaric wolf populations, based on existing genomic resources. We used 158 canid samples to select a panel of highly informative single nucleotide polymorphisms (SNPs) to (i) distinguish wolves in the three regions from domestic dogs (C. l. familiaris) and golden jackals (C. aureus), and (ii) identify their first two hybrid generations. The resulting 192 SNPs correctly identified the five canid groups, all simulated first-generation (F1) hybrids (0.482≤Qi≤0.512 between their respective parental groups) and all first backcross (BC1) individuals (0.723≤Qi≤0.827 to parental groups). An assay design and test with invasive and non-invasive canid samples performed successfully for 178 SNPs. By separating natural population admixture from inter-specific hybridization, our reduced panel can help advance evolutionary research, monitoring, and timely conservation management.

Voles are maintenance hosts of Mycobacterium microti. In line with the goal to eradicate tuberculosis (TB) in livestock, the role of this mycobacteria needs to be assessed since it might interfere with current M. bovis / M. caprae surveillance strategies. To better understand the pathogenesis of TB in voles, an experimental infection model was set up to reproduce M. microti infection in laboratory Bank voles (Myodes glareolus). Two infection routes (intragastric and intraperitoneal) and doses (10 5 and 10 6 CFU/0.1 mL) were assessed. Voles were culled at different post-infection time points. Serology, histopathology, acid-fast bacilli staining, qPCR, and mycobacterial culture from tissues were performed. In addition, qPCR from feces and oral swabs were conducted to assess bacterial shedding. The model allowed us to faithfully reproduce the disease phenotype described in free-ranging voles and characterize the pathogenesis of the infection. Most animals showed multifocal and diffuse granulomatous lesions in the liver and spleen, respectively. Less frequently, granulomas were observed in lungs, lymph nodes, muscles, and salivary gland. Mycobacterial DNA was detected in feces from a few animals but not in oral swabs. However, one contact uninfected vole seroconverted and showed incipient TB compatible lesions, suggesting horizontal transmission between voles.

[EN] Intra- and inter-specific gene flow are natural evolutionary processes. However, human-induced hybridization is a global conservation concern across taxa, and the development of discriminant genetic markers to differentiate among gene flow processes is essential. Wolves (Canis lupus) are affected by hybridization, particularly in southern Europe, where ongoing recolonization of historic ranges is augmenting gene flow among divergent populations. Our aim was to provide diagnostic canid markers focused on the long-divergent Iberian, Italian and Dinaric wolf populations, based on existing genomic resources. We used 158 canid samples to select a panel of highly informative single nucleotide polymorphisms (SNPs) to (i) distinguish wolves in the three regions from domestic dogs (C. l. familiaris) and golden jackals (C. aureus), and (ii) identify their first two hybrid generations. The resulting 192 SNPs correctly identified the five canid groups, all simulated first-generation (F1) hybrids (0.482≤Qi≤0.512 between their respective parental groups) and all first backcross (BC1) individuals (0.723≤Qi≤0.827 to parental groups). An assay design and test with invasive and non-invasive canid samples performed successfully for 178 SNPs. By separating natural population admixture from inter-specific hybridization, our reduced panel can help advance evolutionary research, monitoring, and timely conservation management.

OBJECTIVE: To ensure the safety and quality of vaccines, especially the newest RNA-vaccines against COVID-19, is one of the World Health Organization's current highest priorities. DESIGN: Case description. STUDY SAMPLE: We report three cases of sudden unilateral tinnitus following BNT162b2 mRNA-vaccine injection, which rapidly resolved in 2 out of 3 cases. RESULTS: The mechanism responsible for its development remains unclear. A hypersensitivity reaction with an abnormal autoimmune response or a vasculitic event may be implicated. CONCLUSIONS: Large-scale and well-designed studies are needed to improve surveillance of the COVID-19 vaccine and better define possible adverse reactions involving the cochleo-vestibular system and/or immunisation anxiety-related reactions.

Hybridisation between wild and domestic taxa raises complex questions for conservation. Genetic advances offer new methods for hybrid identification, yet social and cultural factors can influence study design, and the interpretation, application, and communication of results. A relevant illustration is hybridisation between domestic dogs (Canis lupus familiaris) and wild canids, such as grey wolves (C. lupus). For regional European monitoring programs in areas with expanding wolf populations, priorities include shared genetic markers and inclusion of all relevant reference populations to ensure dispersing wolves are identified as such and not classified as wolf-dog hybrids, which may cause harmful management decisions. Beyond technical developments, hybrid research and conservation management can benefit from improved integration of legal and policy perspectives, recognition of phenotypic traits as broadly unreliable for identification, and attention to the drivers of, and responses to, evolution in human-dominated landscapes. Additionally, the proliferation of unsubstantiated reports about hybrids in popular and social media shows that communication based on verified findings of hybridisation is essential. Hybridisation requires more constructive discussion on how to balance potentially competing conservation objectives, and the integration of multidisciplinary perspectives. These encompass the welfare of individual animals and preservation of historical predator-prey relationships. Conservation measures centred on preserving the ecological function of wild canids likely offer the most sustainable prospects but require improved understanding of the extent to which their behavioural ecology might differ from that of hybrids. Accurate genetic identification is required to fill this critical knowledge gap, advance public discourse, and initiate relevant conservation actions.

Small particles in natural sources are a subject of interest for their potential role in intercellular, inter-organism, and inter-species interactions, but their harvesting and assessment present a challenge due to their small size and transient identity. We applied a recently developed interferometric light microscopy (ILM) to assess the number density and hydrodynamic radius (Rh) of isolated small cellular particles (SCPs) from blood preparations (plasma and washed erythrocytes) (B), spruce needle homogenate (S), suspension of flagellae of microalgae Tetraselmis chuii (T), conditioned culture media of microalgae Phaeodactylum tricornutum (P), and liposomes (L). The aliquots were also assessed by flow cytometry (FCM), dynamic light scattering (DLS), ultraviolet-visible spectrometry (UV-vis), and imaging by cryogenic transmission electron microscopy (cryo-TEM). In Rh, ILM showed agreement with DLS within the measurement error in 10 out of 13 samples and was the only method used here that yielded particle density. Cryo-TEM revealed that representative SCPs from Tetraselmis chuii flagella (T) did not have a globular shape, so the interpretation by Rh of the batch methods was biased. Cryo-TEM showed the presence of thin filaments in isolates from Phaeodactylum tricornutum conditioned culture media (P), which provides an explanation for the considerably larger Rh obtained by batch methods than the sizes of particles observed by cryo-TEM images. ILM proved convenient for assessment of number density and Rh of SCPs in blood preparations (e.g., plasma); therefore, its use in population and clinical studies is indicated.

Surface analysis techniques have rapidly evolved in the last decade. Some of these are already routinely used in forensics, such as for the detection of gunshot residue or for glass analysis. Some surface analysis approaches are attractive for their portability to the crime scene. Others can be very helpful in forensic laboratories owing to their high spatial resolution, analyte coverage, speed, and specificity. Despite this, many proposed applications of the techniques have not yet led to operational deployment. Here, we explore the application of these techniques to the most important traces commonly found in forensic casework. We highlight where there is potential to add value and outline the progress that is needed to achieve operational deployment. We consider within the scope of this review surface mass spectrometry, surface spectroscopy, and surface X-ray spectrometry. We show how these tools show great promise for the analysis of fingerprints, hair, drugs, explosives, and microtraces., Green Open Access added to TU Delft Institutional Repository 'You share, we take care!' - Taverne project https://www.openaccess.nl/en/you-share-we-take-care Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public., ChemE/Advanced Soft Matter

[EN] Intra- and inter-specific gene flow are natural evolutionary processes. However, human-induced hybridization is a global conservation concern across taxa, and the development of discriminant genetic markers to differentiate among gene flow processes is essential. Wolves (Canis lupus) are affected by hybridization, particularly in southern Europe, where ongoing recolonization of historic ranges is augmenting gene flow among divergent populations. Our aim was to provide diagnostic canid markers focused on the long-divergent Iberian, Italian and Dinaric wolf populations, based on existing genomic resources. We used 158 canid samples to select a panel of highly informative single nucleotide polymorphisms (SNPs) to (i) distinguish wolves in the three regions from domestic dogs (C. l. familiaris) and golden jackals (C. aureus), and (ii) identify their first two hybrid generations. The resulting 192 SNPs correctly identified the five canid groups, all simulated first-generation (F1) hybrids (0.482≤Qi≤0.512 between their respective parental groups) and all first backcross (BC1) individuals (0.723≤Qi≤0.827 to parental groups). An assay design and test with invasive and non-invasive canid samples performed successfully for 178 SNPs. By separating natural population admixture from inter-specific hybridization, our reduced panel can help advance evolutionary research, monitoring, and timely conservation management.

Giant planets can interact with multiple and chemically diverse environments in protoplanetary discs while they form and migrate to their final orbits. The way this interaction affects the accretion of gas and solids shapes the chemical composition of the planets and of their atmospheres. Here we investigate the effects of different chemical structures of the host protoplanetary disc on the planetary composition. We consider both scenarios of molecular (inheritance from the pre-stellar cloud) and atomic (complete chemical reset) initial abundances in the disc. We focus on four elemental tracers of different volatility: C, O, N, and S. We explore the entire extension of possible formation regions suggested by observations by coupling the disc chemical scenarios with N-body simulations of forming and migrating giant planets. The planet formation process produces giant planets with chemical compositions significantly deviating from that of the host disc. We find that the C/N, N/O, and S/N ratios follow monotonic trends with the extent of migration. The C/O ratio shows a more complex behaviour, dependent on the planet accretion history and on the chemical structure of the formation environment. The comparison between S/N* and C/N* (where * indicates normalisation to the stellar value), constrains the relative contribution of gas and solids to the total metallicity. Giant planets whose metallicity is dominated by the contribution of the gas are characterised by N/O* > C/O* > C/N* and allow for constraining the disc chemical scenario. When the planetary metallicity is instead dominated by the contribution of the solids we find that C/N* > C/O* > N/O*., Comment: 27 pages, 10 figures, 1 table. Published in The Astrophysical Journal

The forensic scenario, on which the round robin study was based, simulated a suspected intentional manipulation of a real estate rental agreement consisting of a total of three pages. The aims of this study were to (i) establish the amount and reliability of information extractable from a single type of evidence and to (ii) provide suggestions on the most suitable combination of compatible techniques for a multi-modal imaging approach to forgery detection. To address these aims, seventeen laboratories from sixteen countries were invited to answer the following tasks questions: (i) which printing technique was used? (ii) were the three pages printed with the same printer? (iii) were the three pages made from the same paper? (iv) were the three pages originally stapled? (v) were the headings and signatures written with the same ink? and (vi) were headings and signatures of the same age on all pages? The methods used were classified into the following categories: Optical spectroscopy, including multispectral imaging, smartphone mapping, UV-luminescence and LIBS; Infrared spectroscopy, including Raman and FTIR (micro-)spectroscopy; X-ray spectroscopy, including SEM-EDX, PIXE and XPS; Mass spectrometry, including ICPMS, SIMS, MALDI and LDIMS; Electrostatic imaging, as well as non-imaging methods, such as non-multimodal visual inspection, (micro-)spectroscopy, physical testing and thin layer chromatography. The performance of the techniques was evaluated as the proportion of discriminated sample pairs to all possible sample pairs. For the undiscriminated sample pairs, a distinction was made between undecidability and false positive claims. It was found that none of the methods used were able to solve all tasks completely and/or correctly and that certain methods were a priori judged unsuitable by the laboratories for some tasks. Correct results were generally achieved for the discrimination of printer toners, whereas incorrect results in the discrimination of inks. For th

The market of falsified or sub-standard medical products is a global scale phenomenon. This issue affects a wide range of medications, including life-saving medical products. In high-income countries the most falsified products are those defined “lifestyle”, which include foremost anabolic steroids and phosphodiesterase 5 inhibitors. The spread of these products in the last years has been possible also because of their online purchase, since they can be bought anonymously and without any medical supervision or prescription. Their use can pose a serious threat for public health, especially because often are manufactured without adherence to quality standards. This leads to final products containing active ingredients different from those declared, at the wrong or unknown dose and contaminated with metals, synthesis by-products and other chemical substances. In this work, we present results on characterisation of illegal pharmaceutical products and doping agents by combining different techniques: chromatography coupled to mass spectrometry for organic analysis and accelerator-based nuclear analytical techniques, such as ion beam analysis (IBA), for elemental analysis. Three IBA techniques, namely PIXE (particle induced X-ray emission), PIGE (particle induced gamma-ray emission) and EBS (elastic backscattering spectrometry) were used in external beam mode to provide an elemental characterisation of the as-is material, placed simply in front of the proton beam, thus avoiding the need of preparing them with pre-analytical steps and greatly enhancing the measurement throughput. Several elements (F, Mg, Al, Si, P, S, Cl, K, Ca, Ti, V, Mn, Fe, Co, Ni, Cu, Zn, Br and Sr) were identified in the analysed products. External beam IBA measurements provided the quantitative elemental characterisation of the illegal pharmaceutical products and doping agents under study, complementary to the organic analysis results by chromatography and mass spectrometry thus allowing a rapid (a fe

Background: Understanding the processes that lead to hybridization of wolves and dogs is of scientific and management importance, particularly over large geographical scales, as wolves can disperse great distances. However, a method to efficiently detect hybrids in routine wolf monitoring is lacking. Microsatellites offer only limited resolution due to the low number of markers showing distinctive allele frequencies between wolves and dogs. Moreover, calibration across laboratories is time-consuming and costly. In this study, we selected a panel of 96 ancestry informative markers for wolves and dogs, derived from the Illumina CanineHD Whole-Genome BeadChip (174 K). We designed very short amplicons for genotyping on a microfluidic array, thus making the method suitable also for non-invasively collected samples. Results: Genotypes based on 93 SNPs from wolves sampled throughout Europe, purebred and non-pedigree dogs, and suspected hybrids showed that the new panel accurately identifies parental individuals, first-generation hybrids and first-generation backcrosses to wolves, while second- and third-generation backcrosses to wolves were identified as advanced hybrids in almost all cases. Our results support the hybrid identity of suspect individuals and the non-hybrid status of individuals regarded as wolves. We also show the adequacy of these markers to assess hybridization at a European-wide scale and the importance of including samples from reference populations. Conclusions: We showed that the proposed SNP panel is an efficient tool for detecting hybrids up to the thirdgeneration backcrosses to wolves across Europe. Notably, the proposed genotyping method is suitable for a variety of samples, including non-invasive and museum samples, making this panel useful for wolf-dog hybrid assessments and wolf monitoring at both continental and different temporal scales.

Marmora’s Warbler (Curruca sarda) and Balearic Warbler (C. balearica) are allopatric sibling species and were recently split mostly based on morphological and ethological characteristics. Here we provide the first phylogenetic and phylogeographic analyses of this species complex to support the taxonomic status of C. sarda and C. balearica in light of integrative taxonomy. We sampled the two taxa in most of their breeding ranges and we sequenced three mitochondrial and one nuclear gene region. All C. balearica individuals had private haplotypes for the four markers and formed monophyletic clades. Genetic distances between the two taxa were comparable with those found between other species belonging to the Curruca genus. Furthermore, most of the genetic variance was expressed at the interspecific level, rather than between different populations within taxa or between individuals within populations. Our results strongly support the current taxonomic status of these two warblers as distinct species.

Background: Understanding the processes that lead to hybridization of wolves and dogs is of scientific and management importance, particularly over large geographical scales, as wolves can disperse great distances. However, a method to efficiently detect hybrids in routine wolf monitoring is lacking. Microsatellites offer only limited resolution due to the low number of markers showing distinctive allele frequencies between wolves and dogs. Moreover, calibration across laboratories is time-consuming and costly. In this study, we selected a panel of 96 ancestry informative markers for wolves and dogs, derived from the Illumina CanineHD Whole-Genome BeadChip (174 K). We designed very short amplicons for genotyping on a microfluidic array, thus making the method suitable also for non-invasively collected samples. Results: Genotypes based on 93 SNPs from wolves sampled throughout Europe, purebred and non-pedigree dogs, and suspected hybrids showed that the new panel accurately identifies parental individuals, first-generation hybrids and first-generation backcrosses to wolves, while second- and third-generation backcrosses to wolves were identified as advanced hybrids in almost all cases. Our results support the hybrid identity of suspect individuals and the non-hybrid status of individuals regarded as wolves. We also show the adequacy of these markers to assess hybridization at a European-wide scale and the importance of including samples from reference populations. Conclusions: We showed that the proposed SNP panel is an efficient tool for detecting hybrids up to the thirdgeneration backcrosses to wolves across Europe. Notably, the proposed genotyping method is suitable for a variety of samples, including non-invasive and museum samples, making this panel useful for wolf-dog hybrid assessments and wolf monitoring at both continental and different temporal scales.

Background: Understanding the processes that lead to hybridization of wolves and dogs is of scientific and management importance, particularly over large geographical scales, as wolves can disperse great distances. However, a method to efficiently detect hybrids in routine wolf monitoring is lacking. Microsatellites offer only limited resolution due to the low number of markers showing distinctive allele frequencies between wolves and dogs. Moreover, calibration across laboratories is time-consuming and costly. In this study, we selected a panel of 96 ancestry informative markers for wolves and dogs, derived from the Illumina CanineHD Whole-Genome BeadChip (174 K). We designed very short amplicons for genotyping on a microfluidic array, thus making the method suitable also for non-invasively collected samples. Results: Genotypes based on 93 SNPs from wolves sampled throughout Europe, purebred and non-pedigree dogs, and suspected hybrids showed that the new panel accurately identifies parental individuals, first-generation hybrids and first-generation backcrosses to wolves, while second- and third-generation backcrosses to wolves were identified as advanced hybrids in almost all cases. Our results support the hybrid identity of suspect individuals and the non-hybrid status of individuals regarded as wolves. We also show the adequacy of these markers to assess hybridization at a European-wide scale and the importance of including samples from reference populations. Conclusions: We showed that the proposed SNP panel is an efficient tool for detecting hybrids up to the third-generation backcrosses to wolves across Europe. Notably, the proposed genotyping method is suitable for a variety of samples, including non-invasive and museum samples, making this panel useful for wolf-dog hybrid assessments and wolf monitoring at both continental and different temporal scales.

Background: Understanding the processes that lead to hybridization of wolves and dogs is of scientific and management importance, particularly over large geographical scales, as wolves can disperse great distances. However, a method to efficiently detect hybrids in routine wolf monitoring is lacking. Microsatellites offer only limited resolution due to the low number of markers showing distinctive allele frequencies between wolves and dogs. Moreover, calibration across laboratories is time-consuming and costly. In this study, we selected a panel of 96 ancestry informative markers for wolves and dogs, derived from the Illumina CanineHD Whole-Genome BeadChip (174 K). We designed very short amplicons for genotyping on a microfluidic array, thus making the method suitable also for non-invasively collected samples. Results: Genotypes based on 93 SNPs from wolves sampled throughout Europe, purebred and non-pedigree dogs, and suspected hybrids showed that the new panel accurately identifies parental individuals, first-generation hybrids and first-generation backcrosses to wolves, while second- and third-generation backcrosses to wolves were identified as advanced hybrids in almost all cases. Our results support the hybrid identity of suspect individuals and the non-hybrid status of individuals regarded as wolves. We also show the adequacy of these markers to assess hybridization at a European-wide scale and the importance of including samples from reference populations. Conclusions: We showed that the proposed SNP panel is an efficient tool for detecting hybrids up to the third-generation backcrosses to wolves across Europe. Notably, the proposed genotyping method is suitable for a variety of samples, including non-invasive and museum samples, making this panel useful for wolf-dog hybrid assessments and wolf monitoring at both continental and different temporal scales.

Background: Understanding the processes that lead to hybridization of wolves and dogs is of scientific and management importance, particularly over large geographical scales, as wolves can disperse great distances. However, a method to efficiently detect hybrids in routine wolf monitoring is lacking. Microsatellites offer only limited resolution due to the low number of markers showing distinctive allele frequencies between wolves and dogs. Moreover, calibration across laboratories is time-consuming and costly. In this study, we selected a panel of 96 ancestry informative markers for wolves and dogs, derived from the Illumina CanineHD Whole-Genome BeadChip (174 K). We designed very short amplicons for genotyping on a microfluidic array, thus making the method suitable also for non-invasively collected samples. Results: Genotypes based on 93 SNPs from wolves sampled throughout Europe, purebred and non-pedigree dogs, and suspected hybrids showed that the new panel accurately identifies parental individuals, first-generation hybrids and first-generation backcrosses to wolves, while second- and third-generation backcrosses to wolves were identified as advanced hybrids in almost all cases. Our results support the hybrid identity of suspect individuals and the non-hybrid status of individuals regarded as wolves. We also show the adequacy of these markers to assess hybridization at a European-wide scale and the importance of including samples from reference populations. Conclusions: We showed that the proposed SNP panel is an efficient tool for detecting hybrids up to the thirdgeneration backcrosses to wolves across Europe. Notably, the proposed genotyping method is suitable for a variety of samples, including non-invasive and museum samples, making this panel useful for wolf-dog hybrid assessments and wolf monitoring at both continental and different temporal scales.

Marmora’s Warbler (Curruca sarda) and Balearic Warbler (C. balearica) are allopatric sibling species and were recently split mostly based on morphological and ethological characteristics. Here we provide the first phylogenetic and phylogeographic analyses of this species complex to support the taxonomic status of C. sarda and C. balearica in light of integrative taxonomy. We sampled the two taxa in most of their breeding ranges and we sequenced three mitochondrial and one nuclear gene region. All C. balearica individuals had private haplotypes for the four markers and formed monophyletic clades. Genetic distances between the two taxa were comparable with those found between other species belonging to the Curruca genus. Furthermore, most of the genetic variance was expressed at the interspecific level, rather than between different populations within taxa or between individuals within populations. Our results strongly support the current taxonomic status of these two warblers as distinct species.

In forensic investigation of firearm-related cases, determination of the residual amount of volatile compounds remaining inside a cartridge could be useful in estimating the time since its discharge. Published approaches are based on following the decrease of selected target compounds as a function of time by using solid phase micro-extraction (SPME). Naphthalene as well as an unidentified decomposition product of nitrocellulose (referred to as "TEA2”) are usually employed for this purpose. However, reliability can be brought into question given their high volatility and low reproducibility of the extracted procedure. In order to identify alternatives and therefore develop improved dating methods, an extensive study on the composition and variability of volatile residues in nine different types of cartridges was carried out. Analysis was performed using headspace sorptive extraction (HSSE), which is a more exhaustive technique compared to SPME. One hundred sixty six compounds were identified (several of which for the first time), and it was observed that the final compositional characteristics of each residue were strongly dependent on its source. Variability of single identified compounds within and between different types of cartridge as well as their evolution over time were also studied. Many explosion products containing up to four aromatic rings were found to be globally present in high proportions. Twenty-seven of them (excluding naphthalene) also presented detectable decreases during the first 24h. Therefore, they could be used as complementary target analytes in future dating methods.

The unconventional infectious agents of transmissible spongiform encephalopathies (TSEs) are prions. Their infectivity co-appears with PrPSc, aberrant depositions of the host’s cellular prion protein (PrPC). Successive heat treatment in the presence of detergent and proteolysis by a keratinase from Bacillus licheniformis PWD-1 was shown before to destroy PrPSc from bovine TSE (BSE) and sheep scrapie diseased brain, however data regarding expected reduction of infectivity were still lacking. Therefore, transgenic Tgbov XV mice which are highly BSE susceptible were used to quantify infectivity before and after the bovine brain treatment procedure. Also four immunochemical analyses were applied to compare the levels of PrPSc. After heating at 115 °C with or without subsequent proteolysis, the original BSE infectivity of 106.2–6.4 ID50 g−1 was reduced to a remaining infectivity of 104.6–5.7 ID50 g−1 while strain characteristics were unaltered, even after precipitation with methanol. Surprisingly, PrPSc depletion was 5–800 times higher than the loss of infectivity. Similar treatment was applied on other prion strains, which were CWD1 in bank voles, 263 K scrapie in hamsters and sheep PG127 scrapie in tg338 ovinized mice. In these strains however, infectivity was already destroyed by heat only. These findings show the unusual heat resistance of BSE and support a role for an additional factor in prion formation as suggested elsewhere when producing prions from PrPC. Leftover material in the remaining PrPSc depleted BSE preparation offers a unique substrate for searching additional elements for prion infectivity and improving our concept about the nature of prions.

[EN] Aim of the paper is the historical-critical study of Castel Camponeschi (aka Castello di Prata), a village at about 25 km from L’Aquila city (Italy), with peculiar urban characteristics –a herringbone pattern– probably founded in the thirteenth century and characterized by important phenomena of modification and stratification. The study roots on the direct analysis of the built heritage through the architectural surveying, realized at multiple scales from the urban settlement to the masonry structures. According to the morphology of the historic center, it could be traced to the process of new settlement foundation that characterized the territory –the border between the Papal State and the Frederic’s Empire first, and then the Angevin kingdom–. The analysis of the village requires both a correlation with the natural and anthropic historical context in which it is inserted and, a specific study of masonry equipments, characterized by the re-used of stone elements from the nearby Vestino-Roman town of Peltuinum.

[EN] Aim of the paper is the historical-critical study of Castel Camponeschi (aka Castello di Prata), a village at about 25 km from L’Aquila city (Italy), with peculiar urban characteristics –a herringbone pattern– probably founded in the thirteenth century and characterized by important phenomena of modification and stratification. The study roots on the direct analysis of the built heritage through the architectural surveying, realized at multiple scales from the urban settlement to the masonry structures. According to the morphology of the historic center, it could be traced to the process of new settlement foundation that characterized the territory –the border between the Papal State and the Frederic’s Empire first, and then the Angevin kingdom–. The analysis of the village requires both a correlation with the natural and anthropic historical context in which it is inserted and, a specific study of masonry equipments, characterized by the re-used of stone elements from the nearby Vestino-Roman town of Peltuinum.

[EN] Aim of the paper is the historical-critical study of Castel Camponeschi (aka Castello di Prata), a village at about 25 km from L’Aquila city (Italy), with peculiar urban characteristics –a herringbone pattern– probably founded in the thirteenth century and characterized by important phenomena of modification and stratification. The study roots on the direct analysis of the built heritage through the architectural surveying, realized at multiple scales from the urban settlement to the masonry structures. According to the morphology of the historic center, it could be traced to the process of new settlement foundation that characterized the territory –the border between the Papal State and the Frederic’s Empire first, and then the Angevin kingdom–. The analysis of the village requires both a correlation with the natural and anthropic historical context in which it is inserted and, a specific study of masonry equipments, characterized by the re-used of stone elements from the nearby Vestino-Roman town of Peltuinum.

[EN] Aim of the paper is the historical-critical study of Castel Camponeschi (aka Castello di Prata), a village at about 25 km from L’Aquila city (Italy), with peculiar urban characteristics –a herringbone pattern– probably founded in the thirteenth century and characterized by important phenomena of modification and stratification. The study roots on the direct analysis of the built heritage through the architectural surveying, realized at multiple scales from the urban settlement to the masonry structures. According to the morphology of the historic center, it could be traced to the process of new settlement foundation that characterized the territory –the border between the Papal State and the Frederic’s Empire first, and then the Angevin kingdom–. The analysis of the village requires both a correlation with the natural and anthropic historical context in which it is inserted and, a specific study of masonry equipments, characterized by the re-used of stone elements from the nearby Vestino-Roman town of Peltuinum.

[EN] Aim of the paper is the historical-critical study of Castel Camponeschi (aka Castello di Prata), a village at about 25 km from L’Aquila city (Italy), with peculiar urban characteristics –a herringbone pattern– probably founded in the thirteenth century and characterized by important phenomena of modification and stratification. The study roots on the direct analysis of the built heritage through the architectural surveying, realized at multiple scales from the urban settlement to the masonry structures. According to the morphology of the historic center, it could be traced to the process of new settlement foundation that characterized the territory –the border between the Papal State and the Frederic’s Empire first, and then the Angevin kingdom–. The analysis of the village requires both a correlation with the natural and anthropic historical context in which it is inserted and, a specific study of masonry equipments, characterized by the re-used of stone elements from the nearby Vestino-Roman town of Peltuinum.

[EN] Aim of the paper is the historical-critical study of Castel Camponeschi (aka Castello di Prata), a village at about 25 km from L’Aquila city (Italy), with peculiar urban characteristics –a herringbone pattern– probably founded in the thirteenth century and characterized by important phenomena of modification and stratification. The study roots on the direct analysis of the built heritage through the architectural surveying, realized at multiple scales from the urban settlement to the masonry structures. According to the morphology of the historic center, it could be traced to the process of new settlement foundation that characterized the territory –the border between the Papal State and the Frederic’s Empire first, and then the Angevin kingdom–. The analysis of the village requires both a correlation with the natural and anthropic historical context in which it is inserted and, a specific study of masonry equipments, characterized by the re-used of stone elements from the nearby Vestino-Roman town of Peltuinum.

Illegal trafficking of pharmaceutical products by criminal organisations is a global threat for public health. Drugs for erectile dysfunction such as phosphodiesterase type 5 inhibitors are the most commonly counterfeited medicines in Europe. The search of possible toxic chemical substances in seized products is needed to provide early warning for public health. Furthermore, the elemental profile of the seized products can be useful in criminal investigations. For the first time an ion beam analysis (IBA) procedure to characterise authentic Viagra® tablets and sildenafil-based illegal products is described. Moreover, results are compared with the ones obtained by instrumental neutron activation analysis (INAA) on authentic Viagra® tablets in two reactors. IBA results showed that a combination of particle-induced X-ray emission (PIXE) and secondary ion mass spectrometry using primary ions with energies in the range of several MeV (MeV-SIMS) is a powerful tool to characterise different products in a straightforward manner, allowing discrimination between legal and illegal products. INAA allowed accurate elemental quantification and also showed a great potential for the future implementation of an inter-laboratory classification system., Accepted Author Manuscript, Technici Pool

To withstand the local wave climate, coastal structure design relies on our understanding of the wave characteristics at the structure wall and the forces they impose onto the wall. However, despite the vast number of protective structures along our coasts, detailed field observations of waves measured directly at the structure wall are virtually absent. In this study, rare observations of wave characteristics at a vertical wall are presented. Contrary to linear wave theory, results suggest that the standing waves are highly nonlinear. Waves at the wall have considerably larger kurtosis and skewness compared to the incoming wave field and the wave steepness at the wall is observed to exceed the theoretical wave steepness limit of standing waves.

BACKGROUND: It has become clear that healthcare workers are at high risk, and otolaryngology has been theorized to be among the highest risk specialties for coronavirus disease 2019 (COVID-19). The purpose of this study was to detail the international impact of COVID-19 among otolaryngologists, and to identify instructional cases. METHODS: Country representatives of the Young Otolaryngologists-International Federation of Otolaryngologic Societies (YO-IFOS) surveyed otolaryngologists through various channels. Nationwide surveys were distributed in 19 countries. The gray literature and social media channels were searched to identify reported deaths of otolaryngologists from COVID-19. RESULTS: A total of 361 otolaryngologists were identified to have had COVID-19, and data for 325 surgeons was available for analysis. The age range was 25 to 84 years, with one-half under the age of 44 years. There were 24 deaths in the study period, with 83% over age 55 years. Source of infection was likely clinical activity in 175 (54%) cases. Prolonged exposure to a colleague was the source for 37 (11%) surgeons. Six instructional cases were identified where infections occurred during the performance of aerosol-generating operations (tracheostomy, mastoidectomy, epistaxis control, dacryocystorhinostomy, and translabyrinthine resection). In 3 of these cases, multiple operating room attendees were infected, and in 2, the surgeon succumbed to complications of COVID-19. CONCLUSION: The etiology of reported cases within the otolaryngology community appear to stem equally from clinical activity and community spread. Multiple procedures performed by otolaryngologists are aerosol-generating procedures (AGPs) and great care should be taken to protect the surgical team before, during, and after these operations.

OBJECTIVE: To investigate the occurrence of olfactory and gustatory dysfunctions in patients with laboratory-confirmed COVID-19 infection. METHODS: Patients with laboratory-confirmed COVID-19 infection were recruited from 12 European hospitals. The following epidemiological and clinical outcomes have been studied: age, sex, ethnicity, comorbidities, and general and otolaryngological symptoms. Patients completed olfactory and gustatory questionnaires based on the smell and taste component of the National Health and Nutrition Examination Survey, and the short version of the Questionnaire of Olfactory Disorders-Negative Statements (sQOD-NS). RESULTS: A total of 417 mild-to-moderate COVID-19 patients completed the study (263 females). The most prevalent general symptoms consisted of cough, myalgia, and loss of appetite. Face pain and nasal obstruction were the most disease-related otolaryngological symptoms. 85.6% and 88.0% of patients reported olfactory and gustatory dysfunctions, respectively. There was a significant association between both disorders (p < 0.001). Olfactory dysfunction (OD) appeared before the other symptoms in 11.8% of cases. The sQO-NS scores were significantly lower in patients with anosmia compared with normosmic or hyposmic individuals (p = 0.001). Among the 18.2% of patients without nasal obstruction or rhinorrhea, 79.7% were hyposmic or anosmic. The early olfactory recovery rate was 44.0%. Females were significantly more affected by olfactory and gustatory dysfunctions than males (p = 0.001). CONCLUSION: Olfactory and gustatory disorders are prevalent symptoms in European COVID-19 patients, who may not have nasal symptoms. The sudden anosmia or ageusia need to be recognized by the international scientific community as important symptoms of the COVID-19 infection.

Auditory neuropathy spectrum disorder (ANSD) refers to a range of hearing impairments characterized by deteriorated speech perception, despite relatively preserved pure-tone detection thresholds. Affected individuals usually present with abnormal auditory brainstem responses (ABRs), but normal otoacoustic emissions (OAEs). These electrophysiological characteristics have led to the hypothesis that ANSD may be caused by various dysfunctions at the cochlear inner hair cell (IHC) and spiral ganglion neuron (SGN) levels, while the activity of outer hair cells (OHCs) is preserved, resulting in discrepancies between pure-tone and speech comprehension thresholds. The exact prevalence of ANSD remains unknown; clinical findings show a large variability among subjects with hearing impairment ranging from mild to profound hearing loss. A wide range of prenatal and postnatal etiologies have been proposed. The study of genetics and of the implicated sites of lesion correlated with clinical findings have also led to a better understanding of the molecular mechanisms underlying the various forms of ANSD, and may guide clinicians in better screening, assessment and treatment of ANSD patients. Besides OAEs and ABRs, audiological assessment includes stapedial reflex measurements, supraliminal psychoacoustic tests, electrocochleography (ECochG), auditory steady-state responses (ASSRs) and cortical auditory evoked potentials (CAEPs). Hearing aids are indicated in the treatment of ANSD with mild to moderate hearing loss, whereas cochlear implantation is the first choice of treatment in case of profound hearing loss, especially in case of IHC presynaptic disorders, or in case of poor auditory outcomes with conventional hearing aids.