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Abstract Chromosomal Instability (CIN) is a common and evolving feature in breast cancer. Large-scale Transitions (LSTs), defined as chromosomal breakages leading to gains or losses of at least 10 Mb, have recently emerged as a metric of CIN due to their standardized definition across platforms. Herein, we report the feasibility of using low-pass Whole Genome Sequencing to assess LSTs, copy number alterations (CNAs) and their relationship in individual circulating tumor cells (CTCs) of triple-negative breast cancer (TNBC) patients. Initial assessment of LSTs in breast cancer cell lines consistently showed wide-ranging values (median 22, range 4–33, mean 21), indicating heterogeneous CIN. Subsequent analysis of CTCs revealed LST values (median 3, range 0–18, mean 5), particularly low during treatment, suggesting temporal changes in CIN levels. CNAs averaged 30 (range 5–49), with loss being predominant. As expected, CTCs with higher LSTs values exhibited increased CNAs. A CNA-based classifier of individual patient-derived CTCs, developed using machine learning, identified genes associated with both DNA proliferation and repair, such as RB1, MYC, and EXO1, as significant predictors of CIN. The model demonstrated a high predictive accuracy with an Area Under the Curve (AUC) of 0.89. Overall, these findings suggest that sequencing CTCs holds the potential to facilitate CIN evaluation and provide insights into its dynamic nature over time, with potential implications for monitoring TNBC progression through iterative assessments.

Bicuspid aortic valve (BAV) is the most common congenital heart malformation in adults, but it can also cause childhood-onset complications. The presentation and clinical course of young adults who present due to BAV complications are relatively uncharacterized. In a multicenter study, we found that young people who experience significant complications related to BAV disease before age 30 are distinguished from the majority of BAV cases that manifest after age 50 by a relatively severe clinical course, with higher rates of surgical interventions, more frequent second interventions, and a greater burden of congenital heart malformations. These observations highlight the need for prompt recognition, regular lifelong surveillance, and targeted interventions to address the significant health burdens of patients with early-onset BAV complications.

Objective: This article reviews clinical implications and costs associated with orthopedic surgeries performed in inpatient versus outpatient settings. Methods: Studies spanning the publication period between 2000 and 2023 were included, exploring a variety of study designs. Results: Results indicate that outpatient surgeries are associated with lower rates of postoperative complications, shorter recovery times, and greater patient satisfaction, as well as with significantly lower direct and indirect costs. Analysis of postoperative complications suggests that factors such as the home recovery environment and early mobilization may contribute to improved clinical outcomes, corroborating existing literature on the safety and efficacy of outpatient surgeries. Faster recovery after outpatient surgeries was consistently observed, with patients returning to normal activities an average of two weeks earlier compared with those undergoing inpatient surgeries. The greater patient satisfaction with outpatient surgeries reflects the convenience of avoiding hospitalization, with less disruption to daily life and reduced anxiety associated with the hospital environment. In economic terms, outpatient surgeries have been shown to be a financially advantageous alternative, with reduced direct costs due to the lower hospital fees and nursing services required, as well as lower indirect costs due to reduced productivity loss and transportation expenses. Conclusion: Findings are particularly relevant in a context of increasing pressure to contain spending in the health system. Level of Evidence I; Systematic review of level I studies.

Objective Subjects with subclinical psoriatic arthritis (PsA), defined as the presence of arthralgia in psoriasis (PsO), are at higher risk of PsA but scant real-world data exist. Our aims were to (1) estimate the probability of PsA development in subclinical PsA, (2) characterise subclinical PsA symptoms and (3) determine the clinical patterns at PsA diagnosis.Methods Patients with PsO, mainly subclinical PsA, were evaluated longitudinally in two European cohorts. The key outcome was new-onset PsA. Musculoskeletal symptoms including inflammatory and non-inflammatory symptoms before PsA diagnosis were collected. Occurrence of PsA was analysed with survival analysis and cumulative incidence functions (CIFs).Results 384 patients with PsO were included with a mean follow-up of 33.0 (±20.9) months. 311 of 384 (80.9%) had subclinical PsA with a PsA incidence rate of 7.7 per 100 patient-years. Subclinical PsA displayed a higher risk of PsA development compared with PsO (HR=11.7 (95% CI 1.57 to 86.7), p=0.016). The probability of new-onset PsA estimated by the CIF was 9.4% (95% CI 4.7% to 10.6%) at month 12 and 22.7% (95% CI 17.2% to 28.6%) at month 36. 58.9% of cases reported inflammatory symptoms in the months immediately prior to PsA diagnosis but prior non-inflammatory symptoms were evident in 83.9% prior to PsA diagnosis. Peripheral joint swelling was the predominant PsA presentation pattern (82.1%).Conclusions The probability of PsA development among subclinical PsA was relatively high, emphasising the importance of emergent musculoskeletal symptoms when aiming for PsA prevention. Joint swelling was the dominant feature in new-onset PsA, likely reflecting clinical confidence in recognising joint swelling.

Aims To investigate the prevalence and distribution of bone erosions in an early psoriatic arthritis (PsA) population using conventional radiography (CR) and to explore the agreement between CR and ultrasound (US) detected bone erosions.Methods Newly diagnosed, treatment naïve PsA patients fulfilling the ClASsification for Psoriatic Arthritis (CASPAR) classification criteria of ≤5 years symptom duration were recruited as part of the Leeds Spondyloarthropathy Register for Research and Observation and underwent CR and US examination of hands and feet.Results Overall, 4655 hand and feet joints were assessed in 122 patients. CR erosions were detected in 24.6% (n=30) with lowest prevalence seen below 8 months of symptoms (17.5% vs 24.3%>24 months). The number of erosions was higher on CR (1.55% (63/4,655); US 1.04% (34/3,270)), with 5th metatarsophalangeal (MTP) joint being the most affected site in both CR (5.21% (11/211)) and US (7.14% (15/210)). Erosions in CR were more evenly distributed compared with US where three-quarters of the total number of bone erosions were detected in wrists, second metacarpophalangeal (MCP) and fifth MTP joints. Most joints had almost perfect prevalence-adjusted bias-adjusted kappa values ranging from 0.91 to 1.Conclusions Erosions were seen in a quarter of patients with newly diagnosed, untreated PsA with a declining trend around the 8-month symptom duration cut-off. High levels of agreement between CR and US were seen with CR detecting more erosions. A focused US assessment of the wrist, second MCP and fifth MTP joints may be useful to detect bone erosions in early PsA.

Background: Monkeypox (Mpox) is a zoonotic disease caused by monkeypox virus (MPXV), an Orthopoxvirus (OPXV). Since we are observing the first MPXV outbreak outside the African continent, the general population probably does not have a pre-existing memory response for MPXV but may have immunity against the previous smallpox vaccine based on a live replicating Vaccinia strain (VACV). Using a whole blood platform, we aim to study the MPXV- T-cell-specific response in Mpox-cured subjects. Methods: We enrolled 16 subjects diagnosed with Mpox in the previous 3–7 months and 15 healthy donors (HD) with no recent vaccination history. Whole blood was stimulated overnight with MPXV and VACV peptides to elicit CD4 and CD8 T-cell-specific responses, which were evaluated by ELISA and multiplex assay. Results: Mpox-cured subjects showed a significant IFN-γ T-cell response to MPXV and VACV. Besides IFN-γ, IL-6, IP-10, IL-8, IL-2, G-CSF, MCP-1, MIP1-α, MIP-1β, IL-1Rα, and IL-5 were significantly induced after specific stimulation compared to the unstimulated control. The specific response was mainly induced by the CD4 peptides MPX-CD4-E and VACV-CD4. Conclusions: We showed that MPXV-specific responses have a mixed Th1- and Th2-response in a whole blood platform assay, which may be useful for monitoring the specific immunity induced by vaccination or infection.

Bicuspid aortic valve (BAV), the most common congenital heart defect, is a major cause of aortic valve disease requiring valve interventions and thoracic aortic aneurysms predisposing to acute aortic dissections. The spectrum of BAV ranges from early onset valve and aortic complications (EBAV) to sporadic late onset disease. Rare genomic copy number variants (CNVs) have previously been implicated in the development of BAV and thoracic aortic aneurysms. We determined the frequency and gene content of rare CNVs in EBAV probands (n = 272) using genome-wide SNP microarray analysis and three complementary CNV detection algorithms (cnvPartition, PennCNV, and QuantiSNP). Unselected control genotypes from the Database of Genotypes and Phenotypes were analyzed using identical methods. We filtered the data to select large genic CNVs that were detected by multiple algorithms. Findings were replicated in a BAV cohort with late onset sporadic disease (n = 5040). We identified 3 large and rare (< 1,1000 in controls) CNVs in EBAV probands. The burden of CNVs intersecting with genes known to cause BAV when mutated was increased in case-control analysis. CNVs intersecting with GATA4 and DSCAM were enriched in cases, recurrent in other datasets, and segregated with disease in families. In total, we identified potentially pathogenic CNVs in 9% of EBAV cases, implicating alterations of candidate genes at these loci in the pathogenesis of BAV.

Abstract Tropical forests are among the most biodiverse biomes on the planet. Nevertheless, quantifying the abundance and species richness within megadiverse groups is a significant challenge. We designed a study to address this challenge by documenting the variability of the insect fauna across a vertical canopy gradient in a Central Amazonian tropical forest. Insects were sampled over two weeks using 6-m Gressitt-style Malaise traps set at five heights (0 m–32 m–8 m intervals) on a metal tower in a tropical forest north of Manaus, Brazil. The traps contained 37,778 specimens of 18 orders of insects. Using simulation approaches and nonparametric analyses, we interpreted the abundance and richness of insects along this gradient. Diptera, Hymenoptera, and Coleoptera had their greatest abundance at the ground level, whereas Lepidoptera and Hemiptera were more abundant in the upper levels of the canopy. We identified species of 38 of the 56 families of Diptera, finding that 527 out of 856 species (61.6%) were not sampled at the ground level. Mycetophilidae, Tipulidae, and Phoridae were significantly more diverse and/or abundant at the ground level, while Tachinidae, Dolichopodidae, and Lauxaniidae were more diverse or abundant at upper levels. Our study suggests the need for a careful discussion of strategies of tropical forest conservation based on a much more complete understanding of the three-dimensional distribution of its insect diversity.

Abstract To assess the usefulness of a theoretical postural instability discrimination index (PIth) in amyotrophic lateral sclerosis (ALS). Prospective regression analyzes were performed to identify the biomechanical determinants of postural instability unrelated to lower limb motor deficits from gait initiation factors. PIth was constructed using a logit function of biomechanical determinants. Discriminatory performance and performance differences were tested. Backward displacement of the pression center (APAamplitude) and active vertical braking of the mass center (Braking-index) were the biomechanical determinants of postural instability. PIth = − 0.13 × APAamplitude − 0.12 × Braking-index + 5.67, (P

Abstract The aetiology of Amyotrophic Lateral Sclerosis (ALS) is still poorly understood. The discovery of genetic forms of ALS pointed out the mechanisms underlying this pathology, but also showed how complex these mechanisms are. Excitotoxicity is strongly suspected to play a role in ALS pathogenesis. Excitotoxicity is defined as neuron damage due to excessive intake of calcium ions (Ca2+) by the cell. This study aims to find a relationship between the proteins coded by the most relevant genes associated with ALS and intracellular Ca2+ accumulation. In detail, the profile of eight proteins (TDP-43, C9orf72, p62/sequestosome-1, matrin-3, VCP, FUS, SOD1 and profilin-1), was analysed in three different cell types induced to raise their cytoplasmic amount of Ca2+. Intracellular Ca2+ accumulation causes a decrease in the levels of TDP-43, C9orf72, matrin3, VCP, FUS, SOD1 and profilin-1 and an increase in those of p62/sequestosome-1. These events are associated with the proteolytic action of two proteases, calpains and caspases, as well as with the activation of autophagy. Interestingly, Ca2+ appears to both favour and hinder autophagy. Understanding how and why calpain-mediated proteolysis and autophagy, which are physiological processes, become pathological may elucidate the mechanisms responsible for ALS and help discover new therapeutic targets.

Abstract X-ray dark-field imaging is a widely researched imaging technique, with many studies on samples of very different dimensions and at very different resolutions. However, retrieval of three-dimensional (3D) information for human thorax sized objects has not yet been demonstrated. We present a method, similar to classic tomography and tomosynthesis, to obtain 3D information in X-ray dark-field imaging. Here, the sample is moved through the divergent beam of a Talbot–Lau interferometer. Projections of features at different distances from the source seemingly move with different velocities over the detector, due to the cone beam geometry. The reconstruction of different focal planes exploits this effect. We imaged a chest phantom and were able to locate different features in the sample (e.g. the ribs, and two sample vials filled with water and air and placed in the phantom) to corresponding focal planes. Furthermore, we found that image quality and detectability of features is sufficient for image reconstruction with a dose of 68 μSv at an effective pixel size of $$0.357 \times {0.357}\,\mathrm{mm}^{2}$$ 0.357 × 0.357 mm 2 . Therefore, we successfully demonstrated that the presented method is able to retrieve 3D information in X-ray dark-field imaging.

Background: Multiple methods have been implemented to limit the impact of radiotherapy on patients affected by mediastinal lymphoma, including breathing control techniques, image-guided radiotherapy (IGRT) and intensity-modulated radiotherapy (IMRT), although the actual diffusion of such techniques is unclear. No surveys have been published to date evaluating the techniques adopted at different centers. Methods: A survey with a dedicated questionnaire was submitted to 195 Italian radiotherapy centers, assessing items regarding the characteristics of the center and clinical practice in the treatment of mediastinal lymphomas. Results: A total of 43 centers (22%) responded, the majority of which were university hospitals (37.2%) or cancer care centers (27.9%). In 95.4% of the centers, IMRT was used in the clinical practice, and the most frequently employed techniques were VMAT (48.8% of centers) and non-rotational IMRT (31.7%). Comparison of multiple plans was performed by 66.7% of the responding centers. Dose constraints for organs at risk were consistently prescribed. IGRT techniques were adopted by 93% of the centers, while breathing control or gating techniques were routinely used by only 25.6% of the centers. A necessity to standardize OAR constraints and define guidelines was perceived by almost all participants. Conclusions: Modern radiotherapy techniques are widely used in the Italian centers, although with heterogeneous characteristics.

Abstract We present a method to generate synthetic thorax radiographs with realistic nodules from CT scans, and a perfect ground truth knowledge. We evaluated the detection performance of nine radiologists and two convolutional neural networks in a reader study. Nodules were artificially inserted into the lung of a CT volume and synthetic radiographs were obtained by forward-projecting the volume. Hence, our framework allowed for a detailed evaluation of CAD systems’ and radiologists’ performance due to the availability of accurate ground-truth labels for nodules from synthetic data. Radiographs for network training (U-Net and RetinaNet) were generated from 855 CT scans of a public dataset. For the reader study, 201 radiographs were generated from 21 nodule-free CT scans with altering nodule positions, sizes and nodule counts of inserted nodules. Average true positive detections by nine radiologists were 248.8 nodules, 51.7 false positive predicted nodules and 121.2 false negative predicted nodules. The best performing CAD system achieved 268 true positives, 66 false positives and 102 false negatives. Corresponding weighted alternative free response operating characteristic figure-of-merits (wAFROC FOM) for the radiologists range from 0.54 to 0.87 compared to a value of 0.81 (CI 0.75–0.87) for the best performing CNN. The CNN did not perform significantly better against the combined average of the 9 readers (p = 0.49). Paramediastinal nodules accounted for most false positive and false negative detections by readers, which can be explained by the presence of more tissue in this area.

Abstract Chronic headache is particularly prevalent in migraineurs and it can progress to a condition known as medication overuse headache (MOH). MOH is a secondary headache caused by overuse of analgesics or other medications such as triptans to abort acute migraine attacks. The worsening of headache symptoms associated with medication overuse (MO) generally ameliorates following interruption of regular medication use, although the primary headache symptoms remain unaffected. MO patients may also develop certain behaviors such as ritualized drug administration, psychological drug attachment, and withdrawal symptoms that have been suggested to correlate with drug addiction. Although several reviews have been performed on this topic, to the authors best knowledge none of them have examined this topic from the addiction point of view. Therefore, we aimed to identify features in MO and drug addiction that may correlate. We initiate the review by introducing the classes of analgesics and medications that can cause MOH and those with high risk to produce MO. We further compare differences between sensitization resulting from MO and from drug addiction, the neuronal pathways that may be involved, and the genetic susceptibility that may overlap between the two conditions. Finally, ICHD recommendations to treat MOH will be provided herein.

Introduzione: La pandemia da Covid-19 ha inevitabilmente avuto un impatto significativo sul nostro stile di vita, stravolgendo e mutando modalità di lavoro, tempo libero e abitudini quotidiane. La più grande e grave pandemia degli ultimi 100 anni ha messo in luce la fragilità dei sistemi sanitari di tutto il mondo. Questo in Italia ha determinato, oltre alle migliaia di morti causate dal virus, anche i “danni collaterali” dovuti al fermo dei ricoveri ospedalieri e delle prestazioni ambulatoriali per tutti gli altri malati. Materiali e Metodi: Attraverso il motore di ricerca Pubmed, con filtro di ricerca la parola “lockdown” AND “Italy” e considerando il periodo di tempo tra l’01/02/2020 e il 6/11/2020, compaiono 591 risultati. Sono stati presi in considerazione esclusivamente gli articoli e le citazioni che contenessero dati ospedalieri riferiti al periodo gennaio - maggio 2020, per un confronto con lo stesso periodo del 2019 Analisi dei dati: Gli studi selezionati mostrano uniformemente in tutto il territorio italiano un calo di ingressi e di ricoveri per patologie diverse dal Covid-19. Su 40 studi presi in cosiderazione, solo una voce ha mostrato un aumento delle prestazioni rispetto all’anno precedente. Le prestazioni sono calate poiché, durante l’era del Coronavirus, si sono ridotte le risorse per tante patologie croniche e di urgenza e per la loro prevenzione, risorse che sono state invece destinate alla lotta al Coronavirus; inoltre sono calati gli accessi al Pronto Soccorso. Conclusione: per evitare eventuali future riduzioni di prestazioni ospedaliere causa pandemia occorre trovare soluzioni alternative all’ospedale, come il potenziamento della prevenzione e della medicina nel territorio. Inoltre è indispensabile l’informazione alla cittadinanza su un uso adatto del Pronto Soccorso, anche se il calo di ricoveri per patologie a rischio vita è un chiaro segno di paura al servirsi di tale servizio in caso di emergenza.

Abstract High sensitivity troponin T (hsTnT) is a strong predictor of adverse outcome during SARS-CoV-2 infection. However, its determinants remain partially unknown. We aimed to assess the relationship between severity of inflammatory response/coagulation abnormalities and hsTnT in Coronavirus Disease 2019 (COVID-19). We then explored the relevance of these pathways in defining mortality and complications risk and the potential effects of the treatments to attenuate such risk. In this single-center, prospective, observational study we enrolled 266 consecutive patients hospitalized for SARS-CoV-2 pneumonia. Primary endpoint was in-hospital COVID-19 mortality. hsTnT, even after adjustment for confounders, was associated with mortality. D-dimer and CRP presented stronger associations with hsTnT than PaO2. Changes of hsTnT, D-dimer and CRP were related; but only D-dimer was associated with mortality. Moreover, low molecular weight heparin showed attenuation of the mortality in the whole population, particularly in subjects with higher hsTnT. D-dimer possessed a strong relationship with hsTnT and mortality. Anticoagulation treatment showed greater benefits with regard to mortality. These findings suggest a major role of SARS-CoV-2 coagulopathy in hsTnT elevation and its related mortality in COVID-19. A better understanding of the mechanisms related to COVID-19 might pave the way to therapy tailoring in these high-risk individuals.

Abstract Evidences of an association between air pollution and Covid-19 infections are mixed and inconclusive. We conducted an ecological analysis at regional scale of long-term exposure to air-borne particle matter and spread of Covid-19 cases during the first wave of epidemics. Global air pollution and climate data were calculated from satellite earth observation data assimilated into numerical models at 10 km resolution. Main outcome was defined as the cumulative number of cases of Covid-19 in the 14 days following the date when > 10 cumulative cases were reported. Negative binomial mixed effect models were applied to estimate the associations between the outcome and long-term exposure to air pollution at the regional level (PM10, PM2.5), after adjusting for relevant regional and country level covariates and spatial correlation. In total we collected 237,749 Covid-19 cases from 730 regions, 63 countries and 5 continents at May 30, 2020. A 10 μg/m3 increase of pollution level was associated with 8.1% (95% CI 5.4%, 10.5%) and 11.5% (95% CI 7.8%, 14.9%) increases in the number of cases in a 14 days window, for PM2.5 and PM10 respectively. We found an association between Covid-19 cases and air pollution suggestive of a possible causal link among particulate matter levels and incidence of COVID-19.

Abstract Worldwide, tropospheric ozone (O3) is a potential threat to wood production, but our understanding of O3 economic impacts on forests is still limited. To overcome this issue, we developed an approach for integrating O3 risk modelling and economic estimates, by using the Italian forests as a case study. Results suggested a significant impact of O3 expressed in terms of stomatal flux with an hourly threshold of uptake (Y = 1 nmol O3 m−2 leaf area s−1 to represent the detoxification capacity of trees), i.e. POD1. In 2005, the annual POD1 averaged over Italy was 20.4 mmol m−2 and the consequent potential damage ranged from 790.90 M€ to 2.85 B€ of capital value (i.e. 255–869 € ha−1, on average) depending on the interest rate. The annual damage ranged from 31.6 to 57.1 M€ (i.e. 10–17 € ha−1 per year, on average). There was also a 1.1% reduction in the profitable forest areas, i.e. with a positive Forest Expectation Value (FEV), with significant declines of the annual national wood production of firewood (− 7.5%), timber pole (− 7.4%), roundwood (− 5.0%) and paper mill (− 4.8%). Results were significantly different in the different Italian regions. We recommend our combined approach for further studies under different economic and phytoclimatic conditions.

This case report describes a 50-year-old woman who developed an allergic granulomatous reaction after a tattoo touch-up.

Abstract Parasites alter many traits of their hosts. In particular, parasites known as “manipulative” may increase their probability of transmission by inducing phenotypic alterations in their intermediate hosts. Although parasitic-induced alterations can modify species’ ecological roles, the proximate factors modulating this phenomenon remain poorly known. As temperature is known to affect host–parasite associations, understanding its precise impact has become a major challenge in a context of global warming. Gammarids are ecologically important freshwater crustaceans and serve as intermediate hosts for several acanthocephalan species. These parasites induce multiple effects on gammarids, including alterations of their behavior, ultimately leading to modifications in their functional role. Here, experimental infections were used to assess the effect of two temperatures on several traits of the association between Gammarus pulex and its acanthocephalan parasite Pomphorhynchus laevis. Elevated temperature affected hosts and parasites in multiple ways (decreased host survival, increased gammarids activity, faster parasites development and proboscis eversion). However, behavioral manipulation was unaffected by temperature. These results suggest that predicted change in temperature may have little consequences on the trophic transmission of parasites through changes in manipulation, although it may modify it through increased infection success and faster parasites development.

Lo scopo di questo articolo è quello di discutere il tema del miglioramento della qualità delle cure primarie. L’obiettivo è quello di sottolineare l’efficacia dell’utilizzo del Significant Event Audit, uno strumento che risponde a logiche qualitative, diffuso prevalentemente in Gran Bretagna.

Detection of Premature Ventricular Contractions (PVC) is of crucial importance in the cardiology field, not only to improve the health system but also to reduce the workload of experts who analyze electrocardiograms (ECG) manually. PVC is a non-harmful common occurrence represented by extra heartbeats, whose diagnosis is not always easily identifiable, especially when done by long-term manual ECG analysis. In some cases, it may lead to disastrous consequences when associated with other pathologies. This work introduces an approach to identify PVCs using machine learning techniques without feature extraction and cross-validation techniques. In particular, a group of six classifiers has been used: Decision Tree, Random Forest, Long-Short Term Memory (LSTM), Bidirectional LSTM, ResNet-18, MobileNetv2, and ShuffleNet. Two types of experiments have been performed on data extracted from the MIT-BIH Arrhythmia database: (i) the original dataset and (ii) the balanced dataset. MobileNetv2 came in first in both experiments with high performance and promising results for PVCs' final diagnosis. The final results showed 99.90% of accuracy in the first experiment and 99.00% in the second one, despite no feature detection techniques were used. The approach we used, which was focused on classification without using feature extraction and cross-validation techniques, allowed us to provide excellent performance and obtain better results. Finally, this research defines as first step toward understanding the explanations for deep learning models' incorrect classifications.

X-ray directional dark-field imaging is a recent technique that can reveal a sample's small-scale structural properties which are otherwise invisible in a conventional imaging system. In particular, directional dark-field can detect and quantify the orientation of anisotropic structures. Here, we present an algorithm that allows for the extraction of a directional dark-field signal from X-ray speckle-based imaging data. The experimental setup is simple, as it requires only the addition of a diffuser to a full-field microscope setup. Sandpaper is an appropriate diffuser material in the hard x-ray regime. We propose an approach to extract the mean scattering width, directionality, and orientation from the recorded speckle images acquired with the technique. We demonstrate that our method can detect and quantify the orientation of fibres inside a carbon fibre reinforced polymer (CFRP) sample within one degree of accuracy and show how the accuracy depends on the number of included measurements. We show that the reconstruction parameters can be tuned to increase or decrease accuracy at the expense of spatial resolution.

(1) Background: The aim of our study is to investigate the main oral lesion patterns in patients with oral graft-versus-host disease and to describe and validate the use of endoscopy enhanced with narrow-band imaging (NBI) as a personalized, reliable and user-friendly tool for the early detection of oral potentially diseases. (2) Methods: We retrospectively evaluated the medical records of 20 patients with chronic GVHD and with oral manifestations, who were referred to our “Interdisciplinary Center for Oropharyngeal Pathology (CIPO)” from January 2017 to July 2022. (3) Results: Data on GVHD, oral localization and NBI endoscopic evaluation are collected. A total of six mucositis, one mucosal erythematous change, ten lichenoid-like changes, eight erosive lesions, one leukoplakia, two erythroplakia and two case of blisters were observed. Two vascular abnormalities were seen with NBI, leading to one excisional biopsy. The patient was diagnosed with squamous cell carcinoma. (4) Conclusion: Our study is the first to highlight the relevance of the routine use of endoscopy with NBI in patients with oral chronic GVHD. We highlighted its role as a reliable, reproducible, easy-to-use and tailor-made tool in the follow-up of those patients and to allow an earlier identification of aberrant neoangiogenesis related to oral potentially malignant disorders and oral cancer.

Despite being usually delivered in oncological patients, radiotherapy can be used as a successful treatment for several non-malignant disorders. Even though this use of radiotherapy has been scarcely investigated since the 1950s, more recent interest has actually shed the light on this approach. Thus, the aim of this narrative review is to analyze the applications of non-oncological radiotherapy in different disorders. Key references were derived from a PubMed query. Hand searching and clinicaltrials.gov were also used. This review contains a narrative report and a critical discussion of non-oncological radiotherapy approaches. In conclusion, non-oncological radiotherapy is a safe and efficacious approach to treat several disorders that needs to be further investigated and used in clinical practice.

Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been shown to cause long-term pulmonary sequelae. Objects: The aim of this study was to evaluate the consequences of the SARS-CoV-2 infection on pulmonary function and on the 6-min walk test related to the severity of the disease. Methods: A cross-sectional study was conducted at the “Policlinico Tor Vergata” Academic Hospital (Rome, Italy), including 75 patients evaluated in post-COVID clinics at the Respiratory Units between November 2020 and September 2021. Complete pulmonary function tests, 6-min walk tests and persistence of symptoms were performed. Results: Of the 75 subjects, 23 had mild, 16 moderate, 26 severe and 10 very severe COVID-19, classified according to WHO. Very severe patients had a lower FVC (100 ± 10%pr) compared to the other groups (116 ± 16%pr, 116 ± 13%pr, 122 ± 20%pr from mild to severe; p < 0.05) and a lower TLC (94 ± 13%pr) compared to the others (102 ± 10%pr, 108 ± 15%pr, 108 ± 12%pr from mild to severe; p < 0.05). DLco and DLco/VA were similar among groups. At the 6MWT, distance, rest and nadir SpO2 were similar among groups, but all groups presented a significant decrease in SpO2 from rest to nadir (Rest SpO2: 97.0 ± 1.0% vs. Nadir SpO2: 93.6 ± 2.7%, p < 0.01). A positive correlation was found between desaturation and delta SpO2 (rest—nadir) (R: 0.29, p < 0.05) and the Distance Desaturation Product (R: 0.39, p < 0.01). Conclusions: These results showed that, although the PFTs are within the normal range, there is still a mild restrictive spirometric pattern after six months in very severe subjects. Moreover, the only persistent pathological sequalae of SARS-CoV-2 infection were a mild desaturation at 6MWT, despite the severity of the infection.

Il metodo più utilizzato per diffondere all’interno della comunità scientifica gli studi e i risultati ottenuti dai gruppi di ricerca è rappresentato dalla pubblicazione di un articolo su riviste peer-review. Il processo di pubblicazione però è sottoposto a regole da seguire non solo da parte di chi scrive l’articolo (Autore), ma anche da parte di chi lo giudica (Revisore). Oggi il numero di articoli presentati alle diverse riviste scientifiche è enorme e i Revisori hanno un ruolo essenziale nell’aiutare gli Editori nel controllo e nella verifica di questa massa di dati e informazioni scientifiche. Da qui dunque l’importanza della regolamentazione dell’attività di revisione in termini di competenza, consapevolezza dei propri doveri verso la rivista e gli Autori, correttezza nell’approccio con gli Autori e, non ultimo, conoscenza delle implicazioni etiche della ricerca e del proprio operato.

Abstract Grating-based X-ray dark-field imaging is a novel imaging modality which has been refined during the last decade. It exploits the wave-like behaviour of X-radiation and can nowadays be implemented with existing X-ray tubes used in clinical applications. The method is based on the detection of small-angle X-ray scattering, which occurs e.g. at air-tissue-interfaces in the lung or bone-fat interfaces in spongy bone. In contrast to attenuation-based chest X-ray imaging, the optimal tube voltage for dark-field imaging of the thorax has not yet been examined. In this work, dark-field scans with tube voltages ranging from 60 to 120 kVp were performed on a deceased human body. We analyzed the resulting images with respect to subjective and objective image quality, and found that the optimum tube voltage for dark-field thorax imaging at the used setup is at rather low energies of around 60 to 70 kVp. Furthermore, we found that at these tube voltages, the transmission radiographs still exhibit sufficient image quality to correlate dark-field information. Therefore, this study may serve as an important guideline for the development of clinical dark-field chest X-ray imaging devices for future routine use.

Abstract Background Perturbation of neuronal excitability contributes to migraine. Neurosteroids modulate the activity of γ-aminobutyric acid A and N-methyl-d-aspartate receptors, and might be involved in the pathogenesis of migraine. Here, we measured plasma levels of four neurosteroids, i.e., allopregnanolone, epiallopregnanolone, dehydroepiandrosterone and deydroepiandrosterone sulfate, in patients affected by episodic migraine, chronic migraine, or cluster headache. Methods Nineteen female patients affected by episodic migraine, 51 female patients affected by chronic migraine, and 18 male patients affected by cluster headache were recruited to the study. Sex- and age-matched healthy control subjects (31 females and 16 males) were also recruited. Patients were clinically characterized by using validated questionnaires. Plasma neurosteroid levels were measured by liquid chromatography-tandem mass spectrometry. Results We found disease-specific changes in neurosteroid levels in our study groups. For example, allopregnanolone levels were significantly increased in episodic migraine and chronic migraine patients than in control subjects, whereas they were reduced in patients affected by cluster headache. Dehydroepiandrosterone and dehydroepiandrosterone sulfate levels were reduced in patients affected by chronic migraine, but did not change in patients affected by cluster headache. Conclusion We have shown for the first time that large and disease-specific changes in circulating neurosteroid levels are associated with chronic headache disorders, raising the interesting possibility that fluctuations of neurosteroids at their site of action might shape the natural course of migraine and cluster headache. Whether the observed changes in neurosteroids are genetically determined or rather result from exposure to environmental or intrinsic stressors is unknown. This might also be matter for further investigation because stress is a known triggering factor for headache attacks in both migraineurs and cluster headache patients.

ABSTRACT BACKGROUND: Sports participation plays an important role in bone gain during childhood and adolescence. The aim here was to identify sex-related determinants of bone mineral density (BMD) differences between male and female adolescents, with emphasis on the role of sports participation. DESIGN AND SETTING: Longitudinal study conducted in a public university in Presidente Prudente, Brazil. METHODS: The sample comprised 48 adolescents aged 11-17 years, of both sexes, who were matched according to sex, age and sports participation. BMD was the main outcome, while muscle mass, sports participation, calendar age and biological maturation were treated as covariates. Participants were followed up after nine months. RESULTS: At baseline, BMD values were similar between the sexes. However, adjustment for covariates showed that BMD was higher among girls at all sites, with a contribution from lean soft tissue (LST) in the model (partial eta-squared, ES-r = 0.619 in upper limbs; 0.643 in lower limbs; 0.699 in spine; and 0.599 in whole body). Sports participation only explained the upper-limb variance (ES-r = 0.99). At the follow-up, the results resembled the baseline except in the lower limbs (P = 0.109), in which BMD was similar between the groups. BMD gain over time was similar between girls and boys in all segments, and baseline LST affected upper-limb and whole-body BMD accrual (ES-r = 0.396 and 0.107, respectively). CONCLUSION: Whole-body and specific-site BMD differed between baseline and follow-up. However,BMD accrual was similar between the sexes, given that muscle mass constituted the most relevant determinant of the difference between them.

Abstract Background Case-finding tools, such as the Identify Chronic Migraine (ID-CM) questionnaire, can improve detection of CM and alleviate its significant societal burden. We aimed to develop and validate the Italian version of the ID-CM (ID-EC) in paper and as a smart app version in a headache clinic-based setting. Methods The study investigators translated and adapted to the Italian language the original ID-CM questionnaire (ID-EC) and further implemented it as a smart app. The ID-EC was tested in its paper and electronic version in consecutive patients referring to 9 Italian tertiary headache centers for their first in-person visit. The scoring algorithm of the ID-EC paper version was applied by the study investigators (case-finding) and by patients (self-diagnosis), while the smart app provided to patients automatically the diagnosis. Diagnostic accuracy of the ID-EC was assessed by matching the questionnaire results with the interview-based diagnoses performed by the headache specialists during the visit according to the criteria of International Classification of Headache Disorders, III edition, beta version. Results We enrolled 531 patients in the test of the paper version of ID-EC and 427 in the validation study of the smart app. According to the clinical diagnosis 209 patients had CM in the paper version study and 202 had CM in the smart app study. 79.5% of patients returned valid paper questionnaires, while 100% of patients returned valid and complete smart app questionnaires. The paper questionnaire had a 81.5% sensitivity and a 81.1% specificity for case-finding and a 30.7% sensitivity and 90.7% specificity for self-diagnosis, while the smart app had a 64.9% sensitivity and 90.2% specificity. Conclusions Our data suggest that the ID-EC, developed and validated in tertiary headache centers, is a valid case-finding tool for CM, with sensitivity and specificity values above 80% in paper form, while the ID-EC smart app is more useful to exclude CM diagnosis in case of a negative result. Further studies are warranted to assess the diagnostic accuracy of the ID-EC in general practice and population-based settings.

Background: The novel SARS-CoV-2 vaccines partially exploit intrinsic DNA or RNA adjuvanticity, with dysregulation in the metabolism of both these nucleic acids independently linked to triggering experimental autoimmune diseases, including lupus and myositis. Methods: Herein, we present 15 new onset autoimmune myositis temporally associated with SARS-CoV-2 RNA or DNA-based vaccines that occurred between February 2021 and April 2022. Musculoskeletal, pulmonary, cutaneous and cardiac manifestations, laboratory and imaging data were collected. Results: In total, 15 cases of new onset myositis (11 polymyositis/necrotizing/overlap myositis; 4 dermatomyositis) were identified in the Yorkshire region of approximately 5.6 million people, between February 2021 and April 2022 (10 females/5 men; mean age was 66.1 years; range 37–83). New onset disease occurred after first vaccination (5 cases), second vaccination (7 cases) or after the third dose (3 cases), which was often a different vaccine. Of the cases, 6 had systemic complications including skin (3 cases), lung (3 cases), heart (2 cases) and 10/15 had myositis associated autoantibodies. All but 1 case had good therapy responses. Adverse event following immunization (AEFI) could not be explained based on the underlying disease/co-morbidities. Conclusion: Compared with our usual regional Rheumatology clinical experience, a surprisingly large number of new onset myositis cases presented during the period of observation. Given that antigen release inevitably follows muscle injury and given the role of nucleic acid adjuvanticity in autoimmunity and muscle disease, further longitudinal studies are required to explore potential links between novel coronavirus vaccines and myositis in comparison with more traditional vaccine methods.

Background The SARS-CoV-2 pandemic is a global health problem. Beside the specific pathogenic effect of SARS-CoV-2, incompletely understood deleterious and aberrant host immune responses play critical roles in severe disease. Our objective was to summarise the available information on the pathophysiology of COVID-19.Methods Two reviewers independently identified eligible studies according to the following PICO framework: P (population): patients with SARS-CoV-2 infection; I (intervention): any intervention/no intervention; C (comparator): any comparator; O (outcome) any clinical or serological outcome including but not limited to immune cell phenotype and function and serum cytokine concentration.Results Of the 55 496 records yielded, 84 articles were eligible for inclusion according to question-specific research criteria. Proinflammatory cytokine expression, including interleukin-6 (IL-6), was increased, especially in severe COVID-19, although not as high as other states with severe systemic inflammation. The myeloid and lymphoid compartments were differentially affected by SARS-CoV-2 infection depending on disease phenotype. Failure to maintain high interferon (IFN) levels was characteristic of severe forms of COVID-19 and could be related to loss-of-function mutations in the IFN pathway and/or the presence of anti-IFN antibodies. Antibody response to SARS-CoV-2 infection showed a high variability across individuals and disease spectrum. Multiparametric algorithms showed variable diagnostic performances in predicting survival, hospitalisation, disease progression or severity, and mortality.Conclusions SARS-CoV-2 infection affects both humoral and cellular immunity depending on both disease severity and individual parameters. This systematic literature review informed the EULAR ‘points to consider’ on COVID-19 pathophysiology and immunomodulatory therapies.

IntroductionPost-operative CSF leak still represents the main drawback of Endoscopic Endonasal Approach (EEA), and different reconstructive strategies have been proposed in order to decrease its rate.ObjectiveTo critically analyze the effectiveness of different adopted reconstruction strategies in patients that underwent EEA.Materials and methodsAdult patients with skull base tumor surgically treated with EEA were retrospectively analyzed. Data recorded for each case concerned patient demographics, type of surgical approach, histotype, anatomical site of surgical approach, intra-operative CSF leak grade (no leak (INL), low flow (ILFL), high flow (IHFL)), reconstructive adopted strategy, Lumbar Drain positioning, post-operative CSF leak rate and intra/post-operative complications.ResultsA total number of 521 patients (January 2012-December 2019) was included. Intra-operative CSF leak grade showed to be associated with post-operative CSF leak rate. In particular, the risk to observe a post-operative CSF leak was higher when IHFL was encountered (25,5%; Exp(B) 16.25). In particular, vascularized multilayered reconstruction and fat use showed to be effective in lowering post-operative CSF leaks in IHFL (p 0.02). No differences were found considering INL and ILFL groups. Yearly post-operative CSF leak rate analysis showed a significative decreasing trend.ConclusionIntra-operative CSF leak grade strongly affected post-operative CSF leak rate. Multilayer reconstruction with fat and naso-septal flap could reduce the rate of CSF leak in high risk patients. Reconstructive strategies should be tailored according also to the type and the anatomical site of the approach.

Odonata can be sampled following different types of protocols. In Brazil, the most used protocol is the scanning in fixed areas method, where a 100-meter transect is delimited in one of the stream margins, subdivided into 20 segments measuring 5 meters. Despite being universally used, the methodological efficiency or limitations of this protocol for Odonata has never been tested. In this scenario, our objective was to assess the efficiency of the sampling protocol to measure the richness and composition of Odonata in three fundamental aspects: the time of sampling and sampling effort over time and space. We show that the best sampling efficiency was achieved in collections performed at noon, in transects measuring 100 meters, requiring at least two samplings in the same location, supporting the procedures traditionally adopted by many studies with the group. While comparing species composition, we did not see any implication between the different treatments on the capture of the local species pool. However, we highlight and discuss some possible methodological flaws when using this protocol to sample specific Odonata groups. We believe the results obtained are fundamental in the inventory of species and to conduct future studies, as well as to aid conservative measures that use the order Odonata as a tool for environmental monitoring.

Optimizing antithymocyte globulin (rATG) dosage is critical for high immunological risk patients undergoing a repeat kidney transplant. This natural retrospective cohort study compared clinical outcomes of two successive cohorts of consecutive recipients of retransplants receiving 5 x 1 mg/kg (rATG-5, n = 100) or a single 3 mg/kg (rATG-3, n = 110) dose of rATG induction therapy. All patients had negative complement-dependent cytotoxicity crossmatch and no anti-HLA A, B, DR donor-specific antibodies (DSA). The primary endpoint was efficacy failure (first biopsy-proven acute rejection, graft loss, or death) at 12 months. There was no difference in the cumulative incidence of efficacy failure (18.0% vs. 21.8%, HR = 1.22, 95% CI 0.66-2.25), respectively. There were no differences in 3-years freedom from biopsy proven acute rejection, and patient, graft, and death-censored graft survivals. There were no differences in the incidence of surgical complications (25.0% vs. 18.2%; p 0.151), early hospital readmission (27.8% vs. 29.5%; p = 0.877) and CMV infections (49% vs. 40%; p = 0.190). There were also no differences in the incidence (59.6% vs. 58.7%, p = 0.897) and duration of delayed graft function but a stable difference in estimate glomerular filtration rate was observed from month 1 (54.7±28.8 vs. 44.1±25.3 ml/min/1.73 m2, p = 0.005) to month 36 (51.1±27.7 vs. 42.5±24.5, p = 0.019). Mean urinary protein concentration (month 36: 0.38±0.81 vs. 0.70±2.40 g/ml, p = 0.008) and mean chronic glomerular Banff score in for cause biopsies (months 4-36: 0.0±0.0 vs. 0.04±0.26, p = 0.044) were higher in the rATG-3 group. This cohort analysis did not detect differences in the incidence of efficacy failure and in safety outcomes at 12 months among recipients of kidney retransplants without A, B, and DR DSA, receiving induction therapy with a single 3 mg/kg rATG dose or the traditional 5 mg/kg rATG.

Primary hyperlipidaemia in Schnauzer is characterized by increased plasma triglycerides (TG) and/or total cholesterol (TC) concentration and is associated with an increased risk of developing pancreatitis, insulin resistance and seizures. In humans, omega-3 fatty acids in addition to a low-fat diet can be used to reduce TG and TC. This study evaluated the therapeutic efficacy of omega-3 fatty acids associated to a diet management with two different fat content in Schnauzer with primary hyperlipidaemia. Eighteen dogs with primary hyperlipidaemia were divided into two groups: group 1, n = 10, 8 females, 2 males, age (mean ± standard deviation) of 7.13 ± 2.70 years and body weight (BW) (mean ± standard deviation) of 7.25 ± 1.22 kg were treated with fish oil (approximately 730 mg/day of omega-3) associated with a low-fat and low-calorie diet (approximately 24g of fat/1000 kcal) for 90 days (T90); and group 2, n = 8 dogs, 6 females, 2 males, with 7.0 ± 1.77 years old and average BW of 8.36 ± 1.51 kg, treated with fish oil (approximately 730 mg/day of omega-3) and maintenance diet with moderate amount of fat (approximately 33g of fat/1000 kcal) for 90 days. Plasma TG and TC concentrations and lipoprotein (LP) profile (VLDL, LDL, HDL) were evaluated before and after treatment. TG and TC serum concentrations, expressed in mg/dL (mean ± standard deviation), before and after treatment in group 1 were: TG = 391.30 ± 487.86 (T0) and 118.7 ± 135.21 (T90); TC = 308.2 ± 63.06 (T0) and 139 ± 36.91 (T90). As for group 2, TG = 391.63 ± 336.89 (T0) and 250.75 ± 211.56 (T90); TC = 257.25 ± 92.88 (T0) and 207.25 ± 63.79 (T90). A reduction (p

Human Cytomegalovirus (HCMV) infects over half the world's population, is a leading cause of congenital birth defects, and poses serious risks for immuno-compromised individuals. To expand the molecular knowledge governing virion maturation, we analysed HCMV virions using proteomics, and identified a significant proportion of host exosome constituents. To validate this acquisition, we characterized exosomes released from uninfected cells, and demonstrated that over 99% of the protein cargo was subsequently incorporated into HCMV virions during infection. This suggested a common membrane origin, and utilization of host exosome machinery for virion assembly and egress. Thus, we selected a panel of exosome proteins for knock down, and confirmed that loss of 7/9 caused significantly less HCMV production. Saliently, we report that VAMP3 is essential for viral trafficking and release of infectious progeny, in various HCMV strains and cell types. Therefore, we establish that the host exosome pathway is intrinsic for HCMV maturation, and reveal new host regulators involved in viral trafficking, virion envelopment, and release. Our findings underpin future investigation of host exosome proteins as important modulators of HCMV replication with antiviral potential.

IntroductionAmyotrophic lateral sclerosis (ALS) is a fatal progressive neurological disorder characterised by a selective degeneration of motor neurons (MNs). Stem cell transplantation is considered as a promising strategy in neurological disorders therapy and the possibility of inducing bone marrow cells (BMCs) to circulate in the peripheral blood is suggested to investigate stem cells migration in degenerated ALS nerve tissues where potentially repair MN damage. Granulocyte-colony stimulating factor (G-CSF) is a growth factor which stimulates haematopoietic progenitor cells, mobilises BMCs into injured brain and it is itself a neurotrophic factor for MN. G-CSF safety in humans has been demonstrated and many observations suggest that it may affect neural cells. Therefore, we decided to use G-CSF to mobilise BMCs into the peripheral circulation in patients with ALS, planning a clinical trial to evaluate the effect of G-CSF administration in ALS patients compared with placebo.Methods and analysisSTEMALS-II is a phase II multicentre, randomised double-blind, placebo-controlled, parallel group clinical trial on G-CSF (filgrastim) and mannitol in ALS patients. Specifically, we investigate safety, tolerability and efficacy of four repeated courses of intravenous G-CSF and mannitol administered in 76 ALS patients in comparison with placebo (indistinguishable glucose solution 5%). We determine increase of G-CSF levels in serum and cerebrospinal fluid as CD34+ cells and leucocyte count after treatment; reduction in ALS Functional Rating Scale-Revised Score, forced vital capacity, Scale for Testing Muscle Strength Score and quality of life; the adverse events/reactions during the treatment; changes in neuroinflammation biomarkers before and after treatment.Ethics and disseminationThe study protocol was approved by the Ethics Committee of Azienda Ospedaliera Universitaria ‘Città della Salute e della Scienza’, Torino, Italy. Results will be presented during scientific symposia or published in scientific journals.Trial registration numberEudract 2014-002228-28.

INTRODUCTION:Thrombotic microangiopathy (TMA) in post-transplant setting has heterogeneous clinical manifestations. METHODS:We retrospectively studied data of 89 patients with post-transplant TMA, which was characterized by thrombi in at least one glomerulus and/or arteriole. Systemic TMA was defined by thrombocytopenia and microangiopathic anemia and early onset TMA, when occurred less than 90 days post transplant. RESULTS:The cumulative incidence was 0.93%. The majority of the recipients were young (mean age 39 years), female (52%) and Caucasian (48%) with primary kidney disease of unknown etiology (37%). Early TMA occurred in 51% of the patients and systemic TMA, in 25%. Underlying precipitating factors were: infection (54%), acute rejection (34%), calcineurin inhibitor toxicity (13%) and pregnancy (3%). 18% of the patients had several triggers. Glomerular TMA was observed in 50% of the biopsies and endothelial cell activation, in 61%. The 1-year patient survival was 97% and corresponding graft survival, 66%. Allograft survival was inferior when acute antibody mediated rejection (ABMR) occurred (with 41%; without 70%, p = 0.01), however no differences were determined by hemolysis, time of onset, thrombi location or endothelial cell activation. CONCLUSIONS:Our results suggest that post-transplant TMA is a rare but severe condition, regardless of its clinical and histological presentation, mainly when associated to ABMR.

The yeast Komagataella phaffii is widely used as a microbial host for heterologous protein production. However, molecular tools for this yeast are basically restricted to a few integrative and replicative plasmids. Four sequences that have recently been proposed as the K. phaffii centromeres could be used to develop a new class of mitotically stable vectors. In this work, we designed a color-based genetic assay to investigate plasmid stability in K. phaffii and constructed vectors bearing K. phaffii centromeres and the ADE3 marker. These genetic tools were evaluated in terms of mitotic stability by transforming an ade2/ade3 auxotrophic strain and regarding plasmid copy number by quantitative PCR (qPCR). Our results confirmed that the centromeric plasmids were maintained at low copy numbers as a result of typical chromosome-like segregation during cell division. These features, combined with in vivo assembly possibilities, prompt these plasmids as a new addition to the K. phaffii genetic toolbox.

The evaluation of cultivars using multi-environment trials (MET) is an important step in plant breeding programs. One of the objectives of these evaluations is to understand the genotype by environment interaction (GEI). A method of determining the effect of GEI on the performance of cultivars is based on studies of adaptability and stability. Initial studies were based on linear regression; however, these methodologies have limitations, mainly in trials with genetic or statistical unbalanced, heterogeneity of residual variances, and genetic covariance. An alternative would be the use of random regression models (RRM), in which the behavior of the genotypes is characterized as a reaction norm using longitudinal data or repeated measurements and information regarding a covariance function. The objective of this work was the application of RRM in the study of the behavior of common bean cultivars using a MET, based on Legendre polynomials and genotype-ideotype distances. We used a set of 13 trials, which were classified as unfavorable or favorable environments. The results revealed that RRM enables the prediction of the genotypic values of cultivars in environments where they were not evaluated with high accuracy values, thereby circumventing the unbalanced of the experiments. From these values, it was possible to measure the genotypic adaptability according to ideotypes, according to their reaction norms. In addition, the stability of the cultivars can be interpreted as variation in the behavior of the ideotype. The use of ideotypes based on real data allowed a better comparison of the performance of cultivars across environments. The use of RRM in plant breeding is a good alternative to understand the behavior of cultivars in a MET, especially when we want to quantify the adaptability and stability of genotypes.

Many authors have tried to explain the shape of the species abundance distribution (SAD). Some of them have suggested that sampling spatial scale is an important factor shaping SADs. These suggestions, however, did not consider the indirect and well-known effect of sample size, which increases as samples are combined to generate SADs at larger spatial scales. Here, we separate the effects of sample size and sampling scale on the shape of the SAD for three groups of organisms (trees, beetles and birds) sampled in the Brazilian Atlantic Forest. We compared the observed SADs at different sampling scales with simulated SADs having the same richness, relative abundances but comparable sample sizes, to show that the main effect shaping SADs is sample size and not sampling spatial scale. The effect of scale was minor and deviations between observed and simulated SADs were present only for beetles. For trees, the match between observed and simulated SADs was improved at all spatial scales when we accounted for conspecific aggregation, which was even more important than the sampling scale effect. We build on these results to propose a conceptual framework where observed SADs are shaped by three main factors, in decreasing order of importance: sample size, conspecific aggregation and beta diversity. Therefore, studies comparing SADs across sites or scales should use sampling and/or statistical approaches capable of disentangling these three effects on the shape of SADs.

Subjects affected by ulcerative colitis and Crohn’s disease with colonic localization have an increased risk of colorectal cancer (CRC). Surveillance colonoscopy is recommended by international guidelines as it can detect early-stage CRC. Based on previous evidence, in 2015 the Surveillance for Colorectal Endoscopic Neoplasia Detection and Management in Inflammatory Bowel Disease Patients International Consensus indicated dye chromoendoscopy (DCE) as the most effective technique for detecting dysplasia. However, advances in endoscopic technology such as high-definition colonoscopes and dye-less virtual chromoendoscopy (VCE) may change future practice. In this review, we summarize the available evidence on CRC surveillance in IBD, focusing on the emerging role of high-definition white light endoscopy (HD-WLE) and VCE over the standard DCE, and the current role of random biopsies.

The liver fluke Fasciola hepatica is an economically important global pathogen of humans and their livestock. To facilitate host invasion and migration, F. hepatica secretes an abundance of cathepsin peptidases but prevents excessive damage to both parasite and host tissues by co-secreting regulatory peptidase inhibitors, cystatins/stefins and Kunitz-type inhibitors. Here, we report a vaccine strategy aimed at disrupting the parasite’s protease/anti-protease balance by targeting these key inhibitors. Our vaccine cocktail containing three recombinant stefins (rFhStf-1, rFhStf-2, rFhStf-3) and a Kunitz-type inhibitor (rFhKT1) formulated in adjuvant Montanide 61VG was assessed in two independent sheep trials. While fluke burden was not reduced in either trial, in Trial 1 the vaccinated animals showed significantly greater weight gain (p < 0.05) relative to the non-vaccinated control group. In both trials we observed a significant reduction in egg viability (36–42%). Multivariate regression analyses showed vaccination and increased levels of IgG2 antibodies specific for the F. hepatica peptidase inhibitors were positive indicators for increased weight gain and levels of haemoglobin within the normal range at 16 weeks post-infection (wpi; p < 0.05). These studies point to the potential of targeting peptidase inhibitors as vaccine cocktails for fasciolosis control in sheep.

In January 2015, during a 3-week period, 12 captive Tonkean macacques at a sanctuary in Italy died. An orthopoxvirus infection was suspected because of negative-staining electron microscopy results. The diagnosis was confirmed by histology, virus isolation, and molecular analysis performed on different organs from all animals. An epidemiologic investigation was unable to define the infection source in the surrounding area. Trapped rodents were negative by virologic testing, but specific IgG was detected in 27.27% of small rodents and 14.28% of rats. An attenuated live vaccine was administered to the susceptible monkey population, and no adverse reactions were observed; a detectable humoral immune response was induced in most of the vaccinated animals. We performed molecular characterization of the orthopoxvirus isolate by next-generation sequencing. According to the phylogenetic analysis of the 9 conserved genes, the virus could be part of a novel clade, lying between cowpox and ectromelia viruses.

Background: We present immunogenicity data 6 months after the first dose of BNT162b2 in correlation with age, gender, BMI, comorbidities and previous SARS-CoV-2 infection. Methods: An immunogenicity evaluation was carried out among health care workers (HCW) vaccinated at the Istituti Fisioterapici Ospitalieri (IFO). All HCW were asked to be vaccine by the national vaccine campaign at the beginning of 2021. Serum samples were collected on day 1 just prior to the first dose of the vaccine and on day 21 just prior to the second vaccination dose. Thereafter sera samples were collected 28, 49, 84 and 168 days after the first dose of BNT162b2. Quantitative measurement of IgG antibodies against S1/S2 antigens of SARS-CoV-2 was performed with a commercial chemiluminescent immunoassay. Results: Two hundred seventy-four HWCs were analyzed, 175 women (63.9%) and 99 men (36.1%). The maximum antibody geometric mean concentration (AbGMC) was reached at T2 (299.89 AU/mL; 95% CI: 263.53–339.52) with a significant increase compared to baseline (p < 0.0001). Thereafter, a progressive decrease was observed. At T5, a median decrease of 59.6% in COVID-19 negative, and of 67.8% in COVID-19 positive individuals were identified with respect to the highest antibody response. At T1, age and previous COVID-19 were associated with differences in antibody response, while at T2 and T3 differences in immune response were associated with age, gender and previous COVID-19. At T4 and T5, only COVID-19 positive participants demonstrated a greater antibody response, whereas no other variables seemed to influence antibody levels. Conclusions: Overall our study clearly shows antibody persistence at 6 months, albeit with a certain decline. Thus, the use of this vaccine in addressing the COVID-19 pandemic is supported by our results that in turn open debate about the need for further boosts.

We hypothesized that the spread of SARS-CoV-2 in urine during a severe COVID-19 infection may be the expression of the worsening disease evolution. Therefore, the aim of this study was to verify if the COVID-19 disease severity is related to the viral presence in urine samples. We evaluated the clinical evolution in acute COVID-19 patients admitted in the sub-intensive care and intensive care units between 28 of December 2020 and 15th of February 2021 and being positive for SARS-CoV-2 RNA in the respiratory tract, including repeated endotracheal aspirates (ETA), sputum, nasopharyngeal swabs (NPS) and urine. We found that those subjects with SARS-COV-2 in the urine at admittance (8 out of 60 eligible patients) had a more severe disease than those with negative SARS-CoV-2 in urine. Further, they showed an increase in fibrinogen and (C-reactive Protein) CRP serum levels, requiring mechanic ventilation. Of those with positive SARS-CoV-2 in the urine, 50% died. According to our preliminary results, it seems that the presence of SARS-CoV-2 in the urine characterizes patients with a more severe disease and is also related to a higher death rate.