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Several studies highlight the role of inflammatory markers in thrombosis as well as in cancer. However, their combined role in cancer-associated deep vein thrombosis (DVT) and the molecular mechanisms, involved in its pathophysiology, needs further investigations. In the present study, C-reactive protein, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1β), matrix metalloproteases-9 (MMP-9), vascular endothelial growth factor (VEGF), tissue factor (TF), fibrinogen and soluble P-selectin, were analyzed in plasma and in monocyte samples from 385 cancer patients, of whom 64 were concomitantly affected by DVT (+). All these markers were higher in cancer patients DVT+ than in those DVT-. Accordingly, significantly higher NF-kB activity was observed in cancer patients DVT+ than DVT-. Significant correlation between data obtained in plasma and monocyte samples was observed. NF-kB inhibition was associated with decreased levels of all molecules in both cancer DVT+ and DVT-. To further demonstrate the involvement of NF-kB activation by the above mentioned molecules, we treated monocyte derived from healthy donors with a pool of sera from cancer patients with and without DVT. These set of experiments further suggest the significant role played by some molecules, regulated by NF-kB, and detected in cancer patients with DVT. Our data support the notion that NF-kB may be considered as a therapeutic target for cancer patients, especially those complicated by DVT. Treatment with NF-kB inhibitors may represent a possible strategy to prevent or reduce the risk of DVT in cancer patients.

Rehabilitation is the only current intervention that improves sensorimotor function in ischemic stroke patients, similar to task-specific intensive training in animal models of stroke. Bone marrow mesenchymal stem cells (BM-MSCs)-derived extracellular vesicles (EVs) are promising in restoring brain damage and function in stroke models. Additionally, the non-invasive intranasal route allows EVs to reach the brain and target specific ischemic regions. Yet unclear is how handling might enhance recovery or influence other therapies such as EVs after stroke. We used the transient middle cerebral artery occlusion (MCAO) model of stroke in rats to assess how intensive handling alone, in the form of sensorimotor behavioral tests, or in combination with an intranasal treatment of EVs restored neurological function and ischemic damage. Handled rats were exposed to a battery of sensorimotor tests, including the modified Neurological Severity Score (mNSS), beam balance, corner, grid walking, forelimb placement, and cylinder tests, together with Magnetic Resonance Imaging (MRI) at 2, 7, 14, 21, and 28 days post-stroke (dps). Handled MCAO rats were also exposed to an intranasal multidose or single dose of EVs. Non-handled rats were evaluated only by mNSS and MRI at 2, 28, and 56 dps and were treated with a single intranasal dose of EVs. Our results showed that handling animals after MCAO is necessary for EVs to work at the tested dose and frequency, and that a single cumulative dose of EVs further improves the neurological function recovered during handling. These results show the importance of rehabilitation in combination with other treatments such as EVs, and highlight how extensive behavioral testing might influence functional recovery after stroke., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Gomez-Galvez et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background & Aims: Current screening pathways, developed from tertiary care cohorts, underestimate the presence of Metabolic-dysfunction associated steatotic liver disease (MASLD) in patients with type 2 diabetes mellitus (T2DM) in the community. We developed, validated, and assessed cost-effectiveness of a new score for screening the presence of fibrosis due to MASLD in primary care., Methods: Consecutive T2DM patients underwent screening for liver diseases with transient elastography (TE). Based on predictors of significant/advanced fibrosis, we generated the BIMAST score (based on aspartate aminotransferase (AST) and body mass index (BMI)) and validated it internally and externally (Royal Free Hospital, London and Palermo Hospital). For cost-effectiveness analysis, 6 screening strategies were compared against standard of care: BIMAST score, ultrasound plus abnormal liver function tests, FIB-4, NAFLD fibrosis score, ELF and transient elastography (TE). A Markov model was built based on fibrosis status. Cost per quality-adjusted life year (QALY) gained and the incremental cost-effectiveness ratio (ICER) were estimated over a lifetime., Results: Among 300 patients enrolled, 64% (186) had MASLD and 10% (28) other causes of liver disease. In the whole population, patients with significant fibrosis, advanced fibrosis, and cirrhosis due to MASLD were 17% (50/287), 11% (31/287), and 3% (8/287), respectively. In primary care, BIMAST performed better than other non-invasive markers at predicting significant and advanced fibrosis. Moreover, BIMAST reduced false negatives from 54% (ELF) and 38% (FIB-4) to 10%. In both validation cohorts, BIMAST performance was as good as FIB-4. In the cost-utility analysis, ICER was £2,337.92/QALY for BIMAST., Conclusion: The BIMAST predicts the presence of significant fibrosis in the community, reduces false negatives and is cost-effective. The BIMAST score should be included in the holistic assessment of diabetic patients., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Forlano et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: Epithelioid hemangioendothelioma (EHE) is an ultra-rare sarcoma, marked by distinctive molecular and pathological features and with a variable clinical behavior. Its natural history is still partially understood, reliable prognostic and predictive factors are lacking and many questions are still open on the optimal management. In the context of EURACAN, a prospective registry specifically dedicated to EHE was developed and launched with the aim of providing, through high-quality prospective data collection, a better understanding of this disease., Study Design: Registry-based cohort study including only new cases of patients with a pathological and molecularly confirmed diagnosis of EHE., Objectives: To improve the understanding of EHE natural history, validate and identify new prognostic and predictive factors, clarify the activity and efficacy of currently available treatment options, describe treatment pattern., Methods: Settings and participantsIt is an hospital-based registry established in centers with expertise in EHE including adult patients with a new pathological and molecularly confirmed diagnosis of EHE starting from the 1st December 2023. The characteristics of each patient in the facility who meets the above-mentioned inclusion criteria will be collected prospectively and longitudinally with follow-up at cancer progression and / or cancer relapse or patient death. It is a secondary use of data which will be collected from the clinical records. The data collected for the registry will not entail further examinations or admissions to the facility and/or additional appointments to those normally provided for routine patient follow-up. VariablesFull details on patients and disease features, treatment and outcome will be collected, according to common clinical practice guidelines developed and shared with all the contributing centers. In addition, data on potential confounders (e.g. comorbidity; functional status etc.) will also be collected. Statistical methodsThe data analyses will include descriptive statistics and analytical analyses. Multivariable Cox's proportional hazards model and Hazard ratios (HR) for all-cause or cause-specific mortality will be used to determine independent predictors of overall survival, recurrence and progression., Results: The registry has been joined by 21 sarcoma reference centers across EU and UK, covering 10 countries. Patients' recruitment started in December 2023. The estimated completion date is December 2033 upon agreement on the achievement of all the registry objectives. The already established collaboration and participation of EHE patient's associations involved in the project will help in promoting the registry and fostering accrual., Competing Interests: Anna Maria Frezza declares institutional research funding from Advenchen Laboratories, Amgen Dompé, AROG Pharmaceuticals, Ayala Pharmaceuticals, Bayer, Blueprint Medicines, Boehriger Ingelheim, Daiichi Sankyo, Deciphera, Eisai, Eli Lilly, Epizyme Inc, Foghorn Therapeutics Inc., Glaxo, Hutchison MediPharma Limited, Inhibrx, Inc., Karyopharm Pharmaceuticals, Novartis, Pfizer, PharmaMar, PTC Therapeutics, Rain Oncology, SpringWorks Therapeutics. Alexia Bertuzzi declares support for attending meeting and/or travels from Pharmamar and Istituto Gentili; support for scientific activities from Istituto Gentili. Antonella Brunello declares consulting fees or serving on advisory boards for Eli Lilly, Roche, GSK, Eisai, Pharmamar, Boehringer Ingelheim, Deciphera; payment or honoraria for educational events by GSK and Pharmamar; travel grants by Pharmamar, Istituto Gentili. Giacomo G. Baldi declares consulting fees from Eli Lilly, Pharmamar, AboutEvents; honoraria from Pharmamar, Eli Lilly, Glaxo Smith Kline, Merck Sharp & Dome, Eisai, IstitutoGentili; support for attending meetings and/or travels from Novartis, Pharmamar, Eli Lilly; participation on the advisory board from Pharmamar, Eli Lilly, Glaxo Smith Kline, Merck Sharp & Dome, Eisai. Paolo G. Casali declares institutional research funding from Advenchen Laboratories, Amgen Dompé, AROG Pharmaceuticals, Ayala Pharmaceuticals, Bayer, Blueprint Medicines, Boehriger Ingelheim, Daiichi Sankyo, Deciphera, Eisai, Eli Lilly, Epizyme Inc, Foghorn Therapeutics Inc., Glaxo, Hutchison MediPharma Limited, Inhibrx, Inc., Karyopharm Pharmaceuticals, Novartis, Pfizer, PharmaMar, PTC Therapeutics, Rain Oncology, SpringWorks Therapeutics. Virginia Ferraresi declares consultancy fees or honoraria from PharmaMar, Boehringer Ingelheim, Gentili e Serb Pharmaceuticals. Matilde Ingrosso reports institutional research funding from Advenchen Laboratories, Amgen Dompé, AROG Pharmaceuticals, Ayala Pharmaceuticals, Bayer, Blueprint Medicines, Boehriger Ingelheim, Daiichi Sankyo, Deciphera, Eisai, Eli Lilly, Epizyme Inc, Foghorn Therapeutics Inc., Glaxo, Hutchison MediPharma Limited, Inhibrx, Inc., Karyopharm Pharmaceuticals, Novartis, Pfizer, PharmaMar, PTC Therapeutics, Rain Oncology, SpringWorks Therapeutics. Laura Monteleone reports institutional research funding from Advenchen Laboratories, Amgen Dompé, AROG Pharmaceuticals, Ayala Pharmaceuticals, Bayer, Blueprint Medicines, Boehriger Ingelheim, Daiichi Sankyo, Deciphera, Eisai, Eli Lilly, Epizyme Inc, Foghorn Therapeutics Inc., Glaxo, Hutchison MediPharma Limited, Inhibrx, Inc., Karyopharm Pharmaceuticals, Novartis, Pfizer, PharmaMar, PTC Therapeutics, Rain Oncology, SpringWorks Therapeutics. Johanna Szkandera reports participation in advisory boards or invited speaker fees for PharmaMar, Bayer, Roche, Lilly, Amgen; travel expenses coverage from PharmaMar, Roche, Lilly, Amgen, Bristol Myers Squibb; research funding from PharmaMar, Roche, Eisai. Bruno Vincenzi reports consulting fees from Eisai, Lilly, Bayer, Deciphera, PharmaMar, Blueprint, Pfizer, GSK, Accord, Abbott and research support from PharmaMar, Novartis, Lilly Silvia Stacchiotti declares advisory board roles with Bayer, Boehringer, Daiichi, Ikena, Nec Oncology, Pharma Essentia, Regeneron, Servier; invited speaker roles for Bayer, Boehringer, Gentili, Pharmamar; institutional research funding from Advenchen Laboratories, Amgen Dompé, AROG Pharmaceuticals, Ayala Pharmaceuticals, Bayer, Blueprint Medicines, Boehriger Ingelheim, Daiichi Sankyo, Deciphera, Eisai, Eli Lilly, Epizyme Inc, Foghorn Therapeutics Inc., Glaxo, Hutchison MediPharma Limited, Inhibrx, Inc., Karyopharm Pharmaceuticals, Novartis, Pfizer, PharmaMar, PTC Therapeutics, Rain Oncology, SpringWorks Therapeutics. Carlo Cicala, Lorenzo D’Ambrosio, Angelo Paolo Dei Tos, Alessandro Gronchi, Nadia Hindi, Robin Jones, Iwona Lugowska, Julia Ketzer, Anna Mariuk-Jarema, Andrea Napolitano, Andri Papakonstantinou, Sandro Pasquali, Marta Sbaraglia, Federico Scolari, Elisa Crotti, Irene Benzonelli, Jean-Yves Blay, Christophe Bouvier, Javier Martin Broto, Hugh Leonard, Ninna Aggerholm-Pedersen, Andri Papakonstantinou, Giuseppe Bianchi, Federica Zuccheri, Paolo Baili, Simone Bonfarnuzzo, Domenico Campanacci, Armelle Dufresne, Sebastian Bauer, Serena Bazzurri, Stefano Gabellini, Claudia Giani, Vincenzo Giannusa, Melissa Gil-Sanjines, Teresa Grassani, Paolo Lasalvia, Stefan Lindskog, Salvatore Vizzaccaro, Nils Dieckmann, Alessandro Mazzocca, Andrei Ivanescu, Giuseppe Badalamenti, Carlo Morosi, Stephanie Elston, Kyetil Boye, Francesca Nardozza, Elisabetta Neri, Maria Nilsson and Annalisa Trama declares no conflict of interest., (Copyright: © 2024 Frezza et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Silybum marianum (L.) Gaertn. is a multipurpose crop native to the Mediterranean and middle east regions and mainly known for the hepatoprotective properties of fruit-derived silymarin. Despite growing interest in milk thistle as a versatile crop with medicinal value, its potential in agroindustry is hindered by incomplete domestication and limited genomic knowledge, impeding the development of competitive breeding programs. The present study aimed to evaluate genetic diversity in a panel of S. marianum accessions (n = 31), previously characterized for morphological and phytochemical traits, using 5,178 polymorphic DArTseq SNP markers. The genetic structure investigated using both parametric and non-parametric approaches (e.g. PCA, AWclust, Admixture), revealed three distinctive groups reflecting geographical origins. Indeed, Pop1 grouped accessions from Central Europe and UK, Pop3 consisted mainly of accessions of Italian origin, and Pop2 included accessions from different geographical areas. Interestingly, Italian genotypes showed a divergent phenotypic distribution, particularly in fruit oleic and linoleic acid content, compared to the other two groups. Genetic differentiation among the three groups, investigated by computing pairwise fixation index (FST), confirmed a greater differentiation of Pop3 compared to other subpopulations, also based on other diversity indices (e.g. private alleles, heterozygosity). Finally, 22 markers were declared as putatively under natural selection, of which seven significantly affected some important phenotypic traits such as oleic, arachidonic, behenic and linoleic acid content. These findings suggest that these markers, and overall, the seven SNP markers identified within Pop3, could be exploited in specific breeding programs, potentially aimed at diversifying the use of milk thistle. Indeed, incorporating genetic material from Pop3 haplotypes carrying the selected loci into milk thistle breeding populations might be the basis for developing milk thistle lines with higher levels of oleic, arachidonic, and behenic acids, and lower levels of linoleic acid, paving new avenues for enhancing the nutritional and agronomic characteristics of milk thistle., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Puglisi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Quadruplex-Duplex (Q-D) junctions are unique structural motifs garnering increasing interest as drug targets, due to their frequent occurrence in genomic sequences. The viral HIV LTR-III sequence was chosen as a Q-D junction model to study the affinity of the selected compounds BMH-21, namitecan (ST-1968), and doxorubicin (DOXO), all containing a planar polycyclic aromatic moiety, linked to either one short aminoalkyl or an aminoglycosyl group. A multidisciplinary approach that combines NMR spectroscopy, molecular modelling, circular dichroism (CD) and fluorescence spectroscopy was employed. The studied ligands induced moderate but clear stabilization to the Q-D junction by interacting with the interfacial tetrad. DOXO was found to be the best Q-D junction binder. Interestingly, the removal of the aminoglycosyl group significantly changed the pattern of the interactions, indicating that highly polar substituents have a stronger affinity with the exposed regions of the Q-D junction, particularly at the level of the interfacial tetrad., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Mazzini et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Erysiphe corylacearum has recently been reported in northern Italy (Piedmont) and other European countries as the causal agent of a new emerging powdery mildew on hazelnut. This disease is much more dangerous than the common hazelnut powdery mildew caused by Phyllactinia guttata as it significantly reduces yield and quality of hazelnuts. This study aimed to perform morphological and molecular characterization of the fungal isolates from powdery mildew-infected plants in the Piedmont Italian region. Additionally, genetic diversity studies and pathogenicity tests were conducted. Thirty-six fungal isolates originating from symptomatic hazelnut plants exhibiting specific powdery mildew symptoms on the superior leaf side were identified morphologically as E. corylacearum. Single- and multilocus sequence typing of five loci (ITS, rpb2, CaM, GAPDH and GS) assigned all isolates as E. corylacearum. Multilocus and GAPDH phylogenetic studies resulted in the most efficient characterization of E. corylacearum. Studied fungal isolates were able to cause new emerging powdery mildew disease by fulfilling Koch's postulates. The emergence of powdery mildew disease in Italy revealed the E. corylacearum subgrouping, population expansion, and high nucleotide similarity with other recently identified E. corylacearum hazelnut isolates. To contain this harmful disease and inhibit the fungus spread into new geographical zones, it will be necessary to implement more rigorous monitoring in neighboring hazelnut plantations near infected hazelnuts, use sustainable fungicides and search for new biocontrol agents., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Matić et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Multi-layer Complex networks are commonly used for modeling and analysing biological entities. This paper presents the advantage of using COMBO (Combining Multi Bio Omics) to suggest a new role of the chromosomal aberration as a cancer driver factor. Exploiting the heterogeneous multi-layer networks, COMBO integrates gene expression and DNA-methylation data in order to identify complex bilateral relationships between transcriptome and epigenome. We evaluated the multi-layer networks generated by COMBO on different TCGA cancer datasets (COAD, BLCA, BRCA, CESC, STAD) focusing on the effect of a specific chromosomal numerical aberration, broad gain in chromosome 20, on different cancer histotypes. In addition, the effect of chromosome 8q amplification was tested in the same TCGA cancer dataset. The results demonstrate the ability of COMBO to identify the chromosome 20 amplification cancer driver force in the different TCGA Pan Cancer project datasets., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Cosentini et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

A previously undocumented shallow water hydrothermal field from Sicily (Southern Tyrrhenian Sea, Italy) is here described, based on a multidisciplinary investigation. The field, covering an area of nearly 8000 m2 and a depth from the surface to -5 m, was explored in June 2021 to characterise the main physico-chemical features of the water column, describe the bottom topography and features, and identify the main megabenthic and nektonic species. Twenty sites were investigated to characterise the carbonate system. Values of pH ranged between 7.84 and 8.04, ΩCa between 3.68 and 5.24 and ΩAr from 2.41 to 3.44. Geochemical analyses of hydrothermal gases revealed a dominance of CO2 (98.1%) together with small amounts of oxygen and reactive gases. Helium isotope ratios (R/Ra = 2.51) and δ13CCO2 suggest an inorganic origin of hydrothermal degassing of CO2 and the ascent of heat and deep-seated magmatic fluids to the surface. Visual census of fishes and megabenthos (mainly sessile organisms) allowed the identification of 64 species, four of which are protected by the SPA/BIO Protocol and two by the International Union for Conservation of Nature. The macroalgae Halopteris scoparia and Jania rubens and the sponge Sarcotragus sp. were the dominant taxa in the area, while among fishes Coris julis and Chromis chromis were the most abundant species. This preliminary investigation of San Giorgio vent field suggests that the site could be of interest and suitable for future experimental studies of ocean acidification., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 D’Alessandro et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Introduction: To examine electroencephalogram (EEG) as a diagnostic tool for late-onset efavirenz (EFV) neurotoxicity syndrome (LENS), an uncommon but severe and potentially fatal complication of EFV therapy., Methods: We conducted a Retrospective case-control study. EEGs from confirmed cases of LENS (clinical syndrome and plasma EFV >4ug/mL) recorded from June 2016 to May 2021 were compared with control EEGs from the same time-period. Controls were adults (18-70 years) with a similar indication for EEG (eg. encephalopathy or confusion), dysrhythmia generalised grade II, and LENS excluded. EEGs were reviewed by two blinded interpreters given a description of the characteristic EEG changes, ie. persistent, diffuse, high voltage, bisynchronous, monomorphic 4-7 Hz theta frequency waveforms with transient attenuation on eye opening. Interpreters were asked to determine whether EEGs showed definite, probable or no changes., Results: Thirteen LENS cases were compared with 50 control EEGs. Interpreter 1 labelled 11/13 LENS cases as having define or probable changes, and interpreter 2 labelled 10/13. Interpreter 1 labelled probable changes in 1/50 controls and interpreter 2 in 3/50. Neither interpreter labelled any controls as having definite changes. Interrater reliability was good with 95% agreement and a Cohen's kappa of 0.83. Sensitivity of EEG under these conditions for the diagnosis of LENS was 85% and 77% for interpreters 1 and 2 respectively, and specificity was 98% and 94%., Conclusions: EEG is a useful tool in the diagnosis of LENS which can be used to aid clinical decisions while awaiting EFV levels, or in low-resource settings where EFV levels are not available., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Nightingale et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Protein-protein-interactions play an important role in many cellular functions. Quantitative non-invasive techniques are applied in living cells to evaluate such interactions, thereby providing a broader understanding of complex biological processes. Fluorescence fluctuation spectroscopy describes a group of quantitative microscopy approaches for the characterization of molecular interactions at single cell resolution. Through the obtained molecular brightness, it is possible to determine the oligomeric state of proteins. This is usually achieved by fusing fluorescent proteins (FPs) to the protein of interest. Recently, the number of novel green FPs has increased, with consequent improvements to the quality of fluctuation-based measurements. The photophysical behavior of FPs is influenced by multiple factors (including photobleaching, protonation-induced "blinking" and long-lived dark states). Assessing these factors is critical for selecting the appropriate fluorescent tag for live cell imaging applications. In this work, we focus on novel green FPs that are extensively used in live cell imaging. A systematic performance comparison of several green FPs in living cells under different pH conditions using Number & Brightness (N&B) analysis and scanning fluorescence correlation spectroscopy was performed. Our results show that the new FP Gamillus exhibits higher brightness at the cost of lower photostability and fluorescence probability (pf), especially at lower pH. mGreenLantern, on the other hand, thanks to a very high pf, is best suited for multimerization quantification at neutral pH. At lower pH, mEGFP remains apparently the best choice for multimerization investigation. These guidelines provide the information needed to plan quantitative fluorescence microscopy involving these FPs, both for general imaging or for protein-protein-interactions quantification via fluorescence fluctuation-based methods., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Petrich et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Bacterial resistance to antimicrobials is considered a major issue worldwide. This condition may account for treatment failure of urinary tract infections, which are among the most common infections both in community and healthcare settings. Therapy against uropathogens is generally administered empirically, possibly leading to unsuccessful therapy, recurrence and development of antibiotic resistance. The reduction in analytical time to obtain antimicrobial susceptibility test (AST) results could play a key role in reducing the cost of healthcare, providing information about antibiotic efficacy and thus preventing from either exploiting new and expensive antibiotics unnecessarily or using obsolete and ineffective ones. A more rational choice among treatment options would hence lead to more effective treatment and faster resolution. In this paper we evaluated the performance of a new Point Of Care Test (POCT) for the rapid prediction of antimicrobial susceptibility in urine samples performed without the need of a laboratory or specialized technicians. 349 patients were enrolled in two open-label, monocentric, non-interventional clinical trials in partnership with an Emergency Medicine ward and the Day Hospital of two large healthcare facilities in Rome. Antibiogram was carried out on 97 patients. Results from analysis of urine samples with the POCT were compared with those from routine AST performed on culture-positive samples, displaying high accuracy (>90%) for all tested antimicrobial drugs and yielding reliable results in less than 12 hours from urine collection thus reducing analytical and management costs., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Arienzo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The aim of this study was to design a new canine posture estimation system specifically for working dogs. The system was composed of Inertial Measurement Units (IMUs) that are commercially available, and a supervised learning algorithm which was developed for different behaviours. Three IMUs, each containing a 3-axis accelerometer, gyroscope, and magnetometer, were attached to the dogs' chest, back, and neck. To build and test the model, data were collected during a video-recorded behaviour test where the trainee assistance dogs performed static postures (standing, sitting, lying down) and dynamic activities (walking, body shake). Advanced feature extraction techniques were employed for the first time in this field, including statistical, temporal, and spectral methods. The most important features for posture prediction were chosen using Select K Best with ANOVA F-value. The individual contributions of each IMU, sensor, and feature type were analysed using Select K Best scores and Random Forest feature importance. Results showed that the back and chest IMUs were more important than the neck IMU, and the accelerometers were more important than the gyroscopes. The addition of IMUs to the chest and back of dog harnesses is recommended to improve performance. Additionally, statistical and temporal feature domains were more important than spectral feature domains. Three novel cascade arrangements of Random Forest and Isolation Forest were fitted to the dataset. The best classifier achieved an f1-macro of 0.83 and an f1-weighted of 0.90 for the prediction of the five postures, demonstrating a better performance than previous studies. These results were attributed to the data collection methodology (number of subjects and observations, multiple IMUs, use of common working dog breeds) and novel machine learning techniques (advanced feature extraction, feature selection and modelling arrangements) employed. The dataset and code used are publicly available on Mendeley Data and GitHub, respectively., Competing Interests: This research is related to the PhD of Marinara Marcato, which is part of collaborations with the Irish Guide Dogs for the Blind. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2023 Marcato et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: The long-term cardiovascular (CV) outcomes of COVID-19 have not been fully explored., Methods: This was an international, multicenter, retrospective cohort study conducted between February and December 2020. Consecutive patients ≥18 years who underwent a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for SARS-CoV2 were included. Patients were classified into two cohorts depending on the nasopharyngeal swab result and clinical status: confirmed COVID-19 (positive RT-PCR) and control (without suggestive symptoms and negative RT-PCR). Data were obtained from electronic records, and clinical follow-up was performed at 1-year. The primary outcome was CV death at 1-year. Secondary outcomes included arterial thrombotic events (ATE), venous thromboembolism (VTE), and serious cardiac arrhythmias. An independent clinical event committee adjudicated events. A Cox proportional hazards model adjusted for all baseline characteristics was used for comparing outcomes between groups. A prespecified landmark analysis was performed to assess events during the post-acute phase (31-365 days)., Results: A total of 4,427 patients were included: 3,578 (80.8%) in the COVID-19 and 849 (19.2%) control cohorts. At one year, there were no significant differences in the primary endpoint of CV death between the COVID-19 and control cohorts (1.4% vs. 0.8%; HRadj 1.28 [0.56-2.91]; p = 0.555), but there was a higher risk of all-cause death (17.8% vs. 4.0%; HRadj 2.82 [1.99-4.0]; p = 0.001). COVID-19 cohort had higher rates of ATE (2.5% vs. 0.8%, HRadj 2.26 [1.02-4.99]; p = 0.044), VTE (3.7% vs. 0.4%, HRadj 9.33 [2.93-29.70]; p = 0.001), and serious cardiac arrhythmias (2.5% vs. 0.6%, HRadj 3.37 [1.35-8.46]; p = 0.010). During the post-acute phase, there were no significant differences in CV death (0.6% vs. 0.7%; HRadj 0.67 [0.25-1.80]; p = 0.425), but there was a higher risk of deep vein thrombosis (0.6% vs. 0.0%; p = 0.028). Re-hospitalization rate was lower in the COVID-19 cohort compared to the control cohort (13.9% vs. 20.6%; p = 0.001)., Conclusions: At 1-year, patients with COVID-19 experienced an increased risk of all-cause death and adverse CV events, including ATE, VTE, and serious cardiac arrhythmias, but not CV death., Study Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT04359927., Competing Interests: GC received research grants from Boston Scientific, Medis, SMT, Siemens, Insight Lifetech. BB reports that he is a consulting expert, on behalf of the plaintiff, for litigation related to two specific brand models of IVC filters. HGG reports a relationship with Biotronik, Boston Scientific, Medtronic, Abbott, Neovasc, Shockwave, Philips, and CorFlow that includes: funding grants. DJA declares that he has received consulting fees or honoraria from Abbott, Amgen, AstraZeneca, Bayer, Biosensors, Boehringer Ingelheim, Bristol-Myers Squibb, Chiesi, Daiichi-Sankyo, Eli Lilly, Haemonetics, Janssen, Merck, PhaseBio, PLx Pharma, Pfizer, and Sanofi. DJA also declares that his institution has received research grants from Amgen, AstraZeneca, Bayer, Biosensors, CeloNova, CSL Behring, Daiichi-Sankyo, Eisai, Eli Lilly, Gilead, Idorsia, Janssen, Matsutani Chemical Industry Co., Merck, Novartis, Osprey Medical, Renal Guard Solutions, and the Scott R. MacKenzie Foundation. JRC is consultant for and has received institutional research grants from Edwards Lifesciences, Medtronic and Boston Scientific. MS declares that he is a consultant for Abbott Vascular and iVascular. SB declares that he is a consultant from Boston Scientific, iVascular and Teleflex, and he received speaker’s fee from Abbott Vascular, outside the present work. Other authors have nothing to disclose. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2022 Ortega-Paz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Modern society grew rapidly over the last few decades and this led to an alarming increase in air pollutants and a worsening of the human health, especially in relation to the respiratory system. Indeed, chronic respiratory diseases were the third main cause of death in 2017, with over 3 million of deaths. Furthermore, the pollution has considerable consequences both for burden medical expenses and environmental. However, the mechanisms linking pollutants to the onset of these diseases remain unclear. Thus, in this study we addressed this problem through the United Kingdom BioBank database, analyzing 170 genome-wide association studies (103 related to respiratory diseases and 67 related to pollutants). We analyzed the genetic correlations and causal relationships of these traits, leveraging the summary statistics and bioinformatics packages such as Linkage Disequilibrium Score Regression and Latent Causal Variable. We obtained 158 significant genetic correlations and subsequently we analyzed them through the Latent Causal Variable analysis, obtaining 20 significant causal relationships. The most significant were between "Workplace full of chemicals or other fumes: Sometimes" and "Condition that has ever been diagnosed by a doctor: Asthma" and between "Workplace very dusty: Sometimes" and "Condition that has ever been diagnosed by a doctor: Emphysema or chronic bronchitis". Finally, we identified single nucleotide polymorphisms independently associated with sveral pollutants to analyze the genes and pathways that could be involved in the onset of the aforementioned respiratory system disorders and that could be useful clinical target. This study highlighted how crucial are the air condition of the working environments and the type of transport used in the onset of respiratory-related morbidity. Based on that, we also suggested some interventions, in order to improve quality life and develop new and eco-friendly society and life style, such as improving indoor air circulation, the use of public transport and urban reforestation., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 D'Antona et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Multiple models exist for the evaluation of human motor performance; some of these rely on the Index of Difficulty (ID), a measure to evaluate the difficulty associated to simple reaching tasks. Despite the numerous applications of the ID in reaching movements, the existing formulations are functions of the geometrical features of the task and do not consider the motor behaviour of subjects performing repetitive movements in interaction with the environment. Variability of movements, length of trajectories, subject-specific strength and skill, and required interaction with the environment are all factors that contribute to the motor difficulty experienced by a moving agent (e.g., human, robot) as it repeatedly interacts with the environment during a given task (e.g., target-reaching movement, locomotion, etc.). A novel concept of motor difficulty experienced by an agent executing repetitive end-effector movements is presented in this study. A stochastic ID formulation is proposed that captures the abovementioned factors and applies to general three-dimensional motor tasks. Natural motor variability, inherent in the proposed model, is representative of the flexibility in motor synergies for a given agent-environment interaction: the smaller the flexibility, the greater the experienced difficulty throughout the movement. The quantification of experienced motor difficulty is demonstrated for the case of young healthy subjects performing three-dimensional arm movements during which different objects are manipulated. Results show that subjects' experienced motor difficulty is influenced by the type of object. In particular, a difference in motor difficulty is observed when manipulating objects with different grasp types. The proposed model can be employed as a novel tool to evaluate the motor performance of agents involved in repetitive movements, such as in pick and place and manipulation, with application in both industrial and rehabilitation contexts., Competing Interests: The authors have declared that no competing interests exist.

Purpose: Obstructive sleep apnea (OSA) is a common, prevalent, and underdiagnosed disorder. Its lack of diagnosis and treatment is associated with increased morbidity and mortality. Previous screening questionnaires investigated parameters including body mass index, age, neck circumference, and sex, in addition to symptoms. This study aimed to validate a new Italian, self-administered, and easy-to-use six-item questionnaire that evaluates only subjective symptoms., Patients and Methods: The present study included 2622 patients (male, 2011; female, 611). Patients who were at least 18 years old, spoke Italian, referred to our sleep clinic for possible OSA, and completed the self-administered SANReSP questionnaire were recruited for the study. The predictive performance of the questionnaire was also evaluated., Results: Nocturnal study showed 89.9% of OSA patients had apnea-hypopnea index (AHI) ≥ 5/h; 68.7%, AHI ≥ 15/h; and 48.2%, AHI ≥ 30/h. The optimal SANReSP score for AHI ≥ 5/h was >3 with a sensitivity and specificity of 74.76% and 67.92%, respectively, and an area under receiver operating characteristic curve (ROC) of 0.76. For moderate-severe OSA, the optimal SANReSP score was >3 (sensitivity, 78.18%; specificity, 46.53%; ROC, 0.66). For severe OSA, the optimal SANReSP score was >4 (sensitivity, 59.10%; specificity, 64.73%; ROC, 0.65). The probability of OSA increased with higher SANReSP scores (98.7% and 97.9% in men and women, respectively)., Conclusion: The SANReSP questionnaire is a short, easy-to-use, and self-administered screening tool for OSA. Its performance is similar to that of other widely used questionnaires; furthermore, it is advantageous in that it does not require anthropometric measurements., Competing Interests: The authors declared that no competing interests exist.

The study aimed to delineate the robustness of the culture-based and molecular biology methods to assess the total bacterial concentration and Campylobacter jejuni (C. jejuni) quantification in caecal content, analysed as fresh or after being stored immediately at ultra-low (-80°C) temperature at different time points (for 3, 7, 14, 28 and 62 days post collection). The caecal content was collected from birds that were artificially colonised with C. jejuni (in-vivo), and quantification was performed using both colony-forming unit (CFU) and qPCR. The results showed that storage time affected the output of culture-based analyses but mostly did not alter concentration retrieved via qPCR. After an initial ~4.5 log10 reduction in CFU observed from fresh (day 0) to frozen samples, bacterial concentration retrieved with culture-based methods seemed to be constant in samples frozen for 3 to 62 days, indicating a possible threshold for C. jejuni loss of viability due to effect of storage temperature. Ranking order analyses, revealed that the molecular biology technique was able to attribute somewhat the same relative C. jejuni concentrations to the samples analysed via qPCR. However, day 0 measurements from culture-based methods were associated with the absence of or negatively weak correlations with the rest of the time points, but ranking order was maintained from day 3 onwards. On the other hand, ranking order correlations were less constant when measuring total bacterial concentration through qPCR. The study suggests that if biological samples can't be analysed as fresh (immediately after collection) and have to be stored prior to analysis, then storage at -80°C samples be recommended to avoid the temporal-dependent effects on C. jejuni concentrations. In addition, irrespective of the method of analysis, an initial loss of CFU must be factored in when interpreting the results obtained from frozen samples., Competing Interests: The authors have declared that no competing interests exist.

Background: Given the ever-shorter length of hospital stay after surgical procedures, nowadays it is more important than ever to study interventions that may have an impact on surgical patients' wellbeing. According to the ERAS (Enhanced Recovery After Surgery protocols) program, early feeding must be considered one of the key components to facilitate early recovery while improving outcomes and patients' overall experiences. To date, the international literature has reported that early postoperative feeding compared with traditional (or late) timing is safe; nevertheless, small clinical outcomes effects has been reported, also for recovery of gastrointestinal function. Therefore, the effectiveness of early postoperative feeding to reduce postoperative ileus duration remains still debated., Objective: To analyse the effects of early versus delayed oral feeding (liquids and food) on the recovery of intestinal motility after gastrointestinal surgery., Search Methods: Pubmed, Embase, Cinahl, Cochrane Central Register of Controlled Trials (CENTRAL), and the ClincalTrials.gov register will be searched to identify the RCTs of interest., Study Inclusion: Randomized clinical trials (RCTs) comparing the effect of early postoperative versus late oral feeding on major postoperative outcomes after gastrointestinal surgery will be included., Data Collection and Analysis: Two review authors will independently screen titles and abstracts to determine the initially selected studies' inclusion. Any disagreements will be resolved through discussion and consulting a third review author. The research team members will then proceed with the methodological evaluation of the studies and their eligibility for inclusion in the systematic review., Competing Interests: The authors have declared that no competing interests exist

Mycoplasma hyorhinis is an emerging swine pathogen bacterium causing polyserositis and polyarthritis in weaners and finishers. The pathogen is distributed world-wide, generating significant economic losses. No commercially available vaccine is available in Europe. Therefore, besides improving the housing conditions for prevention, antimicrobial therapy of the diseased animals is the only option to control the infection. Our aim was to determine the minimal inhibitory concentrations (MIC) of ten antimicrobials potentially used against M. hyorhinis infection. The antibiotic susceptibility of 76 M. hyorhinis isolates from Belgium, Germany, Hungary, Italy and Poland collected between 2019 and 2021 was determined by broth micro-dilution method and mismatch amplification mutation assay (MAMA). Low concentrations of tiamulin (MIC90 0.312 μg/ml), doxycycline (MIC90 0.078 μg/ml), oxytetracycline (MIC90 0.25 μg/ml), florfenicol (MIC90 2 μg/ml) and moderate concentrations of enrofloxacin (MIC90 1.25 μg/ml) inhibited the growth of the isolates. For the tested macrolides and lincomycin, a bimodal MIC pattern was observed (MIC90 >64 μg/ml for lincomycin, tulathromycin, tylosin and tilmicosin and 5 μg/ml for tylvalosin). The results of the MAMA assay were in line with the conventional method with three exceptions. Based on our statistical analyses, significant differences in MIC values of tiamulin and doxycycline were observed between certain countries. Our results show various levels of antimicrobial susceptibility among M. hyorhinis isolates to the tested antibiotics. The data underline the importance of susceptibility monitoring on pan-European level and provides essential information for proper antibiotic choice in therapy., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: U. Klein and W. Depondt are the employees of Huvepharma® NV, the producer of various veterinary antibiotic products. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Cancer genomic heterogeneity presents significant challenges for understanding oncogenic processes and for cancer's clinical management. Variation in driver mutation frequency between patients with the same tumor type as well as within an individual patients' cancer can shape the use of mutations as diagnostic, prognostic, and predictive biomarkers. We have characterized genomic heterogeneity between and within canine splenic hemangiosarcoma (HSA), a common naturally occurring cancer in pet dogs that is similar to human angiosarcoma (AS). HSA is a clinically, physiologically, and genomically complex canine cancer that may serve as a valuable model for understanding the origin and clinical impact of cancer heterogeneity. We conducted a prospective collection of 52 splenic masses from 43 dogs (27 HSA, 15 benign masses, and 1 stromal sarcoma) presenting for emergency care with hemoperitoneum secondary to a ruptured splenic mass. Multi-platform genomic analysis included matched tumor/normal targeted sequencing panel and exome sequencing. We found candidate somatic cancer driver mutations in 14/27 (52%) HSAs. Among recurrent candidate driver mutations, TP53 was most commonly mutated (30%) followed by PIK3CA (15%), AKT1 (11%), and CDKN2AIP (11%). We also identified significant intratumoral genomic heterogeneity, consistent with a branched evolution model, through multi-region exome sequencing of three distinct tumor regions from selected primary splenic tumors. These data provide new perspectives on the genomic landscape of this veterinary cancer and suggest a cross-species value for using HSA in pet dogs as a naturally occurring model of intratumoral heterogeneity., Competing Interests: The authors have read the journal’s policy and have the following competing interests: William P. D. Hendricks is the Founder and Chief Scientific Officer of Vidium Animal Health, a TGen Subsidiary, and the President of Ethos Discovery. EJ Ehrhart, Jacob Cawley, Keith Richter, and Samuel Stewart are current or prior employees of Ethos Veterinary Health. There are no patents, products in development or marketed products associated with this research to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Background: Limited data are available examining the effects of both moderate- and vigorous-intensity physical activity (MVPA) and sedentary behavior (SB) on longevity among patients with heart failure (HF). This study examined the associations of MVPA and SB with all-cause mortality in HF patients using a nationally representative survey data., Methods: National Health and Nutrition Examination Survey data (2007-2014) were used. 711 adults with self-reported congestive HF, linked to 2015 mortality data were analyzed. Self-reported MVPA and SB minutes were used to create the three MVPA [No-MVPA, insufficient (I-MVPA; <150 min/wk), and sufficient (S-MVPA; ≥150 min/wk)] and two SB (<8 and ≥8 hrs/d) groups. Cox proportional hazard models were constructed to test the associations of MVPA and SB with all-cause mortality., Results: 119 deaths occurred over an average of 4.9 years of follow-up. Lower MVPA and higher SB were independently associated with poor survival (P < .001). Joint and stratified analyses showed that the protective effect of MVPA was most pronounced among patients with SB<8 hrs/d. There was no difference in the mortality risk by SB levels within I-MVPA and S-MVPA groups; however, in the No-MVPA group, those with SB≥8 hrs/d had a greater risk of mortality compared to those with <8 hrs/d (Hazard ratio = 1.60)., Conclusion: In this HF cohort, MVPA and SB were independently and jointly associated with all-cause mortality. The beneficial effect of MVPA is attenuated by excessive SB; however, engaging in some amount of MVPA may provide a protective effect and attenuates the detrimental effects associated with excessive SB., Competing Interests: The authors have declared that no competing interests exist.

The genus Stenella is comprised of five species occurring in all oceans. Despite its wide distribution, genetic diversity information on these species is still scarce especially in the Southwest Atlantic Ocean. Some features of this genus can enhance opportunities for potential introgressive hybridization, e.g. sympatric distibution along the Brazilian coast, mixed known associations among species, karyotype uniformity and genome permeability. In this study we analyzed three genes of the mitochondrial genome to investigate the genetic diversity and occurrence of genetic mixture among eighty specimens of Stenella. All species exhibited moderate to high levels of genetic diversity (h = 0.833 to h = 1.000 and π = 0.006 to π = 0.015). Specimens of S. longirostris, S. attenuata and S. frontalis were clustered into differentiated haplogroups, in contrast, haplotypes of S. coeruleoalba and S. clymene were clustered together. We detected phylogenetic structure of mixed clades for S. clymene and S. coeruleoalba specimens, in the Southwest Atlantic Ocean, and also between S. frontalis and S. attenuata in the Northeast Atlantic Ocean, and between S. frontalis and S. longirostris in the Northwest Atlantic Ocean. These specimes were morphologically identified as one species but exhibited the maternal lineage of another species, by mitochondrial DNA. Our results demonstrate that ongoing gene flow is occurring among species of the genus Stenella reinforcing that this process could be one of the reasons for the confusing taxonomy and difficulties in elucidating phylogenetic relationships within this group., Competing Interests: The authors have read the journal’s policy and have the following competing interests to declare: Chevron Brasil Upstream Frade Ltda and BG Group provided funding support for this study. This does not alter our adherence to PLOS ONE policies on sharing data and materials. There are no patents, products in development or marketed products associated with this research to declare.

The Covid19 pandemic has significantly impacted on our lives, triggering a strong reaction resulting in vaccines, more effective diagnoses and therapies, policies to contain the pandemic outbreak, to name but a few. A significant contribution to their success comes from the computer science and information technology communities, both in support to other disciplines and as the primary driver of solutions for, e.g., diagnostics, social distancing, and contact tracing. In this work, we surveyed the Italian computer science and engineering community initiatives against the Covid19 pandemic. The 128 responses thus collected document the response of such a community during the first pandemic wave in Italy (February-May 2020), through several initiatives carried out by both single researchers and research groups able to promptly react to Covid19, even remotely. The data obtained by the survey are here reported, discussed and further investigated by Natural Language Processing techniques, to generate semantic clusters based on embedding representations of the surveyed activity descriptions. The resulting clusters have been then used to extend an existing Covid19 taxonomy with the classification of related research activities in computer science and information technology areas, summarizing this work contribution through a reproducible survey-to-taxonomy methodology., Competing Interests: The authors have declared that no competing interests exist.

Green roof systems, aimed at reducing anthropic impact on the environment, are considered environmental mitigation technologies and adopted by many countries across the world to strengthen urban ecosystem services. This study evaluates two mixtures of succulent: one of Crassulaceae and the other of Aizoaceae, used in the creation of a continuous and homogenous plant groundcover in Mediterranean environments. To assess the species mixtures, the parameters plant height, growth index, cover percentage and flowering were observed. Hydrological observations were also carried out to evaluate the rainfall retained by the test system in any given month. All data were subjected to analysis of variance. Growth indicators in the study showed trends characteristic of xeric plants, which tend to slow down in dry, summer climate conditions to the point of halting plant vertical growth and ground cover development completely. The Aizocaeae mix, during the initial stage, showed prevalent horizontal growth, confirmed by greater a greater growth index (13,21) and cover percentage (45%) compared to Sedum (Growth index: 3,61; Cover: 36%). In contrast, the Sedum mix recorded greater vertical growth at the beginning (Sedum mixture: 7.53 cm; Aizoaceae mixture: 6,11 cm). During the final stages of observations, however, greater vertical growth in the Aizoaceae (7,88 cm) became apparent together with a recovery in horizontal growth in the Sedum (79%), albeit not sufficient to outperform the Aizoaceae mixture (87%). Flowering in the two mixtures occurred between late spring and late summer. The Sedum mixture guaranteed a longer flowering period (130 days) compared to the Aizoaceae (93 days), with a gradual start followed by steady flower emission. Regarding rainfall water retention, a comparison of the mixtures in late winter/early spring revealed that the Sedum performed best (44.9 L m2 vs 37.4 L m2), whilst the Aizoaceae outperformed the Sedum in Autumn (63 L m2 vs 55 L m2), in conjunction with favorable growth rates in both species mixtures. Both mixtures demonstrated satisfying results and are considered suited to a Mediterranean environment. Furthermore, based on the different growth rates of the species in the two test mixtures, this study suggests that new combinations of Sedum and Aizoaceae together might prove more resilient in Mediterranean environments., Competing Interests: The authors have declared that no competing interests exist.

Background: Fluid bolus therapy (FBT) is common in ICUs but whether it achieves the effects expected by intensivists remains uncertain. We aimed to describe intensivists' expectations and compare them to the actual physiological effects., Methods: We evaluated 77 patients in two ICUs (Sweden and Australia). We included patients prescribed a FBT ≥250 ml over ≤30 minutes. The intensivist completed a questionnaire on triggers for and expected responses to FBT. We compared expected with actual values at FBT completion and after one hour., Results: Median bolus size (IQR) was 300 ml (250-500) given over a median (IQR) of 21 minutes (15-30 mins). Boluses were 57% Ringer´s Acetate and 43% albumin (40-50g/L). Hypotension was the most common trigger (47%), followed by oliguria (21%). During FBT, 55% of patients received noradrenaline and 38% propofol. Intensivists expected a median MAP increase of 2.6 mmHg (IQR: -3.1 to +6.8) at end of bolus and of 1.3 mmHg (-3.5 to + 4.1) after one hour. Intensivist´s' expectations were judged to be accurate if they were within 5% above or below measured values. At FBT completion, 33% of MAP expectations were overestimations and 42% were underestimations. One hour later, 19% were overestimations and 43% were underestimations. Only 8% of expectations of measured urine output (UO) were accurate and 44% were overestimations. Correction for sedation or vasopressors did not modify these findings., Conclusions: The physiological expectations of intensivists after FBT carried a high risk of both over and underestimation. Since the physiological effect FBT was often small and did not meet clinical expectations, a reassessment of its rationale, effect, duration, and role appears justified., Competing Interests: The authors have declared that no competing interests exist.

Aim: The aim of this study was to analyze changes in the incidence, management and mortality of DFU in Sicilian Type 2 diabetic patients hospitalized between two eras, i.e. 2008-2013 and 2014-2019., Methods: We compared the two eras, era1: 2008-13, era2: 2014-19. In era 1, n = 149, and in era 2, n = 181 patients were retrospectively enrolled., Results: In the population hospitalized for DFU in 2008-2013, 59.1% of males and 40.9% of females died, whilst in 2014-2019 65.9% of males and 34.1% of females died. Moderate chronic kidney disease (CKD) was significantly higher in patients that had died than in ones that were alive (33% vs. 43%, p < 0.001), just as CKD was severe (14.5% vs. 4%, p < 0.001). Considering all together the risk factors associated with mortality, at Cox regression multivariate analysis only moderate-severe CKD (OR 1.61, 95% CI 1.07-2.42, p 0.021), age of onset greater than 69 years (OR 2.01, 95% CI 1.37-2.95, p <0.001) and eGFR less than 92 ml/min (OR 2.84, 95% CI 1.51-5.34, p 0.001) were independently associated with risk of death., Conclusions: Patients with DFU have high mortality and reduced life expectancy. Age at onset of diabetic foot ulcer, eGFR values and CKD are the principal risk factors for mortality., Competing Interests: The authors have declared that no competing interests exist.

Background: The first Covid-19 epidemic outbreak has enormously impacted the delivery of clinical healthcare and hospital management practices in most of the hospitals around the world. In this context, it is important to assess whether the clinical management of non-Covid patients has not been compromised. Among non-Covid cases, patients with Acute Myocardial Infarction (AMI) and stroke need non-deferrable emergency care and are the natural candidates to be studied. Preliminary evidence suggests that the time from onset of symptoms to emergency department (ED) presentation has significantly increased in Covid-19 times as well as the 30-day mortality and in-hospital mortality., Methods: We check, in a causal inference framework, the causal effect of the hospital's stress generated by Covid-19 pandemic on in-hospital mortality rates (primary end-point of the study) of AMI and stroke over several time-windows of 15-days around the implementation date of the State of Emergency restrictions for COVID-19 (March, 9th 2020) using two quasi-experimental approaches, regression-discontinuity design (RDD) and difference-in-regression-discontinuity (DRD) designs. Data are drawn from Spedali Civili of Brescia, one of the most hit provinces in Italy by Covid-19 during March and May 2020., Findings: Despite the potential adverse effects on expected mortality due to a longer time to hospitalization and staff extra-burden generated by the first wave of Covid-19, the AMI and stroke mortality rates are overall not statistically different during the first wave of Covid-19 than before the first peak. The obtained results provided by RDD models are robust also when we account for seasonality and unobserved factors with DRD models., Interpretation: The non-statistically significant impact on mortality rates for AMI and stroke patients provides evidence of the hospital ability to manage -with the implementation of a dual track organization- the simultaneous delivery of high-quality cares to both Covid and non-Covid patients., Competing Interests: The founder had no roles in our study design, data collection and analysis, decision to publish and preparation of the manuscript. The authors have declared that no competing interests exist.

The increasing availability of textual corpora and data fetched from social networks is fuelling a huge production of works based on the model proposed by psychologist Robert Plutchik, often referred simply as the "Plutchik Wheel". Related researches range from annotation tasks description to emotions detection tools. Visualisation of such emotions is traditionally carried out using the most popular layouts, as bar plots or tables, which are however sub-optimal. The classic representation of the Plutchik's wheel follows the principles of proximity and opposition between pairs of emotions: spatial proximity in this model is also a semantic proximity, as adjacent emotions elicit a complex emotion (a primary dyad) when triggered together; spatial opposition is a semantic opposition as well, as positive emotions are opposite to negative emotions. The most common layouts fail to preserve both features, not to mention the need of visually allowing comparisons between different corpora in a blink of an eye, that is hard with basic design solutions. We introduce PyPlutchik the Pyplutchik package is available as a Github repository (http://github.com/alfonsosemeraro/pyplutchik) or through the installation commands pip or conda. For any enquiry about usage or installation feel free to contact the corresponding author, a Python module specifically designed for the visualisation of Plutchik's emotions in texts or in corpora. PyPlutchik draws the Plutchik's flower with each emotion petal sized after how much that emotion is detected or annotated in the corpus, also representing three degrees of intensity for each of them. Notably, PyPlutchik allows users to display also primary, secondary, tertiary and opposite dyads in a compact, intuitive way. We substantiate our claim that PyPlutchik outperforms other classic visualisations when displaying Plutchik emotions and we showcase a few examples that display our module's most compelling features., Competing Interests: The authors have declared that no competing interests exist.

Background: Patients presenting with the coronavirus-2019 disease (COVID-19) may have a high risk of cardiovascular adverse events, including death from cardiovascular causes. The long-term cardiovascular outcomes of these patients are entirely unknown. We aim to perform a registry of patients who have undergone a diagnostic nasopharyngeal swab for SARS-CoV-2 and to determine their long-term cardiovascular outcomes., Study and Design: This is a multicenter, observational, retrospective registry to be conducted at 17 centers in Spain and Italy (ClinicalTrials.gov number: NCT04359927). Consecutive patients older than 18 years, who underwent a real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for SARS-CoV2 in the participating institutions, will be included since March 2020, to August 2020. Patients will be classified into two groups, according to the results of the RT-PCR: COVID-19 positive or negative. The primary outcome will be cardiovascular mortality at 1 year. The secondary outcomes will be acute myocardial infarction, stroke, heart failure hospitalization, pulmonary embolism, and serious cardiac arrhythmias, at 1 year. Outcomes will be compared between the two groups. Events will be adjudicated by an independent clinical event committee., Conclusion: The results of this registry will contribute to a better understanding of the long-term cardiovascular implications of the COVID19., Competing Interests: The authors have declared that no competing interests exist.

Neurofibromatosis Type 2 (NF2) is an autosomal dominant genetic syndrome caused by mutations in the NF2 tumor suppressor gene resulting in multiple schwannomas and meningiomas. There are no FDA approved therapies for these tumors and their relentless progression results in high rates of morbidity and mortality. Through a combination of high throughput screens, preclinical in vivo modeling, and evaluation of the kinome en masse, we identified actionable drug targets and efficacious experimental therapeutics for the treatment of NF2 related schwannomas and meningiomas. These efforts identified brigatinib (ALUNBRIG®), an FDA-approved inhibitor of multiple tyrosine kinases including ALK, to be a potent inhibitor of tumor growth in established NF2 deficient xenograft meningiomas and a genetically engineered murine model of spontaneous NF2 schwannomas. Surprisingly, neither meningioma nor schwannoma cells express ALK. Instead, we demonstrate that brigatinib inhibited multiple tyrosine kinases, including EphA2, Fer and focal adhesion kinase 1 (FAK1). These data demonstrate the power of the de novo unbiased approach for drug discovery and represents a major step forward in the advancement of therapeutics for the treatment of NF2 related malignancies., Competing Interests: No authors have competing interests.

Background: Endogenous serine protease inhibitors are associated with anti-inflammatory and pro-survival signaling mediated via Low-density lipoprotein receptor-related protein 1 (LRP1) signaling. SP16 is a short polypeptide that mimics the LRP1 binding portion of alpha-1 antitrypsin., Methods: A pilot phase I, first-in-man, randomized, double blind, placebo-controlled safety study was conducted to evaluate a subcutaneous injection at three dose levels of SP16 (0.0125, 0.05, and 0.2 mg/kg [up to 12 mg]) or matching placebo in 3:1 ratio in healthy individuals. Safety monitoring included vital signs, laboratory examinations (including hematology, coagulation, platelet function, chemistry, myocardial toxicity) and electrocardiography (to measure effect on PR, QRS, and QTc)., Results: Treatment with SP16 was not associated with treatment related serious adverse events. SP16 was associated with mild-moderate pain at the time of injection that was significantly higher than placebo on a 0-10 pain scale (6.0+/-1.4 [0.2 mg/kg] versus 1.5+/-2.1 [placebo], P = 0.0088). No differences in vital signs, laboratory examinations and electrocardiography were found in those treated with SP16 versus placebo., Conclusion: A one-time treatment with SP16 for doses up to 0.2 mg/kg or 12 mg was safe in healthy volunteers., Competing Interests: The authors have read the journal’s policy and have the following competing interests: CG is the owner of Serpin Pharma, LLC, DA is a paid employee of the company, and AA and BVT have been paid consultants. This does not alter our adherence to PLOS ONE policies on sharing data and materials. The authors would like to declare that Serpin Pharma LLC holds the following patents for the design and use of SP16: Australia - 2013208293, 2017258892; Canada - 22859777; Europe - 280238; Hong Kong - HK1203161; Israel - 233481; Japan - 6416627; Mexico - 365403; US - 8975224, 9951104, 10214562, 10899797.

Branchiostomatidae (lancelets or amphioxus) comprises about 30 species, several of which are well-established models in evolutionary development. Our zoological and ecological knowledge of the family is nonetheless limited. Despite evident differences can be found among known populations, the taxonomy of Branchiostoma lanceolatum (type species of the genus Branchiostoma) has never been investigated with modern methods through its range in the northeastern Atlantic and Mediterranean Sea. We address this via a multilocus molecular approach and comparing specimens collected from different European populations. Results obtained here confirm the presence of a single species inhabiting the range between the topotypical localities of B. lanceolatum (Atlantic Ocean) and of its junior synonym B. lubricum (Mediterranean Sea), without evincing geographical structure between populations. This suggests that environment most likely drives the characteristics observed in different geographic areas. The long larval phase and the slow mutation rate in lancelets may have played a key role in the evolutionary history of this iconic species., Competing Interests: The authors have declared that no competing interests exist.

Introduction: The hypothesis of a general psychopathology factor that underpins all common forms of mental disorders has been gaining momentum in contemporary clinical research and is known as the p factor hypothesis. Recently, a semiotic, embodied, and psychoanalytic conceptualisation of the p factor has been proposed called the Harmonium Model, which provides a computational account of such a construct. This research tested the core tenet of the Harmonium model, which is the idea that psychopathology can be conceptualised as due to poorly-modulable cognitive processes, and modelled the concept of Phase Space of Meaning (PSM) at the computational level., Method: Two studies were performed, both based on a simulation design implementing a deep learning model, simulating a cognitive process: a classification task. The level of performance of the task was considered the simulated equivalent to the normality-psychopathology continuum, the dimensionality of the neural network's internal computational dynamics being the simulated equivalent of the PSM's dimensionality., Results: The neural networks' level of performance was shown to be associated with the characteristics of the internal computational dynamics, assumed to be the simulated equivalent of poorly-modulable cognitive processes., Discussion: Findings supported the hypothesis. They showed that the neural network's low performance was a matter of the combination of predicted characteristics of the neural networks' internal computational dynamics. Implications, limitations, and further research directions are discussed., Competing Interests: The authors have declared that no competing interests exist.

Despite the increasing demand of cellular therapies for dogs, little is known on the differences between adult and fetal adnexa canine mesenchymal stem cells (MSCs), and data on their metabolic features are lacking. The present study aimed at comparing the characteristics of canine adipose tissue (AT) and umbilical cord matrix (UC) MSCs. Moreover, for the first time in the dog, the cellular bioenergetics were investigated by evaluating the two main metabolic pathways (oxidative phosphorylation and glycolysis) of ATP production. Frozen-thawed samples were used for this study. No differences in mean cell proliferation were found (P>0.05). However, while AT-MSCs showed a progressive increase in doubling time over passages, UC-MSCs showed an initial post freezing-thawing latency. No differences in migration, spheroid formation ability, and differentiation potential were found (P>0.05). RT-PCR analysis confirmed the expression of CD90 and CD44, the lack of CD14 and weak expression of CD34, mostly by AT-MSCs. DLA-DRA1 and DLA-DQA1 were weakly expressed only at passage 0 by UC-MSCs, while they were expressed at different passages for AT-MSCs. There was no difference (P>0.05) in total ATP production between cell cultures, but the ratio between the "mitochondrial ATP Production Rate" and the "glycolytic ATP Production Rate" was higher (P<0.05) in AT- than in UC-MSCs. However, in both MSCs types the mitochondrial respiration was the main pathway of ATP production. Mitochondrial respiration and ATP turnover in UC-MSCs were higher (P<0.05) than in AT-MSCs, but both had a 100% coupling efficiency. These features and the possibility of increasing the oxygen consumption by a spare respiratory capacity of four (AT-MSCSs) and two (UC-MSCs) order of magnitude greater than basal respiration, can be taken as indicative of the cell propensity to differentiate. The findings may efficiently contribute to select the most appropriate MSCs, culture and experimental conditions for transplantation experiments in mesenchymal stem cell therapy for companion animals., Competing Interests: The authors have declared that no competing interests exist.

Parkinson's disease (PD) is a progressive neurological disorder of the central nervous system that deteriorates motor functions, while it is also accompanied by a large diversity of non-motor symptoms such as cognitive impairment and mood changes, hallucinations, and sleep disturbance. Parkinsonism is evaluated during clinical examinations and appropriate medical treatments are directed towards alleviating symptoms. Tri-axial accelerometers, gyroscopes, and magnetometers could be adopted to support clinicians in the decision-making process by objectively quantifying the patient's condition. In this context, at-home data collections aim to capture motor function during daily living and unobstructedly assess the patients' status and the disease's symptoms for prolonged time periods. This review aims to collate existing literature on PD monitoring using inertial sensors while it focuses on papers with at least one free-living data capture unsupervised either directly or via videotapes. Twenty-four papers were selected at the end of the process: fourteen investigated gait impairments, eight of which focused on walking, three on turning, two on falls, and one on physical activity; ten articles on the other hand examined symptoms, including bradykinesia, tremor, dyskinesia, and motor state fluctuations in the on/off phenomenon. In summary, inertial sensors are capable of gathering data over a long period of time and have the potential to facilitate the monitoring of people with Parkinson's, providing relevant information about their motor status. Concerning gait impairments, kinematic parameters (such as duration of gait cycle, step length, and velocity) were typically used to discern PD from healthy subjects, whereas for symptoms' assessment, researchers were capable of achieving accuracies of over 90% in a free-living environment. Further investigations should be focused on the development of ad-hoc hardware and software capable of providing real-time feedback to clinicians and patients. In addition, features such as the wearability of the system and user comfort, set-up process, and instructions for use, need to be strongly considered in the development of wearable sensors for PD monitoring., Competing Interests: The authors have read the journal’s policy and have the following competing interests: Abbvie Inc. provided partial funding for this study. This does not alter our adherence to PLOS ONE policies on sharing data and materials. There are no patents, products in development or marketed products associated with this research to declare.

The sanitary status of grapevines has not yet been considered sufficiently in vineyards throughout Bosnia and Herzegovina (BiH). An extensive survey of five major grapevine viruses in the country was carried out in 2019. A total of 630 samples from the two dominant autochthonous cultivars, named Žilavka and Blatina, were tested by DAS-ELISA for the presence of grapevine leafroll-associated viruses (GLRaV-1 and 3), grapevine fleck virus (GFkV), grapevine fanleaf virus (GFLV) and Arabis mosaic virus (ArMV). Eighty-eight % of the samples were positive for at least one virus, and all five viruses were detected, thought with different incidence, i.e. GLRaV-3 (84%), GFLV (43%), GLRaV-1 (14%), GFkV (10%) and ArMV (0.2%). The majority of infected plants (about 75%) were asymptomatic. Specific virus symptoms were observed in the remaining infected plants, together with the reported GLRaV vectors, Planococcus ficus and Parthenolecanium corni, while nematodes of the Xiphinema genus were not found in the GFLV- or ArMV-infected vineyards. The GLRaV-3 CP phylogenetic analyses showed 75-100% nucleotide identity between the BiH and reference isolates, and the BiH isolates clustered into the major group. The dNS/dS ratio indicated a negative selection of the virus population, and the lack of geographical structuring within the population was observed. In addition, putative GLRaV-3 recombinants with breakpoints in the 5' of the CP gene were detected, while no recombinant strains were identified for the other four viruses. The obtained results indicate a deteriorated sanitary status of the cultivated grapevines, the prevalence and intraspecies genetic diversity of GLRaV-3 throughout the country. The establishment of certified grapevine material and adequate virus vector control is therefore of primary importance to prevent further spread of these viruses. This study presents the results of the first molecular characterisation of grapevine viruses in Bosnia and Herzegovina., Competing Interests: The authors have declared that no competing interests exist.

Backgrounds: Validated tools for predicting individual in-hospital mortality of COVID-19 are lacking. We aimed to develop and to validate a simple clinical prediction rule for early identification of in-hospital mortality of patients with COVID-19., Methods and Findings: We enrolled 2191 consecutive hospitalized patients with COVID-19 from three Italian dedicated units (derivation cohort: 1810 consecutive patients from Bergamo and Pavia units; validation cohort: 381 consecutive patients from Rome unit). The outcome was in-hospital mortality. Fine and Gray competing risks multivariate model (with discharge as a competing event) was used to develop a prediction rule for in-hospital mortality. Discrimination and calibration were assessed by the area under the receiver operating characteristic curve (AUC) and by Brier score in both the derivation and validation cohorts. Seven variables were independent risk factors for in-hospital mortality: age (Hazard Ratio [HR] 1.08, 95% Confidence Interval [CI] 1.07-1.09), male sex (HR 1.62, 95%CI 1.30-2.00), duration of symptoms before hospital admission <10 days (HR 1.72, 95%CI 1.39-2.12), diabetes (HR 1.21, 95%CI 1.02-1.45), coronary heart disease (HR 1.40 95% CI 1.09-1.80), chronic liver disease (HR 1.78, 95%CI 1.16-2.72), and lactate dehydrogenase levels at admission (HR 1.0003, 95%CI 1.0002-1.0005). The AUC was 0.822 (95%CI 0.722-0.922) in the derivation cohort and 0.820 (95%CI 0.724-0.920) in the validation cohort with good calibration. The prediction rule is freely available as a web-app (COVID-CALC: https://sites.google.com/community.unipa.it/covid-19riskpredictions/c19-rp)., Conclusions: A validated simple clinical prediction rule can promptly and accurately assess the risk for in-hospital mortality, improving triage and the management of patients with COVID-19., Competing Interests: The authors have declared that no competing interests exist.

Background: During ischemia a close relationship exists between sub-endocardial blood flow and myocardial function. Strain parameters can capture an impairment of regional longitudinal function but are load dependent. Recently, a novel non-invasive method to calculate Myocardial Work (MW) showed a strong correlation with invasive work measurements. Our aim was to investigate the ability of non-invasive MW indices to identify the ischaemic risk area during transient acute coronary occlusion (TACO)., Methods and Results: The study population comprises 50 individuals with critical coronary stenosis (CCS). Echocardiography recordings were obtained before coronary angiography, during TACO and after revascularization to measure global longitudinal strain (GLS), Myocardial Work Index (MWI), Myocardial Constructive Work (MCW), Myocardial Wasted work (MWW), Myocardial work efficiency (MWE). Compared to baseline, we found a significant reduction of GLS (p = 0.005), MWI, MCW and MWE (p<0.001) during TACO., Conclusions: The non-invasive measurement of MW parameters is a sensitive and early marker of myocardial ischemia during TACO., Competing Interests: The authors have declared that no competing interests exist.

The physiological role of autophagy in the progression of liver diseases is still debated. To understand the clinical relevance of autophagy in primary e secondary hepatic tumors, we analyzed the expression of mTOR (mammalian target of rapamycin), a key regulator of autophagy; Raptor (regulatory-associated protein of mTOR); ULK1 (Unc-51 like kinase 1) determinant in the autophagy initiation; LC3 (microtubule-associated protein 1A/1B-light chain 3), a specific marker of autophagosomes; and p62, a selective autophagy receptor. Samples from subjects with chronic hepatitis (n.58), cirrhosis (n.12), hepatocellular carcinoma (HCC, n.56), metastases (n.48) from colorectal cancer and hyperplasia or gallbladder stones (n.7), the latter considered as controls, were examined. Gene expression analysis was carried out in n.213 tissues by absolute q-PCR, while protein expression by Western Blot in n.191 lysates, including tumoral, surrounding tumoral and normal tissues. Nonparametric statistical tests were used for comparing expression levels in the above-mentioned groups. Subgroup analysis was performed considering viral infection and chemotherapy treatment. The mTOR transcriptional level was significantly lower in metastases compared to HCC (P = 0.0001). p-mTOR(Ser2448) and LC3II/LC3I protein levels were significantly higher in metastases compared to HCC (P = 0.008 and P<0.0001, respectively). ULK(Ser757) levels were significantly higher in HCC compared to metastases (P = 0.0002) while the HCV- and HBV- related HCC showed the highest p62 levels. Chemotherapy induced a down-regulation of the p-mTOR(Ser2448) in metastases and in non-tumor surrounding tissues in treated patients compared to untreated (P = 0.001 and P = 0.005, respectively). Conclusions: the different expression of proteins considered, owning their interaction and diverse tissue microenvironment, indicate an impairment of the autophagy flux in primary liver tumors that is critical for the promotion of tumorigenesis process and a coexistence of autophagy inhibition and activation mechanisms in secondary liver tumors. Differences in mTOR and LC3 transcripts emerged in tumor-free tissues, therefore particular attention should be considered in selecting the control group., Competing Interests: The authors have declared that no competing interests exist.

Metabolic associated fatty liver disease (MAFLD) is the most prevalent liver disease in Western nations, with high heritability. A recent study of Japanese patients with the disease suggested that TLL1 rs17047200 is associated with fibrosis; whether a similar association is observed in Caucasian patients with MAFLD is unknown. We investigated the association of the TLL1 rs17047200 polymorphism with liver fibrosis in a cohort of Caucasian patients with MAFLD (n = 728). We also investigated whether TLL1 expression is altered during liver injury in humans, in murine models of fibrosis, and in in-vitro. While TLL1 expression is upregulated in the liver of humans with MAFLD and in mice, the rs17047200 variant was not associated with fibrosis or any other histological features, or with hepatic TLL1 expression. In conclusion, the TLL1 rs17047200 variant is not a risk variant for fibrosis in Caucasian patients with MAFLD. However, TLL1 could be involved in the pathogenesis of liver fibrosis., Competing Interests: The authors have declared that no competing interests exist.

The use of mobile communication devices in health care is spreading worldwide. A huge amount of health data collected by these devices (mobile health data) is nowadays available. Mobile health data may allow for real-time monitoring of patients and delivering ad-hoc treatment recommendations. This paper aims at showing how this may be done by exploiting the potentialities of fuzzy clustering techniques. In fact, such techniques can be fruitfully applied to mobile health data in order to identify clusters of patients for diagnostic classification and cluster-specific therapies. However, since mobile health data are full of noise, fuzzy clustering methods cannot be directly applied to mobile health data. Such data must be denoised prior to analyzing them. When longitudinal mobile health data are available, functional data analysis represents a powerful tool for filtering out the noise in the data. Fuzzy clustering methods for functional data can then be used to determine groups of patients. In this work we develop a fuzzy clustering method, based on the concept of medoid, for functional data and we apply it to longitudinal mHealth data on daily symptoms and consumptions of anti-symptomatic drugs collected by two sets of patients in Berlin (Germany) and Ascoli Piceno (Italy) suffering from allergic rhinoconjunctivitis. The studies showed that clusters of patients with similar changes in symptoms were identified opening the possibility of precision medicine., Competing Interests: We state that TPS (Technology Projects & Software) Production srl has patented in Italy (International registration number: PCT / IT2018 / 000119) and it is undergoing the registration process at the European Patent Office with the number: 18804719.5 - 1126). AllergyMonitor is an online service developed by TPS Production. It is a free downloadable app for users with the aim of enabling the recording of clinical symptoms, drug consumption and adherence to specific sublingual and subcutaneous immunotherapy (SLIT, SCIT) by patients suffering from allergic diseases such as rhino-conjunctivitis and asthma. The AllergyMonitor system consists of two parts: the web site for the patient (front end) and the website for the doctor (back office). The AllergyMonitor back office was used only for clinical studies. This does not alter our adherence to PLOS ONE policies on sharing data and materials. Salvatore Tripodi is a co-founder of TPS. Paolo Giordani, Serena Perna, Annamaria Bianchi, Antonio Pizzulli and Paolo Maria Matricardi declare that they have no significant competing financial, professional, or personal interests that might have influenced the performance or presentation of the study described in this manuscript.

Mycoplasma synoviae infection occurs worldwide, leading to considerable economic losses in the chicken and turkey industry due to infectious synovitis, respiratory diseases and eggshell apex abnormalities. Control programs against M. synoviae infection are based on eradication, vaccination and medication with antimicrobial agents. Prudent use of antibiotics can be improved greatly by the determination of antibiotic susceptibility prior to the treatment. However, the conventional broth or agar microdilution is very labor-intensive and time-consuming method. Thus, there is an increasing need for rapid antimicrobial susceptibility tests in order to guide antibiotic therapy more effectively. The aim of this study was to develop mismatch amplification mutation assays (MAMAs) to detect resistance-associated mutations in M. synoviae. M. synoviae strains with previously determined minimal inhibitory concentrations (MICs) and whole genomes (n = 92) were used for target selection and assay specification. For the evaluation of the developed assays, 20 clinical samples and an additional 20 M. synoviae isolates derived from these specimens were also included in this study. MIC values of these 20 isolates were determined by broth microdilution method. Five MAMAs were designed to identify elevated MICs of fluoroquinolones, while three MAMAs were developed to detect decreased susceptibility to macrolides and lincomycin. The sensitivity of the MAMA tests varied between 102-104 template copy number/reaction depending on the assay. Clinical samples showed identical genotype calls with the M. synoviae isolates derived from the corresponding specimens in each case. Supporting the results of conventional in vitro sensitivity tests, our approach provides a feasible tool for diagnostics. Rapidity, robustness and cost-effectiveness are powerful advantages of the developed assays. Supporting prudent antibiotic usage instead of empirical treatment, the use of this method can reduce significantly the economic impact of M. synoviae in the poultry industry and decrease bacterial resistance-related public health concerns., Competing Interests: The authors have declared that no competing interests exist.

The cultivation of walnuts (Juglans sp.) in Europe retains high economic, social, and environmental value. The recent reporting of the Thousand Cankers Disease (TCD) fungus, Geosmithia morbida, and of its vector, Pityophthorus juglandis, in walnut trees in Italy is alarming the whole of Europe. Although Italy is at present the only foothold of the disease outside North America, given the difficulties inherent in traditional identification of both members of this beetle/fungus complex, a rapid and effective protocol for the early detection and identification of TCD organisms is an absolute priority for Europe. Here we report the development of an effective and sensitive molecular tool based on simplex/duplex qPCR assays for the rapid, accurate and highly specific detection of both the bionectriaceous fungal pathogen and its bark-beetle vector. Our assay performed excellently, detecting minute amounts of target DNA without any non-specific amplification. Detection limits from various and heterogeneous matrices were lower than other reported assays. Our molecular protocol could assist in TCD organism interception at entry points, territory monitoring for the early identification and eradication of outbreaks, delineation of quarantine areas, and tracing back TCD entry and dispersal pathways., Competing Interests: The authors have declared that no competing interests exist.

Handwriting is a complex skill to acquire and it requires years of training to be mastered. Children presenting dysgraphia exhibit difficulties automatizing their handwriting. This can bring anxiety and can negatively impact education. 280 children were recruited in schools and specialized clinics to perform the Concise Evaluation Scale for Children's Handwriting (BHK) on digital tablets. Within this dataset, we identified children with dysgraphia. Twelve digital features describing handwriting through different aspects (static, kinematic, pressure and tilt) were extracted and used to create linear models to investigate handwriting acquisition throughout education. K-means clustering was performed to define a new classification of dysgraphia. Linear models show that three features only (two kinematic and one static) showed a significant association to predict change of handwriting quality in control children. Most kinematic and statics features interacted with age. Results suggest that children with dysgraphia do not simply differ from ones without dysgraphia by quantitative differences on the BHK scale but present a different development in terms of static, kinematic, pressure and tilt features. The K-means clustering yielded 3 clusters (Ci). Children in C1 presented mild dysgraphia usually not detected in schools whereas children in C2 and C3 exhibited severe dysgraphia. Notably, C2 contained individuals displaying abnormalities in term of kinematics and pressure whilst C3 regrouped children showing mainly tilt problems. The current results open new opportunities for automatic detection of children with dysgraphia in classroom. We also believe that the training of pressure and tilt may open new therapeutic opportunities through serious games., Competing Interests: The authors have declared that no competing interests exist.

Purpose: To test a new method to calculate the Intraocular Lens (IOL) power, that combines R Factor and ALxK methods, that we called Advance Lens Measurement Approach (ALMA)., Design: Retrospective, Comparative, Observational study., Setting: Department of Medicine and Surgery, University of Salerno, Italy., Methods: Ninety one eyes of 91 patients previously treated with Photorefractive Keratectomy (PRK) or Laser-Assisted in Situ Keratomileusis (LASIK) that underwent phacoemulsification and IOL implantation in the capsular bag were analyzed. For 68 eyes it was possible to zero out the Mean Errors (ME) for each formula and for selected IOL models, in order to eliminate the bias of the lens factor (A-Costant). Main outcome, measured in this study, was the median absolute error (MedAE) of the refraction prediction., Results: In the sample with ME zeroed (68 eyes) both R Factor and ALxK methods resulted in MedAE of 0.67 D. For R Factor 33 eyes (48.53%) reported a refractive error <0.5D, and 53 eyes (77.94%) reported a refractive error <1D, For ALxK method, 32 eyes (47.06%) reported a refractive error <0.5 D, and 53 eyes (77.94%) reported a refractive error <1 D. ALMA method, reported a MedAE of 0.55 D, and an higher number of patients with a refractive error <0.5 D (35 eyes, 51.47%), and with a refractive error <1 D (54 eyes, 79.41%)., Conclusions: Based on the results obtained from this study, ALMA method can improve R Factor and ALxK methods. This improvement is confirmed both by zeroing the mean error and without zeroing it., Competing Interests: The authors have declared that no competing interests exist.

Background: The ongoing pandemic of Novel Coronavirus Disease 2019 (COVID-19) infection has created a global emergency. Despite the infection causes a mild illness to most people, some patients are severely affected, demanding an urgent need to better understand how to risk-stratify infected subjects., Design: This is a meta-analysis of observational studies evaluating cardiovascular (CV) complications in hospitalized COVID-19 patients and the impact of cardiovascular risk factors (RF) or comorbidities on mortality., Methods: Data sources: PubMed, Scopus, and ISI from 1 December 2019 through 11 June 2020; references of eligible studies; scientific session abstracts; cardiology web sites. We selected studies reporting clinical outcomes of hospitalized patients with COVID-19. The main outcome was death. Secondary outcomes were cardiovascular symptoms and cardiovascular events developed during the COVID-19-related hospitalization. Extracted data were recorded in excel worksheets and analysed using statistical software (MedCalc, OpenMetanalyst, R). We used the proportion with 95% CI as the summary measure. A Freeman-Tukey transformation was used to calculate the weighted summary proportion under the random-effects model. Heterogeneity was assessed by using the Cochran Q test and I2 values., Results: Among 77317 hospitalized patients from 21 studies, 12.86% had cardiovascular comorbidities or RF. Cardiovascular complications were registered in 14.09% of cases during hospitalization. At meta-regression analysis, pre-existing cardiovascular comorbidities or RF were significantly associated to cardiovascular complications in COVID-19 patients (p = 0.019). Pre-existing cardiovascular comorbidities or RF (p<0.001), older age (p<0.001), and the development of cardiovascular complications during the hospitalization (p = 0.038) had a significant interaction with death., Conclusions: Cardiovascular complications are frequent among COVID-19 patients, and might contribute to adverse clinical events and mortality, together with pre-existing cardiovascular comorbidities and RF. Clinicians worldwide should be aware of this association, to identifying patients at higher risk., Competing Interests: The authors have declared that no competing interests exist.

Background: Knowledge regarding differences in care for psoriatic patients is limited. The aim of this study was to investigate factors influencing prescription of systemic treatments for patients with psoriasis with a special focus on socioeconomic factors., Methods and Findings: This was a non-interventional, cross-sectional study, conducted in 18 Italian University and/or hospital centers with psoriasis-specialized units. Questionnaires evaluating demographic and socioeconomic characteristics were administered to participants. Overall, 1880 consecutive patients affected by mild-to-severe psoriasis were recruited. Univariate and multivariable logistic regression analyses of systemic therapy prescription, with a special focus on biologics, accounting for the above mentioned characteristics were performed. Our analysis showed that all analyzed patients' characteristics were significantly associated with biological therapy compared to non-biological systemic one. Particularly, women were less likely to receive biologics than men (OR = 0.66; 95% CI, 0.57-0.77). Elderly patients (≥65 years) and subjects with a BMI ≥30 had lower odds to receive biologics respect to adults (≥35-64 years) (OR = 0.33; 95% CI, 0.25-0.40), and subjects with BMI≥25<30 (OR = 0.64; 95% CI, 0.53-0.77), respectively. Northern and Southern patients were both less likely to receive biologics than Central patients (OR = 0.75; 95% CI, 0.63-0.89, and OR = 0.56; 95% CI,0.47-0.68, respectively). Lower economic profile and never reading books were both associated with decreased odds of receiving biological therapy., Conclusions: This study shows that sex, age, comorbidities, and socioeconomic characteristics influence the prescription of systemic treatments in psoriasis, highlighting that there are still unmet needs influencing the therapeutic decision-making process that have to be addressed., Competing Interests: P. Amerio has been a scientific consultant for Abbvie, Eli-Lilly, Jannsen, Celgene, Galderma, LEO-Pharma, Sanofi Genzyme and Alfa Wasserman. F. Bardazzi has been a scientific consultant/speaker/clinical study investigator for AbbVie, Celgene, Janssen, LEOPharma, Eli-Lilly, MSD, Novartis and Pfizer. L. Bianchi has been a scientific consultant/speaker for Abbvie, Janssen, Celgene, Pfizer, UCB, Novartis, Sanofi Genzyme and LEO-Pharma. A. Campanati has been a scientific consultant/speaker/clinical study investigator for AbbVie, Celgene, Janssen, LEO-Pharma, Eli-Lilly, MSD, Novartis, Pfizer and Sanofi. S.P. Cannavò has been a scientific consultant/speaker for AbbVie, Celgene, Eli-Lilly, Janssen, LEO-Pharma, Novartis and Sanofi. A. Chiricozzi has been a scientific consultant/speaker/clinical study investigator for Abbvie, Biogen, Fresenius Kabi, LEO-Pharma, Eli-Lilly, Novartis, UCB, Sanofi and Janssen. A. Conti has been a scientific consultant for Abbvie, Eli-Lilly, Janssen Cilag, LEO-Pharma, UCB and Sandoz. P. Dapavo has been a scientific consultant/speaker for Novartis, Abbvie, Eli-Lilly, Celgene, LEOpharma and Janssen. C. De Simone has been a scientific consultant/speaker/clinical study investigator for AbbVie, Almirall, Amgen, Celgene, Janssen, LEO-Pharma, Eli-Lilly, MSD, Novartis, Pfizer, Sanofi and Sun Pharma. M. Esposito has been a scientific consultant/speaker for Abbvie, Novartis, Eli-Lilly, Biogen, LEO-Pharma. M.C. Fargnoli has been a scientific consultant/speaker/clinical study investigator for AbbVie, Celgene, Janssen, LEO-Pharma, Eli- Lilly, Novartis, Pfizer, Sanofi, Galderma, Almirall, Mylan, UCB and Regeneron. C. Guarneri has been a scientific consultant/speaker/clinical study investigator for Wyeth-Pfizer, Abbott Immunology-Abbvie, Janssen-Cilag, Novartis, LEO-Pharma, Eli-Lilly, Celgene and Merck-Serono. K. Hansel has been a scientific consultant/speaker/clinical study investigator for AbbVie, Eli- Lilly, Novartis, Sanofi and Admiral. P. Malagoli has been a scientific consultant/speaker/clinical study investigator for AbbVie, Amgen, Biogen, Celgene, Janssen, LEO-Pharma, Eli-Lilly, Novartis. G. Micali has been a scientific consultant/clinical study investigator for Abbvie, Eli-Lilly, Janssen- Cilag, LEO-Pharma and Novartis. C. Mugheddu has been a speaker/clinical study investigator for Abbvie, Celgene, Boehringer Ingelheim, Janseen, Novartis and LEO-Pharma. A. Offidani has been a scientific consultant/speaker/clinical study investigator for AbbVie, Celgene, Janssen, LEOPharma, Eli-Lilly, MSD, Novartis, Pfizer, Sanofi, Alfasigma and Almirall. F. Prignano has been a scientific consultant/adviser/clinical study investigator for Eli-Lilly, Abbvie, Novartis, LEOPharma, Abiogen-Pharma, Celgene and Janssen-Cilag. F. Rongioletti has been a scientific consultant/speaker/clinical study investigator for AbbVie, Boehringer, Celgene, Janssen, LEOPharma, Eli-Lilly, Novartis and Sanofi. L. Stingeni has been a scientific consultant/speaker/clinical study investigator for AbbVie, Celgene, Eli-Lilly, Novartis, Almirall and Sanofi. A. Balato has been a scientific consultant/speaker/clinical study investigator for AbbVie, Celgene, Janssen, Eli-Lilly, Novartis, Pfizer, Sanofi and UCB. The remaining authors declare no conflicts of interest. This does not alter our adherence to PLOS ONE policies on sharing data and materials.