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Brucellosis represents a major public health concern worldwide. Human transmission is mainly due to the consumption of unpasteurized milk and dairy products of infected animals. The gold standard for the diagnosis of Brucella spp in ruminants is the bacterial isolation, but it is time-consuming. Polymerase Chain Reaction (PCR) is a quicker and more sensitive technique than bacterial culture. Droplet digital PCR (ddPCR) is a novel molecular assay showing high sensitivity in samples with low amount of DNA and lower susceptibility to amplification inhibitors. Present study aimed to develop a ddPCR protocol for the detection of Brucella abortus in buffalo tissue samples. The protocol was validated using proficiency test samples for Brucella spp by real time qPCR. Furthermore, 599 tissue samples were examined. Among reference materials, qPCR and ddPCR demonstrated same performance and were able to detect up to 225 CFU/mL. Among field samples, ddPCR showed higher sensitivity (100%), specificity and accuracy of 93.4% and 94.15%, respectively. ddPCR could be considered a promising technique to detect B. abortus in veterinary specimens, frequently characterized by low amount of bacteria, high diversity in matrices and species and poor storage conditions.

Abstract Transposable elements (TEs) are dynamic components of eukaryotic genomes that are important in speciation. Evolutionary pressure has led to the evolution of silencing mechanisms, such as the involvement of Krüppel box-associated zinc finger proteins (KRAB-ZFPs) that regulate TE transcription via TRIM28-mediated NuRD complex recruitment in sarcopterygians. The transcriptional activity of genes that encode proteins of the NuRD complex was also detected in actinopterygians, even though genes encoding crucial proteins such as TRIM28 and KRAB-ZNF are missing in this evolutionary lineage. This study hypothesized that TRIM33 could serve as a replacement for TRIM28 in this lineage and suggested an evolutionary relationship between the sarcopterygian KRAB-ZFPs, the actinopterygian KRAB-like ZNFs and the fish N-terminal zinc-associated domain (FiNZ) ZNFs that are specific to cyprinids. Through coimmunoprecipitation analyses conducted in the basal teleost Anguilla marmorata and the cyprinid Danio rerio, we evaluated the interaction of TRIM33 with the KRAB-like and FiNZ ZFPs. Moreover, we explored the expression profiles of TEs and genes that encode proteins that are potentially involved in their silencing during zebrafish development. Our results may provide new evidence for more thoroughly understanding the evolutionary mechanisms underlying the adaptation of ray-finned fish.

IntroductionSalmonella spp. is the second most common bacteria associated with foodborne gastrointestinal outbreaks in humans, with the highest contamination levels in meat, especially poultry. Salmonella enterica subsp. enterica serovar Infantis is the primary serovar isolated from broilers, without causing any symptomatic disease. Conversely, certain human strains can result in symptomatic illness (fever, headache, and diarrhoea). Therefore, reducing S. Infantis colonization in broilers is important before slaughter, to prevent this pathogen carryover along the food chain.MethodsHere, we report the characterization of two S. Infantis virulent phages, isolated from broiler feces. Isolates were phenotypically and genetically characterized.Results and discussionPhages (ɸ) SaI_NFG_5581 and SaI_NFG_5577 were characterized as strictly lytic versus S. Infantis but with different bacteriolytic activities and genetic features. They both belong to the Caudoviricetes class, but ɸSaI_NFG_5581 (genome length 112,970 bp) belongs to the Demerecviridae family while ɸSaI_NFG_5577 (genome length 42,481 bp) to the Guernseyvirinae family. Genomic analysis excluded the presence of lysogeny, toxin, or antimicrobial resistance genes, and for those reasons, the two phages could be considered safe. Phages are stable under a broad range of pH (4-10) and temperature (4°C-50°C) conditions. In vitro, both ɸSaI_NFG_5581 and ɸSaI_NFG_5577 were able to lower Salmonella counts of about 2.2 LOG/mL and 3.4 LOG CFU/mL respectively, at MOI 0.1 after 2 h of treatment. After 24 h, Salmonella counts treated with both phages remained lower than the control (non-phage-treated Salmonella). These newly isolated phages have promising features, which could be exploited and further studied for potential in vivo application.

Listeria monocytogenes is a significant concern in the food industry due to its association with outbreaks of listeriosis, particularly affecting vulnerable populations. High-throughput technologies such as RNA sequencing (RNA-seq) and proteomics offer valuable insights into the molecular responses of L. monocytogenes to stress environments. In this study, a combined transcriptomic and immunoproteomic approach was applied to explore the stress response mechanisms of the L. monocytogenes strain ST7, which was responsible for an outbreak in central Italy. The bacterium was exposed to both optimal conditions and a stress environment representative of pork product matrices (pH 5.5; 7 % NaCl) and thermal abuse prior to consumption (12 °C).Transcriptomic analysis revealed variations in gene expression related to pathogenesis, stress responses, and virulence factors under different environmental conditions. Transcriptomic analysis of Listeria involves studying the complete set of RNA transcripts produced by the bacterium under various conditions or during different stages of its lifecycle. It can provide insights into its pathogenicity and virulence mechanisms. Immunoproteomic analysis identified proteins involved in stress response pathways, including oxidoreductases and DNA repair enzymes, uniquely expressed under stress conditions. Furthermore, the study highlighted proteins linked to antibiotic resistance and cell wall biosynthesis. By delineating specific proteins crucial in the stress response pathways, these findings not only deepen our comprehension of L. monocytogenes biology but also pave the way for designing more targeted mitigation strategies to safeguard food safety and public health effectively.

The interest in characterization of drugs abused started in 1948 with the aim of determining the origin of opium. After 75 years there is still a great interest in this approach, called geo-profiling, chemical or forensic profiling in the following decades. Recently chemical attribution signatures (CAS) were proposed by the authors who studied “synthesis precursors and byproducts, impurities, degradation products, and metabolites in various biological matrices” of fentanyl. Forensic profiling evolved during these decades: new analytical approaches were tested and it was applied to more and more products, which threaten the health and security of citizens worldwide.In substances of natural origins (e.g. opium, cannabis and cocaine), it is possible to exploit the great variability of both elements and organic chemical compounds and to study chemical compounds such as reagents and solvents, by-products, and cutting agents used in the production chain. Profiles can be used to classify products from different seizures into groups of similar samples (tactical intelligence) or to determine the origin of samples (strategic intelligence).Chromatographic approaches coupled to mass spectrometry are very common to determine organic profiles, while elemental profiles are obtained by nuclear activation analysis, inductively coupled plasma mass spectrometry or ion beam analysis. A very important role in the field is played by isotope ratio analysis. Approaches to obtain forensic profiles are available also for chemical warfare agents, explosives, illegal medicines, doping agents, supplements, food. Chemometrics can be particularly useful to establish the authenticity of products and for the interpretation of large amount of forensic data. The future of forensic profiling is a challenge for forensic sciences. Organized crime is involved in the manufacturing of a large number of illegal products and forensic profiling is a very powerful tool to support the health of citizens and the administration of justice worldwide.

Rose Bengal antigen and smooth lipopolysaccharide (s-LPS) were produced from a field strain of Brucella ceti (“homologous” antigens) and from the reference strain B. abortus S99 (“heterologous” antigens); they are currently used for the diagnosis of brucellosis in cattle, water buffaloes, sheep, goats, and pigs, as recommended in the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals of the World Organization for Animal Health (WOAH). “Homologous” and “heterologous” antigens were used in a rapid serum agglutination test (Rose Bengal test, RBT) and a competitive ELISA assay (c-ELISA) to test a panel of sera, blood, and other body fluids (cerebrospinal fluid, pericardial fluid, tracheal fluid, and aqueous humor) collected from 71 individuals belonging to five cetacean species (Stenella coeruleoalba; Tursiops truncatus; Grampus griseus; Globicephala melas; and Ziphius cavirostris), which were found stranded on the Italian coastline. Six animals were positive for Brucella spp. for bacterial isolation and/or PCR, and 55 animals were negative; for the remaining 10 animals, no PCR/isolation data were available. A total of 90 samples were tested. Results obtained from the two tests were compared in order to identify the most suitable antigen for the serological diagnosis of Brucella infection in cetaceans. The RBT performed with the “homologous” antigen showed the best results in comparison with the “heterologous” antigen: diagnostic sensitivity, specificity, and accuracy were 80.0%, 44.1%, and 46.9% for the “homologous” antigen and 80.0%, 17.0%, and 21.9% for the “heterologous” antigen. For the c-ELISA, “homologous” and “heterologous” s-LPS showed similar results (diagnostic sensitivity 66.7%, diagnostic specificity 97.3%, and diagnostic accuracy 95.0%). Therefore, the RBT using the “homologous” antigen and c-ELISA with “homologous” or “heterologous” s-LPS could be used in parallel for the detection of antibodies against Brucella spp. in cetaceans.

Background: Drugged driving is associated with an increased risk of road accidents worldwide. In Italy, driving under the influence (DUI) of alcohol and drugs is a reason for driving disqualification or revocation of the driving license. Drivers charged with driving under the influence of alcohol and drugs must attend a Local Medical Commission (LMC) to undergo mandatory examinations to regain the suspended license. Our study mainly aims to report on the analysis performed on hair samples collected from 7560 drivers who had their licenses suspended for drugged or drunk driving between January 2019 and June 2024. Methods: A rapid, sensitive, and selective method for the determination of ethyl glucuronide in hair by UPLC/MS-MS was developed and fully validated. Results: The most frequently detected substances were cocaine (ecgonine methyl ester, norcocaine, and benzoylecgonine) and cannabinoids (Δ9-tetrahydrocannabinol, cannabidiol, and cannabinol), followed by opiates (codeine, morphine, and 6-MAM), methadone (EDDP), and amphetamines (amphetamine, methamphetamine, MDA, MDMA, and MDEA). To perform a more in-depth analysis, we also compared hair color with the drug classes that tested positive. The results showed a significant prevalence of dark hair that tested positive for one or more substances, followed by gray/white hair and light hair. Conclusions: Our study provides an interesting and alarming insight into drug exposure in the general population with serious public health threats, discussing the main aspects of hair matrix analysis and focusing on its advantages and reliability in the interpretation of results.

Il contributo presenta la trascrizione dei documenti più significativi riguardanti Mariano di ser Austerio ad Ancona. Il pittore perugino, attivo nella città per un decennio, realizzò una tavola per la chiesa di Sant’Agostino, inizialmente richiesta da Medea Montiferis nel 1521 e successivamente modificata per volere di Girolamo Ferretti. Alla pala, citata da Giorgio Vasari nell’edizione delle Vite del 1568, si aggiunge l’inedita commissione all’artista della decorazione della cappella di Gabriele Bonarelli a San Francesco alle Scale, della quale rimane il dettagliato contratto del 1524. Entrambe le opere sono andate disperse con i rifacimenti settecenteschi delle due chiese, ma i documenti dimostrano che Mariano intrattenne rapporti commerciali con la città fino agli anni Trenta del Cinquecento. The contribution presents the most significant documents regarding Mariano di ser Austerio in Ancona. The Perugian painter, active in the city for a decade, painted a panel for the church of Sant'Agostino, initially commissioned by Medea Montiferis in 1521 and subsequently modified at the behest of Girolamo Ferretti. To the altarpiece, mentioned by Giorgio Vasari in the 1568 edition of the Vite, is added the unpublished assignment to the artist for the decoration of the chapel of Gabriele Bonarelli in San Francesco alle Scale, for which the detailed contract of 1524 remains. Both works were dispersed with the eighteenth-century renovations of the two churches, but documents show that Mariano maintained commercial relations with the city until the 1530s.

OBJECTIVE: Proposing a Diagnostic Therapeutic Assistance Pathway (PDTA) that enhances the preventive role of the Speech Therapist within Birth Support Courses (CAN), aiming to raise awareness and enhance parental skills regarding the communicative-linguistic, oro-facial-swallowing, and feeding development of newborns, thereby preventing potential disorders and difficulties in these areas. MATERIALS AND METHODS: A narrative review of the literature concerning Guidelines and PDTAs published in the Marche Region related to the Birth Pathway and Prevention in Early Childhood was conducted. The objective was to synthesize information regarding care models organized to promote and monitor the well-being of future parents during pregnancy and in the months following childbirth, as well as prevention programmes concerning the child's first 1000 days of life. The project is implemented by hypothetically involving the Territorial Health Companies (AST) of the Marche Region to develop a PDTA addressed to parental couples being comprised according to specific inclusion criteria (age over 18 years, residing in the Marche Region, physiological pregnancy, Italian native speakers or with at least B1 level of Italian language proficiency, consent to participation expressed during recruitment) and exclusion criteria (pregnant women in the period between 0-5 months). Within the various proposed meetings in the CAN, to integrate the activities of the Speech Therapist with those of other professionals, timetables and flowcharts are hypothesized concerning preventive actions for psychomotor and communicative-linguistic development in children aged 0 - 3 years. The healthcare programme also involves defining indicators of structure, process, and outcome. RESULTS: Conceiving a comprehensive and active clinical network, from prevention to empowerment, through a PDTA (Diagnostic Therapeutic Assistance Pathway) tailored on the real needs of parents and developmental age, which involves an interdisciplinary approach to guide the selection of training and informative contents towards understanding oro-sensory experiences and communicative-linguistic aspects, integrating them within family's environment. DISCUSSION and CONCLUSION: The role of the Speech Therapist in Child Protection Services (CAN) has a preventive objective, supporting the communicative-linguistic and oro-facial-nutritional development of children. Building trust with the various professional figures involved in the PDTA (Diagnostic Therapeutic Assistance Pathway) increases parents' sense of security and adherence during pregnancy, childbirth, and the postnatal period. The PDTA serves as an appropriate tool for preventing oro-facial and communicative-linguistic issues in developmental age, facilitating the work of the Speech Therapist in interdisciplinary teams, and supporting active participation of families throughout the regional territory of Marche Region. The creation of a comprehensive therapeutic and care pathway aimed at implementing the preventive role of the Speech and Language Pathologist within Childbirth Accompaniment Courses has the objective of enhancing parental awareness and skills as far as communicative-linguistic, oro-facial, swallowing and nutrition development is concerned.

OBIETTIVO: proporre un Percorso Diagnostico Terapeutico Assistenziale (PDTA) che amplifichi il ruolo preventivo del Logopedista all’interno di Corsi di Accompagnamento alla Nascita (CAN), al fine di sensibilizzare e valorizzare le competenze dei genitori, per ciò che riguarda lo sviluppo comunicativo-linguistico, facio-oro-deglutitorio e alimentare dei nascituri, prevenendo eventuali disturbi e difficoltà in tali aree. MATERIALI E METODI: revisione narrativa della letteratura relativa a Linee Guida e PDTA pubblicati nella Regione Marche e inerenti al Percorso Nascita e alla Prevenzione nella Prima Infanzia. L’obiettivo è stato quello di sintetizzare le informazioni relative ai modelli di assistenza, organizzati per promuovere e monitorare il benessere dei futuri genitori durante la gravidanza e nei mesi successivi al parto, e ai progetti di prevenzione riguardanti i primi 1000 giorni di vita del bambino. Il progetto viene realizzato ipotizzando il coinvolgimento delle Aziende Sanitarie Territoriali (AST) della Regione Marche, per realizzare un PDTA, rivolto alle coppie genitoriali, coinvolte secondo precisi criteri di inclusione (età superiore ai 18 anni, residenti nella Regione Marche, gravidanza con decorso fisiologico, madrelingua italiana o con livello di conoscenza della lingua italiana almeno B1, consenso alla partecipazione espresso nella fase di reclutamento) e di esclusione (donne in gravidanza nel periodo tra 0-5 mesi). All’interno dei vari incontri proposti nei CAN, per integrare l’attività del Logopedista con le altre figure professionali, vengono ipotizzati cronoprogrammi e flow chart, inerenti alle azioni preventive per lo sviluppo psicomotorio e comunicativo-linguistico nel bambino 0 - 3 anni. Il progetto prevede anche la definizione di indicatori di struttura, di processo e di esito. RISULTATI: una rete clinica completa e attiva, dalla prevenzione all’empowerment, tramite un PDTA mirato alle reali esigenze genitoriali e dell’età evolutiva, che prevede un approccio interdisciplinare per indirizzare la selezione dei contenuti formativi e informativi alla conoscenza delle esperienze oro-sensoriali e agli aspetti comunicativo-linguistici, integrandoli con le routine presenti in ambito familiare. DISCUSSIONE e CONCLUSIONI: il ruolo del Logopedista nei CAN ha finalità preventiva e supporta lo sviluppo comunicativo-linguistico e facio-oro-alimentare dei bambini. Un rapporto di fiducia con le varie figure professionali presenti nel PDTA aumenta il senso di sicurezza e di aderenza dei genitori durante la gravidanza, il parto e il periodo post-natale. Il PDTA rappresenta uno strumento adatto per la prevenzione delle problematiche oro-facciali e comunicativo-linguistiche in età evolutiva, favorendo il lavoro del Logopedista in équipe interdisciplinare e supportando la partecipazione attiva delle famiglie in tutto il territorio regionale.

Lightweight, metal‐free, sustainable, and reusable adsorbent materials are of paramount significance in addressing the challenges of wastewater treatment. Accordingly, semi‐crystalline nanocellulose (NC) is extracted from tissue paper waste and used to modify polyacrylonitrile (PAN) to produce electrospun nanowebs with strand diameters from ≈180–300 nm. The incorporation of NC into PAN is confirmed by infrared and Raman spectroscopy and X‐Ray diffraction. When tested for crystal violet (CV) adsorption, NC‐modified PAN (20% NC@PAN) exhibits the highest CV removal capacity, achieving 91–94% removal over three cycles each, demonstrating exceptional recyclability. In contrast, unmodified PAN significantly decreases in CV adsorption capacity (from 59% to 48% in the third cycle), possibly due to an increased (≈36%) nanofiber diameter. The adsorption kinetics, exhibiting pseudo‐second order, interparticle (in between nanofibers) diffusion, and Elovich kinetic models emphasize the role of multilayer CV adsorption through reversible chemical interactions. Confocal micro‐Raman spectroscopy unveils a multifaceted CV adsorption mechanism, suggesting both surface and multilayer diffusion, with NC‐enhancing interactions. These findings demonstrate the potential of NC‐modified PAN nanowebs as effective and environmentally sustainable adsorbents for removing CV from aqueous solutions, suggesting promising practical applications.

The maned wolf Chrysocyon brachyurus, the fourth largest canid, is categorized as Near Threatened on the IUCN Red List. The objectives of this study were to update information on the occurrence of this species in Argentina, compare the current distribution to that previously described for 1988–2008, and indicate the areas in which the species is exposed to particular hazards. We created a database that included 1,051 new records of C. brachyurus for 2009-2021, in addition to records for 1988–2008 that had not been reported previously. We calculated the area of occupancy of the species in Argentina to be 500,000 km2. Most of the new records of C. brachyurus are in the Chaco Seco, Espinal and Pampas ecoregions. Our findings indicate a current area of occupancy 61% higher than that calculated from records for 1988–2008. However the location and frequency of reported threats and the intense environmental transformation that characterizes the areas for which we obtained new records suggest counterintuitive implications for the conservation of the species because of increased human–wildlife interactions. Our study reveals the need for systematic research to improve the understanding of the ecological processes that affect the maned wolf in Argentina, and will aid in the development of conservation strategies for the species.

IntroductionThe increasing survival of patients with breast cancer has prompted the assessment of mortality due to all causes of death in these patients. We estimated the absolute risks of death from different causes, useful for health-care planning and clinical prediction, as well as cause-specific hazards, useful for hypothesis generation on etiology and risk factors.Materials and methodsUsing data from population-based cancer registries we performed a retrospective study on a cohort of women diagnosed with primary breast cancer. We carried out a competing-cause analysis computing cumulative incidence functions (CIFs) and cause-specific hazards (CSHs) in the whole cohort, separately by age, stage and registry area.ResultsThe study cohort comprised 12,742 women followed up for six years. Breast cancer showed the highest CIF, 13.71%, and cardiovascular disease was the second leading cause of death with a CIF of 3.60%. The contribution of breast cancer deaths to the CIF for all causes varied widely by age class: 89.25% in women diagnosed at age

Preventing L. monocytogenes infection is crucial for food safety, considering its widespread presence in the environment and its association with contaminated RTE foods. The pathogen's ability to persist under adverse conditions, for example, in food processing facilities, is linked to virulence and resistance mechanisms, including biofilm formation. In this study, the protein expression patterns of two L. monocytogenes 1/2a strains, grown under environmental stressors (mild acidic pH, thermal abuse, and high concentration of NaCl), were investigated. Protein identification and prediction were performed by nLC-ESI-MS/MS and nine different bioinformatic software programs, respectively. Gene enrichment analysis was carried out by STRING v11.05. A total of 1,215 proteins were identified, of which 335 were non-cytosolic proteins and 265 were immunogenic proteins. Proteomic analysis revealed differences in protein expression between L. monocytogenes strains in stressful conditions. The two strains exhibited unique protein expression profiles linked to stress response, virulence, and pathogenesis. Studying the proteomic profiles of such microorganisms provides information about adaptation and potential treatments, highlighting their genetic diversity and demonstrating the utility of bioinformatics and proteomics for a broader analysis of pathogens.

The use of benzodiazepines is strongly associated with an increased risk of traffic accidents due to their side effects of sedation and drowsiness, which can significantly impair driving performance. The main aim of our study was to investigate the trend of benzodiazepine use over nine years (2015–2023) in a population of 15,988 subjects who had their license suspended for driving under the influence (DUI) of alcohol or drugs. Among the 15,988 users accessed to our laboratory, 924 tested positive for at least one benzodiazepine. An increase in the number of positive-testing users was observed in the period 2015–2018, followed by a slight decrease in 2019. Overall, the trend of benzodiazepine use was stable over the next four years (2020–2023), with the highest incidence in 2022. The most common benzodiazepines, and/or metabolites, found in urine samples were α-OH-alprazolam (28.66%; n = 366) and oxazepam (27.25%; n = 348). Several cases of mixed positivity were observed in the study population. The main substances taken with benzodiazepines were cocaine and Δ9-tetrahydrocannabinol. Our findings suggest that people taking benzodiazepines should be monitored, as these have a relevant impact on driving ability in addition to significant interindividual differences in the behavioral effects of benzodiazepines on driving performance.

Food production systems in protected conditions are probably the most intensive agricultural systems causing biodiversity loss. Experimental sites in different European areas were established to investigate whether an agroecological approach to organic greenhouse production allows preserving or increasing biodiversity. Soil nematode communities were characterized by DNA-metabarcoding, and some biodiversity indices were calculated. A general increase in absolute number of genera indicates that the innovative treatments potentially positively influence biodiversity. A multidimensional analysis revealed that the change in nematode community compositions is caused more by country and sampling year than by the farming system. However, regionally, some shifts in biodiversity are obvious and the impact by a farming practice likely. This study is the first indication that the implementation of some agroecological measures in European protected cropping systems is feasible as at least they did not worsen (nor improve) soil biodiversity. [ABSTRACT FROM AUTHOR]

Alcohol is responsible for an ever-increasing number of deaths worldwide, and many road accidents are caused by irresponsible drinking and driving. The use of biomarkers that can support a diagnosis of alcohol abuse is a very important tool that can improve the prevention of many alcohol-related diseases and serious traffic accidents. The main aim of our study was the full validation of a rapid and simple method by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to detect ethyl glucuronide in hair (hEtG). The method was successfully applied to n = 171 real hair samples collected from drivers convicted of driving while impaired by alcohol or drugs. A comparison of hEtG and serum Carbohydrate-Deficient Transferrin percentages (% CDT) was also performed to carefully evaluate the data in relation to the specific detection windows of the two different biomarkers. Most of the drivers with hEtG > 30 pg/mg were males in their thirties. None of the hEtG-positives had a serum % CDT above the cutoff (≥2%). Although some researchers suggest caution until solid data are available on the possible effects of interindividual variability that may influence EtG incorporation and metabolism, hEtG is a very useful biomarker of long-term alcohol exposure that shows greater reliability than traditional blood markers. [ABSTRACT FROM AUTHOR]

Rose Bengal antigen and smooth lipopolysaccharide (s-LPS) were produced from a field strain of Brucella ceti ("homologous" antigens) and from the reference strain B. abortus S99 ("heterologous" antigens); they are currently used for the diagnosis of brucellosis in cattle, water buffaloes, sheep, goats, and pigs, as recommended in the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals of the World Organization for Animal Health (WOAH). "Homologous" and "heterologous" antigens were used in a rapid serum agglutination test (Rose Bengal test, RBT) and a competitive ELISA assay (c-ELISA) to test a panel of sera, blood, and other body fluids (cerebrospinal fluid, pericardial fluid, tracheal fluid, and aqueous humor) collected from 71 individuals belonging to five cetacean species (Stenella coeruleoalba; Tursiops truncatus; Grampus griseus; Globicephala melas; and Ziphius cavirostris), which were found stranded on the Italian coastline. Six animals were positive for Brucella spp. for bacterial isolation and/or PCR, and 55 animals were negative; for the remaining 10 animals, no PCR/isolation data were available. A total of 90 samples were tested. Results obtained from the two tests were compared in order to identify the most suitable antigen for the serological diagnosis of Brucella infection in cetaceans. The RBT performed with the "homologous" antigen showed the best results in comparison with the "heterologous" antigen: diagnostic sensitivity, specificity, and accuracy were 80.0%, 44.1%, and 46.9% for the "homologous" antigen and 80.0%, 17.0%, and 21.9% for the "heterologous" antigen. For the c-ELISA, "homologous" and "heterologous" s-LPS showed similar results (diagnostic sensitivity 66.7%, diagnostic specificity 97.3%, and diagnostic accuracy 95.0%). Therefore, the RBT using the "homologous" antigen and c-ELISA with "homologous" or "heterologous" s-LPS could be used in parallel for the detection of antibodies against Brucella spp. in cetaceans. [ABSTRACT FROM AUTHOR]

The suitability of two different low-grade kaolinitic clays and limestone locally available in Switzerland to produce a limestone calcined-clay cement at industrial scale was investigated. Firstly, the clays characteristics related to reactivity were presented. Then, the effect of calcined clays on cement performance was explored according to EN 197-5 and EN 197-1 requirements. Finally, the new cement was produced at industrial scale by using a Swiss cement plant equipped with a Lepol kiln for clay calcination and clinker production and used to manufacture 3 different concretes mixes for 3 different environmental exposure classes (EN 206). The concrete mixes were tested for mechanical and durability performance. The results show that the two calcined low-grade clays are suitable to produce a limestone calcined-clay cement classified as CEM II/C-M (Q-LL). Moreover, all the 3 concretes manufactured with the new cement meet the minimum mechanical and durability standard requirements of the corresponding exposure classes. [ABSTRACT FROM AUTHOR]

The significance of Trypanosoma equiperdum as the causative agent of dourine cannot be understated, especially given its high mortality rate among equids. International movement of equids should be subject to thorough health checks and screenings to ensure that animals are not infected with Trypanosoma equiperdum. This involves the implementation of quarantine protocols, testing procedures, and the issuance of health certificates to certify the health status of the animals. Three proteins, the peptidyl-prolyl cis-trans isomerase (A0A1G4I8N3), the GrpE protein homolog (A0A1G4I464) and the transport protein particle (TRAPP) component, putative (A0A1G4I740) (UniProt accession numbers SCU68469.1, SCU66661.1 and SCU67727.1), were identified as unique to T. equiperdum by bioinformatics analysis. The proteins were expressed as recombinant proteins and tested using an indirect ELISA and immunoblotting test with a panel of horse positive and negative sera for dourine. The diagnostic sensitivity, specificity and accuracy of the i-ELISAs were 86.7%, 53.8% and 59.0% for A0A1G4I8N3; 53.3%, 58.7% and 57.9% for A0A1G4I464; and 73.3%, 65.0% and 66.3% for A0A1G4I740, respectively, while the diagnostic sensitivity, specificity and accuracy of immunoblotting were 86.7%, 92.5% and 91.6% for A0A1G4I8N3; 46.7%, 81.3% and 75.8% for A0A1G4I464; and 80.0%, 63.8% and 66.3% for A0A1G4I740. Among the three proteins evaluated in the present work, A0A1G4I8N3 provided the best results when tested by immunoblotting; diagnostic application of this protein should be further investigated using a greater number of positive and negative sera.

Cancer vaccines present a promising avenue for treating immune checkpoint blockers (ICBs)-refractory patients, fostering immune responses to modulate the tumor microenvironment. We revisit a phase I/II trial using Tumor Antigen-Presenting Cells (TAPCells) (NCT06152367), an autologous antigen-presenting cell vaccine loaded with heat-shocked allogeneic melanoma cell lysates. Initial findings showcased TAPCells inducing lysate-specific delayed-type hypersensitivity (DTH) reactions, correlating with prolonged survival. Here, we extend our analysis over 15 years, categorizing patients into short-term (pos patients exhibit a 53.1% three-year survival compared to 16.1% in DTHneg patients. Extended remissions are observed in long-term survivors, particularly DTHpos/M1cneg patients. Younger age, stage III disease, and moderate immune events also benefit short-term survivors. Immunomarkers like increased C-type lectin domain family 2 member D on CD4+ T cells and elevated interleukin-17A were detected in long-term survivors. In contrast, toll-like receptor-4 D229G polymorphism and reduced CD32 on B cells are associated with reduced survival. TAPCells achieved stable long remissions in 35.2% of patients, especially M1cneg/DTHpos cases. Conclusions: Our study underscores the potential of vaccine-induced immune responses in melanoma, emphasizing the identification of emerging biological markers and clinical parameters for predicting long-term remission.

PurposeThe aim of this study was to compare the functional characteristics of two computer-based systems for quality control of cancer registry data through analysis of their output differences.MethodsThe study used cancer incidence data from 22 of the 49 registries of the Italian Network of Cancer Registries registered between 1986 and 2017. Two different data checking systems developed by the WHO International Agency for Research on Cancer (IARC) and the Joint Research Center (JRC) with the European Network of Cancer Registries (ENCR) and routinely used by registrars were used to check the quality of the data. The outputs generated by the two systems on the same dataset of each registry were analyzed and compared.ResultsThe study included a total of 1,305,689 cancer cases. The overall quality of the dataset was high, with 86% (81.7-94.1) microscopically verified cases and only 1.3% (0.03-3.06) cases with a diagnosis by death certificate only. The two check systems identified a low percentage of errors (JRC-ENCR 0.17% and IARC 0.003%) and about the same proportion of warnings (JRC-ENCR 2.79% and IARC 2.42%) in the dataset. Forty-two cases (2% of errors) and 7067 cases (11.5% of warnings) were identified by both systems in equivalent categories. 11.7% of warnings related to TNM staging were identified by the JRC-ENCR system only. The IARC system identified mainly incorrect combination of tumor grade and morphology (72.5% of warnings).ConclusionBoth systems apply checks on a common set of variables, but some variables are checked by only one of the systems (for example, checks on patient follow-up and tumor stage at diagnosis are included by the JRC-ENCR system only). Most errors and warnings were categorized differently by the two systems, but usually described the same issues, with warnings related to “morphology” (JRC-ENCR) and “histology” (IARC) being the most frequent. It is important to find the right balance between the need to maintain high standards of data quality and the workability of such systems in the daily routine of the cancer registry.