Search Results (18,642)

Barley grain provides an ecological niche in which fungal microorganisms can develop. The presence of mycotoxigenic genera can result in the accumulation of fungal secondary metabolites. The present study aims to survey barley grain samples collected from Northern and Central Italy by assessing: (1) the presence of fungal microorganisms using high-throughput sequencing of the internal transcribed spacer region; (2) the presence and quantification, by real-time quantitative PCR (RT-qPCR), of the main Fusarium species associated with Fusarium head blight; and (3) the presence and quantification of secondary metabolites produced by Fusarium and Alternaria using liquid chromatography–tandem mass spectrometry. Both potentially pathogenic genera (mainly within the Ascomycota division) and non-pathogenic genera (within both Ascomycota and Basidiomycota divisions) were detected. Alternaria and Cryptococcus were the dominant genera within Ascomycota and Basidiomycota divisions, respectively, in both macro-areas. Some genera, such as Aureobasidium and Cryptococcus, showed interesting negative correlations with several pathogenic genera. Two important mycotoxigenic genera (Alternaria and Fusarium) were detected, with RT-qPCR that revealed the predominance of Fusarium graminearum in the Northern macro-area. Fusarium secondary metabolites exhibited generally higher accumulation in the North, whereas Alternaria secondary metabolites were more evenly distributed across Italy, reflecting the abundance of the two genera. The co-occurrence of Fusarium and Alternaria secondary metabolites within single barley grain samples was also observed. Full article

Leishmaniasis is an emerging zoonotic disease recognized by WHO as a priority neglected tropical disease (NTD) targeted for elimination by 2030. The disease is transmitted through bites of infected female phlebotomine sand flies and presents in three main forms: cutaneous, mucocutaneous, and visceral leishmaniasis. Visceral leishmaniasis (VL) has two distinct epidemiological cycles: anthroponotic visceral leishmaniasis (AVL) cycle, caused by L. donovani and transmitted between humans, and zoonotic visceral leishmaniasis (ZVL) cycle, caused by L. infantum, with domestic dogs as the main reservoirs. ZVL represents the most severe clinical form and poses a public health risk due to a high fatality rate. In Kenya, the epidemiology and transmission dynamics of Leishmania infection among the canine population in endemic areas such as Baringo are poorly characterized. A cross-sectional epidemiological study was conducted in Baringo County, Kenya, one of the endemic foci for human visceral leishmaniasis. Blood samples were collected from 140 dogs in Rabai, Sabor, Mbechot, Kapkuikui, and Chemolingot and analyzed using Rapid Diagnostic Test (RDT) and Enzyme-linked immunosorbent assay (ELISA). Data on potential epidemiological factors and clinical status for dogs were collected using a questionnaire. Statistical analysis was performed using Pearson’s chi-square (χ²) test or Fisher’s exact test to assess associations between Leishmania seropositivity and potential risk factors. A stepwise logistic regression procedure with an entry significance level of p < 0.20 was employed to identify factors associated with leishmaniasis seroprevalence. Cohen’s kappa (κ) agreement was used to assess the diagnostic performance and accuracy of the two serological tests in the absence of a gold standard. The overall seropositivity in dogs was 7.79% and 12.14% at 95% CI for the RDT and ELISA, respectively. Multivariate analysis identified breed and clinical signs as significant factors associated with Leishmania seropositivity in dogs. Local-breed dogs were more likely to be Leishmania seropositive than mixed-breed dogs (OR = 11.92; 95% CI: 1.79–79.37; p = 0.010). Dogs without clinical signs exhibited significantly lower odds of ELISA seropositivity (OR = 0.06; 95% CI: 0.01–0.29; p < 0.001). Dogs with proper care and housing conditions had reduced risk of Leishmania seropositivity compared with poorly managed dogs. Significant association with seropositivity was observed for sex, age, origin, and reason for keeping dogs. Diagnostic evaluation showed that the RDT had an excellent specificity and Positive Predictive Value (100%), with good overall agreement with ELISA, although sensitivity was 64.7%. Hence, RDT can be a reliable alternative screening tool compared with ELISA as the reference standard. The study demonstrated the presence of Leishmania infection in the dog population in Baringo County, Kenya, a significant public health risk. Early detection and diagnosis of infections in dogs through an integrated One Health approach during routine surveillance are essential. This will facilitate treatment and/or culling of infected dogs and vector control, subsequently reducing the risk of human transmission. Hence, it will go a long way to achieving WHO’s roadmap of eliminating leishmaniasis by 2030. Full article

The remote detection of nitro compounds typically requires complex analytical procedures and expensive instrumentation, often resulting in prolonged analysis times. Fluorescence-based methods, which exploit the quenching of organic fluorophores in the presence of nitro analytes, offer a more practical and efficient alternative. Particularly promising is the use of excimer emission generated by bis-pyrene species (bis-Pyr), formed either through supramolecular interactions or covalent linkages. Regarding the latter, the integration of covalently linked bis-Pyr sensors onto polymer backbones, such as commercially available polyvinyl chloride (PVC), represents a highly attractive strategy. In this manuscript, we wish to report our recent findings on the mechanosynthesis of bis-pyrene-based PVC-supported chemosensors, and on studies of its fluorescence response towards common nitro-compound/explosive components, such as 2,4-dinitrotoluene (DNT), 2,4,6-trinitrotoluene (TNT), taggant 2,3-dimethyl-2,3-dinitrobutane (DMDNB), pentaerythritol tetranitrate (PETN) and 2,4,6-trinitrophenol (picric acid, PA). For DNT, a selective fluorescence “turn-off” response was observed with a Stern–Volmer quenching constant as high as 1.15 × 10⁴ M⁻¹. Full article

Environmental DNA (eDNA) is a highly sensitive tool widely used in biodiversity studies and for detecting organisms of biosecurity or conservation importance. However, the persistence of eDNA, even after the removal of an organism, can complicate interpretation and lead to false indications of ongoing presence. In contrast, environmental RNA (eRNA) is short-lived and may offer improved spatiotemporal precision for detecting living organisms. In this study, we evaluated the viability of environmental messenger RNA (emRNA) for targeted detection of aquatic weeds, using Amazon frogbit (Hydrocharis laevigata) as a model species. A highly sensitive qPCR assay, targeting the chloroplast rpoB transcript, was used alongside positive and negative controls to ensure workflow reliability. Our results showed that emRNA was undetectable at low density (0.56 plants per litre) and was consistently detectable at very high abundance (over eight plants per litre), although copy numbers were very low. These findings suggest that the effective detection threshold for emRNA may exceed densities typically encountered during early invasion. Consequently, the qPCR-based emRNA approaches evaluated in this study showed limited applicability for biosecurity surveillance, where reliable detection at low abundance is essential. Future research should focus on improving detection sensitivity, including eRNA enrichment. Full article

Implant-associated infections remain a major challenge in bone-related biomedical applications, where bacterial colonization and biofilm formation can compromise tissue integration and clinical performance. This study reports the mechanochemical synthesis, physicochemical characterization, antimicrobial activity, antibiofilm performance, and short-term fibroblast cytocompatibility of a TiO₂-containing bovine-derived biogenic hydroxyapatite ceramic composite (BHAp-TiO₂). The composite was prepared by high-energy mechanical milling using 10 wt% TiO₂ and characterized by X-ray diffraction, Rietveld refinement, Raman spectroscopy, Fourier-transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. XRD/Rietveld analysis identified a multiphase ceramic composite composed of hydroxyapatite, whitlockite, and rutile TiO₂, with no evidence of Ti⁴⁺ substitution into the hydroxyapatite lattice or detectable anatase within the XRD/Rietveld detection limit. SEM-EDS confirmed the granular agglomerated morphology of the powders and the elemental presence of Ti in BHAp-TiO₂. Compared with pristine BHAp, BHAp-TiO₂ produced a concentration-dependent reduction in AlamarBlue^®-derived bacterial metabolic activity against five clinically relevant planktonic strains. At 200 µg/mL, residual metabolic activity decreased to 10.90–32.90%, depending on the bacterial species, with the strongest response observed for Escherichia coli. In crystal violet assays, BHAp-TiO₂ markedly inhibited Pseudomonas aeruginosa biofilm biomass, reaching 91.9 ± 3.4% inhibition at 200 µg/mL. In NIH/3T3 fibroblasts, BHAp-TiO₂ preserved short-term cytocompatibility after 24 h of direct exposure within the 0.1–100 µg/mL range, with MTT- and AlamarBlue^®-derived responses remaining close to or above the 80% cytotoxicity limit. Overall, BHAp-TiO₂ is best interpreted as a rutile TiO₂-containing biogenic calcium phosphate ceramic composite with enhanced antimicrobial and antibiofilm performance while maintaining short-term fibroblast cytocompatibility under the evaluated conditions. Full article

Insufficient vascularization remains a major limitation in tissue engineering, restricting the survival and maturation of larger bioengineered constructs. While candidate hydrogels are commonly characterized with regard to physicochemical properties, gelation behavior, mechanical performance, and cytocompatibility, simple functional assays that assess their capacity to support vascular sprouting are less frequently integrated into early-stage biomaterial evaluation. Here, we investigated the established ex vivo aortic ring assay (ARA) as an exploratory functional approach for the initial comparison of selected hydrogel formulations. Murine aortic rings were embedded in collagen I (Col I), alginate (Alg), or gelatin methacryloyl (GelMA) and cultured under control conditions or with vascular endothelial growth factor A (VEGF-A) stimulation. These hydrogels were intentionally selected as a proof-of-concept panel of representative, non-equivalent material classes with distinct expected cell-interactive properties. After five days, Col I supported robust capillary-like outgrowth that was further enhanced by VEGF-A, whereas the tested GelMA formulation supported only limited cellular migration and the tested unmodified Alg formulation showed no detectable sprouting under the conditions examined. Cluster of differentiation 31 (CD31) immunostaining supported the presence of an endothelial component within the Col I-supported sprouting structures. These findings demonstrate that the ARA can detect pronounced formulation-dependent differences among the specific hydrogels tested using straightforward morphological and immunostaining readouts. Within the scope of the formulations tested, these findings support the ARA as a complementary functional readout alongside conventional biomaterial characterization before more complex tissue engineering or biofabrication studies are performed. Full article

Healthcare workers (HCWs) and medical providers are regularly exposed to a wide range of infectious diseases in the course of their duties. This occupational risk is significant in the context of highly contagious infections such as measles, which can spread rapidly in healthcare institutions. This study aimed to assess the seroprevalence of protective anti-measles and anti-rubella IgG antibodies and factors influencing immunity among HCWs across all 28 districts in Bulgaria. Methods: A total of 1854 individuals, including 540 (29%) doctors and 1314 (71%) medical specialists from all 28 districts in Bulgaria, were screened for protective immunity against measles and rubella. Serum samples were analyzed for the presence of protective IgG antibodies by ELISA (Enzyme-Linked Immunosorbent Assay) using commercial EUROIMMUN Anti-Measles/Anti-Rubella Virus IgG ELISA kits. According to the manufacturer’s instructions, the results were interpreted semi-quantitatively as positive, negative, and equivocal. Results: Anti-measles protective IgG antibodies were detected in 86.1% of participants for measles and in 95.2% for rubella. Our study found a statistically significant difference when analyzing demographic factors (age) for measles: the samples from older participants (over 50-year-old persons) were characterized by higher immunity levels; staff working in non-infectious-disease wards (n+ = 95.7%) were more frequently positive for rubella IgG antibodies compared to staff employed in infectious wards; and region locations varied for both groups. Gender and professional roles were not significant predictors. Conclusions: HCWs are at a greater risk of contracting infections than the general public because they have contact with sick patients or infectious material. Their vaccination or IgG immunity status must be strictly monitored, especially to limit the spread of nosocomial infections in hospital settings. Full article

Historical art materials have always been a starting point for better understanding artists’ artwork composition, the long-term stability of used formulations and for designing appropriate strategies for conservation. It is important to consider how artists have always been involved over the centuries in the selection and preparation of the materials they used. This changed with the advent of industrial formulations, which were produced and marketed on a large scale, leading to significant changes from the 19th century onwards. In this study, the study of a set of 30 green paint tubes from the Munch Museum collection (Oslo) is presented. The selected investigation method allows for a progressive acquisition of complementary and increasingly detailed information through three multi-analytical steps, involving elemental, spectroscopic and chromatographic techniques, with the aim of characterising these materials in their various inorganic and organic components. Results achieved allow the identification of green pigments’ chemical composition and classification of them in distinct groups, presence of inert additives and characterisation of organic binding media. Degradation processes were also highlighted through the detection of metal soap formations. Full article

Background: The fibrovascular stalk is characteristic of papillary bladder tumors and may be visualized on MRI as the inchworm sign. Contrast-enhanced ultrasound (CEUS) may depict the same structure dynamically by demonstrating pedicle perfusion before tumor enhancement. We hypothesized that CEUS would be more sensitive than the MRI inchworm sign for detecting a histologically confirmed fibrovascular stalk and that stalk presence would be associated with non-muscle-invasive bladder cancer without completely excluding detrusor muscle invasion. Methods: This retrospective study assessed 80 consecutive patients with one focal bladder lesion; 69 patients with technically adequate imaging and an adequate TURBT reference standard were included in the final paired analysis. Dedicated retrospective rereads of anonymized stored CEUS cine loops and complete mpMRI datasets were independently performed by two experienced readers per modality, who were blinded to the other imaging modality, histopathology, and each other’s assessments. A blinded pathologist assessed fibrovascular stalk presence and detrusor muscle invasion. Paired performance was evaluated using exact McNemar testing. Results: Histopathology identified a stalk in 48 lesions. CEUS detected 46 of 48 stalks, and MRI detected 36, corresponding to sensitivities of 95.8% and 75.0%, specificities of 90.5% and 81.0%, and accuracies of 94.2% and 76.8%, respectively. Interobserver agreement was 94.2% for both signs, with almost-perfect agreement for the CEUS vascular stalk sign (κ = 0.86) and the MRI inchworm sign (κ = 0.88). For MIBC detection, overall CEUS impression and VI-RADS 4–5 showed sensitivities of 66.7% and 87.5% (paired p = 0.063), specificities of 88.9% and 68.9% (paired p = 0.004), and accuracies of 81.2% and 75.4% (paired p = 0.424), respectively. Conclusions: However, the two signs are not operationally equivalent because the MRI inchworm sign additionally requires preservation of the underlying muscular layer, a criterion that may have contributed to its lower observed sensitivity. The stalk strongly favored non-muscle-invasive disease but did not exclude detrusor invasion. CEUS provides complementary perfusion information, whereas MRI-based VI-RADS remains central to local staging. Full article

The presence of mistletoe in pine stands has expanded in recent decades, currently threatening Mediterranean forests. Mistletoe outbreaks can make the host trees more vulnerable to intense droughts, which are expected to increase due to climate change. We use multispectral (MS) and LiDAR UAV-derived data to classify Viscum album L. ssp. austriacum infestation levels at individual tree level in Scots pine (Pinus sylvestris L.) forests. First, spectral and structural differences between three infestation levels were assessed employing Kruskal–Wallis test with Benjamini–Hochberg correction and post hoc Dunn’s test for individual tree crowns. Second, classification algorithms were applied to evaluate infestation levels at the individual tree scale by combining UAV-derived datasets. The outcomes revealed significant differences between infestation levels in canopy cover and height based on LiDAR-derived metrics. Significant changes in vegetation vigor were also found through spectral and textural metrics. The highest classification accuracy (0.87) was achieved by the spectral metrics CIRE and NDVI in an SVM model, outperforming models based on the combination of sensor metrics (0.82) or solely LiDAR variables (0.69, MLR). This approach demonstrates their potential for detecting and characterizing morphological changes in up to three levels of mistletoe infestation at individual trees in Mediterranean Scots pine forests, lending support to forest management monitoring. Full article

Background: Human respiratory exposure to microplastics (MPs) has recently been demonstrated, but their distribution within thoracic tissues and association with lung malignancy remain poorly characterised. We investigated the presence, burden, morphology, and spectroscopic characteristics of MPs in thoracic tissue specimens and their relationship with lung malignancy. Methods: In this prospective observational study, 50 adults undergoing diagnostic thoracic surgical procedures for suspected lung malignancy were enrolled. Lung, pleural, and mediastinal lymph node specimens underwent contamination-controlled processing, microscopy, and representative particle analysis by confocal micro-Raman spectroscopy. Associations between MPs and clinicopathological variables were analysed. Results: Sixty-two morphologically identified microplastic particles were detected in 50 specimens, with MPs detected in 32 patients (64%). Fragments accounted for 97% of particles, with a median burden of one particle per specimen (IQR, 2). MP detection was significantly more frequent in patients with primary lung malignancy than in patients without primary lung malignancy, including those with metastatic malignancy and benign disease (81.8% vs. 50.0%, p = 0.020), who also exhibited a higher tissue MP burden (p = 0.024). Primary lung malignancy remained independently associated with MP detection after adjustment for age and smoking exposure (OR 4.09, 95% CI 1.06–15.79; p = 0.041). Particle morphology varied by biopsy site, with significantly smaller particles identified in mediastinal lymph nodes (p = 0.006). Synthetic anthropogenic spectral signatures were identified in 71% of particles. Conclusions: MPs are frequently present in human thoracic tissues and are associated with primary lung malignancy. Compartment-specific particle characteristics suggest distinct patterns of respiratory MP deposition and retention, supporting multicentre mechanistic studies. Full article

This study presents an integrated surface diagnostic investigation and a diagnostic-driven restoration of the marble sculpture Real Infante Carlo Tito di Borbone (1775), attributed to Giuseppe Sanmartino and preserved at the Royal Palace of Caserta (Italy). Despite the artistic relevance of the sculptor, scientific investigations of his works remain limited. The aim of this work is to characterize surface coatings, identify alteration processes, and support conservation strategies through a multi-analytical approach. The results reveal that the sculpture is composed of calcitic marble covered by an altered organic surface film. Spectroscopic and chromatographic data indicate the presence of lipidic compounds consistent with natural wax, likely associated with previous conservation treatments. An epoxy-based adhesive was also identified, suggesting past restoration interventions. Microbiological analyses detected fungal and bacterial species potentially involved in surface alteration processes. The integration of analytical data enabled the development of a targeted restoration strategy. Cleaning procedures were defined based on the chemical nature of the surface coatings, while structurally stable previous restorations were preserved. A biocide treatment was applied to mitigate biological colonization. This study highlights the importance of surface-oriented diagnostics in understanding coating materials and guiding conservation interventions, providing a methodological framework applicable to marble artworks. Full article

Winter survival of crops is influenced by multiple environmental stress factors, including low temperature, excessive soil moisture, and pathogen pressure. With climate change, the role of flooding during cold acclimation has become increasingly important. This study examined the effect of cold flooding on frost tolerance of two winter rye (Secale cereale L.) lines differing in resistance to snow mould. Plants were subjected to cold acclimation for three weeks at 4 °C, followed by ten days of flooding at the same cold temperature. Cold flooding improved frost tolerance only in the snow mould-resistant line (343), which maintained the presence of a 50 kDa β-1,3-glucanase isoform and exhibited a flooding-induced decrease in cytosolic sucrose synthase (SuS) activity in leaves, while maintaining higher SuS activity in roots. In contrast, the less resistant line (620) lacked the 50 kDa isoform in roots and showed substantially lower SuS activity than line 343; however, leaf SuS activity increased during prolonged flooding to levels comparable with those of line 343. The 35 kDa isoform was no longer detectable in the leaves of either line after one or ten days of flooding, whereas in the roots of both lines, its accumulation was maintained after one day but was abolished after ten days of flooding. Additionally, line 620 exhibited prolonged flooding-induced upregulation of glu-8 and TLP3 genes, which, in the absence of protein accumulation, suggests post-transcriptional or translational constraints associated with prolonged stress. These findings demonstrate that higher frost tolerance under cold flooding involves the accumulation of specific pathogenesis-related (PR) proteins, maintaining a high reduced and oxidised glutathione ratio and metabolic adjustment of sucrose synthesis. Furthermore, cold acclimation alone did not differentiate frost tolerance between rye genotypes differing in snow mould resistance, whereas flooding under low-temperature conditions enhanced freezing tolerance only in the resistant line, indicating that flooding may function as a positive signal involved in frost tolerance induction. Full article

Planted and unplanted recirculating constructed wetland mesocosms maintained under non-aerated (reducing) and aerated (oxidizing) conditions were compared in their ability to remove 1 mg/L As(V) from simulated domestic wastewater over a 33-week period. Each week, one day prior to the addition of arsenic, non-aerated mesocosms removed 70% of the nitrate and 95% of the organic carbon likely through denitrification while aerated mesocosms removed organic carbon only (97%). Upon arsenic addition and treatment for the remainder of the week, non-aerated mesocosms removed more arsenic (72%) than aerated mesocosms (43%). For the first time in the literature, we estimated the proportion of arsenic found in biofilms detached from gravel and incorporated this aspect into mass balance calculations. We discovered that biofilms from non-aerated mesocosms contained a higher proportion of arsenic (28%) than biofilms from aerated mesocosms (5%). Plants from both types of mesocosms removed a negligible amount of arsenic (<0.41%). Through X-ray fluorescence (XRF) analysis, it was discovered that biofilms from non-aerated mesocosms had the highest iron concentrations (35,000 mg/kg), and this iron was identified as pyrite (2–15 wt%) through X-ray diffraction (XRD) analysis. The iron concentration of aerated biofilms was no different from that found in clean gravels (17,500 mg/kg), and no iron minerals were detectable in these samples using XRD analysis. Although some of the arsenic removal in both types of wetlands was thought to be attributable to adsorption onto substrates and biofilms, as well as uptake into biofilms, removal of arsenic in the non-aerated mesocosms was likely enhanced by the presence of pyrite because pyrite can adsorb arsenic directly or indirectly through the formation of iron oxides that can adsorb or coprecipitate with the iron oxides. Full article

The incidence of fungal infections, particularly those caused by Candida species, has significantly risen among immunocompromised patients in recent years. Aim: This study evaluated the species distribution, antifungal susceptibility patterns, and virulence gene profiles of Candida species isolated from critically ill ICU patients while comparing the diagnostic performance of conventional phenotypic isolation media with molecular genotypic identification methods. Materials and Methods: A total of 211 clinical specimens were obtained from 200 hospitalized patients at Menoufia University Hospitals, Egypt. Candida species were phenotypically identified using cornmeal agar supplemented with Tween 80 (CTA) and CHROMagar^TM Candida (CMA) and were molecularly confirmed at the species level by sequencing PCR-amplified ITS regions. Antifungal susceptibility profiles to fluconazole, voriconazole, and amphotericin B were evaluated using the disk diffusion method. Furthermore, real-time PCR was deployed to detect genes encoding adherence factors and Secreted Aspartyl Proteinase. Results: Candida species were recovered from 45 out of 211 clinical specimens. The most frequently isolated species was Candida albicans (15/45, 33.3%), followed by C. parapsilosis (9/45, 20.0%), C. glabrata (7/45, 15.5%), C. tropicalis (7/45, 15.5%), C. krusei (4/45, 8.8%), and C. famata (3/45, 6.7%). Among the recovered Candida isolates, the percentage rates of the ALS1, ALS3, HWP1, and SAP4 genes were 40%, 42.22%, 44.4%, and 37.78%, respectively. HWP1 was the most frequently detected pathogenicity factor, whereas SAP4 was the least prevalent. Conclusions: In the studied immunocompromised cohort, non-albicans Candida species constituted a substantial proportion of clinical isolates. Notable antifungal resistance patterns were also observed, emphasizing the value of integrated phenotypic and genotypic identification for tailored management. Furthermore, virulence-associated genes (ALS1, ALS3, HWP1, and SAP4) were detected in isolates of several Candida species, highlighting their presence in clinical strains. Full article