Search Results (50,676)

Background/Objective: Timely identification of bloodstream pathogens is critical for sepsis management; however, PCR inhibitors such as sodium polyanetholesulfonate (SPS) in blood culture broth compromise nucleic acid recovery and long read sequencing. We assessed whether coupling a benzyl alcohol SPS-removal step to the fully automated LabTurbo AIO extractor improves Oxford Nanopore-based pathogen detection. Methods: Thirteen positive blood culture broths were pre-treated with benzyl alcohol and divided: half volumes were purified on the LabTurbo AIO; paired aliquots underwent manual QIAamp extraction. DNA purity was evaluated by NanoDrop and Qubit. Barcoded libraries were sequenced on MinION R9.4.1 flow cells for 6 h. Results: Automated eluates showed a median A₂₆₀/A₂₈₀ of 1.92 and A₂₆₀/A₂₃₀ of 1.96, versus 1.80 and 1.48 for manual extracts. The automated workflow generated 1.69 × 10⁶ total reads compared with 3.9 × 10⁵ reads for manual extraction. The median N50 read length increased from 5.9 kb to 8.7 kb, and the median proportion of reads classified to species increased from 62% to 84%. The hands-on time was <5 min and the sample-to-answer turnaround was <8 h, compared with >9 h and 90 min for the manual protocol, respectively. Conclusions: Benzyl alcohol SPS removal integrated into the LabTurbo AIO extractor yielded purer, longer, and higher read counts, enhancing nanopore sequencing depth and accuracy while compressing diagnostic turnaround to a single working day. This represents a practical advance for rapid blood culture pathogen identification in critical care settings. Full article

Objective: This study aimed to validate the clinical utility of a salivary molecular platform (Oral Predict^®) for periodontal pathogen detection across preventive, therapeutic, and maintenance settings. Methods: A longitudinal randomized study was conducted involving 78 adults who provided saliva samples at baseline, one month, and three months after professional dental hygiene. Participants were randomized into two groups: control group (n = 39) and probiotic group with Oral Predict^® probiotic supplementation (n = 39). Crude saliva was processed directly without nucleic acid extraction and analyzed by multiplex real-time PCR using either the compact Real-time PCR system or standard thermocyclers. Results: At baseline, Fusobacterium nucleatum was the most prevalent pathogen (84.6%), followed by Tannerella forsythia (53.8%) and Porphyromonas gingivalis (46.2%). The Total Pathogen Burden Score (TPBS) showed progressive increases with age, smoking, and poor oral hygiene, and was significantly higher in participants with gingival bleeding. Among individual pathogens, no significant associations were observed with periodontitis staging or grading. Professional hygiene induced mean reductions of 1–2 logs across all pathogens, with TPBS decreasing from 8.7 ± 3.2 to 4.1 ± 2.8 (p < 0.001). At three months, 69.2% of the control group experienced bacterial rebound, whereas 85% of probiotic users sustained or improved bacterial reductions. Conclusions: Salivary molecular testing provides a robust, non-invasive approach for periodontal pathogen detection, treatment monitoring, and long-term maintenance assessment. The flexibility of the Oral Predict^® platform across point-of-care and laboratory settings, combined with automated interpretation, supports integration into preventive protocols and personalized periodontal care. These findings demonstrate the potential of saliva-based molecular diagnostics to shift periodontal management from reactive to predictive and precision-based strategies. Full article

Congenital hearing loss, frequently resulting from defective hair cells, remains poorly understood due to the incomplete identification of key pathogenic genes. Oncomodulin (OCM) is a kind of calcium-binding protein (CaBP) that regulates diverse cellular processes and is thought to play crucial roles in auditory function. In teleost fish, parvalbumin 8 (pvalb8) and parvalbumin 9 (pvalb9) belong to the oncomodulin lineage and are highly expressed in hair cells. In this study, we first reported the oncomodulin lineage function in fish and identified pvalb8 as an essential regulator of hair cell development. Single-cell RNA sequencing (scRNA-seq) and whole-mount in situ hybridization (WISH) revealed that pvalb8 is highly and specifically expressed in supporting cells and hair cells. Functional loss of pvalb8, achieved via CRISPR/Cas9 knockout or morpholino knockdown, resulted in reduced neuromast size and a significant decrease in neuromast hair cell number, leading to auditory behavioral deficits. In addition, pvalb9 mutants exhibited hair cell defects similar to those observed in pvalb8 mutants, including a significant reduction in hair cell number. Moreover, pvalb8 loss strongly inhibited the proliferation of supporting cells, which likely accounts for the reduced number of differentiated hair cells. The expression levels of Wnt target genes, axin2, ccnd1, and myca, were all significantly reduced in pvalb8 mutants compared to control zebrafish, while activation of the Wnt signaling pathway rescued the hair cell loss observed in pvalb8 mutants, indicating that pvalb8 promotes hair cell development via Wnt-dependent proliferative signaling. These findings highlight pvalb8 as a critical factor in the regulation of auditory hair cell formation and function in zebrafish, offering new insights into the role of oncomodulin lineage in sensory cell development. Full article

Restrictions on adding antibiotics to animal diets have posed challenges in managing gut pathogens, emphasizing the significance of effective non-antibiotic growth promoters to maintain animal health and productivity. This study assessed the efficacy of hydroxamic acid (HA), derived from local maize varieties, as a non-antibiotic growth promoter in broilers. Among 10 different maize varieties, the Azam variety yielded the highest HA concentration (35 ± 7 μg/g of roots), as quantified by high-performance liquid chromatography (HPLC). In vitro antimicrobial assays demonstrated the lowest minimum inhibitory concentration (MIC) of 0.022 mg for Azam-derived HA against pathogenic E. coli. To further assess in vivo efficacy, 108 birds were allocated at random to six treatment groups. The treatments include birds fed a basal diet without an E. coli challenge (negative control); an antibiotic-treated group challenged with E. coli and treated with enrofloxacin at a dosage of 5 milligrams (mg) per kilogram (kg), administered orally once daily from day 5 post-infection (dpi) for 7 consecutive days (standard); broilers challenged with E. coli and supplemented with a basal diet with HA at concentrations of 1, 10, or 100 mg/kg of feed from 5 dpi for one week (HA 1 mg, HA 10 mg, and HA 100 mg, respectively); and broilers challenged with E. coli without enrofloxacin/HA (positive control). The results demonstrated that birds fed a diet supplemented with the HA-100 mg improved the body weight (BW) and feed conversion ratio (FCR) compared to the positive control group. There were no significant differences (p > 0.05) observed for BW and FCR observed for the broilers fed on the standard and HA 100 mg groups. The addition of HA at 100 mg improved (p < 0.05) the hemoglobin (Hb) and packed cell volume (PCV) and reduced (p < 0.05) levels of malondialdehyde (MDA) compared to positive control group. A significantly low carcass weight (p < 0.05) was shown for positive control birds compared to other groups. Our findings indicate that maize-derived HA presents a phytogenic alternative to antibiotics by controlling enteric pathogens and improving health and performance affected by E. coli infection in broilers. Full article

Antimicrobial resistance (AMR) remains a public health problem in European Union countries, and elderly patients represent a vulnerable category due to aging and its associated risk factors. In this research, we investigated the trend of the antimicrobial resistance pattern of Gram-negative pathogens isolated in samples collected from elderly patients (over 65 years) hospitalized in the intensive care unit (ICU) between 2022 and 2024. A total of 2510 samples, including blood, tracheal aspirate, sputum, urine, pus/wound swabs, exudates, intravascular catheters, cerebrospinal fluid, and sterile fluids, were collected from 1864 elderly patients. Almost two-thirds of clinical specimens were harvested from the respiratory tract. The most frequently reported pathogens from 3094 Gram-negative bacterial isolates were Klebsiella spp., Acinetobacter spp., and Escherichia coli. During the studied period, almost 40% of all the Klebsiella spp. strains were multidrug-resistant (MDR)/extensively drug-resistant (XDR), with a significant increase in the resistance to cephalosporins (p ≤ 0.05), fluoroquinolones (p ≤ 0.05), and carbapenems (imipenem—(p ≤ 0.05), ertapenem—p < 0.001). The proportion of carbapenem-non susceptible Klebsiella spp. rose from 24.41% in 2023 to 32.48% in 2024, p = 0.01. Two-thirds of Acinetobacter spp. isolates were MDR/XDR, and over 80% were carbapenem-non-susceptible in 2023–2024. The results draw attention to the need to quickly adopt measures to reduce the prevalence, limit the transmission of MDR/XDR pathogens, and improve therapeutic protocols in this age category. Full article

Chronic wounds, such as diabetic ulcers, remain a significant clinical challenge due to high infection risk and delayed healing. This study presents a comprehensive evaluation of a novel wound dressing incorporating curcumin-functionalized silver–zinc oxide (Ag-ZnO) nanoparticles. The formulation was rationally designed based on molecular docking simulations that identified curcumin as a high-affinity ligand for Staphylococcus aureus Protein A. The synthesized nanoparticles demonstrated potent, broad-spectrum antibacterial activity, achieving complete inhibition of multidrug-resistant pathogens, including MRSA, within 60 s. A critical comparative assessment, incorporating an unloaded Ag-ZnO nanoparticle control group, was conducted in both a rabbit wound model and a randomized clinical trial (n = 75 patients). This design confirmed that the enhanced wound-healing efficacy is specifically attributable to the synergistic effect of curcumin combined with the nanoparticles. The curcumin-loaded Ag-ZnO treatment group showed a statistically significant reduction in healing time compared to both standard care and unloaded nanoparticle controls (e.g., medium wounds: 19.6 days vs. 90.6, p < 0.001). These findings demonstrate that curcumin-functionalized Ag-ZnO nanoparticles offer a safe and highly effective therapeutic strategy, providing robust antibacterial action and significantly accelerated wound healing. Full article

The colon plays a crucial role in energy metabolism and intestinal health of ruminants during various physiological stages. Plateau ruminants have long been subjected to extreme environments characterized by hypoxia, cold, and nutritional scarcity, which makes their dependence on energy metabolism particularly pronounced. However, existing research on the regulatory effects of dietary energy levels on the colonic function of plateau ruminants is still quite limited. This study involved 60 healthy male Pamir yaks with consistent body conditions, which were randomly divided into three groups: a low-energy diet group (YG, Neg 1.53 MJ/kg), a medium-energy diet group (QG, Neg 2.12 MJ/kg), and a high-energy diet group (RG, Neg 2.69 MJ/kg). Each yak was provided with 5 kg of mixed feed daily over a 170-day feeding trial. The results indicated that a high-energy diet enhanced growth performance in yaks (p < 0.05). However, it also induced local colonic inflammation, decreased levels of immune factors (IgA, IgG, and IL-10), and increased the abundance of potentially pathogenic bacteria, such as Klebsiella and Campylobacter (p < 0.05). Conversely, a medium-energy diet fostered the proliferation of beneficial bacteria such as Bradymonadales, Parabacteroides, and Mogibacterium (p < 0.05), and preserved immune homeostasis. Additionally, multi-omics analysis revealed that the QG group was significantly enriched in key metabolic pathways, including pyruvate metabolism and glycine, serine, and threonine metabolism and panto-thenate and CoA biosynthesis pathways, among others (p < 0.05), demonstrating a synergistic regulatory effect among the microbiome, metabolism, and host. In summary, a moderate-energy diet can promote the proliferation of beneficial bacteria in the extreme environment of the plateau. By regulating pathways such as Amino acid, Nucleotide, and Lipid metabolism, it coordinates the expression of key host genes and metabolite levels, effectively balancing immune signals and energy metabolism. This interaction establishes a beneficial microbial-metabolism-host pattern that supports colon health. Full article

Background/Objectives: In response to the COVID-19 pandemic, we developed a vaccine candidate against SARS-CoV-2. Spike Ferritin Nanoparticle (SpFN) comprises 24 identical prefusion-stabilized spike proteins anchored to a self-assembled nanoparticle. Organized along the three-fold axis of the ferritin particle, eight SARS-CoV-2 spike trimers are presented per nanoparticle. Methods: Here, we describe the CGMP processes for manufacturing SpFN using transient transfection of Expi293F cells. Results: The final yield of SpFN was ~10 mg per liter of media supernatant. The resulting protein is stable in cold storage for two years at −20 °C, as well as for a month at room temperature, and can withstand multiple freeze/thaw cycles. SpFN material produced using the CGMP protocols adjuvanted with Army Liposomal Formulation-QS-21 (ALFQ) elicited potent neutralizing antibodies against WA-1, Alpha, Beta, and Delta variants in mice as measured by a pseudovirus neutralization assay. Conclusions: This work demonstrates rapid development and scaled-up production of clinical-grade SARS-CoV-2 vaccine protein material, allowing permissive storage and transport conditions, and serves as a framework for recombinant protein production for future emergent pathogens. Full article

Neurodegenerative diseases (NDDs) are a group of progressive diseases affecting neuronal cells in specific areas of the brain, causing cognitive decline and movement impairment. Nowadays, NDDs play a significant role in the global burden of disease, and their incidence is increasing, particularly due to population aging. NDD onset is multi-factorial; based on the current knowledge, genetic, environmental, and cellular factors are believed to contribute to their occurrence and progression. Taking into account that at an early stage, the symptoms are not clearly defined, and diagnosis may be delayed, the development of innovative and non-invasive methodological approaches for early diagnosis of NDDs is strategic for timely and tailored disease management, as well as for the overall improvement of patients’ quality of life. The present review aims to provide, in the first part, an overview based on the current level of knowledge on the environmental risk factors that can explicate a role in the onset of the most common NDDs and on the main pathogenic mechanisms involved in disease initiation and progression. The second part aims to define the current state of the art regarding the significance of Volatile Organic Compounds (VOCs) in the volatome of different human biological matrices (exhaled breath, feces, and skin sebum) as candidate biomarkers of specific NDDs, with the aim of developing non-invasive diagnostic approaches for the early diagnosis and personalized management of the patients. A critical synthesis and discussion on the applied methodological approaches and on the relevant outcomes obtained across the studies is reported. Full article

Nontuberculous mycobacteria are opportunistic pathogens increasingly associated with human disease. Within this group, the Mycobacterium avium complex (MAC), which includes M. avium, M. intracellulare and M. intracellulare subsp. chimaera, is the most frequent cause of infection. The increase in MAC cases worldwide has made it crucial to understand their population structure, clinical relevance and resistance mechanisms. Recent advances in whole-genome sequencing (WGS) and molecular approaches have improved the knowledge of taxonomy, population structure and genetic diversity, while also enabling the investigation of transmission and epidemiology. Clinically, MAC most often causes chronic pulmonary disease, but extrapulmonary forms, including disseminated disease, also occur. Presentation can vary by infecting species, while host factors such as pre-existing lung disease or immunosuppression further increase the risk. Treatment outcomes remain less favourable than desired, in part due to antimicrobial resistance involving de novo-acquired mutations. Pathogenesis is also influenced by interactions between MAC and host cells, including mechanisms of immune evasion and inflammatory modulation. In addition, emerging evidence suggests that gut–lung axis dysbiosis may influence susceptibility to MAC infection. This review outlines current knowledge on the population structure, clinical significance, resistance and host–pathogen interactions of MAC. Full article

The interaction between microorganisms in the dental microfilm (plaque) at the gingival margin, the immune system, and the brain is vital for gingival health. The brain constantly receives information regarding microbial composition and inflammation status through afferent nerves and the bloodstream. It modulates immune responses via efferent nerves and hormonal systems to maintain homeostasis. This relationship determines whether the gingiva remains healthy or develops into gingivitis (non-destructive inflammation) or periodontitis (a destructive condition), collectively referred to as periodontal disease. Factors associated with severe periodontitis heighten the responsiveness of this homeostatic system, diminishing the adaptive immune system’s defence against symbiotic microorganisms with pathogenic properties, known as pathobionts. This leads to excessive innate immune system activation, effectively preventing infection but damaging the periodontium. Consequently, investigating the microbiota–brain axis is vital for understanding its impact on periodontal health and disease. Herein, we examine recent advancements in how the defence against pathobionts is organised within the brain, and how it regulates and adapts the pro-inflammatory and anti-inflammatory immune balance, controlling microbiota composition. It also discussed how pathobionts and emotional stress can trigger neurodegenerative diseases, and how inadequate coping strategies for managing daily stress and shift work can disrupt brain circuits linked to immune regulation, weakening the adaptive immune response against pathobionts. Full article

Helicobacter pylori infects the human stomach and causes various gastrointestinal diseases. Saucerneol D is a type of lignan, which is a polyphenol compound that exists naturally in plants, and it is abundant in flaxseed, sesame seeds, whole grains, vegetables, and fruits. Saucerneol D is found in Saurus chinensis extract and has been reported to exert a variety of effects, such as antioxidant and anti-inflammatory abilities. However, its antibacterial effect against H. pylori has not been reported; therefore, we analyzed the effect of saucerneol D on H. pylori in the present study. Changes in the expression of pathogenic factors and gene transcription in H. pylori were observed after treatment with saucerneol D using Western blotting and RT-PCR. It was confirmed that saucerneol D suppressed the growth of H. pylori by decreasing the expression of the genes dnaN and polA, which are required for bacterial replication. Saucerneol D also reduced the secretion of the major pathogenic toxin protein, CagA, by downregulating the expression of type IV secretion system-composing proteins. Furthermore, saucerneol D reduced ammonia production by inhibiting the expression of urease proteins, which are essential for the survival of H. pylori in the acidic gastric environment. Additionally, saucerneol D decreased the expression of flaB, potentially reducing motility. Finally, it was confirmed that the expression of the sabA gene, associated with cell adhesion, was reduced. These results suggest that saucerneol D inhibits the growth of H. pylori and the expression of several pathogenic factors, indicating that saucerneol D has an antimicrobial effect against H. pylori. Full article

Background: Odontogenic abscesses may significantly affect maternal health during pregnancy. Aim: This study analyzes three cases of pregnant patients with odontogenic infections, comparing them to a control group of non-pregnant women, and reviews recent literature. Materials and Methods: Between January 2020 and April 2025, 3 pregnant and 70 non-pregnant women with odontogenic abscesses were treated. Clinical presentation, pathogens, therapy, and outcomes were compared. Results: Severe sequelae, such as rapid abscess spread and systemic inflammation, were more frequent in pregnant women, though not statistically significant (p = 0.068). Pregnant patients also tended toward prolonged intubation (p = 0.194) and targeted antibiotic use (p = 0.133). Antibiotic selection was based on gestational age, with beta-lactams preferred. Surgical interventions were more extensive, often involving multiple neck spaces. Hospitalization was longer (≥4 days in most cases) due to maternal–fetal monitoring. Conclusions: Odontogenic abscesses in pregnancy require individualized management and gestation-adjusted antibiotic therapy. Full article

Xylella fastidiosa was first detected in Portugal in 2019 in Lavandula dentata. In response, the national plant health authorities promptly established a Demarcated Zone in the affected area and implemented a series of eradication and control measures, including the systematic removal and destruction of infected and host plants. This study analyzes the economic and operational impacts of these eradication efforts in the northern region of Portugal, with a focus on Demarcated Zones such as the Porto Metropolitan Area, Sabrosa, Alijó, Baião, Mirandela, Mirandela II, and Bougado between 2019 and June 2023. During this period, about 412,500 plants were uprooted. The majority were Pteridium aquilinum (bracken fern), with 360,324 individuals (87.3%), reflecting its wide distribution and the large area affected. Olea europaea (olive tree) was the second most common species removed, with 7024 plants (1.7%), highlighting its economic relevance. Other notable species included Quercus robur (3511; 0.85%), Pelargonium graveolens (3509; 0.85%), and Rosa spp. (1106; 0.27%). Overall, destruction costs were estimated at about EUR 1.04 million, with replanting costs of roughly EUR 6.81 million. In parallel, prospection activities—conducted to detect early signs of infection and monitor disease spread—generated expenses of roughly EUR 5.94 million. While prospecting represents a significant financial investment, the results show that it is considerably more cost-effective than large-scale eradication. Prospection enables early detection and containment, preventing the widespread destruction of vegetation and minimizing disruption to agricultural production, biodiversity, and local communities. Importantly, our findings reveal a sharp decline in confirmed cases in the initial outbreak area—the Porto Demarcated Zone—from 124 cases in 2019 to just 5 in 2023, indicating the effectiveness of the eradication and monitoring measures implemented. However, the presence of 20 active Demarcated Zones across the country as of 2023 highlights the continued risk of spread and the need for sustained vigilance. The complexity of managing Xylella fastidiosa across ecologically and logistically diverse territories justifies the high costs associated with surveillance and targeted interventions. This study reinforces the strategic value of prospection as a proactive and sustainable tool for plant health management. Effective surveillance requires the integration of advanced methodologies aligned with the phenological stages of host plants and the biological cycle of vectors. Targeting high-risk locations, optimizing sample numbers, ensuring diagnostic accuracy, and maintaining continuous training for field teams are critical for improving efficiency and reducing costs. Ultimately, the findings underscore the need to refine and adapt monitoring and eradication strategies to contain the pathogen, safeguard agricultural systems, and prevent Xylella fastidiosa from becoming endemic in Portugal. Full article

Background: Autosomal dominant intellectual developmental disorder 51 (MIM #617788) is caused by pathogenic variants in KMT5B, a histone methyltransferase essential for transcriptional repression and central nervous system development. The disorder manifests as a complex neurodevelopmental syndrome with variable neurological and systemic features. Methods: Two adolescents with nonsense KMT5B variants underwent detailed clinical, neuropsychological, and neuroimaging evaluations, including MRI and ¹⁸FDG PET/CT, analyzed with Statistical Parametric Mapping against matched controls. RNA sequencing was performed, and the literature was reviewed to assess genotype–phenotype correlations. Results: Both patients showed global developmental delay, progressing to autism spectrum disorder (ASD) and developmental coordination disorder (DCD), without intellectual disability (ID). The MRI was normal, but neuropsychological testing revealed executive function impairment, expressive language deficits, and behavioral disturbances. PET/CT consistently demonstrated cerebellar and temporal lobe hypometabolism, correlating with symptom severity. RNA sequencing identified shared dysregulated pathways, notably DDIT4 upregulation, linked to synaptic dysfunction and neuronal atrophy in animal models. Conclusions: The findings highlight cerebellar involvement in DCD and ASD, medial temporal lobe contribution to ASD and executive dysfunction, and DDIT4 as a possible molecular signature of KMT5B loss-of-function. An integrative multimodal approach refined genotype–phenotype correlations and revealed novel brain regions and pathways implicated in KMT5B-related disorders. Full article