Search Results (289)

The interspecies transmission of avian influenza viruses remains a significant public health concern. H6 viruses have gained attention following the first human infection by a chicken-origin H6N1 virus (A/Taiwan/02/2013, Hu/13), highlighting their zoonotic potential. To understand the evolutionary trajectory and mammalian adaptation of this Taiwan lineage, we compared two avian isolates (A/Chicken/Taiwan/CF19/2009, Ck/09; A/Chicken/Taiwan/2267/2012, Ck/12) and Hu/13 in vitro and in vivo. Hu/13 exhibited enhanced replication in MDCK cells, producing larger plaques and higher viral titers than Ck/09 and Ck/12. In BALB/c mice, Hu/13 demonstrated the highest pathogenicity and mortality, followed by Ck/12, while Ck/09 induced minimal morbidity. Hu/13 and Ck/12 replicated efficiently in respiratory tissues, eliciting robust cytokine responses and severe pulmonary lesions. In ferrets, Hu/13 showed relatively efficient transmission, infecting all direct physical-contact and two out of three airborne-contact ferrets, whereas Ck/09 failed to transmit. Histopathology confirmed escalating lung pathology from Ck/09 to Ck/12 and Hu/13. Whole-genome sequencing identified adaptive mutations in Hu/13 during ferret replication, though no canonical mammalian-adaptive changes (e.g., PB2-E627K or HA-Q226L) were detected. These findings demonstrate progressive mammalian adaptation, replication efficiency, and transmissibility within the Taiwan H6N1 lineage. Enhanced surveillance is crucial to monitor mammalian-adaptive mutations, informing pandemic preparedness and public health strategies. Full article

Neurological symptoms involving the central nervous system (CNS) and peripheral nervous system (PNS) are common complications of acute COVID-19 as well as post-COVID conditions. Most research into these neurological sequalae focuses on the CNS, disregarding the PNS. Guinea pigs were previously shown to be useful models of disease during the SARS-CoV-1 epidemic. However, their suitability for studying SARS-CoV-2 has not been experimentally demonstrated. To assess the suitability of guinea pigs as models for SARS-CoV-2 infection and the impact of SARS-CoV-2 infection on the PNS, and to determine routes of CNS invasion through the PNS, we intranasally infected wild-type Dunkin-Hartley guinea pigs with ancestral SARS-CoV-2 USA-WA1/2020. We assessed PNS sensory neurons (trigeminal ganglia, dorsal root ganglia), autonomic neurons (superior cervical ganglia), brain regions (olfactory bulb, brainstem, cerebellum, cortex, hippocampus), lungs, and blood for viral RNA (RT-qPCR), protein (immunostaining), and infectious virus (plaque assay) at three- and six-days post infection. We show that guinea pigs, which have previously been used as a model of SARS-CoV-1 pulmonary disease, are not susceptible to intranasal infection with ancestral SARS-CoV-2, and are not useful models in assessing neurological impacts of infection with SARS-CoV-2 isolates from the early pandemic. Full article

This study aims to isolate and characterize the lytic phage XC1 targeting Acinetobacter nosocomialis and systematically analyze its biological properties and genomic structure, providing theoretical support for developing novel treatments against antibiotic-resistant infections. Phage XC1 was isolated and purified from lake water. Its morphology, optimal multiplicity of infection (MOI), thermal stability, and pH tolerance were analyzed. Genomic sequencing and functional annotation were performed to identify its lysis-associated genes. Phage XC1 demonstrated a short latent period (20 min) and high burst size (310 plaque-forming units per cell, PFU/cell). It remained stable under temperatures of 50–60 °C and at pH 7, indicating good environmental stability. Genomic analysis revealed a 45,324 bp genome with a GC content of 38.21%, including 84 open reading frames (ORFs), without any lysogenic, virulence, or antibiotic-resistance genes, confirming its safety. Average Nucleotide Identity (ANI) analysis shows that the ANI values between phage XC1 and other phages range from 80% to 95%. As the ANI value between strains of the same species is typically ≥95%, this suggests that phage XC1 may be a previously undiscovered new phage. Classified within the genus Obolenskvirus (class Caudoviricetes), phage XC1 is a virulent bacteriophage with rapid lytic activity and extreme environmental tolerance. Its therapeutic potential against multidrug-resistant infections, either as a monotherapy or in synergy with antibiotics, warrants further investigation. Full article

Handling cultured isolates and clinical, environmental, or wildlife surveillance samples containing Risk Group 3 and 4 pathogens presents considerable biosafety challenges in minimizing human exposure during processing and transport. Safe handling typically requires high- or maximum-containment facilities, demanding substantial logistical planning and resources. We evaluated PrimeStore Molecular Transport Medium (PS-MTM), a guanidine-based solution created to kill pathogens and preserve nucleic acids at ambient temperatures, for inactivating Crimean-Congo hemorrhagic fever, eastern equine encephalitis, Ebola, Hendra, Japanese encephalitis, Lassa, Marburg, Nipah, Rift Valley fever, and West Nile viruses. To mimic diagnostic conditions, human whole blood spiked with any of these viruses was incubated with PS-MTM for 20-, 30-, or 60-min. Samples with titers up to 10⁷ PFU/mL exposed to PS-MTM at all time points resulted in complete loss of infectivity judged by plaque assays. A 30-min incubation provided a 50% safety margin over the minimum inactivation time and was used for quantification with the tissue culture infectious dose (TCID₅₀) assay, enabling evaluation of PS-MTM’s activity for viruses that do or do not produce well-defined plaques. Results confirmed that PS-MTM inactivated all tested viruses at titers up to 10⁷ TCID₅₀/mL, underscoring its reliability for enhancing biosafety in diagnostics, outbreak management, and surveillance. Full article

Background: Celastrol, a pentacyclic triterpenoid active component isolated from the root bark of the traditional medicinal plant Tripterygium wilfordii, displays significant anti-inflammatory, antioxidant, and immunomodulatory properties. However, its clinical application remains limited due to inadequate bioavailability. Methods: Regarding these issues, we innovatively developed a novel peanut cultivar (cel-peanut) enriched with celastrol through distant hybridization combined with metabolomics screening. Guided by the research concept of “natural anti-inflammatory diets for metabolic disease management”, we established a high-fat diet-induced ApoE^−/− atherosclerotic mouse model to systematically evaluate the anti-atherosclerosis effects and mechanisms of cel-peanut. Results: Our results revealed that cel-peanut significantly reduced serum levels of triglycerides (TGs) and increased high-density lipoprotein cholesterol (HDL-C). Concurrently, cel-peanut markedly decreased the atherosclerotic lesion area and enhanced collagen content within plaques. Mechanistic investigations demonstrated that cel-peanut reduced serum malondialdehyde (MDA) levels and suppressed the concentration of pro-inflammatory cytokine IL-6 in atherosclerotic lesions. Furthermore, cel-peanut promoted intestinal health by modulating the composition and functionality of gut microbiota, thereby attenuating atherosclerosis progression. Conclusions: Overall, these findings indicate that cel-peanut exerts therapeutic effects against atherosclerosis through its anti-inflammatory, antioxidant, and gut microbiota-modulating properties. This study proposes a novel nutritional intervention strategy for atherosclerosis and provides a promising adjuvant strategy for clinical atherosclerosis treatment. Full article

Flow virometry (FVM) offers a promising approach for monitoring viruses and virus-like particles (VLPs) in environmental samples. This study compares levels of non-specific VLPs across a wastewater treatment plant (WWTP) with levels of somatic coliphage, (F+) specific coliphage, Pepper Mild Mottle Virus (PMMoV), CrAssphage (CrAss), and Tomato Brown Rugose Fruit Virus (ToBRFV). All targets were quantified in influent, secondary-treated effluent, and tertiary-treated effluent at the University of California, Davis Wastewater Treatment Plant (UCDWWTP) over 11 weeks. We established an FVM-gating boundary for VLPs using bacteriophages T4 and ϕ6 as well as four phages isolated from wastewater. We then utilize T4 alongside three submicron beads as quality controls in the FVM assay. Coliphage was measured by standard plaque assays, and genome copies of PMMoV, CrAss, and ToBRFV were measured by digital droplet (dd)PCR. FVM results for wastewater revealed distinct microbial profiles at each treatment stage. However, correlations between VLPs and targeted viruses were poor. Trends for virus inactivation and removal, observed for targeted viruses during wastewater treatment, were consistent with expectations. Conversely, VLP counts were elevated in the WWTP effluent relative to the influent. Additional sampling revealed a decrease in VLP counts during the filtration treatment step following secondary treatment but a substantial increase in VLPs following ultraviolet disinfection. Defining application boundaries remain crucial to ensuring meaningful data interpretation as flow cytometry and virometry take on greater significance in water quality monitoring. Full article

Staphylococci are prevalent in dogs’ microbiota, with commensal strains being able to exhibit resistance and virulence traits, complicating secondary infection management. As antimicrobial resistance is a global threat, particularly in middle-income countries like Angola, surveillance of resistant bacteria is crucial. We analyzed the prevalence, antimicrobial resistance, and virulence profiles of staphylococci from dogs in Angola. Isolates were identified using VITEK^® 2 Compact (bioMérieux© SA, Marcy l’Etoile, France), and their genetic diversity was assessed via PCR fingerprinting. Isolates’ susceptibility to relevant antimicrobials was determined by disk diffusion, and their virulence profiles were evaluated using plaque assays. The relationship between antibiotic resistance and animal-related factors was also assessed by statistical analysis. Isolates were identified as Mammaliicoccus sciuri (former Staphylococcus sciuri, 38%), Staphylococcus xylosus (30%), Staphylococcus equorum (13%), Mammaliicoccus vitulinus (former Staphylococcus vitulinus, 7%), Mammaliicoccus lentus (former Staphylococcus lentus, 5%), Staphylococcus aureus (2%), and Staphylococcus spp. (5%). Of these, 86% were resistant to at least one of the antimicrobials tested, and 30% were classified as multidrug-resistant, being more common in females, dogs with clinical signs of disease, and vaccinated animals. Moreover, 93% of the isolates were able to produce biofilm, 46% could produce lecithinase and gelatinase, and 23% could produce hemolysins. Companion dogs from Angola can carry resistant staphylococci able to express several virulence factors, potentially representing a One Health risk. Full article

Background: This study aimed to assess the effectiveness of a modified entire papilla preservation technique (MEPPT) for treating isolated intrabony defects in patients with stage III periodontitis. Material and Methods: Fifteen patients with 15 interdental intrabony defects were treated with a MEPPT using enamel matrix derivative and allogenic bone. Their probing pocket depth (PPD), clinical attachment level (CAL), gingival recession (GR), keratinized tissue width (KTW), defect depth (DD), full-mouth plaque score (FMPS), full mouth bleeding score (FMBS), radiographic images (radiographic angles, BF and LDF) and intrasurgical parameters were assessed at baseline and 3 years postsurgery. Standardized measurements were taken to evaluate the defect characteristics and treatment outcomes. Results: At 3 years, significant improvements from baseline were maintained. Probing pocket depth (PPD) decreased from 7.03 ± 1.61 mm to 3.33 ± 0.89 mm (p < 0.0001), clinical attachment level (CAL) improved to 3.08 ± 1.16 mm (p < 0.001) and defect depth (DD) decreased from 4.59 ± 1.24 mm to 0.38 ± 0.31 mm (p < 0.001). The changes in gingival recession and keratinized tissue were not statistically significant. The results demonstrate sustained clinical stability over a 3-year period. Conclusions: Within the limitations of this study, the findings suggest that the modified entire papilla preservation technique (MEPPT) in conjunction with enamel matrix proteins and allogenic bone grafting is an effective approach for the treatment of intrabony defects, leading to statistically significant and sustained clinical improvements over a 3-year period. The study protocol was registered in ClinicalTrials.gov ID NCT05029089. Full article

Background: Since 2011, re-emerging pseudorabies virus (PRV) variant strains have been widespread in swine herds immunized with the classical PRV vaccine in China, suggesting that it is necessary to develop a new vaccine against these PRV variant strains. Methods: Here, based on a PRV mutant strain isolated in Jinmen (JM), two recombinant strains were constructed using CRISPR/Cas9 technology, including PRV-JM-ΔEK with the deletion of the gE and TK genes and PRV-JM-ΔEI92K with the deletion of the gE, gI, US2, US9, and TK genes. Results: A one-step growth curve and plaque assay revealed that the cell-to-cell transmission ability of PRV-JM-ΔEI92K was lower than that of PRV-JM-ΔEK. However, the replication ability of PRV-JM-ΔEI92K was approximately 10 times higher than that of PRV-JM-ΔEK, similar to wild-type PRV-JM. The intramuscular injection of 10⁶ TCID₅₀ of PRV-JM-ΔEK or PRV-JM-ΔEI92K could not cause death in mice, and both could produce specific antibodies against gB and gD. The survival rate of mice immunized with both recombinant viruses was 100% when the mice were challenged by the PRV-JM strain. Histopathological sections from the PRV-JM-ΔEK group showed milder pathological changes compared to the PRV-JM-ΔEI92K group, proving that PRV-JM-ΔEK provided more effective protection. In pigs injected with 10⁶ TCID₅₀ of PRV-JM-ΔEK or PRV-JM-ΔEI92K, their body temperature did not rise, and their weight gain was not affected. Both recombinant viruses could induce the production of gB- and gD-specific antibodies and neutralizing antibodies. After the challenge of the PRV-JM virus, neutralizing antibody production was rapidly induced and lasted for at least 3 weeks. Pigs immunized with both PRV-JM-ΔEI92K and PRV-JM-ΔEK had a 100% survival rate, demonstrating that both recombinant viruses could provide effective protection. Conclusions: Compared with PRV-JM-ΔEK, PRV-JM-ΔEI92K had better safety. In conclusion, we constructed two PRV recombinant viruses, which have the potential to be used as a live carrier vaccine. Full article

The ideal bacteriophages (phages) for the treatment of bacterial disease (phage therapy) would bypass bacterial evolution to phage resistance. However, this feature (called a hyper-aggression feature) has never been observed to our knowledge. Here, we microbiologically characterize, fractionate, genomically classify, and perform electron microscopy of the newly isolated Bacillus thuringiensis phage 0524phi7-1, which we find to have this hyper-aggression feature. Even visible bacterial colonies are cleared. Phage 0524phi7-1 also has three other features classified under hyper-aggression (four-feature-hyper-aggressive phage). (1) Phage 0524phi7-1 forms plaques that, although sometimes beginning as semi-turbid, eventually clear. (2) Clear plaques continue to enlarge for days. No phage-resistant bacteria are detected in cleared zones. (3) Plaques sometimes have smaller satellite plaques, even in gels so concentrated that the implied satellite-generating phage motion is not bacterial host generated. In addition, electron microscopy reveals that phage 0524phi7-1 (1) is a myophage with an isometric, 91 nm-head (diameter) and 210 nm-long contractile tail, and (2) undergoes extensive aggregation, which inhibits typical studies of phage physiology. The genome is linear double-stranded DNA, which, by sequencing, is 157.103 Kb long: family, Herelleviridae; genus, tsarbombavirus. The data suggest the hypothesis that phage 0524phi7-1 undergoes both swimming and hibernation. Techniques are implied for isolating better phages for phage therapy. Full article

Background: The spontaneous rupture of the internal iliac artery (IIA) is an exceedingly rare vascular event, typically associated with congenital anomalies or degenerative conditions. This report details an unprecedented case of isolated IIA rupture in an elderly patient with evidence of plaque rupture but devoid of congenital vascular pathology. Case Presentation: An 81-year-old Caucasian male presented to the Emergency Department following a syncopal episode and acute right iliac fossa pain. His significant medical history was atrial fibrillation managed with anticoagulation (Apixaban), non-insulin-dependent diabetes mellitus, and recent hospitalization for multidrug-resistant Klebsiella pneumoniae pneumonia. Initial imaging with contrast-enhanced computed tomography revealed an aneurysmatic dilatation of the right IIA, indicative of rupture. An endovascular repair was performed, employing a combination of stent grafts to achieve proximal and distal sealing and to restore vascular continuity. Outcome: The patient exhibited hemodynamic stability throughout the perioperative period and was transferred to the general ward postoperatively. However, he suffered a recurrent rupture on the 30th postoperative day, prompting a second endovascular intervention to extend the graft landing zone into the common iliac artery. Intraoperative findings confirmed localized plaque rupture as the underlying trigger for the initial vessel rupture. He ultimately achieved clinical stability and was discharged on the 35th postoperative day. Discussion: This case illustrates the critical importance of recognizing spontaneous IIA rupture as a potential complication in elderly patients, particularly in the context of recent severe infections. While the relationship between the rupture and the Klebsiella pneumoniae infection remains speculative, this report underscores the necessity of further research into the role of infectious processes in vascular integrity and susceptibility to rupture. Conclusions: The successful management of this rare and complex vascular emergency using endovascular techniques underscores the evolving landscape of minimally invasive interventions. This case contributes to the limited existing literature on spontaneous IIA rupture and highlights the need for increased clinical vigilance regarding atypical presentations in similar patient populations. Full article

Background/Objectives: Several gene targets were identified for psoriasis. Some are currently being explored as potential therapeutic targets, including CCL11. Our task was to prove a possible association of single-nucleotide polymorphisms +67 G/A and -426 T/C in the eotaxin gene (CCL11, 17q 21.3) with the development and clinical aspects of psoriasis as an immune-based dermatological disease and evaluate its relationship to potential comorbidities. Material and Methods: In total, 460 patients with psoriasis were included in the case–control and genotype–phenotype study together with 167 control persons of similar age and sex distributions without a personal and/or family history of chronic disease of the skin. Two eotaxin gene polymorphisms were detected from isolated DNA via standard PCR, restriction analysis methods, and horizontal electrophoresis. Results: No significant case–control differences in the frequency of the CCL11 genotype in both polymorphisms were observed. In polymorphism +67 G/A, a significant increase in the AA genotype in patients with psoriasis guttata compared to plaque psoriasis was found (p = 0.006). A significant association of the A allele in psoriatic patients with a personal history of allergy was found (p = 0.02). The A alle was also significantly associated with a family history of psoriasis (p = 0.00008). In men, a higher risk of a delayed start of psoriasis (later than 40 years) associated with the T allele of -426 T/C polymorphism (p = 0.0007) was found. When double genotypes of both polymorphisms were evaluated, we observed significant differences in double genotype distribution between men with and without a family history of allergy (Pdg = 0.0005) and between those with and without affected siblings (Pdg = 0.03). In women with psoriasis, a higher risk of the TT genotype of -426 T/C polymorphism in patients with a personal history of diabetes (p = 0.001) as well as in patients with both a personal history of cardiovascular disease and diabetes (p = 0.00005) was proved. When double genotypes of both polymorphisms were evaluated, the significance of double genotype difference between those with and without personal history of diabetes was very high (Pdg = 0.0002). Similarly, the significance of the double genotype difference between those with and without personal history of cardiovascular diseases and diabetes was very high (Pdg = 0.000001). Conclusions: CCL11 is considered one of the basic chemokines responsible for the origin and development of immune-based reactions. Based on our results, we suggest that the +67 G/A CCL11 polymorphism should be considered as a gene modulator of psoriasis in specific subgroups of patients. Full article

Periodontitis is one of the most common diseases associated with the lifestyle habits of adults and is caused by the formation of biofilms, called dental plaques, in periodontal pockets by oral bacteria, such as Porphyromonas gingivalis. Bangle, Zingiber purpureum Rosc. (Indonesian ginger), a native Indonesian plant, has been traditionally consumed as food and medicine across Southeast Asia. The cis- and trans-banglenes, components of the rhizomes of Z. purpureum, have been reported to possess neurotrophic activity. Hexane extract of bangle exhibited antibacterial activity against P. gingivalis, with a minimum inhibitory concentration of 8 μg/mL. We isolated several compounds from the active fractions through the bioassay-guided isolation of hexane extract. Further, we found that c- and t-banglene inhibited the growth of P. gingivalis at 4 µg/mL; however, these compounds showed no antibacterial effects against oral microorganisms. We also observed that c- and t-banglenes resulted in 47% and 40% reductions in biofilm formation. In conclusion, our results suggest that banglene has specific antibacterial effects against the periodontopathogen P. gingivalis, with minimal impact on oral microorganisms. Thus, banglene has potential applications in the prevention of periodontitis without the risk of substituted microbisms. Full article

Background/Objectives: Extracts of the plant Centella asiatica can enhance mitochondrial function, promote antioxidant activity and improve cognitive deficits. Asiatic acid (AA) is one of the constituent triterpene compounds present in the plant. In this study, we explore the effects of AA on brain mitochondrial function, antioxidant response and cognition in a beta-amyloid (Aβ)-overexpressing 5xFAD mouse line. Methods: Six- to seven-month-old 5xFAD mice were treated with 1% AA for 4 weeks. In the last week of treatment, associative memory was assessed along with mitochondrial bioenergetics and the expression of mitochondrial and antioxidant response genes from isolated cortical synaptosomes. The Aβ plaque burden was also evaluated. Results: AA treatment resulted in improvements in associative memory in female 5xFAD mice without altering the Aβ plaque burden. Cortical mitochondrial function and mitochondrial gene expression were increased in the AA-treated female 5xFAD mice, as was the expression of antioxidant genes. More modest effects of AA on cortical mitochondrial function and mitochondrial and antioxidant gene expression were observed in male 5xFAD mice. Conclusions: Oral AA treatment improved cognitive and mitochondrial function and activated antioxidant in Aβ-overexpressing mice. These changes occurred independent of alterations in Aβ plaque burden, suggesting that AA could have translational therapeutic relevance in later-stage AD when plaques are well established. Full article

Fusobacterium necrophorum subspecies necrophorum, a resident of the rumen, is the causative agent of bovine liver abscesses. Currently, tylosin, a macrolide, is used in the feed to reduce liver abscesses. Because macrolides are medically important antibiotics, their use in food animal production is of public health concern. There is significant interest in finding antimicrobial alternatives. Bacteriophages that lyse subsp. necrophorum have the potential to replace tylosin. Our objective was to isolate bacteriophages lytic to subsp. necrophorum. Pooled ruminal fluid from slaughtered cattle and pooled sewage samples were collected and incubated overnight with lysine, and subsp. necrophorum strains and filtrates were spotted on F. necrophorum lawns. Phage plaques were harvested and purified. Bacteriophage isolation frequencies were compared between sample types, sampling dates, and necrophorum strains. Overall relative frequency of isolated bacteriophages lytic to subsp. necrophorum was 17.1%. The frequency of bacteriophage isolation ranged from 0 to 25.4% for ruminal fluid, and from 13.7 to 32.0% for sewage. Isolation frequency was significantly higher in sewage than in ruminal fluid samples (p < 0.01). Isolation rates varied significantly between necrophorum strains. Sewage was a rich source of bacteriophages lytic to necrophorum, which have the potential to be used to prevent liver abscesses. Full article