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Fusarium Keratitis is a corneal infection that causes blindness if left untreated. Immune profiling of Fusarium-infected corneas may facilitate the development of an effective immunotherapy. The ex vivo caprine (goat) cornea model examines immune cell and cytokine activity in response to Fusarium keratitis. Dissected caprine corneas maintained in ex vivo culture conditions were infected with the clinical Fusarium isolate (CSH_1) to study local immune responses. Immuno-phenotyping was performed using immune cell markers CD45, CD11b, and HLA-DR/DQ, followed by quantification of IL-1ß, IL-6, IFN-γ, TGF-ß, IL-4, IL-10, and MCP-1 through ELISA. The efficacy of antifungals was evaluated by sectioning drug-treated infected corneas by H&E staining and cytokine profiling to assess the immunomodulatory effect of antifungal treatment. Results show that CD45⁺ lymphocytes, recognized as a universal marker for immune cells, were abundant in the cornea. Activated myeloid cells (CD 11b⁺HLA DR/DQ⁺) and cytokines IL-1ß, IL-6, IFN-γ, TGF-ß, IL-4, IL-10, and MCP-1 were significantly elevated in infected corneas relative to uninfected samples. Among the drugs tested, posaconazole showed complete inhibition of Fusarium, while natamycin and amphotericin B showed the least inhibition and moderate inhibition was shown by voriconazole. The ex vivo model offers a valuable framework for evaluating pre-clinical efficacy of different treatment protocols for fungal keratitis on local tissue.

Pseudomonas aeruginosa is a prominent uropathogen associated with biofilm-related urinary tract infections. It can coexist in a biofilm environment with other pathogens, including Staphylococcus aureus, and the availability of metabolites in urine may influence interspecies interactions that can be cooperative or competitive. Here, a dual-species biofilm model consisting of uropathogenic and reference strains of P. aeruginosa and S. aureus was used to understand their coexistence under different nutritional conditions using synthetic urine supplemented with creatinine, glucose, albumin, and haem. A dual-species biofilm was developed using equal initial cell densities of pathogens. Biofilm intensity was quantified by crystal violet staining. Biofilm biomass, growth, ureolysis were estimated and compared with mono-species cultures. P. aeruginosa formed higher biofilm than S. aureus (p < 0.01) under mono-species culture. Overall, dual-species biofilm intensity was significantly higher than mono-species biofilm (p < 0.01), except in albumin, where S. aureus mono-species biofilm was higher. Biofilm biomass enumeration indicates near-equilibrium coexistence of species within the biofilm matrix. The ureolytic activity correlated with growth and biofilm observations. Our study highlights the complex interactions in dual-species biofilms and emphasizes the need for further studies involving diverse uropathogenic strains to assess metabolite influences on biofilm structural dynamics and its effect on antibiotic responses for targeted therapeutics.

Biofilms are structured communities of microorganisms embedded in extracellular polymeric substances (EPSs) that adhere to surfaces or interfaces. These biofilms are resistant to antimicrobial agents and are responsible for persistent infections. An effective strategy is needed to inhibit the biogenesis of polymeric substances by Enterococcus faecalis. This study explored a synergistic approach using an indole terpenoid molecule, rhodethrin, combined with chloramphenicol to disrupt EPS production by E. faecalis. TGA revealed three stages of decomposition: moisture content (15% at 35°C–200°C), pyrolysis temperature (40%–60% at 200°C–400°C), and polysaccharide crystals (< 10% at < 600°C). X-ray diffraction analysis indicated that the EPS consisted of 40%–60% crystalline and 60%–75% amorphous domains. FTIR analysis also identified various functional groups, like aliphatic CH₂, hydroxyl groups, aliphatic methyl groups, asymmetrical C=H stretching, and amine groups. LCMS analysis provided insights into the molecular mass of EPS constituents. SEM analysis revealed a condensed matrix characterized by polymeric carbohydrates and filamentous structures. The results showed a significant therapeutic importance of their combined effects on the biosynthesis of EPSs in E. faecalis. Further research on their molecular mechanisms and clinical applications is essential to harness their benefits and develop effective treatment against biofilm-associated infections caused by E. faecalis.

Intraoperative margin evaluation is very important in the management of breast cancer patients, specifically in breast conserving therapy in which attaining negative margins is a prerequisite. Microscopic involvement of margins shows two to three times increase in local recurrence. Frozen section is mostly employed for intraoperative assessment of margins. It has, however, the inherent limitations of being expensive, more time-consuming, with difficult serial sectioning, associated technical issues regarding adipose tissue freezing and tissue consumption during the procedure that could leave limited tissue for the permanent sections, thus compromising the final diagnosis, in addition to its intrinsic procedural standardization deficits. A variety of other techniques and methodologies have been developed that can complement frozen section or be employed independently for increased diagnostic efficacy and accuracy like intraoperative touch preparation cytology, assessment by margin probes, flow cytometry, optical coherence tomography (OCT), integrated OCT with microscopy or with dye-enhanced field polarization, polarization-sensitive multimodal imaging, and nonlinear microscopy. This review article states what defines free margins, highlights the different histological options for margin evaluation, and reflects on the evolving methodologies to diagnose the margins intraoperatively.

Colistin (COL) belongs to the polymyxin group of drugs, which possesses a positive charge and interacts with lipopolysaccharide (LPS) of Gram-negative bacterial outer membranes. Acinetobacter baumannii, a bacterium in the “ESKAPE” group of “priority pathogens,” has acquired resistance against the majority of available antibiotics, including the last resort antibiotic COL. Though plasmid-encoded acquisition of mcr-genes has been associated with clinical resistance, efflux, loss of LPS by inactivation of the biosynthetic pathway (lpxACD), and modifications of target LPS by products of chromosomal pmrCAB genes has been ascribed to resistance evolution. Systemic characterization of trade-offs and traits accompanying the evolution of COL resistance in the bacteria remains unaccomplished. Here we report adaptive evolution of extreme COL resistance of the reference strain A. baumannii ATCC19606. Systemic phenotypic characterization of the mutants revealed hyperbiofilm formation and a striking decrease in fitness as the major evolution-associated attributes. Comprehensive antibiotic susceptibility profiling indicated collateral sensitivity against vancomycin and fosfomycin. Whole genome sequencing of the resistant strains led to the identification of mutations associated with COL resistance. Phenotypic characterization of three COL-resistant clinical isolates of A. baumannii revealed similarity with experimentally evolved resistant mutants in one of the isolates, in which none of the mcr-genes could be detected.

Atrial fibrillation (AF) ranks among the most prevalent cardiovascular diseases, and its prevalence is increasing over the years. Atrial fibrosis is one of the key causes of AF. This study aimed to investigate miR-490-3p levels in AF and the regulatory effect of the miR-490-3p/TGFBR1 axis on atrial fibrosis. In this study, RT-qPCR was utilized to detect miR-490-3p and TGFBR1 levels in AF patients. ROC was utilized to assess the diagnostic value of miR-490-3p in AF. Logistic regression was utilized to analyze left atrial fibrosis risk factors in AF patients. The AF cell model was established using human cardiac fibroblasts (HCFs). In vitro experiments included CCK-8 viability assay and luciferase reporter assay. The levels of fibrotic factors were detected using western blot. miR-490-3p expression in AF patients was clearly lower. miR-490-3p had good diagnostic ability for AF and was a risk factor. Luciferase assays confirmed TGFBR1 as a target of miR-490-3p. In vitro cell experiments confirmed that TGFBR1 overexpression can rescue the inhibitory effect of miR-490-3p on HCFs proliferation and fibrotic factor expression. In conclusion, miR-490-3p may act as a potential AF marker and inhibit the occurrence of atrial fibrosis through the miR-490-3p/TGFBR1 axis.

Long non-coding RNAs (lncRNAs) are crucial regulators of hepatocellular carcinoma (HCC) development. The objective of the present investigation was to examine the lncRNA HOXC-AS1 levels in HCC and to explore the function of the HOXC-AS1/miR-876-3p/NR3C1 axis in HCC. Relative HOXC-AS1, miR-876-3p, and NR3C1 levels in tumor and paracancerous specimens were assessed by quantitative RT-PCR. sh-HOXC-AS1 was transfected into HCC cells to analyze its effects on HCC. The detection of cell growth, migration, and invasiveness was conducted by the CCK-8 assay along with Transwell analysis. Flow cytometry was employed to assess cell apoptosis. To evaluate the prognostic significance of HOXC-AS1, a Kaplan–Meier survival analysis was carried out. HOXC-AS1 was increased in HCC specimens in comparison with paracancerous tissues. Patients with high HOXC-AS1 levels had lower disease-free survival and overall survival rates than cases with low HOXC-AS1 levels (p < 0.001). Silencing HOXC-AS1 reduced the growth, migration, and invasiveness of HCC cells while promoting their apoptosis. As the direct target gene, miR-876-3p is negatively related to HOXC-AS1 levels. miR-876-3p could reverse the functions of HOXC-AS1 for HCC cells by targeting NR3C1. Upregulation of HOXC-AS1 promotes HCC progression through the miR-876-3p/NR3C1 axis.