Introduction/objective Retinal diseases are an important focus of in vivo gene therapy, for which adeno-associated virus (AAV) is a preferred vector. Intravitreal injections are a highly desirable delivery route because it is simpler, safer, and less expensive than the main alternative, subretinal injection. However, existing capsids have low efficiency, require high doses, and are susceptible to neutralization by antibodies that are present in the vitreous. Methods We developed a novel AAV capsid with improved properties to address these issues. Using AAV5 as starting material, we applied a combination of directed evolution and rational design to develop PT1, a novel capsid capable of transducing the retina after intravitreal injection. Results Compared to its parent capsid AAV5, PT1 has 15 surface amino acid substitutions in five variable regions and has its VP1-unique region swapped with that of AAV8. Its production yield is similar to that of another AAV5-derived control capsid and superior to that of AAV2. PT1 could transduce the retina-derived cell line ARPE-19 as efficiently as control retinal capsids and with better specificity than AAV2. PT1 performed exceptionally well in an in vitro immune evasion assay, largely surpassing control capsids. In vivo transduction following intravitreal injection in mice suggested strong specificity for Müller glia cells. Discussion PT1’s most striking property is its ability to evade preexisting neutralizing antibodies (NABs), apparently thanks to its multiple mutations spread over large areas of the capsid surface, disrupting several epitopes. Another interesting property is its apparent specificity for Müller glial cells, an important therapeutic target. Both of these properties are based on limited evidence and will require further experimental confirmation. Conclusion The results of our preliminary study suggest that PT1 could become a useful addition to the current retinal AAV capsid toolkit, should future studies confirm our findings.
Introduction Spontaneous abortion causes approximately 50,000 maternal deaths annually, primarily due to microbial infection and hemorrhage, accounting for 8–13% of global maternal deaths. This study aimed to investigate the association between microbial infections, bacterial diversity, and patterns related to spontaneous abortion in two groups of women. Methods A total of 156 bacterial isolates were recovered from 48 patient samples, while 67 isolates were obtained from 48 healthy controls. All samples were cultured under aerobic and anaerobic conditions on enrichment and selective media, including Blood agar, MacConkey agar, and Chocolate agar. Results Gram-positive bacteria predominated among total isolates (53.84%) compared to Gram-negative bacteria (46.15%). The primary Gram-positive pathogens were S. aureus (29.16%), L. acidophilus (27.08%), Peptostreptococcus spp. (22.91%), and C. perfringens 20.83%. Among Gram-negative isolates, E. coli was most prevalent (47.91%), followed by P. aeruginosa (18.75%) and K. pneumonia (16.6%). In the control group, L. acidophilus (43.75%) and E. coli (18.75%) were most common. Diversity analyses showed no significant difference in α-diversity, but β-diversity revealed clear distinctions between groups. Linear Discriminant Analysis identified specific biomarkers differentiating the microbial compositions of the two cohorts. Discussion These findings indicate a Gram-positive predominance, with Staphylococcus aureus and E. coli identified as the primary pathogenic drivers in the study groups. Although species richness remained consistent across cohorts, β- diversity and Linear Discriminant Analysis biomarkers confirmed distinct microbial shifts and bacterial signatures between the examined groups. Conclusion L. acidophilus appears to help maintain vaginal health and reduce the risk of spontaneous abortion, as β- diversity and biomarker analyses revealed a shift from beneficial to pathogenic bacteria in women with spontaneous abortion.
Introduction The growing application of biosides post-COVID-19 has heightened concern over the development and spread of antiseptic resistance determinants, especially qac genes in different types of bacteria. This study aimed to evaluate antiseptic resistance determinants in clinical P. aeruginosa isolates and to determine their response to antimicrobial resistance and biofilm-forming ability. Methods Fifty clinical P. aeruginosa isolates were evaluated for antimicrobial susceptibility using the Kirby-Bauer disk diffusion method, Biofilm formation using the microtiter plate crystal violet assay, and biocide tolerance by broth microdilution. PCR was performed to identify qacE and qacEΔ1 genes in selected highly resistant isolates. Statistical analysis was applied to identify the correlation among variables. Results The findings showed a high rate of MDR (64.0%) and XDR (6.0%). Biofilm assays showed strong (38.0%), moderate (30.0%), and weak (30.0%) production. Furthermore, strong /moderate biofilm formation was significantly associated with MDR/XDR phenotypes ( p < 0.0001 ). Reduced antiseptic sensitivity was detected in (60.0%) of isolates and strongly correlated with both antimicrobial resistance status and biofilm strength ( p < 0.0001 ). Molecular profiling of selected resistance isolates showed a universal presence of the qacEΔ1 gene (100%) and high prevalence of the qacE gene (73.3%). Discussion The significant correlation between qac genes and strong biofilm ability demonstrates a key mechanism of antibiotic and biocide tolerance in P. aeruginosa. Conclusion Qac genes are highly prevalent in clinical P. aeruginosa isolates and strongly correlated with multidrug resistance and biofilm-forming ability. Ongoing molecular monitoring of the biocide resistance genes in clinical setups is vital to prevent the propagation of exceptionally stubborn strains in clinical environments.
PPCPs have emerged as a significant class of environmental contaminants due to their extensive global consumption and persistence in natural ecosystems. Conventional wastewater treatment plants are often unable to completely remove these compounds, resulting in their continued release into aquatic and terrestrial environments. The bioaccumulation and long-term persistence of PPCPs raise serious concerns regarding potential ecological risks, disruption of microbial communities, and adverse impacts on human health through food chains and water reuse. This review provides a comprehensive synthesis of current advances in the biodegradation of PPCPs, with a particular focus on microbial degradation pathways. The roles of diverse microbial groups, including bacteria, fungi, and algae, as well as specific enzymatic systems, in the transformation and mineralization of PPCPs are critically examined. Special emphasis is placed on ligninolytic fungi, whose extracellular oxidative enzymes, such as laccases, peroxidases, and oxidoreductases, show high potential for degrading structurally complex PPCPs. Furthermore, the application of genetically engineered microorganisms is discussed as a promising strategy to enhance biodegradation efficiency by tailoring metabolic pathways. The review also evaluates environmental and physicochemical factors such as pH, temperature, redox potential, and the molecular properties of PPCPs that influence degradation outcomes. Beyond laboratory findings, advanced biotechnological innovations and decentralized on-site treatment technologies for soil and water systems are highlighted as practical approaches for sustainable remediation. By consolidating recent research findings, this work not only identifies existing challenges and knowledge gaps but also outlines future directions and integrated strategies toward effective, eco-friendly, and scalable PPCP mitigation.
Background Both chronic periodontitis and atherosclerotic cardiovascular disease are highly prevalent inflammatory diseases. The potential biomarker for these conditions is lactate dehydrogenase. The purpose of this study was to investigate salivary lactate dehydrogenase concentrations in patients with CP with and without ATH. Methods This was a cross-sectional study involving 87 participants aged 30-60 years, divided into four groups: the ATH-only group (n=25), the CP-only group (n=25), the ATH+CP group (n=25), and the normal control group (n=12). The participants underwent periodontal examination and sampling of unstimulated saliva. Salivary LDH was analyzed by ELISA. Statistical analysis included one-way ANOVA, post hoc tests, Pearson correlation, and ROC curve analysis. Results The salivary LDH values were significantly different among the groups ( p < 0.001). The ATH+CP group had the highest value of salivary LDH (24.55 ± 2.55 U/L), followed by the CP group (13.44 ± 4.27 U/L), the ATH group (11.68 ± 3.86 U/L), and the control group (1.53 ± 0.31 U/L). The values of salivary LDH were positively correlated with the clinical parameters of periodontal disease in the diseased groups. The diagnostic accuracy of the ROC analysis was excellent, with an AUC = 0.987. Discussion The results obtained in this study suggest that salivary LDH is potentially useful as a noninvasive biomarker for estimating inflammation in patients with periodontitis and cardiovascular disease. The increased levels of LDH, particularly among patients with combined conditions, suggest pathophysiological connections and support emerging evidence concerning the bidirectionality of associations between oral and systemic health. Conclusion Salivary LDH is a potential noninvasive biomarker of periodontal disease and may reflect the inflammatory burden in patients with concomitant ATH. The presence of elevated LDH in patients with both conditions indicates pathophysiological overlap.
