Search

Identification of antimicrobial resistance markers in urinary tract infections could provide a more targeted approach in the diagnosis and treatment of UTIs while reducing overall public health burdens. We describe a molecular assay as a diagnostic tool for antibiotic resistance characterization to promote faster diagnosis of antibiotic regimens compared to standard microbiology techniques. Targeted antibiotic usage for pathogenic infections remains a main goal for effective antibiotic treatment protocols and reducing the overall public health burden. Rapid identification of the pathogen(s) causing the infection and harboring the antibiotic resistance gene is also a main area of exploration for antibiotic appropriation and stewardship. Urinary tract infections are a common clinical disease and reservoir for pathogenic infection and the development of antibiotic resistance, especially in hospital- and community-acquired settings. Standard methods require urine culture, which is time consuming and relies on phenotypic characterization. A genetic diagnostic method is warranted for the rapid molecular characterization of antibiotic resistance genes to reduce inappropriate exposure to antibiotics while improving the overall treatment model for urinary tract infections. The purpose of this study is to demonstrate logical viability for real-time molecular diagnostics for early identification, active surveillance and overall targeted antibiotic stratification that is proposed as an in vitro rapid and comprehensive tool for assessing proper antibiotic stewardship in UTIs. Here, we describe a multiplex real-time fluorescence polymerase chain reaction (PCR) for probe-based detection of the top 24 antibiotic resistance genes with targeted relationships to target molecular drug classes and administered antibiotics. Multiplexed analysis based on molecular features enables rapid testing while shifting the diagnostic detection paradigm from monocentric infections towards polymicrobial infections. We utilized 366 samples from the FDA-CDC Antimicrobial Resistance Isolate Bank to test the efficacy of the assay and propose a model to infer the identity of bacterial isolates. We found that, in addition to a high level of accuracy in predicting bacterial genus classification, the assay was mostly in agreement with CDC-tested genotypic and phenotypic results. This study provides evidence for using genetic diagnostic methods, such as multiplex qPCR, in the rapid identification of antibiotic resistance (ABR) genes for the characterization and treatment of urinary tract infections.

Recent studies have reported several beneficial effects of natural compounds on cancerous cells, highlighting their use for future treatments. These preliminary findings have encouraged experiments with natural substances, such as plant extracts, to examine both cytotoxic and mitogenic effects and find alternative treatments for diseases such as breast cancer. This study examines the effects of microwave-assisted and ethanol maceration of marjoram (Origanum majorana) on MCF-7 breast cancer cell lines and normal breast tissue cell lines used as controls. Marjoram extracts displayed a cytotoxic effect on the MCF-7 cell lines and a mitogenic effect on the control cell lines at the MTS test. The metabolic profiles of MCF-7 and control cell lines were also assessed using the Biolog Phenotype Mammalian Metabolic (PM-M) platform and revealed statistically significant differences in the utilization of energy sources, metabolic activity in the presence of certain ionic species, and responses to metabolic effectors, such as stimulant/catabolic compounds and steroid hormones. Exposure to marjoram extracts exerted positive effects on the MCF-7 cells on the abnormal utilization of energy sources and the responses to metabolic effectors, while no major effects were detected on control cells. These effects were compared to the metabolic impact of the chemotherapeutic agent doxorubicin, which showed profound cytotoxic effects on both cancerous and normal breast cells. In conclusion, our in vitro evidence indicates that marjoram extracts are a promising alternative to chemotherapy in breast cancer since they can successfully eliminate cancerous cells by affecting their metabolic capacity to proliferate without inducing noticeable adverse effects on normal breast tissue.

The overprescribing and misuse of antibiotics have led to the rapid development of multidrug-resistant bacteria, such as those that cause UTIs. UTIs are the most common outpatient infections and are mainly caused by Escherichia coli and Klebsiella spp., although some Gram-positive bacteria, such as Pseudomonas aeruginosa, have been isolated in many cases. The rise of antimicrobial-resistant bacteria is a major public health concern, as it is predicted to lead to increased healthcare costs and poor patient outcomes and is expected to be the leading cause of global mortality by 2050. Antibiotic resistance among bacterial species can arise from a myriad of factors, including intrinsic and acquired resistance mechanisms, as well as mobile genetic elements, such as transposons, integrons, and plasmids. Plasmid-mediated resistance is of major concern as drug-resistance genes can quickly and efficiently spread across bacterial species via horizontal gene transfer. The emergence of extended-spectrum β-lactamases (ESBLs) such as NDM-1, OXA, KPC, and CTX-M family members has conferred resistance to many commonly used antibiotics in the treatment of UTIs, including penicillins, carbapenems, cephalosporins, and sulfamethoxazole. This review will focus on plasmid-mediated bacterial genes, especially those that encode ESBLs, and how they contribute to antibiotic resistance. Early clinical detection of these genes in patient samples will provide better treatment options and reduce the threat of antibiotic resistance.