Background: Extrapulmonary tuberculosis (TB) presents significant diagnostic challenges, particularly in the context of multidrug-resistant (MDR) strains. This study assessed the utility of the WHO-recommended rapid molecular assays, originally validated for pulmonary TB, in diagnosing extrapulmonary TB and detecting the MDR Mycobacterium tuberculosis complex (MTBC). Materials and Methods: A total of 6274 clinical samples, including 4891 pulmonary and 1383 extrapulmonary samples, were analyzed between 2019 and 2022 using the BD MAX™ MDR-TB assay (BD MAX), the Xpert^® MTB/RIF assay (Xpert MTB/RIF), the Xpert^® MTB/XDR assay (Xpert MTB/XDR), FluoroType MTB, and phenotypic drug susceptibility testing (DST). Results: MTBC was detected in 426 samples using BD MAX (376 pulmonary and 50 extrapulmonary), of which 277 were culture-confirmed. Phenotypic testing confirmed 299 positive cultures on Löwenstein–Jensen (LJ) medium and 347 in BD BACTEC™ MGIT™ (BACTEC MGIT) mycobacterial growth indicator tube (BBL) liquid culture. BD MAX showed high sensitivity and specificity for extrapulmonary TB detection (93.1% and 98.4%, respectively). Resistance to isoniazid or rifampicin was identified in 11% of MTBC-positive cases, whereas 3.69% were confirmed as MDR-TB. The molecular assays effectively detected resistance-associated mutations (katG, inhA, and rpoB), with high concordance to phenotypic tests (DST) (κ = 0.69–0.89). Conclusions: This study demonstrates that molecular assays, although validated for pulmonary TB, are also reliable for extrapulmonary TB detection and drug resistance profiling. Their rapid turnaround and robust accuracy support broader implementation in routine diagnostics, especially for challenging extrapulmonary specimens where early detection is critical for targeted therapy.

Ureaplasma species (Ureaplasma spp.) are commonly found as commensals in the human urogenital tracts, although their overgrowth can lead to infection in the urogenital tract and at distal sites. Furthermore, ureaplasmas lack a cell wall and do not synthesize folic acid, which causes all β-lactam and glycopeptide antibiotics, and sulfonamides and diaminopyrimidines, to be of no value. The antibiotics used in therapy belong to the fluoroquinolone, tetracycline, chloramphenicol and macrolide classes. However, the growing incidence of antibiotic-resistant Ureaplasma spp. in the population becomes a problem. Thus, there is a need to search for new drugs effective against these bacteria. Since 1951, the FDA-approved, well-tolerated, inexpensive, orally administered drug disulfiram (DSF) has been used in the treatment of chronic alcoholism, but recently, its antimicrobial effects have been demonstrated. The main biological metabolite of DSF, i.e., N,N-diethyldithiocarbamate (DDC), is generally believed to be responsible for most of the observed pharmacological effects of DSF. In the presented studies, the effect of DDC at concentrations of 2 µg/mL, 20 µg/mL and 200 µg/mL on the growth and survival of Ureaplasma urealyticum and Ureaplasma parvum was tested for the first time. The results indicated that all the used DDC concentrations showed both bacteriostatic and bactericidal activity against both tested strains.

The growing global burden of antimicrobial resistance (AMR) highlights the urgency of identifying new therapeutic options or adjuvant agents capable of limiting microbial growth. This study aimed to evaluate the potential applicability of selected essential oils (EOs) as alternative antimicrobial agents by assessing their biocidal activity against clinically relevant microorganisms and determining their cytotoxicity toward human cell lines. Serial dilutions of the studied EOs were prepared to determine the minimum bactericidal (MBC), minimum fungicidal (MFC), and minimum biofilm eradication concentrations (MBEC) against Staphylococcus aureus ATCC^® 25923, Escherichia coli ATCC^® 25922, Candida albicans ATCC^® 90028, and Nakaseomyces glabratus ATCC^® 15545. Cytotoxicity towards cell lines was evaluated using the MTT assay. The tested EOs displayed significant biocidal effects against studied microorganisms in both planktonic and biofilm forms. Cinnamon bark oil and cinnamaldehyde showed the highest biocidal activity against S. aureus ATCC^® 25923, E. coli ATCC^® 25922, C. albicans ATCC^® 90028, and N. glabratus ATCC^® 15545, except for S. aureus in biofilm form. For thymol, yeast strains exhibited eight-fold lower susceptibility compared to bacterial strains. Cytotoxicity of the tested EOs towards the studied cell lines was significant. EOs proved to be effective against microorganisms but at the same time showed substantial cytotoxicity, thus potentially limiting their application. Further research on formulation strategies, selective delivery, and dose optimization is required to enhance their safety profile and clinical feasibility.

Tuberculosis is still one of the leading infectious causes of morbidity and mortality worldwide. Rapid diagnosis is essential for effective treatment and control of tuberculosis transmission. In recent years, nucleic acid amplification tests (NAATs), such as GeneXpert MTB/RIF, BD MAX™, Xpert MTB/RIF-Ultra, have significantly improved tuberculosis diagnostics. However, they mainly require expensive and advanced equipment. The aim of our study was to assess the usefulness of the novel RITA MTBC assay in this diagnostic context. A total of 61 clinical specimens were tested using the RITA MTBC assay in comparison with established molecular diagnostic platforms (GeneXpert and BD MAX™), used as molecular reference assays. Culture and microscopy were performed as part of initial clinical assessment, but the comparative analysis focused on molecular assays to provide a relevant evaluation of diagnostic performance. Among 31 samples previously identified as positive for M. tuberculosis DNA, the assay correctly detected 30 (LOT HPA01/20230601) and 29 (LOT HPA01/20230602). Of 30 negative samples, 28 and 30 were confirmed negative for the respective lots. These results correspond to an average sensitivity of 95% and an average specificity of 97%. The kit demonstrated diagnostic performance that meets requirements for molecular testing in tuberculosis, with sensitivity and specificity comparable to established platforms, although further validation on larger sample sets is necessary. Nevertheless, its excellent specificity, rapid turnaround time, and operational simplicity, make it especially well-suited for decentralized or resource-limited settings. These findings underscore the potential of RITA MTBC as a valuable diagnostic tool in both routine clinical settings and in populations with limited access to healthcare.

The work provides novel insight into the development of advanced antibacterial surfaces using the combination of essential oils, cinnamon oil, thyme oil, and tea tree oil, as well as their active compounds, including cinnamaldehyde, thymol, and terpinene-4-ol, embedded in the chitosan and sodium alginate matrix. All coatings obtained in a two-stage electrophoretic deposition process on stainless steel and titanium substrates were characterized by high adhesion strength. The microstructural differences between the coatings were mainly related to the size and location of the additives. Structural investigation showed the impact of individual oil components on intermolecular bonds between polysaccharide chains and the formation of molecular interactions in a specific spatial conformation. The surface of all coatings was minimally rough and had a hydrophilic character. A clear matrix-dependent trade-off between antibacterial efficacy and cytocompatibility was observed: alginate-based coatings achieved strong anti-Staphylococcus aureus activity (2.81 log CFU/mL) at the expense of increased cytotoxicity, while chitosan-based systems provided a more favorable cytocompatibility profile, maintaining cell viability above 70% for selected formulations. This work provides insight into the development of natural antibacterial surfaces by the combination of active compounds and shows the distinctions on many levels between the coatings with various polysaccharide matrices.

Ureaplasma species (Ureaplasma spp.) are commonly found as commensals in the human urogenital tracts , but their overgrowth can lead to infection in the urogenital tract and at distal sites. Treatment of Ureaplasma spp. infections is difficult. The lack of a cell wall confers resistance to all β-lactam and glycopeptide antibiotics whereas the lack of de novo synthesis of folic acid makes them resistant to sulfonamides and diaminopyrimidines. Thus, there is a need to search for new drugs effective against these bacteria. In the presented studies, the effect of N,N-diethyldithiocarbamate (DDC) at a concentration of 2 µg/ml, 20 µg/ml and 200 µg/ml on the growth and survival of Ureaplasma urealyticum and Ureaplasma parvum was tested. Our results indicated that all DDC concentrations used showed both bacteriostatic and bactericidal activity against both tested strains In brief, for the first time we showed the bacteriostatic and bactericidal activity of DDC against Ureplasma urealyticum and Ureaplasma parvum. Based on literature data, we conclude that this DDC activity is related to the inhibition of urease activity, which is an indispensable enzyme for these bacteria. Further research is needed and the results obtained in the present study should be verified in in vivo models of infection.