A series of new unique acetylene derivatives of 8-hydroxy- and 8-methoxyquinoline- 5-sulfonamide 3a-f and 6a-f were prepared by reactions of 8-hydroxy- and 8-methoxyquinoline- 5-sulfonyl chlorides with acetylene derivatives of amine. A series of new hybrid systems containing quinoline and 1,2,3-triazole systems 7a-h were obtained by reactions of acetylene derivatives of quinoline-5-sulfonamide 6a-d with organic azides. The structures of the obtained compounds were confirmed by 1H and 13C NMR spectroscopy and HR-MS spectrometry. The obtained quinoline derivatives 3a-f and 6a-f and 1,2,3-triazole derivatives 7a-h were tested for their anticancer and antimicrobial activity. Human amelanotic melanoma cells (C-32), human breast adenocarcinoma cells (MDA-MB-231), and human lung adenocarcinoma cells (A549) were selected as tested cancer lines, while cytotoxicity was investigated on normal human dermal fibroblasts (HFF-1). All the compounds were also tested against reference strains Staphylococcus aureus ATCC 29213 and Enterococcus faecalis ATCC 29212 and representatives of multidrug-resistant clinical isolates of methicillin-resistant S. aureus (MRSA) and vancomycin-resistant E. faecalis. Only the acetylene derivatives of 8-hydroxyquinoline-5-sulfonamide 3a-f were shown to be biologically active, and 8-hydroxy-N-methyl-N-(prop-2-yn-1-yl)quinoline-5-sulfonamide (3c) showed the highest activity against all three cancer lines and MRSA isolates. Its efficacies were comparable to those of cisplatin/doxorubicin and oxacillin/ciprofloxacin. In the non-cancer HFF-1 line, the compound showed no toxicity up to an IC50 of 100 μM. In additional tests, compound 3c decreased the expression of H3, increased the transcriptional activity of cell cycle regulators (P53 and P21 proteins), and altered the expression of BCL-2 and BAX genes in all cancer lines. The unsubstituted phenolic group at position 8 of the quinoline is the key structural fragment necessary for biological activity.
- MeSH
- antibakteriální látky * farmakologie chemie chemická syntéza MeSH
- chinoliny * chemie farmakologie chemická syntéza MeSH
- Enterococcus faecalis účinky léků MeSH
- lidé MeSH
- mikrobiální testy citlivosti * MeSH
- molekulární struktura MeSH
- nádorové buněčné linie MeSH
- protinádorové látky * farmakologie chemie chemická syntéza MeSH
- racionální návrh léčiv MeSH
- Staphylococcus aureus účinky léků MeSH
- sulfonamidy * farmakologie chemie chemická syntéza MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Antimicrobial resistance remains a global issue, hindering the control of bacterial infections. This study examined the antimicrobial properties of 2,3-N,N-diphenyl quinoxaline derivatives against Gram-positive, Gram-negative, and Mycobacterium species. Two quinoxaline derivatives (compounds 25 and 31) exhibited significant activity against most strains of Staphylococcus aureus, Enterococcus faecium, and Enterococcus faecalis tested, with MIC values ranging from 0.25 to 1 mg/L. These compounds also showed effective antibacterial activity against methicillin-resistant S. aureus (MRSA) and vancomycin-resistant E. faecium/E. faecalis (VRE) strains. They demonstrated comparable or superior activity to four current antibiotics (vancomycin, teicoplanin, daptomycin, and linezolid) against a wide range of clinically relevant isolates. Additionally, they were more effective in preventing S. aureus and E. faecalis biofilm formation compared to several other antibiotics. In summary, these two quinoxaline derivatives have potential as new antibacterial agents.
- MeSH
- antibakteriální látky * farmakologie MeSH
- biofilmy * účinky léků růst a vývoj MeSH
- chinoxaliny * farmakologie MeSH
- Enterococcus faecalis * účinky léků MeSH
- lidé MeSH
- methicilin rezistentní Staphylococcus aureus * účinky léků MeSH
- mikrobiální testy citlivosti * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
The MLST scheme currently used for Enterococcus faecium typing was designed in 2002 and is based on putative gene functions and Enterococcus faecalis gene sequences available at that time. As a result, the original MLST scheme does not correspond to the real genetic relatedness of E. faecium strains and often clusters genetically distant strains to the same sequence types (ST). Nevertheless, typing has a significant impact on the subsequent epidemiological conclusions and introduction of appropriate epidemiological measures, thus it is crucial to use a more accurate MLST scheme. Based on the genome analysis of 1,843 E. faecium isolates, a new scheme, consisting of 8 highly discriminative loci, was created in this study. These strains were divided into 421 STs using the new MLST scheme, as opposed to 223 STs assigned by the original MLST scheme. The proposed MLST has a discriminatory power of D = 0.983 (CI95% 0.981 to 0.984), compared to the original scheme's D = 0.919 (CI95% 0.911 to 0.927). Moreover, we identified new clonal complexes with our newly designed MLST scheme. The scheme proposed here is available within the PubMLST database. Although whole genome sequencing availability has increased rapidly, MLST remains an integral part of clinical epidemiology, mainly due to its high standardization and excellent robustness. In this study, we proposed and validated a new MLST scheme for E. faecium, which is based on genome-wide data and thus reflects the tested isolates' more accurate genetic similarity. IMPORTANCE Enterococcus faecium is one of the most important pathogens causing health care associated infections. One of the main reasons for its clinical importance is a rapidly spreading resistance to vancomycin and linezolid, which significantly complicates antibiotic treatment of infections caused by such resistant strains. Monitoring the spread and relationships between resistant strains causing severe conditions represents an important tool for implementing appropriate preventive measures. Therefore, there is an urgent need to establish a robust method enabling strain monitoring and comparison at the local, national, and global level. Unfortunately, the current, extensively used MLST scheme does not reflect the real genetic relatedness between individual strains and thus does not provide sufficient discriminatory power. This can lead directly to incorrect epidemiological measures due to insufficient accuracy and biased results.
In this study, we have focused on a multiparametric microbiological analysis of the antistaphylococcal action of the iodinated imine BH77, designed as an analogue of rafoxanide. Its antibacterial activity against five reference strains and eight clinical isolates of Gram-positive cocci of the genera Staphylococcus and Enterococcus was evaluated. The most clinically significant multidrug-resistant strains, such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant S. aureus (VRSA), and vancomycin-resistant Enterococcus faecium, were also included. The bactericidal and bacteriostatic actions, the dynamics leading to a loss of bacterial viability, antibiofilm activity, BH77 activity in combination with selected conventional antibiotics, the mechanism of action, in vitro cytotoxicity, and in vivo toxicity in an alternative animal model, Galleria mellonella, were analyzed. The antistaphylococcal activity (MIC) ranged from 15.625 to 62.5 μM, and the antienterococcal activity ranged from 62.5 to 125 μM. Its bactericidal action; promising antibiofilm activity; interference with nucleic acid, protein, and peptidoglycan synthesis pathways; and nontoxicity/low toxicity in vitro and in vivo in the Galleria mellonella model were found to be activity attributes of this newly synthesized compound. In conclusion, BH77 could be rightfully minimally considered at least as the structural pattern for future adjuvants for selected antibiotic drugs. IMPORTANCE Antibiotic resistance is among the largest threats to global health, with a potentially serious socioeconomic impact. One of the strategies to deal with the predicted catastrophic future scenarios associated with the rapid emergence of resistant infectious agents lies in the discovery and research of new anti-infectives. In our study, we have introduced a rafoxanide analogue, a newly synthesized and described polyhalogenated 3,5-diiodosalicylaldehyde-based imine, that effectively acts against Gram-positive cocci of the genera Staphylococcus and Enterococcus. The inclusion of an extensive and comprehensive analysis for providing a detailed description of candidate compound-microbe interactions allows the valorization of the beneficial attributes linked to anti-infective action conclusively. In addition, this study can help with making rational decisions about the possible involvement of this molecule in advanced studies or may merit the support of studies focused on related or derived chemical structures to discover more effective new anti-infective drug candidates.
- MeSH
- antibakteriální látky farmakologie chemie MeSH
- antiinfekční látky * farmakologie MeSH
- Enterococcus MeSH
- methicilin rezistentní Staphylococcus aureus * MeSH
- mikrobiální testy citlivosti MeSH
- rafoxanid farmakologie MeSH
- Staphylococcus aureus MeSH
- Staphylococcus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The beneficial influence of bacteriocin-producing, probiotic, mostly non-autochthonous bacteria has already been reported in various animals. However, their use in horses provides limited information, and results with autochthonous bacteria have not been reported. Therefore, the main objective of this model study was to test the effect of autochthonous, bacteriocin-producing faecal strain Enterococcus faecium EF 412 application in horses. One gram of freeze-dried EF 412 strain (109 CFU/mL for 21 days) was applied to horses in a small feed ball. Clinically healthy horses (12), Slovak warm-blood breed of various ages (5-13 years), were involved in a 35-day-long experiment, also functioning as control for themselves. They were stabled in separate boxes (university property), fed twice a day (hay, whole oats or grazed) with water access ad libitum. Sampling was performed at the start of the experiment, i.e. at days 0/1, 21 (3 weeks of EF 412 application) and at day 35 (2 weeks of EF 412 cessation). EF 412 colonized GIT of horses was 3.54 ± 0.75 CFU/g (log 10) at day 21. The eggs of the nematode Strongylus spp. were not found in horses after EF 412 application, and Eimeria spp. oocysts were similarly not found. The other microbiota were not reduced as evaluated by the use of standard method. Using next-generation sequencing, at phylum level, phyla Bacteroidetes and Firmicutes dominated and at family level, they were Bacteroidales BS11 and S24-7 gut goups and Lentisphaerae. In horses, the increasing tendency in phagocytic activity was noted after EF 412 application. Biochemical parameters were in the physiological range. Total protein value was significantly decreased at day 21 compared with day 0/1 as well as with day 35 (P < 0.05). Cholesterol and triglycerides were influenced (decreased) at day 21 compared with day 0/1 and day 35. Neither nematode eggs Strongylus spp. nor Eimeria spp. oocysts were found in faeces after EF 412 application. Autochthonous, faecal strain E. faecium EF 412 showed promising application potential.
A small receptor molecule composed of a porphyrin core with tetrakis-ammonium glycine pickets (liptin 3e) appears to target anionic phosphatidylglycerol (PG) lipid head groups through multifunctional binding-pocket complementarity. Although a major component of bacterial cell membranes, PG is not widely found in animal cells, making PG potentially selective for bacterial targeting. Growth of microbial isolates was monitored in liquid cultures treated with liptin 3e by dilution plate counts and turbidity. Inhibition of growth by liptin 3e was observed for the ESKAPE human pathogens (Enterobacter aerogenes, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterococcus faecium), Escherichia coli, Mycobacterium smegmatis, Streptococcus sobrinus, and methicillin-resistant S. aureus (MRSA), with certain species suppressed at <1 μg/mL (sub-μM) concentrations. Prolonged lag phases were observed, although cell viability was mainly unaffected, suggesting that liptin treatment caused bacteriostasis. Cultures treated with liptin 3e eventually recovered, resumed growth, and reached the same final densities as untreated cultures. Growth of the fungus Candida albicans was not appreciably inhibited by liptin 3e. If liptins exhibit bacteriostasis through broad extracellular binding to PG head groups, thereby disrupting cellular processes, liptins may be considered for development into preclinical drug candidates or be useful as a targeting system for molecular beacons or antibacterial drugs.
- MeSH
- antibakteriální látky farmakologie MeSH
- Enterococcus faecium * MeSH
- Escherichia coli MeSH
- fosfatidylglyceroly MeSH
- lidé MeSH
- methicilin rezistentní Staphylococcus aureus * MeSH
- mikrobiální testy citlivosti MeSH
- receptory umělé * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Cíl: Zhodnotit soubor pacientů hospitalizovaných pro infekční endokarditidu (IE) v Komplexním kardiovaskulárním centru (KKC) FN Plzeň, posoudit změny mikrobiálního spektra a prognózu nemocných. Metodika: Retrospektivně jsme analyzovali soubor 109 pacientů, kteří byli v období 5/2011-4/2019 hospitalizováni v KKC FN Plzeň a kteří splňovali diagnózu IE podle modifikovaných Duke kritérií. Uvedený 8letý časový úsek jsme dále rozdělili do dvou 4letých období k porovnání vývoje souboru nemocných v čase. Výsledky: V souboru pacientů bylo 85 mužů a 24 žen. U 56 nemocných (51 %) se jednalo o IE nativní chlopně (NVE), u 35 pacientů (32 %) o IE chlopenní protézy (PVE) a u 18 pacientů (17 %) šlo o IE v přítomnosti kardiostimulačního nebo defibrilačního systému (CDRIE). Z 35 případů PVE bylo postiženo 8 mechanických a 27 biologických protéz (včetně 2 případů postižení katetrizačně implantované aortální chlopně - TAVI). Při srovnání období od 5/2011-4/2015s obdobím od 5/2015-4/2019 došlo k vzestupu počtu hospitalizovaných pacientů s IE o 14 %. Významně se snížil počet nemocných s NVE (35 vs. 21, p = 0,001), zatímco došlo k nárůstu počtu nemocných s PVE (10 vs. 25, statisticky významně, p = 0,0131) a CDRIE (6 vs. 12, statisticky nevýznamně, p = 0,3017). Dominantním patogenem u pacientů s NVE a CDRIE zůstává Staphylococcus aureus (32 %, respektive 33 %), u nemocných s PVE byly častěji identifikovány koaguláza negativní stafylokoky (31 %). Celková hospitalizační mortalita byla 23 %, relativně nižší u pacientů s NVE (16 %), naopak výrazně vyšší u pacientů s PVE (31 %) a CDRIE (28 %). Závěr: V této retrospektivní studii jsme potvrdili vzestup celkové incidence infekční endokarditidy. Významně se snížil počet nemocných s NVE, zatímco došlo k nárůstu počtu nemocných s PVE a CDRIE. Pacienti s PVE a CDRIE rovněž vykazují vyšší mortalitu. Dominantními patogeny IE jsou Staphylococcus aureus, koaguláza negativní stafylokoky a Enterococcus faecalis.
Aim: To assess the set of patients with infective endocarditis (IE) in Complex Cardiovascular Center (CCC) in Pilsen and evaluate changes in microbial spectrum and prognosis of patients. Methods: We used a retrospective analysis to identify 109 patients with the diagnosis of IE according to modified Duke criteria, who were hospitalised in CCC in Pilsen from May 2011 to April 2019. This eight-year time period was divided into two four-year intervals to compare the changes in the set of patients. Results: The study included 85 men and 24 women. 56 patients (51 %) had native valve endocarditis (NVE), 35 patients (32 %) had prosthetic valve endocarditis (PVE) and 18 patients (17 %) had cardiac device related infective endocarditis (CDRIE). In the PVE group, 8 mechanical and 27 biological prosthesis (including 2 cases of IE after transcatheter aortic valve implantation - TAVI) were affected. When comparing the periods of May 2011 to April 2015 with May 2015 to April 2019, we found out a 14 % increase in the number of patients with IE. The number of patients with NVE decreased significantly (35 vs. 21, p = 0,001), while the number of patients with PVE (10 vs. 25, statistically significant, p = 0,0131) and CDRIE (6 vs. 12, statistically non-significant, p = 0,3017) increased. Staphylococcus aureus remains the dominant pathogen in NVE and CDRIE group (32 % and 33 %, respectively), while in PVE patients there were more often identified coagulase negative Staphylococci as a causative microbial agent (31 %). The overall in-hospital mortality rate was 23 %, relatively lower in patients with NVE (16 %), on the contrary, significantly higher in patients with PVE (31 %) a CDRIE (28 %). Conclusion: This retrospective study confirms an increase of total incidence of infective endocarditis. The number of patients with NVE decreased significantly, while the number of patients with PVE and CDRIE increased. Patients with PVE and CDRIE also show higher mortality. The dominant pathogens are Staphylococcus aureus, coagulase negative Staphylococci and Enterococcus faecalis.
- MeSH
- bakteriální endokarditida * epidemiologie etiologie mikrobiologie mortalita MeSH
- Enterococcus faecalis MeSH
- hospitalizace MeSH
- interpretace statistických dat MeSH
- klinický obraz nemoci MeSH
- lidé MeSH
- retrospektivní studie MeSH
- senioři MeSH
- srdeční chlopně umělé mikrobiologie MeSH
- srdeční chlopně mikrobiologie MeSH
- Staphylococcus MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
The study aimed to identify colonized patients as a possible source of eventual VRE (vancomycin-resistant enterococci) infection from stool samples positive for glutamate dehydrogenase antigen, as well as for Clostridioides difficile toxins A and B. The study was carried out from 7/2020 to 9/2021. Stool samples were grown in a brain heart infusion medium with a gram-positive non-spore-forming bacteria supplement under aerobic conditions. The samples for VRE identification were grown on CHROMID® VRE agar, and the MICs for vancomycin and teicoplanin were also estimated. The presence of the vanA/vanB genes was tested using the PCR method. The total number of 113 stool samples positive for Clostridioides difficile toxins was analyzed. Of these samples, 44 isolates with VRE characters were identified. The most prevalent isolates in our set of isolates were Enterococcus faecium (27 isolates, 62%), Enterococcus faecalis (9 isolates, 21%), Enterococcus solitarius (4 isolates, 9%), Enterococcus durans (2 isolates, 4%), 1 isolate Enterococcus sulfurous (2%), and Enterococcus raffinosus (2%). In total, 26 isolates were detected in the study in the presence of vanA genes (24 isolates E. faecium, 2 isolates E. faecalis) and 18 isolates detected in the presence of vanB genes (7 isolates E. faecalis, 4 isolates E. solitarius, 3 isolates E. faecium, 2 isolates E. durans, 1 isolate E. sulfurous, and E. raffinosus). The results of this study showed the local dominance character of the vanA gene of hospital VRE isolates that were carriers of genes associated with high resistance to vancomycin, teicoplanin, and occasionally linezolid.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální proteiny genetika MeSH
- Clostridioides difficile * genetika MeSH
- Enterococcus faecium * genetika MeSH
- enterokoky rezistentní vůči vankomycinu * genetika MeSH
- grampozitivní bakteriální infekce * mikrobiologie MeSH
- lidé MeSH
- mikrobiální testy citlivosti MeSH
- teikoplanin farmakologie MeSH
- vankomycin farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Slovenská republika MeSH
Enterococcus faecalis is an important factor in nosocomial infections. The aim of this study was the isolation of the pathogenic genes of Enterococcus faecalis in mouth infection and the study of the expression of these genes by real-time PCR. In this study, 60 isolates of E. faecalis were isolated from oral infections. The presence and frequency of cyl, hyl, and esp genes and their expression by ionic liquid were evaluated using real-time PCR. MIC was determined by broth dilution method and biofilm production was measured, then biofilm inhibition ability and cytotoxicity test were performed by MTT method. The esp, cyl, and hyl genes were observed in 10, 11, and 2 isolates, respectively. cyl gene with the highest frequency of expression in the treated group was reduced by 1.14% under the influence of ionic liquid with methionine base. The results of MIC and Sub MIC concentrations were obtained with the effect of ionic liquid including 125 and 225 μg/mL, respectively. Amino-acid-based ionic liquids can also reduce biofilm production at sub-MIC concentrations (P < 0.05), and changes in cytotoxicity at different concentrations and over time are significant (P-value < 0.001). E. faecalis strains are genetically diverse and this indicates the polyclonal prevalence of strains in clinical specimens. Combination treatment of ionic liquid with common antimicrobial drugs has good antibacterial effects against Enterococcus species, and ionic liquid with a minimum dose can be a good alternative to single-drug treatment of Enterococcus infections.
- MeSH
- aminokyseliny MeSH
- antibakteriální látky farmakologie MeSH
- biofilmy MeSH
- Enterococcus faecalis MeSH
- grampozitivní bakteriální infekce * farmakoterapie mikrobiologie MeSH
- iontové kapaliny * farmakologie MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé MeSH
- methionin genetika farmakologie MeSH
- mikrobiální testy citlivosti MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
