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Despite the urgent need for new antibiotics, very few innovative antibiotics have recently entered clinics or clinical trials. To provide a constant supply of new drug candidates optimized in terms of their potential to select for resistance in natural settings, in vitro resistance-predicting studies need to be improved and scaled up. In this review, the following in vitro parameters are presented: frequency of spontaneous mutant selection (FSMS), mutant prevention concentration (MPC), dominant mutant prevention concentration (MPC-D), inferior-mutant prevention concentration (MPC-F), and minimal selective concentration (MSC). The utility of various adaptive laboratory evolution (ALE) approaches (serial transfer, continuous culture, and evolution in spatiotemporal microenvironments) for comparing hits in terms of the level and time required for multistep resistance to emerge is discussed. We also consider how the hit-to-lead stage can benefit from high-throughput ALE setups based on robotic workstations, do-it-yourself (DIY) continuous cultivation systems, microbial evolution and growth arena (MEGA) plates, soft agar gradient evolution (SAGE) plates, microfluidic chips, or microdroplet technology. Finally, approaches for evaluating the fitness of in vitro-generated resistant mutants are presented. This review aims to draw attention to newly emerged ideas on how to improve the in vitro forecasting of the potential of compounds to select for resistance in natural settings. [ABSTRACT FROM AUTHOR]

Simple Summary: Fluoroquinolones, mainly levofloxacin, are considered an alternative treatment option of Stenotrophomonas maltophilia infections to trimethoprim/sulfamethoxazole. However, an increase in the number of levofloxacin-resistant strains is observed worldwide. The fluoroquinolone resistance in S. maltophilia is usually caused by an overproduction of various multidrug efflux pumps, which are able to extrude antibiotics and chemotherapeutics from the bacterial cells. The purpose of the study was to analyze the contribution of efflux systems to levofloxacin resistance in S. maltophilia clinical strains, isolated in Warsaw, by phenotypic and molecular methods. Previously, the occurrence of genes encoding various ten efflux pumps was shown in 94 studied isolates. Additionally, 44 of 94 isolates demonstrated reduction in susceptibility to levofloxacin. In this study, in the presence of efflux pump inhibitors, an increase in levofloxacin susceptibility was observed in 13 isolates. The overexpression of genes encoding two efflux pump system, such as SmeDEF and Sme VWX (in five and one isolate, respectively), was demonstrated. Sequencing analysis revealed an amino acid change in the local regulators of these efflux pump operons. Our data indicate that the overproduction of the SmeVWX efflux system, unlike SmeDEF, plays a significant role in the levofloxacin resistance of the clinical isolates. Levofloxacin is considered an alternative treatment option of Stenotrophomonas maltophilia infections to trimethoprim/sulfamethoxazole. The fluoroquinolone resistance in S. maltophilia is usually caused by an overproduction of efflux pumps. In this study, the contribution of efflux systems to levofloxacin resistance in S. maltophilia clinical isolates was demonstrated using phenotypic (minimal inhibitory concentrations, MICs, of antibiotics determination ± efflux pump inhibitors, EPIs) and molecular (real-time polymerase-chain-reaction and sequencing) methods. Previously, the occurrence of genes encoding ten efflux pumps was shown in 94 studied isolates. Additionally, 44/94 isolates demonstrated reduction in susceptibility to levofloxacin. Only 5 of 13 isolates (with ≥4-fold reduction in levofloxacin MIC) in the presence of EPIs showed an increased susceptibility to levofloxacin and other antibiotics. The overexpression of smeD and smeV genes (in five and one isolate, respectively) of 5 tested efflux pump operons was demonstrated. Sequencing analysis revealed 20–35 nucleotide mutations in local regulatory genes such as smeT and smeRv. However, mutations leading to an amino acid change were shown only in smeT (Arg123Lys, Asp182Glu, Asp204Glu) for one isolate and in smeRv (Gly266Ser) for the other isolate. Our data indicate that the overproduction of the SmeVWX efflux system, unlike SmeDEF, plays a significant role in the levofloxacin resistance. [ABSTRACT FROM AUTHOR]

A micellar electrokinetic chromatography method for simultaneous assay of ticarcillin and clavulanic acid in Timentin i.v. injection preparation was developed. This method ensures excellent separation of both components of Timentin preparation. The validation of the method was performed, and specificity, reproducibility, precision and accuracy were confirmed. The detection and quantitative limits for Timentin were established in the concentrations 0.04 and 0.08 mg/ml, respectively. The elaborated technique was compared with two methods routinely used-UV and high performance liquid chromatography (HPLC). The obtained results and their statistical analysis proved the same precision of all methods, however, no significant differences were observed between CE and HPLC. [Copyright &y& Elsevier]

A micellar electrokinetic capillary electrophoretic method for determination of sultamicillin in Unasyn oral preparations—tablets and suspension—was evaluated. Phosphate–borate buffer at pH 7.0 containing 1.0% sodium dodecylsulfate was used as a mobile phase. The elaborated method ensures separation of sultamicillin from p-toluenesulfonic acid and the impurities, ampicillin, sulbactam and penicillamine. The method was validated for specificity, reproducibility, precision, accuracy and assay linearity (in a concentration range of sultamicillin of 0.05—1.5 mg/ml). Statistical analysis by Student’s t-test showed no significant differences between the results obtained by micellar electrokinetic chromatography and HPLC, tcalculated 0.519 for suspension assays and 0.284 for tablets assays were smaller then ttabulated. [Copyright &y& Elsevier]

Simple Summary: Nosocomial infections caused by Stenotrophomonas maltophilia have been increasing worldwide. These bacteria are intrinsically resistant to most antibiotics. The underestimated resistance mechanism of Gram-negative rods is an overexpression of multidrug-resistant (MDR) efflux pumps. The aim of this study was to analyze the genetic diversity of isolates derived from various clinical materials, including blood, and the prevalence of MDR efflux pump genes and susceptibility profiles to the anti-S. maltophilia drugs. The research was conducted on 94 S. maltophilia isolates derived from hospitalized patients and outpatients in Warsaw, Poland. All isolates were susceptible to trimethoprim-sulfamethoxazole and minocycline, while 44/94 isolates demonstrated reduction in susceptibility to levofloxacin. A large genetic variation was observed among these isolates. However, a clonal relationship was revealed among two groups of bloodstream isolates from one hospital ward: (1) nine isolates, (2) six isolates. Moreover, the presence of genes encoding ten different efflux pumps from the resistance-nodulation-division family and the ATP-binding cassette family was shown in the majority of the 94 isolates. The obtained knowledge about the prevalence of efflux pump genes in clinical S. maltophilia strains makes it possible to predict the scale of the risk of resistance emergence in strains as a result of gene overexpression. An increase of nosocomial infections caused by Stenotrophomonas maltophilia strains has recently been observed all over the world. The isolation of these bacteria from the blood is of particular concern. In this study we performed the phenotypic and genotypic characterization of 94 S. maltophilia isolates, including isolates from patients hospitalized in a tertiary Warsaw hospital (n = 79) and from outpatients (n = 15). All isolates were found to be susceptible to trimethoprim-sulfamethoxazole and minocycline, while 44/94 isolates demonstrated a reduction in susceptibility to levofloxacin. A large genetic variation was observed among the isolates tested by pulsed-field gel electrophoresis. A clonal relationship with 100% similarity was observed between isolates within two sub-pulsotypes: the first included nine bloodstream isolates and the second involved six. Multilocus sequence typing showed two new sequence types (ST498 and ST499) deposited in public databases for molecular typing. Moreover, the presence of genes encoding ten different efflux pumps from the resistance-nodulation-division family and the ATP-binding cassette family was shown in the majority of the 94 isolates. The obtained knowledge about the prevalence of efflux pump genes in clinical S. maltophilia strains makes it possible to predict the scale of the risk of resistance emergence in strains as a result of gene overexpression. [ABSTRACT FROM AUTHOR]

The efflux pumps, beside the class D carbapenem-hydrolysing enzymes (CHLDs), are being increasingly investigated as a mechanism of carbapenem resistance in Acinetobacter baumannii. This study investigates the contribution of efflux mechanism to carbapenem resistance in 61 acquired blaCHDL-genes-carrying A. baumannii clinical strains isolated in Warsaw, Poland. Studies were conducted using phenotypic (susceptibility testing to carbapenems ± efflux pump inhibitors (EPIs)) and molecular (determining expression levels of efflux operon with regulatory-gene and whole genome sequencing (WGS)) methods. EPIs reduced carbapenem resistance of 14/61 isolates. Upregulation (5–67-fold) of adeB was observed together with mutations in the sequences of AdeRS local and of BaeS global regulators in all 15 selected isolates. Long-read WGS of isolate no. AB96 revealed the presence of AbaR25 resistance island and its two disrupted elements: the first contained a duplicate ISAba1-blaOXA-23, and the second was located between adeR and adeA in the efflux operon. This insert was flanked by two copies of ISAba1, and one of them provides a strong promoter for adeABC, elevating the adeB expression levels. Our study for the first time reports the involvement of the insertion of the ΔAbaR25-type resistance island fragment with ISAba1 element upstream the efflux operon in the carbapenem resistance of A. baumannii. [ABSTRACT FROM AUTHOR]

The potential role of non-antibiotic medicinal products in the treatment of multidrug-resistant Gram-negative bacteria has recently been investigated. It is highly likely that the presence of efflux pumps may be one of the reasons for the weak activity of non-antibiotics, as in the case of some non-steroidal anti-inflammatory drugs (NSAIDs), against Gram-negative rods. The activity of eight drugs of potential non-antibiotic activity, active substance standards, and relevant medicinal products were analysed with and without of efflux pump inhibitors against 180 strains of five Gram-negative rod species by minimum inhibitory concentration (MIC) value determination in the presence of 1 mM MgSO4. Furthermore, the influence of non-antibiotics on the susceptibility of clinical strains to quinolones with or without PAβN (Phe-Arg-β-naphthylamide) was investigated. The impacts of PAβN on the susceptibility of bacteria to non-antibiotics suggests that amitriptyline, alendronate, nicergoline, and ticlopidine are substrates of efflux pumps in Gram-negative rods. Amitriptyline/Amitriptylinum showed the highest direct antibacterial activity, with MICs ranging 100–800 mg/L against all studied species. Significant decreases in the MIC values of other active substances (acyclovir, atorvastatin, and famotidine) tested with pump inhibitors were not observed. The investigated non-antibiotic medicinal products did not alter the MICs of quinolones in the absence and in the presence of PAβN to the studied clinical strains of five groups of species. [ABSTRACT FROM AUTHOR]

The adhesion and aggregation are characteristic attributes of probiotic strains belonging to Lactobacillaceae genus. Due to these properties the host organisms can avoid colonisation of the intestinal tract by enteropathogenic bacteria. The presented research includes a comparison of the properties of various strains belonging to different Lactobacillaceae species and isolated from different sources The aim of this study was to investigate the ability of Lactocaseibacillus rhamnosus, Lactiplantibacillus plantarum, and Lactobacillus strains (L. acidophilus, L. gasseri, L. ultunensis) from probiotic products and clinical specimens to direct and competitive adherence to Caco-2 and HT-29 cell lines. Furthermore, the ability of lactobacilli and enteropathogenic bacteria, E. coli, E. faecalis, and S. Typhimurium, to auto- and co-aggregation was also investigated. The results showed that all tested strains adhered to Caco-2 and HT-29 cell lines. Though, the factor of adhesion depended on the species and origin of the strain. L. rhamnosus strains showed a lowest degree of adherence as compared to L. plantarum and Lactobacillus sp. strains. On the other side both, L. rhamnosus and L. acidophilus strains reduced the pathogenic bacteria in competition adherence test most effectively. All tested lactobacilli strains were characterised by auto- and co-aggregation abilities, to various degrees. The properties of Lactobacillaceae strains analysed in this study, like adhesion abilities, competitive adherence, auto- and co-aggregation, may affect the prevention of colonisation and elimination of pathogenic bacteria in gastrointestinal tract. [ABSTRACT FROM AUTHOR]

Summary The damage of Candida albicans blastoconidia exposed to the lowest concentration of chemical disinfectant and antiseptic passing the EN 1275:2005 at 5 min contact time was examined under transmission electron microscopy and scanning electron microscopy. The blastoconidia of C. albicans 82 exposed to the chemical products showed the following damage: Incidin Liquid Spray (disinfectant) and Spitaderm (antiseptic) - cytoplasm congealing; Incidin Plus and Lysoformine 3000 (disinfectants) - cell wall and cell membrane damage, and leakage of the cytoplasm; Medicarine (disinfectant) - cytoplasm denaturation and rough cytoplasm. The damage of blastoconidia of the strains C. albicans 24 and C. albicans 28, exhibiting different susceptibility to Lysoformine 3000 was compared under scanning electron microscopy. The blastoconidia of the strain C. albicans 28 did not exhibit essential damage at a concentration 16 times lower (0.02%) than the lowest concentration of Lysoformine 3000 (Lc-LYS) for this strain (0.32%). At this concentration (0.02%, corresponding to Lc-LYS for C. albicans 24) the more sensitive isolate 24 showed substantial damage. The conducted study exhibited the effects of the tested products against the viability and structural integrity of the cells. Blastoconidial buds at the early stage of development seem to be very susceptible to the action of the tested products. The budding areas are located mainly at the blastoconidial poles. [ABSTRACT FROM AUTHOR]

Abstract: Ertapenem, a Group 1 carbapenem, is most recently introduced into the market. It is a β-lactam antibiotic that possesses a broad antibacterial spectrum including common community-acquired Gram-positive and Gram-negative aerobic and anaerobic pathogens, but low activity against some nosocomial pathogens such as Pseudomonas aeruginosa, Acinetobacter spp., enterococci and methicillin-resistant staphylococci. The elaborated method of micellar electrokinetic chromatography (MEKC) of ertapenem separation from its impurities was successfully performed using normal stacking mode (NSM) and stacking with reverse migrating micelles (SRMM), followed by UV absorption detection at 214nm. The best results were obtained with 60mM sodium dihydrogen phosphate and 20mM boric acid buffer pH 6.0, as background electrolyte. Uncoated fused-silica capillary and neutral-coated capillary with normal and reverse polarity, and voltage values of +18 and −12kV, respectively, were used throughout the investigation. Sodium dodecyl sulfate was employed as the pseudostationary phase. A comparison of applied techniques, including sensitivity enhancement factors and limits of detection (LOD), is presented. The optimized method was validated in terms of linearity, accuracy and precision. Comparable LOD was obtained using both stacking methods (0.3μg/mL) but better efficiency of ertapenem peak was obtained using NSM. Under the optimum stacking conditions, about 183–4.75-fold and 1289–4.07-fold improvements in peak areas were obtained for NSM and SRMM, respectively, compared to the usual hydrodynamic sample injection (10s). The reproducibility, expressed by relative standard deviations (RSD) of the migration times, for NSM was about 0.96–1.25 and for SRMM was 0.32–0.45. The RSD of corrected peak areas, for NSM was about 1.07–8.14 and for SRMM was 0.74–8.12. The difference in separation time between the two techniques was not obvious. Satisfactory separation was possible after less than 11min of electrophoresis. The evaluated MEKC method was applied in the analysis of medicinal product containing ertapenem: Invanz—ertapenem for injection. [Copyright &y& Elsevier]

Abstract: Linezolid is the first compound of a truly new class of antibiotics—the oxazolidinones. The elaborated method of capillary electrophoresis (CE) of linezolid separation from its achiral impurities was successfully performed using sweeping preconcentration, followed by UV absorption detection at 254nm. The best results were obtained with 125mM Tris buffer, pH 2.0, with addition of 20% (v/v) methanol as background electrolyte. Sodium dodecyl sulfate (150mM) was added to the electrolyte in the inlet vial as the sweeping agent. The separation was carried out at negative polarity. Then, the optimized method was validated in terms of linearity, accuracy and precision. Sweeping preconcentration of linezolid provides detection limit at 0.05μg/ml level. The evaluated CE method was applied in the analysis of medicinal product containing linezolid–linezolid solution for infusion. [Copyright &y& Elsevier]

Abstract: A method for the enantioseparation of linezolid, the first compound of a truly new class of antibiotics—the oxazolidinones, was developed. The elaborated method of linezolid enantiomers separation was successfully performed using an anionic single-isomer cyclodextrin—heptakis-(2,3-diacetyl-6-sulfato)-β-cyclodextrin (HDAS-β-CD) as a resolving agent with the help of the charged resolving agent migration model (CHARM model). The best results were obtained with 27.5mM HDAS-β-CD dissolved in 50mM borate buffer, pH 9.0, 15°C, normal polarity. The facile strategies for the reversal of the enantiomers elution order are also described. Afterwards, the optimized method was validated in terms of sensitivity, linearity, accuracy and precision. [Copyright &y& Elsevier]

Abstract: The application of capillary electrophoresis for separation of penicillin V and its impurities was investigated. The phosphate–borate buffer supplemented with sodium dodecyl sulfate (SDS) 20.0g/L (69mM) and pentanesulfonic acid sodium salt (PS) 2.2g/L (12.5mM) adjusted to pH 6.3, and current voltage 15kV seem to provide optimal conditions for this aim. The resolution between penicillin V and each impurity was very good. The statistical analysis of phenoxymethylpenicillin V assay showed no significant differences between the results obtained by CE and HPLC methods. [Copyright &y& Elsevier]

Capillary zone electrophoresis (CZE) has been elaborated for separation, identification and determination of ciprofloxacin and its impurities. The separation, phosphate buffer pH 6.0 was supplemented with 0.075 M pentane-1-sulfonic acid sodium salt. The elaborated method was validated. The selectivity, linearity, limits of detection (LOD) and quantification (LOQ), precision, and accuracy of capillary zone electrophoresis were evaluated. The results obtained by CZE were also compared with those obtained by liquid chromatography. Regarding the validation results the CE method fulfils the current European Pharmacopoeia (Eur. Ph.) requirements. The evaluated CE method could be applicable to the analysis of different medicinal products containing ciprofloxacin. [Copyright &y& Elsevier]

The application of capillary electrophoresis for separation of benzylpenicillin, procaine, benzathine and clemizole was investigated. Phosphate–borate buffer supplemented with sodium dodecyl sulphate 14.4 g/l and electrophoresis voltage 18 kV seem to provide optimal conditions for micellar electrokinetic chromatographic assay of penicillin salts. This method is selective and precise. The results obtained from CE method recovery assay (above 98% for all but procaine—97% substances) and from determination of benzylpenicillin by CE compared with HPLC results, confirmed good accuracy. [Copyright &y& Elsevier]

Capillary electrophoresis method for identification and simultaneous determination of procaine, dihydrostreptomycin and penicillin G, present in multiantibiotic veterinary preparations, was elaborated. The influence of pH (5.0–9.75) and concentration of disodium tetraborate decahydrate in running buffers (0.02–0.1 M) as well as temperatures (25–40 °C) on separation efficacy were analyzed. For quantitative analysis, 0.08 M borate buffer (pH 8.0) at 35 °C and 15 kV were chosen. Method was validated, selectivity, precision, linearity, LOD, LOQ, accuracy and specificity of capillary zone electrophoresis (CZE) were evaluated. [Copyright &y& Elsevier]

A rapid, capillary electrophoresis method was evaluated for determination of amoxicillin and clavulanic acid in Augmentin as well as ampicillin and sulbactam in Unasyn preparations for injections. Phosphate–borate buffer at pH 8.66 containing 14.4% sodium dodecyl sulfate was used as a mobile phase. The method was validated. Reproducibility, precision, accuracy and assay linearity in concentration of amoxicillin 0.05–3.03 mg/ml and ampicillin 0.05–3.08 mg/ml, as well as clavulanic acid 0.02–2.02 mg/ml and sulbactam 0.05–2.08 mg/ml were established. This new method is fast, inexpensive and limits consumption of organic solvents when compared with alternative high performance liquid chromatography (HPLC) method, used for drug analysis. Statistical analysis by Student''s t-test showed no significant differences between the results obtained by the two methods tcalculated 0.32 and 1.69 for amoxicillin and clavulanic acid and 0.67 and 1.93 for ampicillin and sulbactam were smaller than ttabulated. [Copyright &y& Elsevier]

Acinetobacter baumannii is an important cause of nosocomial infections worldwide. The elucidation of the carbapenem resistance mechanisms of hospital strains is necessary for the effective treatment and prevention of resistance gene transmission. The main mechanism of carbapenem resistance in A. baumannii is carbapenemases, whose expressions are affected by the presence of insertion sequences (ISs) upstream of blaCHDL genes. In this study, 61 imipenem-nonsusceptible A. baumannii isolates were characterized using phenotypic (drug-susceptibility profile using CarbaAcineto NP) and molecular methods. Pulsed field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) methods were utilized for the genotyping. The majority of isolates (59/61) carried one of the following acquired blaCHDL genes: blaOXA-24-like (39/59), ISAba1-blaOXA-23-like (14/59) or ISAba3-blaOXA-58-like (6/59). Whole genome sequence analysis of 15 selected isolates identified the following intrinsic blaOXA-66 (OXA-51-like; n = 15) and acquired class D β-lactamases (CHDLs): ISAba1-blaOXA-23 (OXA-23-like; n = 7), ISAba3-blaOXA-58-ISAba3 (OXA-58-like; n = 2) and blaOXA-72 (OXA-24-like; n = 6). The isolates were classified into 21 pulsotypes using PFGE, and the representative 15 isolates were found to belong to sequence type ST2 of the Pasteur MLST scheme from the global IC2 clone. The Oxford MLST scheme revealed the diversity among these studied isolates, and identified five sequence types (ST195, ST208, ST208/ST1806, ST348 and ST425). CHDL-type carbapenemases and insertion elements upstream of the blaCHDL genes were found to be widespread among Polish A. baumannii clinical isolates, and this contributed to their carbapenem resistance. [ABSTRACT FROM AUTHOR]

Cross-infection involves the transmission of microorganisms through secretions, bodily fluids and excreta, as well as undisinfected surfaces and medical equipment. In the dental office, diseases are transmitted via various routes, e.g. from patient to dentist or other member of dental team, from doctor or dental team member to patient, from patient to another patient, from dental office to community and from community to patient. The study was conducted to evaluate the effectiveness of infection control in dental practices based on the qualitative and quantitative assessment of microbiological contaminants detected on the surface of filling material packaging used in dental offices. The material for research were 9 packages containing dental materials during their use in 3 dental settings. The packages were placed in sterile flasks and rinsed to wash microorganisms from the surfaces. The washes were filtered through membrane filters and cultured under proper aerobic and anaerobic conditions, and at elevated CO2 concentration. Microbial growth on TIO and TSB media was observed. The contamination of most samples remained low as indicated by the growth from one to a maximum of five colonies on TSA. The contamination remained at the level of 10-50 CFU/package, i.e. <100 CFU/single package. The tests evaluating the contamination of dental package surfaces with aerobic bacteria confirmed high hygiene standards observed in dental offices from which the packages were brought. [ABSTRACT FROM AUTHOR]

Ferrocene‐amino acid ester‐uracil conjugates, derived from four amino acids (l‐alanine, glycine, β‐alanine and γ‐aminobutyric acid), were synthesized via the Cu(I)‐catalyzed azide‐alkyne cycloaddition as the key step. The new compounds were characterized spectroscopically and by single‐crystal X‐ray crystallography in the case of the derivatives of glycine (monoclinic, P21/c) and γ‐aminobutyric acid (orthorhombic, Pca21). Electrochemical measurements showed subtle influence of the compounds' structure on the redox potential of the Fc/Fc+ couple. Among the four compounds tested for antimicrobial activity, the conjugate obtained from l‐alanine possessed weak antifungal activity against Candida guillermondii. [ABSTRACT FROM AUTHOR]

A series of 22 benzosiloxaboroles, silicon analogues of strong antimicrobial agents - benzoxaboroles, have been synthesized and tested against β-lactamases KPC- and pAmpC-producing strains of Gram-negative rods. Comprehensive structural-property relationship studies supported by molecular modelling as well as biological studies reveal that 6-B(OH) 2 -substituted derivative 27 strongly inhibits the activity of cephalosporinases (chromosomally encoded AmpC and plasmid encoded CMY-2) and KPC carbapenemases. It also shows strong ability to inhibit growth of the strains producing KPC-3 when combined with meropenem. In addition, halogen-substituted (mono-, di- or tetra-) benzosiloxaboroles demonstrate high antifungal activity (MIC 1.56–6.25 mg/L) against C. tropicalis , C. guilliermondii and S. cerevisiae. The highest activity against pathogenic yeasts (C. albicans , C. krusei and C. parapsilosis - MICs 12.5 mg/L) and against Gram-positive cocci (S. aureus and E. faecalis - 6.25 mg/L and 25 mg/L respectively) was displayed by 6,7-dichloro-substituted benzosiloxaborole. The studied systems exhibit low cytotoxity toward human lung fibroblasts. Image 1 • Synthesis and characterization of a series of 22 functionalized benzosiloxaboroles. • Strong antifungal activity of fluorinated derivatives. • KPC/AmpC inhibitory activity of selected benzosiloxaboroles. • Docking and molecular dynamics studies on the binding to the KPC-2 β-lactamase. [ABSTRACT FROM AUTHOR]

Peri-implant infective diseases (PIIDs) in oral implantology are commonly known as peri-implant mucositis (PIM) and periimplantitis (PI). While PIM is restricted to the peri-implant mucosa and is reversible, PI also affects implant-supporting bone and, therefore, is very difficult to eradicate. PIIDs in clinical outcome may resemble gingivitis and periodontitis, as they share similar risk factors. However, recent study in the field of proteomics and other molecular studies indicate that PIIDs exhibit significant differences when compared to periodontal diseases. This review aims to elucidate the current knowledge of PIIDs, their etiopathology and diversified microbiology as well as the role of molecular studies, which may be a key to personalized diagnostic and treatment protocols of peri-implant infections in the near future. [ABSTRACT FROM AUTHOR]

Ceftaroline fosamil, the prodrug of ceftaroline, is an advanced-generation cephalosporin antibacterial agent approved for treatment in the European Union in 2012. The drug is dedicated to curing complicated skin and soft tissue infections and community-acquired pneumonia. The developed analytical method of high performance liquid chromatography (HPLC) followed by diode array detector (DAD) set at 243 nm was used to identify and determine degradation products of ceftaroline fosamil. The elaborated method demonstrated good selectivity and linearity [r > 0.998 for the range 0.8–1.2 mg/mL (80–120%) and r > 0.997 for the range 0.005–0.015 mg/mL (0.5–1.5%)]. Limit of detection (LOD) and limit of quantification (LOQ) values were equal to 0.15 μg/mL and 0.5 μg/mL, respectively. The forced decomposition of ceftaroline fosamil carried out under acidic, basic, oxidative, photolytic and thermal conditions revealed the high sensitivity of ceftaroline on photodegradation and thermolysis processes. Representative ceftaroline samples were selected and analysed by LC coupled with electrospray ionisation time-of-flight mass spectrometry (LC-ESI-Q-TOF-MS) to identify the related substances that appeared under stress conditions. The LC-DAD method was transferred without any modification to LC–MS/MS system, allowing it to correlate results back to the LC-DAD method. Eight unknown signals were detected in the photolytic and thermal stress solutions for ceftaroline fosamil. The evaluated method was applied to the analysis of a medicinal product containing ceftaroline fosamil − Zinforo™, 600 mg, powder for concentrate for solution for infusion. Moreover, the optimised conditions allowed for the successful separation of eight cephalosporin antibiotics possessing a molecular structure similar to ceftaroline (cefepime, cefapirin, ceftazidime, cefpirome, cefalonium, cefotaxime, cefquinome, cefalotin). [ABSTRACT FROM AUTHOR]

Knowledge of the prevalence of ESBL enzymes among P. aeruginosa strains compared to the Enterobacteraiceae family is limited. The phenotypic tests recommended by EUCAST for the detection of ESBL-producing Enterobacteriaceae are not always suited for P. aeruginosa strains. This is mainly due to the presence of other families of ESBLs in P. aeruginosa isolates more often than in Enterobacteriaceae, production of natural AmpC cephalosporinase and its overexpression, and co-production of metallo-β-lactamases. The aim of this study was to determine the occurrence of ESBLs in P. aeruginosa isolated from patients from hospitals in Warsaw, to evaluate the ESBL production of these isolates using currently available phenotypic tests, their modifications, multiplex PCR and molecular typing of ESBL-positive isolates by PFGE. Clinical isolates of P. aeruginosa were collected in 2000-2014 from four Warsaw hospitals. Based on the data obtained in this study, we suggest using three DDST methods with inhibitors, such as clavulanic acid, sulbactam and imipenem, to detect ESBL-producing P. aeruginosa strains. Depending on the appearance of the plates, we suggest a reduction in the distance between discs with antibiotics to 15 mm and the addition of boronic acid at 0.4 mg per disc. The analysed isolates carried genes encoding ESBL from the families VEB (69 isolates with VEB-9), GES (6 with GES-1, 1 GES-5, 5 GES-13 and 2 with GES-15), OXA-2 (12 with OXA-15, 1 OXA-141, 1 OXA-210, 1 OXA-543 and 1 with OXA-544) and OXA-10 (5 isolates with OXA-74 and one with OXA-142). The most important result of this study was the discovery of three new genes, blaGES-15, blaOXA-141 and blaOXA-142; their nucleotide sequences have been submitted to the NCBI GenBank. It is also very important to note that this is the first report on the epidemiological problem of VEB-9-producing bacterial strains, not only in Poland but also worldwide. [ABSTRACT FROM AUTHOR]

Background: Most patients with bacterial infections suffer from fever and various pains that require complex treatments with antibiotics, antipyretics, and analgaesics. The most common drugs used to relieve these symptoms are non-steroidal anti-inflammatory drugs (NSAIDs), which are not typically considered antibiotics. Here, we investigate the effects of NSAIDs on bacterial susceptibility to antibiotics and the modulation of bacterial efflux pumps. Methodology: The activity of 12 NSAID active substances, paracetamol (acetaminophen), and eight relevant medicinal products was analyzed with or without pump inhibitors against 89 strains of Gram-negative rods by determining the MICs. Furthermore, the effects of NSAIDs on the susceptibility of clinical strains to antimicrobial agents with or without PAβN (Phe-Arg-β-naphtylamide) were measured. Results: The MICs of diclofenac, mefenamic acid, ibuprofen, and naproxen, in the presence of PAβN, were significantly (≥4-fold) reduced, decreasing to 25–1600 mg/L, against the majority of the studied strains. In the case of acetylsalicylic acid only for 5 and 7 out of 12 strains of P. mirabilis and E. coli, respectively, a 4-fold increase in susceptibility in the presence of PAβN was observed. The presence of Aspirin resulted in a 4-fold increase in the MIC of ofloxacin against only two strains of E. coli among 48 tested clinical strains, which included species such as E. coli, K. pneumoniae, P. aeruginosa, and S. maltophilia. Besides, the medicinal products containing the following NSAIDs, diclofenac, mefenamic acid, ibuprofen, and naproxen, did not cause the decrease of clinical strains’ susceptibility to antibiotics. Conclusions: The effects of PAβN on the susceptibility of bacteria to NSAIDs indicate that some NSAIDs are substrates for efflux pumps in Gram-negative rods. Morever, Aspirin probably induced efflux-mediated resistance to fluoroquinolones in a few E. coli strains. [ABSTRACT FROM AUTHOR]

The production of β-lactamases is the most important mechanism of Gram-negative rod resistance to β-lactams. Resistance to ceftazidime and cefepime in clinical isolates of Enterobacteriaceae (especially ESβL-positive E. coli and K. pneumoniae) and P. aeruginosa is life-threatening. However, all strains of the above mentioned species possess chromosomally encoded RND efflux pump systems in addition to β-lactamase production. The main goal of this study was to assess the role of efflux pump systems in cefepime and/or ceftazidime resistant phenotypes of ESβL-positive clinical strains of Enterobacteriaceae and P. aeruginosa. The influence of the efflux pump inhibitor PAβN on the minimum inhibitory concentration (MIC) values of tested cephalosporins was species-dependent. Generally, a significant reduction (at least four-fold) of β-lactam MICs was observed in the presence of PAβN only in the case of P. aeruginosa clinical isolates as well as the ESβL-producing transformant PAO1161 ΔampC. The usage of this agent resulted in the restoration of susceptibility to cefepime and/or ceftazidime in the majority of the P. aeruginosa ESβL-positive strains with low and moderate resistance to the above cephalosporins. Moreover, an outer membrane permeabilizing effect in the presence of PAβN was identified. Strain-dependent β-lactamase leakage upon PAβN or β-lactam treatment was demonstrated. The most important observation was the restoration of susceptibility of P. aeruginosa WUM226 to cefepime (MIC decrease from 32 to 4 mg/L) and ceftazidime (MIC decrease from 128 to 4 mg/L) in the presence of PAβN, which occurred despite an almost complete lack of β-lactamase leakage from bacterial cells. In conclusion, these data indicate that RND efflux pumps can modify the susceptibility to β-lactams in Gram-negative rods producing ESβLs. However, this phenomenon occurs only in P. aeruginosa strains and was not observed among E. coli and K. pneumoniae strains, representing the Enterobacteriaceae family. [ABSTRACT FROM AUTHOR]

Abstract: The elaborated method of micellar electrokinetic chromatography (MEKC) used to separate biapenem from its related substances was successfully implemented using sweeping under an enhanced electric field, followed by UV absorption detection at 200nm. The best results were obtained with formic buffer (22.5mM) pH 4.3 and sodium dodecyl sulfate (150mM) added to the electrolyte as the sweeping agent. Neutral capillary (60/50cm; 50μm ID) with reverse polarity and voltage values of 22kV, were used throughout the investigation. The optimized method of biapenem determination, validated in terms of linearity, accuracy and precision, provides a detection limit of 0.5μg/mL at S/N=3 for biapenem. The repeatability of the CE system, expressed by relative standard deviations (RSD) in the migration times, for biapenem and its degradation products varied from 0.14 to 1.48%, whereas for the corrected peak areas RSD were about 0.68–8.43%. Satisfactory separation was achieved within 20min of electrophoresis; moreover all carbapenems (imipenem, meropenem, ertapenem, doripenem and biapenem) were separated from each other during analysis. The evaluated MEKC method was applied to the analysis of a medicinal product containing biapenem – Omegacin® 0.3g for intravenous drip infusion. [Copyright &y& Elsevier]

Abstract: The oxazolidinones are a new and potent class of antimicrobial agents with activity mainly against Gram-positive strains. The commercial success of linezolid, the only FDA-approved oxazolidinone, has prompted many pharmaceutical companies to devote resources to this area of investigation. Until now, four types of chemical modifications of linezolid and oxazolidinone-type antibacterial agents, including modification on each of the A-(oxazolidinone), B-(phenyl), and C-(morpholine) rings as well as the C-5 side chain of the A-ring substructure, have been described. Division into sections according to side chain modification or the type of ring will be used throughout this review, although the process of synthesis usually involves the simultaneous modification of several elements of the linezolid substructure; therefore, assignment into the appropriate section depends on the structure–activity relationships (SAR) studies. This review makes an attempt to summarise the work carried out in the period from 2006 until mid-2012. [Copyright &y& Elsevier]

The aim of the study was to evaluate the activity of biocides against wide spectrum of clinical fungal strains and to examine whether disinfectants applied in hospital areas are active against the selected most resistant strains. Material and methods: The activity of the 4 disinfectants and antiseptic against 192 clinical and 3 reference fungal strains was tested according to the standard PN-EN 1275:2006. 189 of clinical isolates were susceptible to all tested biocides. Medicarine passed the standard against all tested clinical fungal strains. Results: Incidin LiquidSpray did not pass the test against two strains of Trichophyton mentagrophytes 7 and 187 and Aspergillus niger ATCC 16404; Spitaderm did not pass the test against Candida tropicalis 176 and T mentagrophytes 187;lncidin Plus showed activity against A. niger ATCC 16404 only at high concentration. Evaluation of the biocides activity against the selected resistant fungi was conducted in accordance with the standard PN-EN 13697:2003. All analyzed clinical strains were susceptible to disinfectants in parameters recommended by manufacturers. Lysoformine 3000 displayed no activity against A. niger ATCC 16404. Conclusions:A. niger ATCC 16404, C albicans ATCC 10231 and clinical isolates C. tropicalls 176, Penicillium citrinum 170, T mentagrophytes 7 and T mentagrophytes 187 should be used to evaluate the antifungal activity of biocides. The study provides data about various efficiency of the biocides against fungal strains. Selection of the most resistant strains should be used to evaluate the antifungal activity of biocides and to investigate the mechanisms of fungal resistance to biocides. [ABSTRACT FROM AUTHOR]

Abstract: A collection of 185 Streptococcus pneumoniae isolates was tested for their susceptibility to antipneumococcal drugs, with a focus on the distribution of tetracycline resistance determinants tet(M) and tet(O). Resistance patterns were compared with established correlates of multidrug resistance, and tetracycline-resistant isolates were tested for clonality and allelic variation within tet(M). Resistance to tetracyclines, penicillins and macrolides were all strongly related to multidrug resistance. Over one-quarter of the strains were tetracycline resistant, all via the tet(M)-mediated mechanism. Restriction fragment length polymorphism analysis revealed a high degree of allelic variation within tet(M) and gave evidence of a clonal and horizontal spread of selected alleles. A tet(M) variant that emerged with the onset of epidemic multidrug-resistant strains was replacing old alleles in the population. [Copyright &y& Elsevier]

• 12 new degradation products and three new by-products were identified. • fragmentation pathways were proposed for related compounds. • structure elucidation was performed by high resolution mass spectrometry (HRMS). • the kinetics and shelf-life values in solution-state studies were determined. • activation energies were calculated for alkaline and thermolytic degradation. The aim of the study was to investigate the intrinsic stability and to identify potential degradation products of tedizolid disodium phosphate (TED-OPO 3 Na 2), which belongs to the antimicrobial agents of the oxazolidinone class. Tedizolid, as disodium phosphate (prodrug), is registered under the trade name SIVEXTRO®, at a dose of 200 mg, in the form of powder for injection or infusion. The stability-indicating assay method was optimised using HPLC with diode array detection and with electrospray ionisation time-of-flight mass spectrometry. In solution-state studies, the forced decomposition of TED-OPO 3 Na 2 carried out under acidic, basic, oxidative, photocatalytic, and thermal conditions revealed the lability of TED-OPO 3 Na 2 to acidic, basic, and photocatalytic (UV) conditions, while it was relatively stable in oxidative conditions and during thermolysis processes. The kinetics of degradation and shelf-life values in solution-state studies were determined, and activation energies were calculated for alkaline and thermolytic degradation. In contrast, in the solid state degradation study, TED-OPO 3 Na 2 was stable under thermal conditions at high humidity and in visible light, while moderate degradation was observed under thermal conditions of low humidity and ultraviolet light. The developed method enabled the identification of 12 new degradation products and 3 new by-products. [ABSTRACT FROM AUTHOR]