INTRODUCTION: It is hypothesized that systemically administered antibiotics penetrate wound sites more effectively during negative pressure wound therapy (NPWT). However, there is a lack of clinical data from patients who receive NPWT for deep sternal wound infection (DSWI) after open-heart surgery. Here, we evaluated vancomycin penetration into exudate in this patient group. PATIENTS AND METHODS: For this prospective observational study, we enrolled 10 consecutive patients treated with NPWT for post-sternotomy DSWI. On the first sampling day, serum and exudate samples were synchronously collected at 0 (pre-dose), 0.5, 1, 2, 3 and 6 h after vancomycin administration. On the following three consecutive days, additional samples were collected, only before vancomycin administration. RESULTS: The ratio of average vancomycin concentration in wound exudate to in serum was higher for free (unbound) (1.51 ± 0.53) than for total (bound + unbound) (0.91 ± 0.29) concentration (p = 0.049). The percentage of free vancomycin was higher in wound exudate than serum (0.79 ± 0.19 vs. 0.46 ± 0.16; p = 0.04). Good vancomycin wound penetration was maintained on the following three days (vancomycin trough exudate-to-serum concentration ratio > 1). The total hospital stay was significantly longer in patients with DSWI (46 ± 11.6 days) versus without DSWI (14 ± 11.7 days) (p < 0.001). There was no in-hospital or 90-day mortality. Two patients experienced late DSWI recurrence. All-cause mortality was 4.8% during a median follow-up of 2.5 years. CONCLUSION: Vancomycin effectively penetrates wound exudate in patients receiving NPWT for DSWI after open-heart surgery.The protocol for this study was registered at ClinicalTrials.gov on July 16, 2024 (NCT06506032).

• MeSH
• Anti-Bacterial Agents * pharmacokinetics   administration & dosage   MeSH
• Exudates and Transudates metabolism   microbiology   MeSH
• Surgical Wound Infection * MeSH
• Cardiac Surgical Procedures * adverse effects   MeSH
• Middle Aged MeSH
• Humans MeSH
• Prospective Studies MeSH
• Aged MeSH
• Sternotomy * adverse effects   MeSH
• Sternum surgery   MeSH
• Negative-Pressure Wound Therapy * methods   MeSH
• Vancomycin * administration & dosage   pharmacokinetics   MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Observational Study MeSH

Ceramides are key components of the skin's permeability barrier. In atopic dermatitis, pathological hydrolysis of ceramide precursors - glucosylceramides and sphingomyelin - into lysosphingolipids, specifically glucosylsphingosine (GS) and sphingosine-phosphorylcholine (SPC), and free fatty acids (FFAs) has been proposed to contribute to impaired skin barrier function. This study investigated whether replacing ceramides with lysosphingolipids and FFAs in skin lipid barrier models would exacerbate barrier dysfunction. When applied topically to human stratum corneum sheets, SPC and GS increased water loss, decreased electrical impedance, and slightly disordered lipid chains. In lipid models containing isolated human stratum corneum ceramides, reducing ceramides by ≥ 30% significantly increased permeability to four markers, likely due to loss of long-periodicity phase (LPP) lamellae and phase separation within the lipid matrix, as revealed by X-ray diffraction and infrared spectroscopy. However, when the missing ceramides were replaced by lysosphingolipids and FFAs, no further increase in permeability was observed. Conversely, these molecules partially mitigated the negative effects of ceramide deficiency, particularly with 5%-10% SPC, which reduced permeability even compared to control with "healthy" lipid composition. These findings suggest that while ceramide deficiency is a key factor in skin barrier dysfunction, the presence of lysosphingolipids and FFAs does not aggravate lipid structural or functional damage, but may provide partial compensation, raising further questions about the behavior of lyso(sphingo)lipids in rigid multilamellar lipid environments, such as the stratum corneum, that warrant further investigation.

• MeSH
• Models, Biological MeSH
• Ceramides * metabolism   MeSH
• Phosphorylcholine analogs & derivatives   MeSH
• Skin * metabolism   MeSH
• Fatty Acids, Nonesterified metabolism   MeSH
• Humans MeSH
• Lysophospholipids metabolism   MeSH
• Permeability drug effects   MeSH
• Sphingosine analogs & derivatives   metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

The biosynthesis of the lincosamide antibiotics lincomycin A and celesticetin involves the pyridoxal-5'-phosphate (PLP)-dependent enzymes LmbF and CcbF, which are responsible for bifurcation of the biosynthetic pathways. Despite recognizing the same S-glycosyl-L-cysteine structure of the substrates, LmbF catalyses thiol formation through β-elimination, whereas CcbF produces S-acetaldehyde through decarboxylation-coupled oxidative deamination. The structural basis for the diversification mechanism remains largely unexplored. Here we conduct structure-function analyses of LmbF and CcbF. X-ray crystal structures, docking and molecular dynamics simulations reveal that active-site aromatic residues play important roles in controlling the substrate binding mode and the reaction outcome. Furthermore, the reaction selectivity and oxygen-utilization of LmbF and CcbF were rationally engineered through structure- and calculation-based mutagenesis. Thus, the catalytic function of CcbF was switched to that of LmbF, and, remarkably, both LmbF and CcbF variants gained the oxidative-amidation activity to produce an unnatural S-acetamide derivative of lincosamide.

• MeSH
• Catalytic Domain MeSH
• Crystallography, X-Ray MeSH
• Lincosamides * chemistry   biosynthesis   metabolism   MeSH
• Pyridoxal Phosphate * metabolism   chemistry   MeSH
• Molecular Dynamics Simulation MeSH
• Molecular Docking Simulation MeSH
• Publication type
• Journal Article MeSH

• MeSH
• Cell Tracking methods   MeSH
• Fluorine chemistry   MeSH
• Fluorocarbons MeSH
• Contrast Media MeSH
• Cell Movement MeSH
• Fluorine-19 Magnetic Resonance Imaging MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• radiologie, nukleární medicína a zobrazovací metody
• NML Publication type
• kolektivní monografie

The spread of multidrug-resistant Escherichia coli in healthcare facilities is a global challenge. Hospital-acquired infections produced by Escherichia coli include gastrointestinal, blood-borne, urinary tract, surgical sites, and neonatal infections. Therefore, novel approaches are needed to deal with this pathogen and its rising resistance. The concept of attenuating virulence factors is an alternative strategy that might lead to low levels of resistance and combat this pathogen. A sub-inhibitory concentration (1⁄4 MIC) of sitagliptin and nitazoxanide was used for phenotypic assessments of Escherichia coli virulence factors such as biofilm production, swimming motility, serum resistance, and protease production. Moreover, qRT-PCR was used to determine the impact of sub-MIC of the tested drugs on the relative expression levels of papC, fimH, fliC, kpsMTII, ompT_m, and stcE genes encoding virulence factors in Escherichia coli. Also, an in vivo model was conducted as a confirmatory test. Phenotypically, our findings demonstrated that the tested strains showed a significant decrease in all the tested virulence factors. Moreover, the genotypic results showed a significant downregulation in the relative expression levels of all the tested genes. Besides, the examined drugs were found to be effective in protecting mice against Escherichia coli pathogenesis. Sitagliptin and nitazoxanide exhibited strong anti-virulence activities against Escherichia coli. In addition, it is recommended that they might function as adjuvant in the management of Escherichia coli infections with either conventional antimicrobial agents or alone as alternative treatment measures.

• MeSH
• Anti-Bacterial Agents * pharmacology   MeSH
• Biofilms drug effects   MeSH
• Nitro Compounds MeSH
• Escherichia coli * drug effects   pathogenicity   genetics   MeSH
• Virulence Factors genetics   metabolism   MeSH
• Escherichia coli Infections * drug therapy   microbiology   MeSH
• Humans MeSH
• Microbial Sensitivity Tests MeSH
• Drug Resistance, Multiple, Bacterial MeSH
• Mice MeSH
• Escherichia coli Proteins genetics   MeSH
• Sitagliptin Phosphate * pharmacology   MeSH
• Thiazoles * pharmacology   MeSH
• Virulence drug effects   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Biopriming seeds with beneficial bacteria has potential to enhance seed germination. Therefore, in this investigation, five sulphur-oxidizing bacterial cultures, viz., Pantoea dispersa SOB2, Bacillus velezensis SN06, Bacillus cereus SN20, Bacillus tropicus SN16, and Bacillus megaterium SN11, were evaluated for different plant growth-promoting traits and their impact on Vigna radiata L. (mung bean) and Brassica juncea L. (mustard) seed germination. Among these, three bacterial cultures Pantoea dispersa SOB2, Bacillus velezensis SN06, and Bacillus megaterium SN11 evinced potential for mineral solubilization on solid medium where Pantoea dispersa SOB2 had the maximum solubilization indices-3.06, 5.14, and 2.48 for phosphate, zinc, and potassium respectively. The culture also displayed higher indole acetic acid (113.12 μg/mL), gibberellic acid (162.66 μg/mL), ammonia (5.23 μg/mL), and siderophore (69.53%) production than other bacterial cultures whereas Bacillus cereus SN20 showed maximum exopolysaccharide production (9.26 g/L). Bacterial culture Pantoea dispersa SOB2 significantly ameliorated the germination rate (3.73 no./day) and relative seed germination (208%) of Brassica juncea L., while Bacillus velezensis SN06 and Bacillus cereus SN20 followed with germination rate and relative seed germination of 2.86 no./day and 207%, respectively. Pantoea dispersa SOB2 displayed lowest mean germination time 2.91 days followed by Bacillus megaterium SN11 with 3.19 days. Biopriming with sulphur-oxidizing bacterial cultures, germination parameters of Vigna radiata L. were also markedly improved. Pantoea dispersa SOB2 demonstrated the highest germination rate (6.72 no./day), relative seed germination (115.56%), and minimum mean generation time (1.73 days). Bacillus velezensis SN06 inoculation had a beneficial effect on the seedling growth of Vigna radiata L., whereas Pantoea dispersa SOB2 greatly aided the seedling growth of Brassica juncea L. Results corroborated a prominent positive correlation between seed germination and plant growth-promoting traits. This is the first study on Pantoea dispersa as sulphur oxidizer, displaying plant growth promoting traits and seed germination potential. The potent sulphur-oxidizing bacterial cultures possessing plant growth promoting activities enhanced seed germination under in vitro conditions that could be further explored in field as biofertilizers to enhance the growth and yield of Brassica juncea L. and Vigna radiata L. crop.

• MeSH
• Bacillus * metabolism   MeSH
• Bacteria * metabolism   MeSH
• Mustard Plant * growth & development   microbiology   MeSH
• Germination * MeSH
• Indoleacetic Acids metabolism   MeSH
• Oxidation-Reduction MeSH
• Pantoea metabolism   MeSH
• Plant Growth Regulators metabolism   MeSH
• Seeds * microbiology   growth & development   MeSH
• Sulfur * metabolism   MeSH
• Vigna * growth & development   microbiology   MeSH
• Publication type
• Journal Article MeSH

Roztroušená skleróza (RS) je závažné autoimunitní zánětlivé a neurodegenerativní onemocnění centrálního nervového systému. Počet pacientů s autoimunitními chorobami globálně dramaticky narůstá, nemocní s několika různými autoimunitními komorbiditami dnes nejsou vzácností. Ozanimod je léčivo ze skupiny modulátorů sfingosin-1- -fosfátových receptorů, které je v současnosti schváleno ve dvou indikacích: u pacientů s aktivní relabující-remitující RS (RR-RS) a také pacientů s kolonoskopicky a biopticky verifikovanou středně těžkou až těžkou aktivní formou ulcerózní kolitidy (UC). Uvádíme kazuistiku pacientky, u níž byla pět let po stanovení diagnózy RS diagnostikována UC, a díky terapii ozanimodem se podařilo dosáhnout jak klinické a radiologické stabilizace RS, tak i klinické a endoskopické remise UC.

Multiple sclerosis (MS) is a serious autoimmune inflammatory and neurodegenerative disease of the central nervous system. The number of patients with autoimmune diseases is dramatically increasing globally, and patients with several different autoimmune comorbidities are no longer considered as rare. Ozanimod is a drug from the group of sphingosine-1-phosphate receptor modulators, which is currently approved for two indications - in patients with active reiapsing-remitting MS (RR-MS), and also in patients with coionoscopicaiiy and biopsy-verified moderate to severe active form of ulcerative colitis (UC). We present a case report of a patient who was diagnosed with UC five years after the diagnosis of MS, and thanks to the initiation of ozanimod, it was possible to achieve both ciinicai and radiological stabiiization of MS, as weii as ciinicai and endoscopic remission of UC.

OBJECTIVES: Stability of concentrations of urinary stone-related metabolites was analyzed from samples of recurrent urinary stone formers to assess necessity and effectiveness of urine acidification during collection and storage. METHODS: First-morning urine was collected from 20 adult calcium-stone forming patients at Tomas Bata Hospital in the Czech Republic. Urine samples were analyzed for calcium, magnesium, inorganic phosphate, uric acid, sodium, potassium, chloride, citrate, oxalate, and urine particles. The single-voided specimens were collected without acidification, after which they were divided into three groups for storage: samples without acidification ("NON"), acidification before storage ("PRE"), or acidification after storage ("POST"). The analyses were conducted on the day of arrival (day 0, "baseline"), or after storage for 2 or 7 days at room temperature. The maximum permissible difference (MPD) was defined as ±20 % from the baseline. RESULTS: The urine concentrations of all stone-related metabolites remained within the 20 % MPD limits in NON and POST samples after 2 days, except for calcium in NON sample of one patient, and oxalate of three patients and citrate of one patient in POST samples. In PRE samples, stability failed in urine samples for oxalate of three patients, and for uric acid of four patients after 2 days. Failures in stability often correlated with high baseline concentrations of those metabolites in urine. CONCLUSIONS: Detailed procedures are needed to collect urine specimens for analysis of urinary stone-related metabolites, considering both patient safety and stability of those metabolites. We recommend specific preservation steps.

• MeSH
• Urinalysis methods   MeSH
• Adult MeSH
• Hydrogen-Ion Concentration MeSH
• Uric Acid urine   MeSH
• Middle Aged MeSH
• Humans MeSH
• Urinary Calculi * urine   MeSH
• Specimen Handling methods   MeSH
• Recurrence * MeSH
• Urine Specimen Collection methods   MeSH
• Aged MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

BackgroundOn 29 January 2024, the European Centre for Disease Prevention and Control distributed an alert about a metronidazole-resistant Clostridioides difficile outbreak of PCR ribotype (RT) 955 in England.AimWe aimed to investigate the presence of RT955 in Czech, Slovak and Polish C. difficile isolates and evaluate different culture media for detecting its metronidazole resistance.MethodsIsolates with binary toxin genes identified as 'unknown' by the WEBRIBO PCR ribotyping database up to 2023 were re-analysed after adding the RT955 profile to the database. The RT955 isolates were characterised by whole genome sequencing and tested for susceptibility to 15 antimicrobials.ResultsWe did not find RT955 in Czech (n = 6,661, 2012-2023) and Slovak (n = 776, 2015-2023) isolates, but identified 13 RT955 cases (n = 303, 2021-2023) in three hospitals in Poland. By whole genome multilocus sequence typing, 10 isolates clustered into one clonal complex including a sequence of United Kingdom strain ERR12670107, and shared similar antimicrobial resistance genes/mutations. All 13 isolates were resistant to ciprofloxacin/moxifloxacin, erythromycin/clindamycin and ceftazidime. All isolates had a mutation in the nimB gene promoter and in NimB (Tyr130Ser and Leu155Ile). The metronidazole resistance was detected in all isolates using brain-heart-infusion agar supplemented with haemin and Chocolate agar. Results were discrepant with the European Committee on Antimicrobial Susceptibility Testing-recommended Fastidious anaerobe agar and Brucella blood agar.ConclusionThe identification of clonally related haem-dependent metronidazole-resistant C. difficile RT955 in multiple hospitals indicates a need for prospective surveillance to estimate its prevalence in Europe.

• MeSH
• Anti-Bacterial Agents * pharmacology   MeSH
• Drug Resistance, Bacterial * genetics   MeSH
• Clostridioides difficile * genetics   drug effects   isolation & purification   classification   MeSH
• Disease Outbreaks MeSH
• Clostridium Infections * epidemiology   microbiology   drug therapy   MeSH
• Humans MeSH
• Metronidazole * pharmacology   MeSH
• Microbial Sensitivity Tests MeSH
• Multilocus Sequence Typing MeSH
• Polymerase Chain Reaction MeSH
• Ribotyping * MeSH
• Whole Genome Sequencing MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic MeSH
• Poland MeSH
• Slovakia MeSH

N-Methyl-d-aspartate receptors (NMDARs) play a crucial role in excitatory neurotransmission, with numerous pathogenic variants identified in the GluN subunits, including their ligand-binding domains (LBDs). The prevailing hypothesis postulates that the endoplasmic reticulum (ER) quality control machinery verifies the agonist occupancy of NMDARs, but this was tested in a limited number of studies. Using microscopy and electrophysiology in the human embryonic kidney 293 (HEK293) cells, we found that surface expression of GluN1/GluN2A receptors containing a set of alanine substitutions within the LBDs correlated with the measured EC50 values for glycine (GluN1 subunit mutations) while not correlating with the measured EC50 values for l-glutamate (GluN2A subunit mutations). The mutant cycle of GluN1-S688 residue, including the pathogenic GluN1-S688Y and GluN1-S688P variants, showed a correlation between relative surface expression of the GluN1/GluN2A receptors and the measured EC50 values for glycine, as well as with the calculated ΔGbinding values for glycine obtained from molecular dynamics simulations. In contrast, the mutant cycle of GluN2A-S511 residue did not show any correlation between the relative surface expression of the GluN1/GluN2A receptors and the measured EC50 values for l-glutamate or calculated ΔGbinding values for l-glutamate. Coexpression of both mutated GluN1 and GluN2A subunits led to additive or synergistic alterations in the surface number of GluN1/GluN2A receptors. The synchronized ER release by ARIAD technology confirmed the altered early trafficking of GluN1/GluN2A receptors containing the mutated LBDs. The microscopical analysis from embryonal rat hippocampal neurons (both sexes) corroborated our conclusions from the HEK293 cells.

• MeSH
• Glycine metabolism   MeSH
• HEK293 Cells MeSH
• Hippocampus cytology   metabolism   MeSH
• Rats MeSH
• Glutamic Acid metabolism   MeSH
• Humans MeSH
• Ligands MeSH
• Mutation genetics   MeSH
• Protein Domains MeSH
• Nerve Tissue Proteins MeSH
• Receptors, N-Methyl-D-Aspartate * metabolism   genetics   chemistry   MeSH
• Protein Transport physiology   genetics   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH