Biology subjects

Czaplewska, P.

Publications and source records attributed to Czaplewska, P..

6 recordsLinked to original sources

bioRxiv2026

Antibacterial activity of a newly identified phage-derived endolysin and its parental bacteriophage against clinical uropathogenic Escherichia coli

The increasing prevalence of antibiotic-resistant uropathogenic Escherichia coli highlights the need for antibacterial strategies that can complement or extend beyond conventional antibiotic treatment. Bacteriophages and phage-derived lytic enzymes represent promising alternatives because of their distinct mechanisms of bacterial killing and their potential activity against drug-resistant pathogens. In this study, we characterized the newly discovered UPEC-infecting bacteriophage vB-EcoS_57-3 and the endolysin 57_3Lys, encoded by this phage, combining genomic, structural, and functional approaches. The phage demonstrated lytic activity against clinical UPEC isolates and retained antibacterial potential under conditions relevant to the urinary tract. Genomic and sequence analyses revealed distinctive features of 57_3Lys associated with signal-anchor-release endolysins and suggested a less common mode of intracellular translocation and activation. Functional experiments supported the involvement of the bacterial secretion machinery in endolysin-mediated lysis. Notably, the purified enzyme also displayed antibacterial activity against intact clinical E. coli cells, despite the intrinsic barrier presented by the Gram-negative cell envelope. Although this activity developed slowly, it significantly reduced both bacterial culture turbidity and viable cell number. Together, these findings provide new insights into the antimicrobial strategies based on the phages and their phage lytic enzymes, supporting further exploration of vB-EcoS_57-3 and 57_3Lys as potential tools against antibiotic-resistant UPEC.

microbiology↗

bioRxiv2026

Novel native serum peptidomics workflow enables the discovery of circulating subtype-specific peptide biomarkers in acute ischemic and haemorrhagic stroke

Novel serum peptidomics offers a direct insight into proteolytic activity, tissue injury, and systemic signaling. Nevertheless, existing workflows suffer from low peptide yields, low throughput, and limited recovery of low-abundance species. Here we present a native serum peptidomics protocol that integrates mild acid treatment, solid-phase extraction with molecular weight cutoff filtration and data-independent acquisition mass spectrometry (DIA-MS). The protocol requires less than 100 {micro}l of serum or plasma, is completed within hours, time-cost-effective and compatible with 96-well formats without specialized equipment. Applied to a proof-of-concept cohort of patients with acute ischemic stroke (AIS), intracranial haemorrhage (ICH), and healthy controls, the workflow identified over 12,000 peptides, exceeding the three-fold threshold of existing peptidomics approaches. DIA-MS analysis across independent batches demonstrated 78-83% peptide overlap and consistent fold-change directionality. We further introduce peptide locus analysis, which aggregates overlapping peptides within defined protein regions. This approach revealed bidirectional regulation within individual precursor proteins such as the fibrinogen alpha chain (FIBA), resolving intraprotein proteolytic dynamics. Three candidate peptides from TYB4, CO4B, and ITIH4 proteins accurately distinguished stroke subtypes and controls, while characteristic shifts in peptide physicochemical properties were observed across strokes. This workflow substantially advances the sensitivity, throughput, and biological resolution of serum peptidomics for quantitative multi-biomarker discovery, validation and its output promises effective implementation of AI/ML models aiming for new dimensions in diagnostics, prognostics, prediction and monitoring.

neuroscience↗

bioRxiv2024

Tailocin-mediated interactions in Soft Rot Pectobacteriaceae

Bacteria carry phage-derived elements within their genomes, some of which can produce phage-like particles (tailocins) used as weapons to kill kin strains in response to environmental conditions. This study investigates the production and activity of tailocins by plant pathogenic bacteria: Pectobacterium, Dickeya, and Musicola genera, which compete for niche, providing an attractive model to study the ecological role of tailocins. Microscopy revealed that most analyzed strains (88%) produced tailocins. Tailocin-mediated killing interactions were assessed across 351 strain pairs, showing that Dickeya spp. had a higher likelihood of killing neighbors (57.1%) than Pectobacterium spp. (21.6%). Additionally, Dickeya spp. strains exhibited broader phylogenetic killing, targeting both Pectobacterium spp. and Musicola sp., while Pectobacterium spp. tailocins were genus-specific. Mutual killing was observed in 33.9% of interactions, predominantly within Dickeya spp. Although tailocins were morphologically indistinguishable between producers, genomic analyses identified conserved clusters having distinct differences between Pectobacterium spp. and Dickeya spp tailocins. This suggests different origins of these particles. Induction experiments demonstrated that tailocin production was boosted by hydrogen peroxide, supporting the role of these particles in bacteria-bacteria competition during infection. Tailocins were detectable in infected potato tissue but not in river water, highlighting the particular ecological relevance of tailocins in plant environments.

microbiology↗

bioRxiv2023

Soft rot pathogen Dickeya dadantii 3937 produces tailocins resembling the tails of Enterobacteria bacteriophage P2

Tailocins are nanomolecular machines with bactericidal activity. They are produced by bacteria to contribute to fitness in mixed communities, and hence, they play a critical role in their ecology in a variety of habitats. Here, we characterized the new tailocin produced by Dickeya dadantii strain 3937, a well-characterized member of plant pathogenic Soft Rot Pectobacteriaceae (SRP). Tailocins induced in D. dadantii were ca. 166 nm long tubes surrounded by contractive sheaths with baseplates having tail fibers at one end. A 22-kb genomic cluster involved in their synthesis and having high homology to the cluster coding for the tail of the Enterobacteriophage P2 was identified. The D. dadantii tailocins, termed dickeyocin P2D1 (phage P2-like dickeyocin 1), were resistant to inactivation by pH (3.5 - 12), temperature (4 - 50 {degrees}C), and elevated osmolarity (NaCl concentration: 0.01 - 1 M). P2D1 could kill a variety of different Dickeya spp. but not any strain of Pectobacterium spp. tested and were not toxic to Caenorhabditis elegans. TeaserTailocins are nanomolecular entities similar to syringes that are produced by various bacteria to fight other microorganisms present in the same environment.

microbiology↗

bioRxiv2023

Being spontaneous has its costs! Characterization of the spontaneous phage ΦD5-resistant mutants of Dickeya solani strain IPO 2222

Lytic bacteriophages able to infect and kill Dickeya spp. can be readily isolated from virtually all Dickeya spp.-containing environments, yet little is known about the selective pressure those viruses exert on their hosts. Here, we identified two spontaneous D. solani IPO 2222 mutants (0.8% of all obtained mutants), DsR34 and DsR207, resistant to infection caused by lytic phage vB_Dsol_D5 ({Phi}D5) that expressed a reduced ability to macerate potato tuber tissues compared to the wild-type, phage-susceptible D. solani IPO 2222 strain. Genome sequencing revealed that genes encoding: secretion protein HlyD (mutant DsR34) and elongation factor Tu (EF-Tu) (mutant DsR207) were altered in these strains. Both mutations impacted the proteomes of cells grown in both rich and minimal media, including the abundance of the cell envelope and transmembrane transport-associated proteins. Furthermore, features essential for the ecological success of these mutants in a plant environment, including their ability to use various carbon and nitrogen sources, produce plant cell wall degrading enzymes, ability to form biofilms, siderophore production, swimming and swarming motility and virulence in planta were assessed. Compared to the wild-type strain, D. solani strain IPO 2222, mutants DsR34 and DsR207 had a reduced ability to macerate chicory leaves and to colonize and cause symptoms in growing potato plants. The implications of the {Phi}D5 resistance on driving traits affecting the ecological performance of D. solani are discussed.

microbiology↗

bioRxiv2022

Resistance of Dickeya solani strain IPO 2222 to lytic bacteriophage vB_Dsol_D5 ({Phi}D5) results in fitness tradeoffs for the bacterium during infection

Resistance to bacteriophage infections protects bacteria in phage-full environments, allowing them to survive and multiply in the presence of their viral predators. However, such resistance may cause direct costs for strains linked with the ecological fitness expressed as reduced competitiveness for resources or reduced virulence or both. Unfortunately, limited knowledge exists about such costs paid by phage-resistant plant pathogenic bacteria in their natural environments. This study analyzed the costs of phage resistance paid by broad host phytopathogenic pectinolytic bacterium Dickeya solani both in vitro and in potato (Solanum tuberosum L.) plants. Thirteen D. solani IPO 2222 Tn5 mutants were identified that exhibited resistance to infection caused by lytic bacteriophage vB_Dsol_D5 ({Phi}D5). The genes disrupted in these 13 mutants encoded proteins involved in the synthesis of the bacterial envelope components (viz. LPS, EPS and capsule). The ability of {Phi}D5-resistant D. solani mutants to colonize and cause symptoms on potato plants as well as other phenotypes that are known to contribute to the ecological fitness of D. solani in-plant environment, including growth rate, production of effectors, swimming and swarming motility, use of various carbon and nitrogen sources and biofilm formation were assessed. Although phage resistance did not affect most of the phenotypes of {Phi}D5-resistant D. solani evaluated in vitro, all phage resistant mutants were significantly compromised in their ability to survive on and colonize and cause disease symptoms in potato plants. This study is, to our knowledge, one of few to show the direct link between phage resistance and the fitness of plant pathogenic bacteria and the first one to assess phage-host associations for D. solani.

microbiology↗