Search Results (1,073)

The emergence of multidrug-resistant Staphylococcus aureus underscores the urgent need for novel therapeutic agents targeting essential bacterial pathways. The lipoteichoic acid synthase (LtaS) is crucial for the synthesis of lipoteichoic acid in the cell wall of Gram-positive bacteria and represents a promising and vulnerable target for antimicrobial drug development. This study employed a comprehensive computational pipeline to identify potent inhibitors of the LtaS enzyme. A library of natural compounds was retrieved from the COCONUT database and screened against the crystal structure of the extracellular domain of LtaS (eLtaS) (PDB ID: 2W5R, obtained from the Protein Data Bank) through a multi-stage molecular docking strategy. This process started with High-Throughput Virtual Screening (HTVS), followed by Standard Precision (SP) docking, and culminated in Extra Precision (XP) docking to refine the selection of hits. The top-ranking compounds from XP docking were subsequently subjected to MM-GBSA binding free energy calculations for further filtration. The stability and dynamic behavior of the resulting candidate complexes were then evaluated using 100 ns molecular dynamics (MD) simulations, which confirmed the structural integrity and binding stability of the ligands. Density Functional Theory calculations revealed that screened ligands exhibit improved electronic stabilization and charge-transfer characteristics compared to a reference compound, suggesting enhanced reactivity and stability relevant for hit identification. Finally, ADMET (Absorption, Distribution, Metabolism, Excretion, and Toxicity) profiling was conducted to assess the drug-likeness and pharmacokinetic safety of the lead compounds. These findings support them as promising orally active leads for further optimization. Our integrated approach shortlisted eight initial hits (A–H) that showed interesting scaffold diversity and finally identified two compounds, herein referred to as Compound A and Compound B, which demonstrated stable binding, favorable free energy, and an acceptable Absorption, Distribution, Metabolism, and Excretion, and Toxicity (ADMET) profile. These candidates emerge as promising starting points for developing novel anti-staphylococcal agents targeting the LtaS enzyme that cand be further proved by experimental validation. Full article

Background: Milk is a highly nutritious food, but its composition makes it an ideal medium for microbial growth, particularly for bacteria like Staphylococcus aureus (S. aureus). In Ecuador, raw milk consumption is culturally rooted, and contamination risks are heightened, especially in informal markets. Staphylococcus aureus, a Gram-positive, coagulase-positive bacterium, commonly colonizes mucous membranes and can cause a range of infections due to its production of thermostable toxins. Its impact extends to bovine mastitis, severely affecting dairy production. Of particular concern is the emergence of methicillin-resistant S. aureus (MRSA) strains, associated with the acquisition of the mecA gene located on the “staphylococcal chromosomal cassette mec” (SCCmec) element and identification of a mecA homologue, mecC, further complicates detection and monitoring efforts. Objectives: This study evaluated the prevalence of S. aureus and MRSA strains in raw milk from Ecuadorian provinces Pichincha and Manabí. Methods: A total of 633 samples were collected and analyzed via real-time PCR (qPCR) and bacterial isolation methods, complemented by endpoint PCR assays for mecA and mecC genes detection. Results: A high prevalence of S. aureus (84%) was observed, with significant differences between regions. MRSA was detected in 23% of all samples, with mecA being more prevalent than mecC among isolates. Sequencing of 16S rDNA confirmed the identity of isolates, while phylogenetic analysis of mecA and mecC genes validated their presence. The findings suggest that suboptimal hygiene practices and varied biosecurity protocols, especially among small and medium dairy producers, may contribute to the persistence of resistant strains. Conclusions: This study highlights the presence of S. aureus and MRSA in raw milk, underscoring the need for strengthened surveillance, improved hygiene practices, the use of molecular diagnostic tools, and proper heat treatments to reduce the public health risks associated with contaminated milk and its derivatives. Full article

Nymphaea ‘Blue Bird’, a tropical water lily prized for its ornamental appeal, has been less explored as a source of bioactive flavonoids. This study developed an efficient extraction and purification protocol for flavonoids from this plant and compared their distribution and bioactivities across different tissues. Supercritical CO₂ fluid extraction (SFE) proved optimal, yielding 2.56% under conditions of 24.3 MPa, 39 °C, 91 min, and a CO₂ flow rate of 16 L/min. Subsequent purification with HPD500 macroporous resin enhanced flavonoid purity from 3.05% to 11.46%. Among the tissues analyzed, petals contained the highest levels of total flavonoids (6.43 mg/g) and total phenolics (45.71 mg/g), and exhibited the most potent antioxidant (as shown by the lowest EC₅₀ values for ABTS⁺ and DPPH scavenging) and broad-spectrum antibacterial activities (indicated by the lowest MIC and MBC against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans). Antibacterial efficacy was generally superior against Gram-positive bacteria. Widely targeted metabolomics identified 560 metabolites, predominantly flavonols and flavonoids. Principal component and cluster analyses revealed tissue-specific metabolite profiles. KEGG enrichment analysis underscored the significance of the flavonoid biosynthetic pathway, and key differential metabolites—such as luteolin, myricetin, taxifolin, and quercetin—were strongly correlated with the observed bioactivities. These results highlight N. ‘Blue Bird’ petals as a promising source of natural antioxidants and antimicrobials, providing a scientific basis for their future functional applications. Full article

The genus Kocuria includes Gram-positive and environmentally versatile bacteria, which are of biotechnological interest due to their ability to synthesize secondary metabolites. In this study, the genome of Kocuria sp. KH4, isolated from alkaline mine tailings (southeastern Mexico), was sequenced and analyzed to determine its taxonomic affiliation and explore its metabolic and adaptive potential. The assembled genome showed a size of 3.89 Mb, a GC content of 73.2%, and 3609 coding genes. Phylogenomic analyses and genomic relationship indices (ANI, AAI, and dDDH) confirmed that strain KH4 represents a novel genomospecies within the genus Kocuria. Functional analysis revealed broad metabolic diversity, with genes associated with the transport and metabolism of amino acids, carbohydrates, and inorganic ions. A total of 165 genes linked to metal resistance and homeostasis mechanisms were identified, including ABC-type transport systems and ATPases, as well as specific genes for Fe, Ni, Zn, Cu, As, and Hg. Forty-eight genomic islands were also identified, encoding a wide variety of functions and mobile genetic elements (MGEs). Furthermore, six biosynthetic gene clusters (BGCs) involved in the production of nonribosomal peptides, type III polyketides, terpenes, and siderophores were detected, suggesting a remarkable potential for the synthesis of bioactive compounds. Taken together, the results highlight this strain as a promising source of secondary metabolites with potential applications in environmental, pharmaceutical, and industrial biotechnology, underscoring the importance of Kocuria genomes as natural reservoirs of new biosynthetic pathways. Full article

The problem of antibiotic resistance is one of the challenges that science and medicine face in the 21st century. Nucleoside analogs have already proven as antiviral and antitumor agents, and, currently, there are more and more reports on their antibacterial and antifungal activity. The substitution of an oxygen atom by a sulfur one leads to the emergence of unique properties. Here, we report the synthesis of eight new 4-thioanalogs of 5-substituted (5-alkyloxymethyl and 5-alkyltriazolylmethyl) derivatives of 2′-deoxyuridine and uridine, which were active against Mycobacterium tuberculosis and Gram-positive bacteria. The novel sulfur-containing nucleosides were synthesized via activation of the pyrimidine C4 position, followed by condensation with thioacetic acid and deblocking. To increase the solubility, oligoglycol carbonate depot forms were obtained via activation of the 3′-hydroxyl group using N,N’-carbonyldiimidazole and condensation with triethylene glycol. The highest inhibitory activity was demonstrated by 3′-triethylene glycol depot forms of 4-thio-5-undecyl- and 5-dodecyloxymethyl-2′-deoxyuridine (4a,b) against two strains of M. smegmatis. The most promising compounds were 5-[4-decyl-(1,2,3-triazol-1-yl)methyl]-4-thio-2′-deoxy- and ribouridine (3c,g) and 5-undecyloxymethyl 4-thiouridine (3e) active toward clinical M. intracellulare isolates. Overall, novel sulfur-containing nucleoside analogs were low toxic, demonstrated better inhibitory activity compared to their C4-oxo ones, and, thus, are promising compounds for the development of new antibacterial agents. Full article

Quitral (Tristerix corymbosus), a Chilean and Argentine parasitic mistletoe, is traditionally used by Mapuche natives to treat stomach ulcers, nervous disorders, and cholesterol reduction, although scientific support is scarce. Methanolic and chloroform extracts from its leaves and stems were prepared. Chemical analysis included antioxidant capacity assays (ORAC-FL and DPPH) and chromatographic determinations. The antimicrobial activity was tested against nine bacteria and two yeast strains. Additionally, cytotoxicity (hemolysis) and toxicity (against Caenorhabditis elegans) assays were performed. The results revealed that the methanolic leaf extracts had the highest ORAC-FL value, with DPPH assays showing solvent-dependent differences. Thirty-one compounds were tentatively identified, of which 61% were phenolic compounds, primarily flavonoids like quercetin and its derivatives. Antimicrobial results showed activity against Gram-positive bacteria (Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, and Enterococcus faecalis), but not against yeast Candida guillermondii and Candida tropicalis. Methanolic extracts induced dose-dependent erythrocyte hemolysis, while chloroform extracts showed no relevant cytotoxicity. Toxicity against Caenorhabditis elegans was also dose-dependent for methanolic extracts; leaf extract reduced survival at 50 mg mL⁻¹ after 24 h. These findings partially validate some traditional uses, highlight the importance of solvent polarity in extraction and biological effects, and establish quitral as a flavonoid source. Full article

Background: Gram-positive bacteria, once considered incapable of producing extracellular vesicles (EVs) due to their thick peptidoglycan layer, are now known to secrete EVs that transport virulence factors and modulate host immunity. These EVs contribute to bacterial pathogenicity by facilitating biofilm formation, immune evasion, and inflammation. Granulicatella adiacens, an oral commensal associated with infective endocarditis, represents a clinically relevant model to study EV-mediated virulence. Objectives: This study’s aim was to investigate whether the proteomic composition and immunomodulatory activity of G. adiacens EVs differ between biofilm and planktonic lifestyles, thereby contributing to distinct pathogenic behaviours. Methods: EVs isolated from G. adiacens CCUG 27809 cultures were characterized using nano LC-ESI-MS/MS, followed by comprehensive bioinformatic and cytokine assays. Results: Quantitative proteomic profiling identified 1017 proteins, revealing distinct signatures between biofilm- and planktonic-derived EVs. Principal component analysis showed clear segregation between the two states, with biofilm EVs enriched in proteins linked to stress adaptation, adhesion, and structural integrity, while planktonic EVs exhibited growth- and metabolism-related proteins. A total of 114 virulence-associated proteins were identified, including several novel candidates. Functionally, EVs from both conditions significantly induced pro-inflammatory cytokines IL-8 and IL-1β in a dose-dependent manner (p < 0.05), whereas IL-17 remained unchanged. Conclusions: G. adiacens EVs exhibit lifestyle-dependent proteomic and immunomodulatory differences, underscoring their role in host–pathogen interactions and endocardial infection. These findings provide a foundation for future mechanistic and in vivo studies exploring EV-mediated virulence and potential therapeutic modulation. Full article

Addressing global food waste challenges, this study investigated plant-based byproducts, spent coffee grounds, apple, and chokeberry pomaces, as sources of phenolic acids and biodegradable carriers for lipase immobilization. The goal was to enhance the lipophilicity and functionality of natural phenolics by enzymatic lipophilization. Microbial lipase from A. oryzae was immobilized on these materials, with native spent coffee grounds (NSCG) showing the highest activity (6.0 U/g hydrolytic; 1036 U/g synthetic). Chlorogenic acid (CGA), predominant in extracts, served as a model substrate. Using response-surface methodology, optimal conditions for butyl-CGA synthesis were determined. This is the first report of CGA lipophilization using food-waste-immobilized biocatalysts, where reaction yield for NSCG increased with alcohol chain length, peaking with dodecanol (34.06%). Among synthesized esters, butyl chlorogenate displayed the highest antioxidant activity, comparable to free CGA and BHT, and increased lipophilicity, though a “cut-off” effect appeared for longer chains. Medium-chain esters (C6, C8) showed selective antimicrobial activity against Gram-positive bacteria. While lipophilization of chokeberry pomace and spent coffee grounds extracts reduced antioxidant activity, short-chain esters (C4–C6) improved rapeseed oil stability. The findings highlight food waste as a sustainable source for developing biocatalysts and value-added bioactives with enhanced functional properties. Full article

Nisin is a widely used natural antibacterial peptide in the food industry. However, its instability limits its practical application in food preservation. In this study, the antibacterial activity of previously built nisin/carboxylic curdlan (C6-Cc) complexes were tested against Gram-positive bacteria: Staphylococcus aureus. The results revealed that when the mass ratio of C6-Cc to nisin was 1:8 (C6-Cc: nisin, w/w), the nisin/C6-Cc complexes exhibited strong antibacterial stability over the pH range of 1–4, with inhibition zone diameters ranging from 11.5 to 13.0 mm and a minimum inhibitory concentration (MIC) of 0.71 mg/mL. Notably, the complexes also maintained good thermal stability even at 121 °C. Furthermore, the complexes with a mass ratio of 1:8 (w/w) showed superior physicochemical stability under low-pH conditions (pH < 5) within the salt concentration of 0–100 mM, and the stable temperature ranged from 25 to 121 °C. Twenty-eight days of storage had no significant impact on its antibacterial or physicochemical stability. These findings suggest that the nisin/C6-Cc complexes will have broad application prospects in food bio-preservation. Full article

Introduction: Suppressive antibiotic therapy (SAT) is a therapeutic alternative for complex infections where a cure is considered unlikely or impossible. SAT involves the prolonged, often indefinite, administration of antibiotics, typically given orally, to control symptoms. However, the increasing incidence of multidrug-resistant microorganisms limits the availability of oral options. Dalbavancin is a parenteral antibiotic with broad coverage against Gram-positive bacteria that offers the advantage of an extended dosing interval. The aim of this study was to describe the characteristics and clinical outcomes of patients with implant-associated osteoarticular infections receiving dalbavancin as SAT. A secondary objective was to identify factors associated with SAT failure with dalbavancin. Materials and Methods: We conducted a multicentre, observational study with retrospective recruitment of patients treated with dalbavancin as (SAT) for complex implant-associated osteoarticular infections, in which curative surgery was either not feasible or insufficient. Cohort characteristics were described, and variables associated with SAT failure under dalbavancin treatment were analysed. Results: A total of 43 patients received dalbavancin as SAT. The most frequent indication was prosthetic joint infection (38 [88.4%]). A total of 28 patients (65.1%) had chronic infections; the remaining cases were acute infections that had failed conservative management. Nine different dosing regimens of dalbavancin were used. Dalbavancin provided adequate symptomatic control in 32 patients (74.4%) over a follow-up period of 836.5 days (IQR 402–1288.5). The antibiotic was well tolerated; only one adverse effect was reported in a patient. Three patients developed resistance during treatment, which accounted for SAT failure. Conclusions: Dalbavancin is shown to be a safe and convenient alternative for SAT for orthopaedic implant infection. Although the development of resistance was infrequent, it can occur and should be monitored. Full article

In the current research a series of new copper(II) complexes with novel acylhydrazone ligands were synthesized and their antibacterial and anticancer activities were determined. The complexes were characterized by molecular spectroscopy (FT-IR and UV-Vis) and conductivity measurements. Additionally, their structure was confirmed by single-crystal X-ray analysis. The crystallographic data revealed that all compounds are mononuclear Cu(II) species. The Cu(II) ion is four-coordinated by the ONO donor set from mono-deprotonated hydrazone ligand and one Cl^¯ anion, forming distorted square-planar geometry. The biological studies revealed that the compounds exhibit high antimicrobial activity, especially against Gram-positive bacteria, in some cases greater than the reference substances, and better activity than free ligands. The tested complexes possessed the lowest MIC and MBC values towards Staphylococcus epidermidis ATCC 12228 and Micrococcus luteus ATCC 10240. Furthermore, they showed no toxicity towards normal cell lines. Full article

Inflammatory bowel diseases (IBDs), including Crohn’s disease (CD) and ulcerative colitis (UC), are chronic immune-mediated disorders characterized by mucosal injury, cycles of inflammation and repair, and tissue damage. Persistent inflammation accelerates epithelial turnover, generates oxidative and replication stress, and remodels the stromal niche, contributing to the risk of colorectal cancer (CRC). Systematic dysplasia surveillance remains essential. Cellular senescence has emerged as a unifying mechanism linking inflammation, impaired epithelial repair, fibrosis, and neoplasia. In UC, p16/p21 upregulation, telomere erosion, and loss of lamin B1 accumulate and adopt a senescence-associated secretory phenotype (SASP) that perpetuates barrier dysfunction. In CD, senescence within stem and stromal compartments limits regeneration, promotes pro-fibrotic remodeling, and sustains cycles of injury and repair via chronic SASP signaling. IBD prevalence continues to rise from environmental factors, dietary changes, antibiotic exposures, and gut microbiota alterations. Pathogenesis integrates genetic factors (e.g., NOD2, IL23R, HLA, and ATG16L1 mutations), environmental modifiers, dysbiosis characterized by loss of short-chain fatty-acid-producing Gram-positive bacteria and expansion of Proteobacteria, and a dysregulated immune system. Therapeutic strategies have shifted toward targeted biologics and small molecules to promote mucosal healing. In this review, we recapitulate the mechanistic axes of inflammation, oxidative stress, and senescence in IBD and then critically evaluate emerging targeted therapies. Topics include anti-TNFα, integrin blockade, IL-12/23 and IL-23 inhibition, JAK inhibitors, S1P receptor modulators, microRNA modulation, senomorphics, mesenchymal cell therapy, and microbiome interventions. We endorse biomarker-guided therapy and propose future directions to break the SASP-driven inflammatory loop and mitigate long-term carcinogenic risk. Full article

Background/Objectives: The global rise in antibiotic resistance necessitates the development of novel antimicrobial agents. Antimicrobial peptides (AMPs), key components of innate immunity, are promising candidates. This study aimed to develop novel therapeutic peptides with enhanced properties through the mutagenesis of natural AMPs and high-throughput screening. Methods: We constructed mutant libraries of three broad-spectrum AMPs—melittin, cecropin, and Hm-AMP2—using mutagenesis with partially degenerate oligonucleotides. Libraries were expressed in Escherichia coli, and antimicrobial activity was assessed through bacterial growth kinetics and droplet serial dilution assays. Candidate molecules were identified by DNA sequencing, and the most promising variants were chemically synthesized. Antimicrobial activity was determined by minimal inhibitory concentration (MIC) against E. coli and Bacillus subtilis, while cytotoxicity was evaluated in human Expi293F cells (IC₉₀) viability. The therapeutic index was calculated as the ratio of an AMP’s cytotoxic concentration to its effective antimicrobial concentration. Results: Mutant forms of melittin (MR1P7, MR1P8) showed significantly reduced cytotoxicity while retaining antimicrobial activity. Cecropin mutants exhibited reduced efficacy against E. coli, but variants CR2P2, CR2P7, and CR2P8 gained activity against Gram-positive bacteria. Mutagenesis of Hm-AMP2 generally decreased activity against E. coli, though two variants (A2R1P5 and A2R3P6) showed retained or enhanced efficacy against B. subtilis while maintaining low cytotoxicity. Conclusions: The proposed strategy successfully generated peptides with improved therapeutic profiles, including reduced toxicity or a broader spectrum of antimicrobial activity, despite not improving all parameters. This approach enables the discovery of novel bioactive peptides to combat antibiotic-resistant pathogens. Full article

Racemosol, a natural phenolic compound, is known for its antimicrobial potential, yet experimental studies remain limited. In this study, two new racemosol derivatives (4 and 5) and four known compounds (1–3, 6) were isolated from the bark of Bauhinia malabarica and structurally elucidated using spectroscopic analyses. Most of isolated compounds exhibited notable activity against Gram-positive bacteria, including Staphylococcus aureus, Bacillus subtilis, and Listeria monocytogenes, while showing limited effects on Gram-negative strains. Racemosol (1) and its derivatives (2, 4, and 6) displayed potent antibacterial activity with MIC values of 0.156–0.625 µg/µL and bactericidal properties confirmed by comparable MBCs. Compound 6 exhibited the highest potency, indicating that specific structural modifications enhance activity. These findings provide new insights into the structure activity relationships of racemosol derivatives and highlight B. malabarica as a promising natural source of phenolic antibacterial agents. Full article

Background: Various thiolactones are known as biologically active compounds, capable of stimulating the development of several human diseases and quorum sensing of Gram–positive bacteria. The enzymatic hydrolysis of thiolactones represents a promising approach to preventing their action. Methods: Thirteen enzymes, including various lactonases and serine hydrolases were studied in this work using several substrates including the homocysteine thiolactone (HTL), and its derivatives the N–acetylhomocysteine thiolactone (C₂–HTL) and the isobutyryl–homocystein thiolactone (i–but–HTL). The potential interactions of the ligands with the surface of enzymes molecules were predicted in silico using computational modeling and checked in wet experiments in vitro. Results: Based on the data obtained several enzymes were selected with localization of the thiolactones near their active sites, indicating the possibility of effective catalysis. The lactonase (AiiA), metallo-β-lactamase (NDM-1) and the organophosphate hydrolase with hexahistidine tag (His₆–OPH) were among them. Determination of catalytic characteristics of enzymes in the hydrolytic reactions with the HTL and the C₂–HTL revealed the maximal value of catalytic efficiency constant for the NDM-1 in the hydrolysis of the HTL (826 M⁻¹ s⁻¹). The maximal activity in the hydrolysis of C₂–HTL was established for AiiA (137 M⁻¹ s⁻¹). The polyaspartic (PLD₅₀) and the polyglutamic (PLE₅₀) acids were used to obtain polyelectrolyte complexes with enzymes. The further combination of these complexes with the clotrimazole and polymyxin B possessing antimicrobial properties resulted in notable improvement of their action in relation to Staphylococcus cells. Conclusions: It was revealed that the antimicrobial activity of the polymyxin B is enhanced by 9–10 times against bacteria and yeast when combined with the His₆–OPH polyelectrolyte complexes. The antimicrobial activity of clotrimazole was increased by ~7 times against Candida tropicalis cells in the case of the AiiA/PLE₅₀/Clotrimazole combination. These results make the obtained biology attractive and promising for their further advancement to practical application. Full article