Timely management of sepsis relies on rapid diagnosis of bloodstream infections (BSIs), including early pathogen identification and antimicrobial susceptibility testing (AST). The Vitek® REVEALTM is an innovative rapid AST platform that detects volatile organic compounds emitted by Gram-negative bacilli (GNB) directly from positive blood cultures. This study assessed the analytical performance and clinical contribution of the BioFire® BCID2 molecular panel and the Vitek® REVEALTM system in the diagnosis of GNB-BSI among intensive care unit (ICU) patients. We analyzed the prospective phase of the BacteREVEAL study, conducted between March and October 2023 at Nîmes University Hospital. Results from BioFire® BCID2 and Vitek® REVEALTM were compared to standard-of-care methods, including MALDI-TOF identification and direct disk diffusion AST. Performance metrics for Vitek® REVEALTM were evaluated according to ISO 20776-2:2007 standards. Among 94 ICU patients with a first episode of GNB-BSI, 50 monomicrobial cases were included. Escherichia coli was the most frequently identified species (n = 30, 60%), followed by Pseudomonas aeruginosa (n = 7, 14%). BioFire® BCID2 provided accurate identification for all isolates. The innovative tools reduced median turnaround times for identification by 19.0 h and for AST by 14.9 h. The mean time to result for Vitek® REVEALTM was 5.7 h. Overall categorical agreement with standard AST was 97.7%, with 8 very major errors, 8 major errors, and 2 minor errors. These findings support the integration of BioFire® BCID2 and Vitek® REVEALTM into routine workflows to accelerate GNB-BSI diagnosis and improve sepsis management.

The rising incidence of Escherichia coli (E. coli) strains harbouring ESBL enzymes in healthcare environment including hospitals poses significant public health concerns. This study sought to assess the occurrence of E. coli strains that carry ESBL enzymes in clinical isolates from patients at hospitals in Tabuk, Saudi Arabia, focusing on the responsible genes and antibiotic susceptibility profiles. The 100 clinical samples positive for these ESBL genes were collected from the different Tabuk hospitals' in-patients for a 6-month period only, with multiplex PCR used to identify genes that encode ESBLs (blaTEM, blaSHV, blaCTX-M-group 1, blaCTX-M-group 9, and blaCTX-M-group 25). These results yielded that 24% of the clinical isolates exhibited positive for ESBL, with the CTX-M-1 gene having the highest prevalence (67% of ESBL-positive samples). This study underscores the high occurrence of the CTX-M-1 gene isolated in ESBL (+) E. coli within the Tabuk region, providing critical insights into local antibiotic resistance patterns. While these findings enhance our understanding of regional dynamics, they also reaffirm existing knowledge regarding global trends in ESBL gene prevalence. Other genes detected included blaSHV (46%), blaTEM (50%), blaCTX-M-9 (63%), and blaCTX-M-25 (50%). Co-occurrence of blaSHV and blaCTX-M-1 genes was observed in 42% of isolates, while blaTEM and blaSHV co-occurred in 12.5%. These ESBL-positive E.coli were primarily isolated from urine (70.8%), wound (12.5%) and blood (16.5%). While these enzymes confer resistance to many beta-lactam classes, they generally remain sensitive to carbapenems, cephamycin and certain β-lactamase inhibitors, while cefoxitin showed 95.8% resistance in this study. Although the study was conducted over a limited six-month duration with a relatively small sample size, the isolation of these specific strains is nonetheless significant. These outcomes highlight the necessity for ongoing surveillance studies and robust infection control programs to combat antibiotic resistance in healthcare settings. To address the study's objectives, it is recommended to develop targeted antibiotic stewardship programs that align with local resistance patterns, enhance surveillance systems to monitor ESBL-producing bacteria, and implement stringent infection control protocols in healthcare settings. Additionally, launching educational initiatives for healthcare professionals and the public, promoting research on resistance mechanisms, and collaborating with public health authorities will ensure that findings inform evidence-based policies. Finally, fostering multidisciplinary approaches will help create comprehensive strategies for effectively managing antibiotic resistance at both local and global levels.

The multidrug resistant Mycobacterium abscessus is an opportunistic pathogen of increasing concern, causing infections of skin and soft tissue as well as pulmonary, it can be life-threatening for immunocompromised individuals. Current treatment strategies are ineffective and new approaches to combat infections are imperative. The antimicrobial potential of Manuka honey has been recently demonstrated to be effective against M. abscessus, but the potential to improve its efficacy as well as reduce demand on a natural resource have not yet been explored. The development of medical honey paves the way for generation of a synthetic honey, but this information is largely hidden behind patents and also not explored from a synergist perspective of combined activity of MGO, H2O2 and defensin-1. Through addition of key enzymes or components and antimicrobial susceptibility testing, we have demonstrated that a vegan honea, containing no bee derived component results in antimicrobial activity against M. abscessus, causing not only growth inhibition but bactericidal activity. Both MGO and H2O2 are demonstrated to be relevant to the killing effect against M. abscessus.

Urinary tract infections (UTIs) are among the most prevalent bacterial infections and are increasingly complicated by the emergence of multidrug-resistant uropathogens. The present study investigated the probiotic potential and anti-uropathogenic activity of Lactobacillus strains isolated from traditional dairy products in western Algeria using combined in vitro screening and an in vivo rat model of UTI. Uropathogenic bacteria recovered from urine samples of female patients were identified as Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, and Proteus vulgaris using standard microbiological methods and the API 20E identification system. Sixteen lactic acid bacteria isolated from raw milk and fermented dairy products were characterized phenotypically and identified by API 50 CHL and 16S rDNA sequencing as Lactobacillus acidophilus (n = 5), Limosilactobacillus fermentum (n = 4), Levilactobacillus brevis (n = 5), and Lactobacillus delbrueckii subsp. lactis (n = 2). In vitro assays demonstrated strain-dependent variations in cell surface hydrophobicity, autoaggregation, and phenol tolerance among the isolates. Most isolates showed susceptibility or intermediate responses to several clinically relevant antibiotics, while resistance was mainly observed to vancomycin, penicillin G, piperacillin, and nalidixic acid. Agar well diffusion assays revealed inhibitory activity against the principal uropathogens, with the strongest inhibition observed against E. coli. Two selected strains (L. acidophilus and L. fermentum) were further evaluated through intravesical administration in rats. The in vivo efficacy was strictly dependent on the frequency of administration; notably, a single-dose treatment was completely ineffective and failed to control leukocyturia or reduce pathogen colonization. In sharp contrast, precolonization followed by repeated instillations led to a dramatic clearance of the pathogen by day 11, reducing urinary E. coli counts down to 49 CFU/mL for L. acidophilus and 356 CFU/mL for L. fermentum. This repeated regimen also significantly controlled the inflammatory host response, with leukocyte counts decreasing from greater than 104 cells/mL to approximately 4 × 103 cells/mL by day 11, while successfully preventing kidney dissemination and minimizing bladder tissue damage. These findings highlight their potential as complementary strategies to prevent or mitigate UTIs.

To study the ″preference″ of bacterial membranes for the secondary structure of antimicrobial peptides (AMPs), we designed a universal palindromic sequence template G(LK)n(KL)nG-NH2 (designated as GnPm, where n = 2-6 and m = 0-3) that can cover various secondary structures of AMPs, with 0 to 3 proline residues uniformly and dispersedly inserted in the middle region to regulate their secondary structures. Results confirmed that both the numbers of LK/KL repeats and proline can regulate secondary structure, thereby controlling the antibacterial activity of the AMPs. The optimized peptide G5P2 exhibits potent antibacterial activity via a typical membrane-disrupting mechanism, is not prone to inducing bacterial drug resistance, can synergize with traditional antibiotics to delay the emergence of antibiotic resistance, and has excellent in vivo antibacterial activity (a 93.52% bacterial clearance) and high safety (LD50 = 89.87 mg/kg), thus providing new chemical strategies and candidate molecules for the clinical management of drug-resistant bacterial infections.

The increasing threat of infections caused by multidrug-resistant Gram-positive pathogens such as MRSA and VRE has driven the structural remodeling and repurposing of traditional antibiotics as a key strategy to combat bacterial resistance. Herein, using bacitracin A as a lead template, we established a site-selective semisynthetic modification strategy via reductive amination of its N-terminal primary amine. 32 derivatives were designed and synthesized, and their structure-activity relationships were systematically evaluated. Optimized derivatives exhibited potent activity against MRSA and VRE, with 4-32-fold enhanced antibacterial potency compared with bacitracin A, together with improved bactericidal properties, safety profiles, and pharmacokinetic characteristics. Mechanistically, hydrophobic N-terminal modification endows bacitracin A with multiple antibacterial modes, including enhanced inhibition of cell wall peptidoglycan biosynthesis, obvious membrane depolarization, and disruption of the purine metabolic pathway. These findings provide a rational strategy for bacitracin optimization and highlight the potential of N-terminal modification for developing improved antibacterial agents.

Tuberculosis remains a grave threat to global public health, and the development of novel and effective therapeutic agents against mycobacterial infections is an urgent priority. Herein, we developed a series of cyclometalated palladium(II) complexes with 2-aminobiphenyl that exhibit antibacterial activities and fully characterized by 1H NMR, ESI-MS, FT-IR, ICP-OES and elemental analysis. All synthesized complexes exhibited potent in vitro antimycobacterial activity with MICs as low as 1.25 μM. Importantly, these complexes exhibit minimal cytotoxicity, robust stability, prominent cell-penetrating bactericidal potency and low propensity to induce drug resistance. Mechanism-of-action study demonstrated that their favorable antimycobacterial activity may be achieved by targeting the transporter MmpL11 and intracellular ROS accumulation. Molecular docking simulations confirmed that cyclometalated palladium complexes inhibit MmpL11 via multiple interaction effect, suggesting MmpL11 is a potential target for developing drug-resistant mycobacterial inhibitors. Taken together, our study identifies cyclometalated palladium complexes as potential compounds with potent antimycobacterial agents.

Methylation of a critical adenine residue of the 23S rRNA residing in the nascent exit tunnel of the bacterial ribosome, by Erythromycin resistant methyltransferases (Erms), reduces binding of macrolide, lincosamide, and streptogramin B (MLSB) class of antibiotics and confers high-level resistance. Using an allosteric site-directed focused virtual screening approach, nucleoside analogues were identified as the initial scaffold. 6-thioinosine, recognized as a significant initial hit molecule, was confirmed through in vitro screening to be a potent scaffold. Further, structure-based inhibitor evolution approach yielded improved analogues that demonstrated an MIC of 8 μg/mL via a combination therapy approach where Erm inhibitors were administered along with standard antibiotics in MLSB resistant clinical isolates. Our findings demonstrate that a strategy where allosteric inhibitors are developed to impede the ability of pathogenic ribosomal methyltransferases to recognize target RNA can serve as a generic approach for reversal of antibiotic resistance in a combination therapy mode.

Endophytic fungi are recognized as prolific sources of structurally diverse secondary metabolites with significant pharmaceutical potential. In the present study, endophytic fungi were isolated from healthy tissues of Atalantia racemosa (Rutaceae), an ethnomedicinal plant traditionally used for respiratory disorders and blood purification. A total of three fungal isolates were obtained and identified as Aspergillus flavus, Pleosporales sp., and Endothia sp. based on morphological and molecular (ITS sequencing) analyses. Crude extracts of the isolates were subjected to phytochemical characterization using GC-MS, FTIR, and LC-MS techniques. GC-MS profiling revealed the presence of fatty acids, sterols, phenolics, and terpenoids, while FTIR analysis confirmed key functional groups such as hydroxyl, carbonyl, and aromatic moieties. Importantly, LC-MS analysis provided high-resolution metabolite profiling, identifying diverse bioactive compounds including alkaloids, flavonoids, quinic acid derivatives, xanthones (mangostin derivatives), phenolic amides (N-feruloyl-octopamine), diterpenoids, chalcones, macrolides, and terpene lactones in both positive and negative ionization modes. Biological evaluation demonstrated notable antioxidant, antibacterial, anti-inflammatory, anticancer, and wound healing activities, with Endothia sp. showing superior anticancer activity (IC50: 19.37 µg/mL) and significant wound healing potential. The integration of LC-MS data highlights the metabolic diversity and pharmacological relevance of these fungal isolates. Overall, the study confirms that endophytic fungi associated with A. racemosa are promising sources of bioactive metabolites with potential applications in drug discovery.

The peel of Citrus hystrix DC. (kaffir lime), an underexploited agro-industrial by-product, may have bioactives. This work tested the ethanolic extract of C. hystrix peel's (EeChP) phytochemicals by GC-MS profiling, and the antidiabetic effect was evaluated using the alpha-amylase inhibitory test, and the anti-inflammatory activity was done using the human red blood cell (HRBC) membrane model. The antibacterial effect was determined using the agar well diffusion method against Staphylococcus aureus and Escherichia coli. Major constituents subjected to pharmacokinetic and ADMET filtering, molecular docking, Gene Ontology (GO), and KEGG pathway analysis for mechanistic plausibility. The GC-MS detected 44 phytoconstituents, including limonene, γ-sitosterol, and β-amyrin. EeChP inhibited α-amylase (IC50 = 80.97 µg/mL) and the stabilized HRBC membrane (IC50 = 64 µg/mL). The extract demonstrated substantial antibacterial action against S. aureus (IZD = 6 mm at 200 µg/well) but failed against E. coli. The β-amyrin and scoparone were found to be non-toxic. Scoparone and carbonic anhydrase 12 were connected with β-amyrin and protein tyrosine phosphatase non-receptor type 1. EeChP has anti-diabetic, multi-target, modulatory, and anti-inflammatory effects. The results recommend C. hystrix peel's bioresource potential, but enzyme kinetics, antimicrobial MIC, and in vivo research are necessary. The research has contributed to accomplishing SDG 3 and SDG 12.

A series of 33 perillyl alcohol based 1,3,4-thiadiazole-thioether compounds were designed and synthesized through active substructure splicing. Their structures were confirmed by [1]H NMR, [13]C NMR, and HRMS, and their antifungal activities were evaluated in vitro. At a concentration of 50 mg/L, the target compounds exhibited varying inhibitory activities against Gibberella zeae (G. zeae), Botrytis cinerea, Rhizoctonia solani (R. solani), Rhizoctonia cerealis (R. cerealis), and Physalospora piricola (P. piricola). Among these, compound V15 (R = 3-FC6H4) demonstrated the strongest effects against G. zeae, R. solani, R. cerealis, and P. piricola, with inhibition rates of 77.8%, 67.3%, 83.9%, and 86.1%, respectively. Notably, its activity against G. zeae (77.8%) surpassed that of the positive control chlorothalonil (73.1%), and its activity against P. piricola (86.1%) was comparable to that of chlorothalonil (89.3%). A preliminary three-dimensional quantitative structure-activity relationship (3D-QSAR) model for R. cerealis inhibition was established using comparative molecular field analysis (CoMFA), yielding a reliable model (r[2] = 0.991, q[2] = 0.547).

Motivated by the promising antimicrobial and ecotoxicological effects of 4-alkylamino-7-chloroquinoline and N-alkylphthalimide derivatives, these privileged nuclei were linked to investigate how lipophilicity influences antibacterial activity and ecotoxicity. For this purpose, seven 4-alkylamino-7-chloroquinolines (2a-g) were prepared, of which four were used as precursors (2a-d) and reacted with different anhydrides. Using a clean synthetic approach, 4-alkylamino-7-chloroquinoline-succinimides (3a-d) and -phthalimides (5a-d) were obtained in yields of 60%-99% after only filtration with distilled water. In vitro assays were performed to determine the minimum inhibitory/bactericidal concentration (MIC/MBC) against Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus mutans. Lipophilicity (log P) and toxicity were predicted using the pkCSM program. In vitro toxicity tests on Artemia salina larvae were used as an indicator of ecotoxicity. 4-Alkylamino-7-chloroquinolines (2b, 2f, 2g) revealed a moderate MIC of 156.25 µg·mL[-1] against Gram-(-) and Gram-(+) bacteria, whose predicted toxicity does not pose a potential risk to human health. Results indicate a correlation between increased lipophilicity and higher ecotoxicity, where less lipophilic compounds were less toxic to A. salina (LC50 133.01-777.50 µg·mL[-1]) than 4-alkylamino-7-chloroquinoline-phthalimides, suggesting further investigation of their cytotoxicity.

Plants are sources of compounds with benefits in human health and animal well-being. Plant-based foods may contain huge numbers of different phytochemicals. This is the case of a Cameroonian medicinal plant Eriosema glomeratum from which phytochemical studies reported so far, the isolation of 22 compounds (1-22) with 4 new ones. Several semisynthesis were done on erioschalcones A and B, two new dihydrochalcones isolated from this plant, to afford 18 (23-38) derivatives from which thirteen were new. Among the compounds isolated and their derivatives, several had significant inhibitory activities against microbial strains Bacillus megaterium, Escherichia coli, Chlorella fusca and Microbotryum violaceum; and strong to moderate cytotoxicities on human cancer cell lines, lung (A549), breast (MCF-7), and cervical (HeLa). Compounds were mostly alkyl caffeate, flavonols, isoflavonoids, and dihydrochalcones. The use of medicinal plants for their benefits in the mankind is daily increasing and their chemotaxonomic categorization and the natural origin of their metabolites are essential for more valorization. In this work, we have summarized all the investigations carried out on E. glomeratum and discussed the relevant correlation on the biosynthetic relationship between the different classes of compounds, the structure-activity relationship (SAR), as well as the chemophenetic importance of isoflavonoids and dihydrochalcones.

To discover novel green antifungal candidates, 20 3-acylaminoisoquinoline derivatives were designed and synthesized by introducing the active fragment phenoxypyridine. Their structures were verified by [1]H NMR, [13]C NMR, and HRMS. The structure of compound X9 was further confirmed via X-ray crystallography. Their antifungal activities against five phytopathogenic fungi were evaluated to develop isoquinoline-based antifungal agents. Among them, compound X6 demonstrated significant antifungal activity against Botrytis cinerea with an EC50 value of 7.55 µg/mL. In vivo assays revealed that compound X6 exhibited strong protective and curative efficacy against B. cinerea with rates of 77.86% and 45.45% at a concentration of 100 µg/mL. Molecular docking studies confirmed that isoquinoline amide compounds possess strong binding affinity to succinate dehydrogenase (SDH). Molecular dynamics (MD) simulations revealed that compound X6 has better pocket spatial compatibility and more stable binding interactions with SDH than boscalid. Furthermore, SDH activity assays showed compound X6 inhibited SDH activity by 66.7% at 4 EC50 concentration, which was comparable to that of the boscalid (75.85%). These findings demonstrated that isoquinoline derivatives incorporating the phenoxypyridine fragment possess potent fungicidal properties for crop safety, with compound X6 emerging as a promising candidate for a novel SDH inhibitor.