Search Results (168)

Background: The treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections is extremely challenging due to its antibiotic resistance, and the combination of plant active ingredients with antibiotics represents a potential strategy to address this issue. Methods: We determined the combinatorial relationship between baicalin (BA) and kanamycin (KM) using the checkerboard dilution method. The antibacterial activity of the baicalin–kanamycin (BA/KM) combination was evaluated through growth curve determination assays and scanning electron microscopy (SEM). The effects of the BA/KM combination on the cell membrane and cell wall of MRSA were analyzed using reactive oxygen species (ROS) detection assays, intracellular protein leakage experiments, alkaline phosphatase (AKP) activity assays, laser scanning confocal microscopy (LSCM) observations, and molecular docking simulations. The antibiofilm activity and related mechanisms of the BA/KM combination were elucidated via crystal violet staining, MTT assay, phenol-sulfuric acid method, congo red staining, staphyloxanthin determination assays, and quantitative real-time polymerase chain reaction (qPCR). The safety of the BA/KM combination was assessed through hemolytic activity analysis, and its anti-MRSA efficacy was evaluated on lettuce. Results: BA/KM combination showed a synergistic antibacterial effect on MRSA USA300. Mechanistic studies revealed that BA may interact with amino acid residues of peptidoglycan synthetase PBP2a to hinder peptidoglycan synthesis, thereby facilitating KM penetration through the cell wall. Subsequently, BA binds to amino acid residues of the membrane transporter NorA, leading to disruption of cell membrane homeostasis and enhancing KM’s ability to induce intracellular ROS accumulation in MRSA. Furthermore, the BA/KM combination reduced MRSA biofilm formation by 77.85% and decreased the metabolic activity of biofilm cells by 42.93% through inhibiting the synthesis of biofilm components EPS and PIA. Additionally, this combination suppressed the synthesis of staphyloxanthin and downregulated the expression of agrA and agrC genes. When 1/8 MIC BA was combined with 1/4 MIC KM, the count of MRSA on lettuce surfaces was reduced by 0.88 log CFU/cm², an effect comparable to that of 0.2% (v/v) hydrogen peroxide. Conclusions: According to these findings, the BA/KM combination may offer a promising option for enhancing antibacterial efficacy through synergism, reducing antibiotic usage concentrations, and limiting MRSA transmission in fresh agricultural products. Full article

Background: Helicobacter pylori is associated with a wide range of gastroduodenal diseases, including chronic gastritis, peptic ulcer disease, and gastric cancer. Eradication efforts are challenged by increasing antimicrobial resistance rates, particularly in Southeast Asia. We sequenced the whole genomes of clinical H. pylori isolates from Southern Thailand to elucidate their resistance profiles, virulence determinants, and evolutionary relationships. Methods: Three clinical H. pylori isolates (004, 117, and 189) were subjected to whole-genome sequencing, phenotypic antimicrobial susceptibility testing, and comparative genomic analyses. Results: All strains exhibited high-level resistance to metronidazole. Additionally, H. pylori 117 was resistant to both amoxicillin and levofloxacin, classifying it as multidrug-resistant. Genomic analysis revealed mutations in rdxA, frxA, and rpoB, as well as in penicillin-binding protein genes (pbp2 and pbp3), supporting the phenotypic findings. While all isolates harboured clarithromycin resistance mutations (A2142G and A2143G in the 23S rRNA gene), they were phenotypically susceptible, highlighting a potential discordance that requires further investigation. Virulence gene profiling identified 115–118 conserved genes per strain, including cagA, vacA, oipA, babA, and flagellar, urease, and lipopolysaccharide biosynthesis genes. Phylogenetic analysis using core-genome single-nucleotide polymorphisms demonstrated that these strains formed a distinct Southern Thai monophyletic clade, suggesting localised clonal expansion driven by regional selective pressures. Conclusions: Region-specific surveillance strategies and treatment guidelines are urgently needed in Thailand. The combination of high-risk virulence genes and rising antimicrobial resistance in H. pylori strains necessitates tailored therapeutic approaches, the integration of genomic surveillance into clinical diagnostics, and expanded studies linking genotype to clinical outcomes in diverse populations. Full article

Penicillin-binding proteins (PBPs) are essential enzymes involved in bacterial cell wall biosynthesis and represent the primary targets of β-lactam antibiotics. However, the efficacy of these agents is threatened by β-lactamase production and PBP alterations, prompting the search for alternative strategies. In this context, boronic acids, long established as potent inhibitors of serine β-lactamases (SBLs), have been proposed as scaffolds for PBP inhibition based on the shared structural and mechanistic features of these enzyme families. This perspective provides a literature-based survey with structural analysis to evaluate emerging evidence on the potential role of boronic acids as PBP-targeting agents, with a particular focus on peptidomimetic boronic acids, repurposed β-lactamase inhibitors, and novel scaffold architectures. While early work showed limited activity against low-molecular-mass PBPs, more recent compounds, particularly certain bicyclic boronates, have demonstrated potent binding and, in some cases, antibacterial activity. Structural analyses reveal diverse binding modes and underscore the role of conformational dynamics in modulating affinity. Despite these advances, significant challenges remain, including target selectivity, membrane permeability, and species-specific differences. Nevertheless, the direct inhibition of PBPs by boronic acids, while still in early development, may offer a viable complement or alternative to β-lactam therapy, warranting further exploration through structure-guided design and comprehensive biological evaluation. Here, we analyze the potential of boronic acid inhibitors (BAIs) to target PBP enzymes, considering their promise as non-β-lactam antimicrobial agents with possible clinical relevance. Full article

The tomato leafminer (Tuta absoluta), a globally invasive pest, poses a major economic threat to tomato production. Although chemical control remains the primary management method, sustainable alternatives are urgently needed. Sex pheromone communication is critical for moth courtship and mating, with pheromone-binding proteins (PBPs) playing a key role in this process. In this study, we identified a PBP gene, TabsPBP2, from the T. absoluta transcriptome. Real-time quantitative PCR (RT-qPCR) revealed that TabsPBP2 is highly expressed in the antennae, with a strong male-biased expression pattern. Ligand-binding assays demonstrated that TabsPBP2 has the highest affinity for the sex pheromones (3E, 8Z, 11Z)-tetradecatrienyl acetate (TDTA) and (3E, 8Z)-tetradecadienyl acetate (TDDA). It also demonstrated a moderate-to-strong binding affinity to several tomato volatiles, including 2-carene, myrcene, α-pinene, cis-3-hexen-l-ol, methyl salicylate, sabinene, and α-terpinene. Molecular docking suggested that hydrophobic interactions predominantly stabilize the TabsPBP2–ligand complexes, with PHE118, PHE12, LEU90, LEU68, and ALA73 identified as key interacting residues. Electroantennogram (EAG) and Y-tube olfactometer assays confirmed that TDTA and TDDA act as strong attractants for male T. absoluta. This study enhances our understanding of the pheromone recognition in T. absoluta and provides a foundation for developing novel, pheromone-based pest control strategies. Full article

Background: Antibacterial resistance (ABR) poses a major challenge to global health, with methicillin-resistant Staphylococcus aureus (MRSA) being one of the prominent multidrug-resistant strains. MRSA has developed resistance through the expression of Penicillin-Binding Protein 2a (PBP2a), a key transpeptidase enzyme involved in bacterial cell wall biosynthesis. Objectives: The objective was to design and characterize a novel small-molecule inhibitor targeting PBP2a as a strategy to combat MRSA. Methods: We synthesized a new indole triazole conjugate (ITC) using eco-friendly and click chemistry approaches. In vitro antibacterial tests were performed against a panel of strains to evaluate the ITC antibacterial potential. Further, a series of in silico evaluations like molecular docking, MD simulations, free energy landscape (FEL), and principal component analysis (PCA) using the crystal structure of PBP2a (PDB ID: 4CJN), in order to predict the mechanism of action, binding mode, structural stability, and energetic profile of the 4CJN-ITC complex. Results: The compound ITC exhibited noteworthy antibacterial activity, which effectively inhibited the selected strains. Binding score and energy calculations demonstrated high affinity of ITC for the allosteric site of PBP2a and significant interactions responsible for complex stability during MD simulations. Further, FEL and PCA provided insights into the conformational behavior of ITC. These results gave the structural clues for the inhibitory action of ITC on the PBP2a. Conclusions: The integrated in vitro and in silico studies corroborate the potential of ITC as a promising developmental lead targeting PBP2a in MRSA. This study demonstrates the potential usage of rational drug design approaches in addressing therapeutic needs related to ABR. Full article

The repurposing potential of Efavirenz (EFV), a clinically established non-nucleoside reverse transcriptase inhibitor, was comprehensively evaluated for its in vitro antibacterial effect either alone or in combination with other antibacterial agents on several Gram-positive clinical strains showing different antibiotic resistance profiles. The binding potential assessed by an in silico study included Penicillin-binding proteins (PBPs) and WalK membrane kinase. Despite the relatively high minimum inhibitory concentration (MIC) limiting the use of EFV as a single antibacterial agent, it exhibits significant synergistic activity at sub-MIC levels when paired with various antibiotics against Enterococcus species and Staphylococcus aureus. EFV showed restored sensitivity of β-lactams against Methicillin-resistant S. aureus (MRSA). It increased the effectiveness of antibiotics tested against Methicillin-sensitive S. aureus (MSSA). It also helped to overcome the intrinsic resistance barrier for several antibiotics in Enterococcus spp. In silico binding studies aligned remarkably with experimental antimicrobial testing results and highlighted the potential of EFV to direct the engagement of PBPs with moderate to strong binding affinities (pK_a 5.2–6.1). The dual-site PBP2 binding mechanism emerged as a novel inhibition strategy, potentially circumventing resistance mutations. Special attention should be paid to WalK binding predictions (pK_a = 4.94), referring to the potential of EFV to interfere with essential regulatory pathways controlling cell wall metabolism and virulence factor expression. These findings, in general, suggest the possibility of EFV as a promising lead for the development of new antibacterial agents. Full article

In personalized nutrition, dietary guidelines must be adapted to the physiological and developmental needs of individuals across the lifespan, especially during childhood and adolescence. These should account for nutritional status, health conditions, and early-life risk factors, including those that emerge during pregnancy. This narrative review synthesizes recent evidence (2020–2025) on the biological value of protein sources in supporting pediatric growth and development. While adequate protein intake is essential for physical and cognitive development in individuals under nineteen, excessive intake may accelerate growth and increase the long-term risks of overweight and obesity. Compared to animal-based proteins (ABPs), plant-based proteins (PBPs) carry a higher risk of nutrient deficiencies in vulnerable populations due to lower digestibility and incomplete amino acid profiles. Although plant-based diets are encouraged for environmental reasons—particularly to reduce the ecological impact of livestock—protein intake must remain appropriate for age, sex, health status, and context. Nutritional strategies must ensure an adequate supply of essential amino acids and proper micronutrient supplementation, regardless of whether children follow diets rich in ABPs, PBPs, or a combination of both. Attention to these factors is vital to balancing nutritional adequacy with long-term health and sustainability goals. Full article

Phycobiliproteins (PBPs), captivating water-soluble proteins found in cyanobacteria, red algae, and cryptophytes, continue to fascinate researchers and industries due to their unique properties and multifaceted applications. These proteins consist of chromophores called phycobilins (PBs), covalently linked to specific protein subunits. Major phycobiliproteins include phycocyanin (PC), allophycocyanin (APC), and phycoerythrin (PE), each distinguished by distinct absorption and emission spectra. Beyond their colorful properties, PBs exhibit a broad spectrum of biological activities, including antibacterial, antifungal, antiviral, and antidiabetic effects, making them valuable for pharmaceutical, biotechnological, and medical purposes. The extraction and purification methods for PBs have been optimized to enhance their bioavailability and stability, opening new avenues for industrial production. For this review, a comprehensive literature search was conducted using scientific databases such as PubMed, Scopus, and Web of Science, prioritizing peer-reviewed articles published between 2000 and 2025, with an emphasis on recent advances from the last five years, using keywords such as “phycobiliproteins”, “phycobilins”, “bioactivities”, “therapeutic applications”, and “industrial use”. Studies were selected based on their relevance to the biological, technological, and pharmacological applications of PBPs and PBs. This review explores the diverse applications of PBs in therapeutic, diagnostic, and environmental fields, highlighting their potential as natural alternatives in the treatment of various diseases. The future perspectives for PBs focus on their incorporation into innovative drug delivery systems, biocompatible materials, and functional foods, presenting exciting opportunities for advancing human health and well-being. Full article

A series of 4-aminoquinoline derivatives were prepared using a microwave-assisted method. The reactions were initially carried out on a small scale and subsequently scaled up using a sealed tube. Heating the reactions to 90–150 °C for 90–120 minutes obtained products with up to 95% yields. Structural analysis and characterization were achieved using FT-IR, ¹H- and ¹³C-NMR spectroscopy and HR-MS. Four compounds displayed low-to-moderate antibacterial activity, with 6-chlorocyclopentaquinolinamine (7b) exhibiting potent inhibition against MRSA (MIC = 0.125 mM) and 2-fluorocycloheptaquinolinamine (9d) showing activity against S. pyogenes (MIC = 0.25 mM). Structure–activity relationship (SAR) docking studies within the Penicillin Binding Protein (PBP2a) binding site (PDB: 4DK1) showed that compounds 7b and 5b (7-chlorophenylquinolinamine) bind through hydrophobic interactions (ALA601, ILE614), hydrogen bonding (GLN521), and halogen contacts (TYR519, THR399). Compound 7b demonstrated enhanced MRSA inhibition due to additional π-alkyl interactions and optimal docking parameters. Conversely, the bulky structure of 9d may explain its weaker activity as it likely hindered binding to the target site. This paper highlights the role of structural features in antibacterial efficacy and guides the future optimization of 4-aminoquinoline derivatives. Full article

Background/Objectives: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the resistant pathogenic microorganisms that poses a global health threat due to its resistance to β-lactam antibiotics where the protein penicillin-binding protein 2a (PBP2a) plays a crucial role in its resistance. This study explores the potential of phytochemicals from Uvaria chamae, a plant with known medicinal properties, to serve as dual-site inhibitors of PBP2a, targeting both the active and allosteric sites. Methods: Phytochemicals previously identified in U. chamae were subjected to molecular docking and molecular dynamics simulations to evaluate their binding affinities and stability at PBP2a’s active and allosteric sites. The compounds’ pharmacokinetic profiles were predicted in silico using SwissADME tools. Root-mean-square deviation (RMSD), radius of gyration, and binding free energy were analyzed for dynamic stability. Results: Among the evaluated compounds, Uvarinol and Dichamanetin demonstrated high binding affinities compared to the co-crystallized ligand and standard antibiotics like ceftaroline. Uvarinol exhibited the highest binding affinity at both sites, with a docking score of −14.94 kcal/mol and a predicted inhibition constant (Ki) of 0.01 nM. Molecular dynamics simulations further confirmed the robust stability of Uvarinol and Dichamanetin, as indicated by consistently lower RMSD values relative to the co-crystallized ligand. Pharmacokinetic predictions revealed favorable drug-likeness and low toxicity, although Uvarinol showed limited gastrointestinal absorption. Conclusions: Uvarinol and Dichamanetin show promise as dual-site PBP2a inhibitors, offering a novel strategy to combat MRSA resistance. Their structural and pharmacokinetic properties make them viable candidates for further development, though experimental validation and formulation optimization are necessary to overcome bioavailability challenges. Full article

We have used single molecule tracking to investigate dynamics of four penicillin-binding proteins (PBPs) in Bacillus subtilis to shed light on their possible modes of action. We show that Pbp2a, Pbp3, Pbp4, and Pbp4a, when expressed at very low levels, show at least two distinct states of mobility: a state of slow motion, likely representing molecules involved in cell wall synthesis, and a mode of fast motion, likely representing freely diffusing molecules. Except for Pbp4, all other PBPs showed about 50% molecules in the slow mobility state, suggesting that roughly half of all molecules are engaged in a substrate-bound mode. We observed similar coefficients for the slow mobility state for Pbp4 and Pbp4a on the one hand, and for Pbp2a and Pbp3 on the other hand, indicating possible joint activities, respectively. Upon induction of osmotic stress, Pbp2a and Pbp4a changed from a pattern of localization mostly at the lateral cell membrane to also include localization at the septum, revealing that sites of preferred positioning for these two PBPs can be modified during stress conditions. While Pbp3 became more dynamic after induction of osmotic stress, Pbp4 became more static, showing that PBPs reacted markedly differently to envelope stress conditions. The data suggest that PBPs could take over functions in cell wall synthesis during different stress conditions, increasing the resilience of cell wall homeostasis in different environmental conditions. All PBPs lost their respective localization pattern after the addition of vancomycin or penicillin G, indicating that patterns largely depend on substrate availability. Our findings show that PBPs rapidly alter between non-targeted motion through the cell membrane and capture at sites of active cell wall synthesis, most likely guided by complex formation with other cell wall synthesis enzymes. Full article

During their lives, insects must cope with a plethora of chemicals, of which a few will have an impact at the behavioral level. To detect these chemicals, insects use several protein families located in their main olfactory organs, the antennae. Inside the antennae, odorant-binding proteins (OBPs), as the most studied protein family, bind volatile chemicals to transport them. Pheromone-binding proteins (PBPs) and general-odorant-binding proteins (GOPBs) are two subclasses of OBPs and have evolved in moths with a putative olfactory role. Predictions for OBP–chemical interactions have remained limited, and functional data collected over the years unused. In this study, chemical, protein and functional data were curated, and related datasets were created with descriptors. Regression algorithms were implemented and their performance evaluated. Our results indicate that XGBoostRegressor exhibits the best performance (R² of 0.76, RMSE of 0.28 and MAE of 0.20), followed by GradientBoostingRegressor and LightGBMRegressor. To the best of our knowledge, this is the first study showing a correlation among chemical, protein and functional data, particularly in the context of the PBP/GOBP family of proteins in moths. Full article

The tea tussock moth (Euproctis pseudoconspersa) is a common tea plantation pest with Type III sex pheromone components (SPCs). However, the olfactory genes involved in the perception of Type III SPCs remain unknown. To identify the olfactory genes involved in E. pseudoconspersa olfactory perception, we sequenced the transcriptomes of different tissues from male and female moths. We identified 27 chemosensory proteins, 39 odorant-binding proteins (OBPs), 28 ionotropic receptors (IRs), and 67 odorant receptors (ORs). Phylogenetic and antennal abundance analyses showed that EpseOR12, EpseOR13, EpseOR15, EpseOR16, and EpseOR18 belonged to the pheromone receptor clades of Type II moths, with predominant expression in male antennae. Besides these EpseORs, EpseOR14 and EpseOR32 were two of the most abundant EpseORs in male antennae, where they were predominantly expressed. Four pheromone-binding proteins (PBPs) were identified, with higher expression in male antennae. EpseORs and EpsePBPs may be involved in Type III SPC detection. Additionally, a few EpseOBPs, EpseIRs, and EpseORs were predominantly expressed in either male or female antennae. These genes may play important roles in olfaction and may be involved in detecting host plant volatiles and pheromones. These results provide a foundation for further exploration of the molecular mechanisms of E. pseudoconspersa olfaction. Full article

The mitochondrial serine β-lactamase-like protein LACTB has emerged as a critical regulator in cancer biology, distinguished by its unique structural and functional attributes. Defined by its conserved penicillin-binding proteins and β-lactamases (PBP-βLs) domain and SXXK catalytic motif, LACTB demonstrates properties distinct from its prokaryotic homologs, including the ability to polymerize into filaments. These structural characteristics enable LACTB to modulate mitochondrial organization and enzymatic activity, influencing lipid metabolism and indirectly affecting cellular proliferation. Importantly, the expression and functional roles of LACTB exhibit cancer-type-specific variation, underscoring its dual function as both a tumor suppressor and an oncogene. Decreased LACTB expression is associated with poor clinical outcomes in cancers such as breast cancer, lung cancer, and colorectal cancer, while specific mutations and regulatory mechanisms have been linked to its oncogenic activity in osteosarcoma and pancreatic adenocarcinoma. Mechanistically, LACTB regulates key processes in cancer progression, including mitochondrial dynamics, epithelial–mesenchymal transition (EMT), and cell death pathways. This duality highlights LACTB as a promising therapeutic target and underscores its relevance in advancing precision oncology strategies. This review provides a comprehensive analysis of expression level, structure–function relationships, and the diverse roles of LACTB in oncogenesis, underscoring its promise as a focal point for precision cancer therapies. Full article

In this study, three fungal steroids (1–3) were isolated from the fruiting bodies of the poisonous mushroom Gymnopilus orientispectabilis, based on bioactivity-guided isolation methods. The chemical structures of the isolates (1–3) were determined using NMR spectroscopic methods. Compounds 1–3 exhibited inhibition activity against E. coli, and their interactions with several bacterial drug targets were studied via in silico molecular docking, where the lowest binding energies were observed for penicillin binding protein 3 (PBP3) (−62.89, −75.89 and −74.47 kcal/mol, for compounds 1, 2 and 3, respectively). An MD simulation was performed to examine the conformational stability, motion and flexibility of protein–ligand complexes. In conclusion, this study investigates fungal steroids from G. orientaspectabilis as potential sources for new antimicrobial agents, encouraging further research to develop novel therapies. Full article