Molecules

Flavonoids have been widely investigated for their antiviral and anti-inflammatory properties, but their mechanisms of action often remain insufficiently defined. In the present study, high-purity flavonoids were evaluated using an integrated workflow combining molecular docking, LigPlot+ interaction mapping, surface plasmon resonance (SPR), fluorescence-based TMPRSS2 inhibition assays, and cell-based viability studies. Docking with AutoDock Vina identified Hesperidin as the strongest overall candidate among the compounds evaluated. Hesperidin showed strong active-site engagement with TMPRSS2, including interactions with catalytic residues His296, Asp345, and Ser441, and stable binding within the SARS-CoV-2 main protease (Mpro) pocket. Comparative docking showed weaker or more peripheral interaction patterns for Rutin and moderate Spike binding for Hesperidin and Rutin. Experimental validation demonstrated dose-dependent inhibition of TMPRSS2 activity with an IC50 of 79.1 {micro}M for Hesperidin and 43.5 {micro}M for Hesperetin, while Rutin showed partial inhibition without a defined IC50 in the tested range. In Calu-3 cells, pre-treatment with Hesperidin or Rutin reduced SARS-CoV-2 Spike-induced cytotoxicity by approximately 30% without detectable intrinsic toxicity at the concentrations tested Docking analysis of Hesperidin and Rutin with the SARS-CoV-2 Spike protein revealed moderate interaction patterns involving residues such as Asn343, Ser371, and Val367. Hydrogen bond distances were generally in the range of approximately 2.9-3.3 [A], indicating moderate stabilization compared with the stronger active-site interactions observed for Hesperidin in TMPRSS2. The resulting binding poses suggest that these flavonoids can associate with structurally relevant regions of the Spike receptor-binding domain; however, they do not strongly overlap with the key residues required for ACE2 interaction. Rutin, in particular, exhibited a more peripheral and distributed binding mode within the Spike-ACE2 complex, indicating limited potential for direct disruption of the binding interface. In addition to SARS-CoV-2 targets, docking analysis extended to influenza viral proteins revealed moderate interaction of Hesperidin with hemagglutinin (HA) and strong catalytic-pocket binding of Rutin to neuraminidase (NA), involving key residues associated with enzymatic activity. These findings broaden the scope of the study to include influenza viral entry and release mechanisms, supporting a multi-virus, multi-target framework.

The importance of human aldehyde oxidase (hAOX1) has increased over the last decades due to its involvement in drug metabolism. Inhibition studies concerning hAOX1 are extensive and a common reducing agent, dithiothreitol (DTT), was recently found to inactivate the enzyme. However, in previous crystallographic studies of hAOX1, DTT was found to be essential for crystallization. To surpass this concern another reducing agent used in crystallization trials. Using tris(2-carboxyethyl)phosphine (TCEP), a sulphur-free reducing agent, it was possible to obtain well-ordered crystals from hAOX1 wild type and variant, hAOX1_6A, which diffracted beyond 2.3 [A]. Instead of the typical star-shaped crystals of hAOX1, at pH 4.7, plates are obtained in the orthorhombic space group (P22121) with two molecules in the asymmetric unit. Activity assays with the enzyme incubated with both reducing agents show that contrary to DTT, TCEP does not lead to irreversible inactivation of the enzyme. The replacement of DTT with TCEP in crystallization of hAOX1 provides a strategy to circumvent enzyme inactivation during crystallographic studies, allowing future applications of new assays, such as time-resolved crystallography.

Asparagus racemosus commonly known as Shatamull, is a medicinal plant with pharmacological applications documented in both Indian and British Pharmacopoeias and various traditional medicinal practices. Previous studies have reported that A. racemosus reduces hyperglycemia by enhancing insulin secretion. The aim of the current study was to assess the antihyperglycemic actions and explore the underlying mechanisms of action of A. racemosus utilizing in vitro carbohydrate digestion, glucose diffusion, glucose uptake, 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and preliminary phytochemical screening. The inhibition of carbohydrate digestion was assessed using -amylase and -glucosidase enzyme assays. The effect on glucose diffusion was evaluated using cellulose ester dialysis tube. Subsequently, glucose uptake was measured in a yeast cell model at different glucose concentrations, and the antioxidant potential was evaluated by measuring DPPH radical scavenging activity. A. racemosus notably reduced (p<0.05, 0.001) glucose release during in vitro starch digestion by 37.69%, whereas glucose absorption decreased significantly by 33.60% (p<0.01-0.001). Additionally, the most significant enhancement (p<0.05, 0.001) in glucose uptake by 67.53%, was observed at 5 mM glucose concentration. Furthermore, it showed significant antioxidant activity by scavenging DPPH (p<0.01-0.001) radicals by 55.06%. Preliminary phytoconstituent screening indicated the existence of flavonoids, tannins, steroids, glycosides and saponins. In conclusion, A. racemosus shows an inhibitory effect on carbohydrate digestion and absorption, enhances glucose uptake and demonstrates significant DPPH radical scavenging activity, potentially due to the presence of naturally occurring phytochemicals. Thus, A. racemosus may contribute as a promising antidiabetic drug for the treatment of diabetes mellitus. More investigations are needed to determine the active compounds in A. racemosus that contribute to its antidiabetic effects.

Antimicrobial resistance (AMR) in plant pathogenic bacteria poses a serious threat to global agriculture, necessitating the development of novel antibacterial agents targeting virulence mechanisms. This study presents an integrated bioinformatics-driven framework for the rational design and computational validation of Solres, a newly designed small molecule targeting key virulence proteins in phytopathogenic bacteria. Approximately 10,000 active compounds from PubChem BioAssay (AID: 588726) were analyzed using structural clustering and scaffold mining to identify conserved molecular motifs associated with antibacterial activity. Guided by high-frequency substructures, Solres was designed de novo and screened for structural novelty against PubChem, ChEMBL, and WIPO databases. Drug-likeness evaluation using Lipinskis Rule of Five confirmed favorable physicochemical properties. Molecular docking was performed against essential virulence regulators, including PhcA, PhcR, HrpB, PehA, and Egl from Ralstonia solanacearum and Xanthomonas spp., with active sites predicted using CaspFold. Docking analyses revealed strong binding affinities and stable interactions with key catalytic and regulatory residues. Complex stability and conformational integrity were further validated through molecular dynamics simulations. Quantum chemical descriptors, including HOMO-LUMO energy gap and dipole moment, supported the electronic suitability and reactivity profile of Solres. Collectively, this study demonstrates the effective integration of cheminformatics, structural bioinformatics, molecular simulations, and quantum chemical analyses for plant-focused antibacterial discovery. The compound Solres represents a promising lead candidate for mitigating bacterial wilt disease and provides a computational framework for future experimental validation and sustainable crop protection strategies against AMR-driven phytopathogens.

The rapid emergence of antimicrobial resistance, particularly among multidrug-resistant (MDR) and extended-spectrum {beta}-lactamase (ESBL)-producing Escherichia coli, necessitates the development of novel therapeutic strategies. In this study, we report the green synthesis and functionalization of silver nanoparticles (AgNPs) using Azadirachta indica leaf extract conjugated with amoxicillin (Amoxicillin-AI-AgNPs) to enhance antibacterial efficacy. The synthesized nanoparticles were characterized using UV-Vis spectroscopy, FTIR, XRD, DLS, SEM, EDAX, and TEM analyses, confirming the formation of stable, spherical, crystalline nanoparticles with an average size of [~]87 nm and a zeta potential of -28.73 mV. High conjugation efficiency ([~]94%) of amoxicillin with AgNPs was achieved after 96 hours of incubation. Antimicrobial activity assessed against 88 clinical MDR and ESBL-producing E. coli isolates demonstrated significantly enhanced efficacy of Amoxicillin-AI-AgNPs compared to amoxicillin alone, with minimum inhibitory concentrations (MIC) ranging from 1.56 to 6.25 {micro}g/mL and minimum bactericidal concentrations (MBC) between 25-100 {micro}g/mL. Cytotoxicity evaluation on HEK-293 cells revealed a relatively high IC50 value (382.14 {+/-} 6.59 {micro}g/mL), indicating low toxicity at antibacterial doses. The synergistic interaction between AgNPs and amoxicillin likely contributes to improved bacterial inhibition and overcoming resistance mechanisms. Overall, this study highlights the potential of plant-mediated antibiotic- nanoparticle conjugates as an effective and biocompatible approach to combat antibiotic-resistant bacterial infections.

Catechinopyranocyanidins (Cpcs) which consist of diastereomers A and B are pigments derived from adzuki beans and are compounds in which the catechin and cyanidin skeletons are condensed to a pyrano ring. While catechins and anthocyanidins possess high antioxidant capacity, the physiological functions of Cpcs remains unclear. In this study, the antioxidant capacity of Cpcs was evaluated by in vitro antioxidant assays and by assessing their cytoprotective activity against oxidative stress in normal human dermal fibroblasts (NHDFs). Antioxidant capacity based on the hydrogen atom transfer (HAT) mechanism, as assessed by the ORAC assay revealed that Cpcs exhibit 14.1 mol TE/mol (Trolox equivalent antioxidant capacity: TEAC). Meanwhile, capacity based on the single electron transfer (SET) mechanism, as assessed by the DPPH, ABTS and CUPRAC assays revealed, they exhibit 2.1-3.6 mol TE/mol. Since TEAC value of Cpcs demonstrated by the HAT based mechanism higher than its SET based oxidative capacity suggesting that the antioxidant capacity of Cpcs is driven by the HAT mechanism. In cell culture experiments, Cpcs ameliorate cell toxicity in rotenone-induced injury model, suggesting to cytoprotective activity against mitochondrial dysfunction-dependent apoptosis. These results reveal novel physiological functions of Cpcs which may serve as a design guideline for elucidating in vivo dynamics based on antioxidant mechanisms.

Marburg virus (MARV) is a highly pathogenic filovirus that causes hemorrhagic fever with a high mortality rate, with very limited treatment options. The urgent need for targeted antiviral agents emphasizes the importance of structure-based drug discovery approaches. The present study aimed to evaluate the antiviral potential of Withaferin A (PubChem CID-265237) against three key proteins of MARV: viral protein 35 (VP35), and nucleoproteins (NP). Three-dimensional structures of these proteins were retrieved from RCSB-Protein Data Bank and docked with Withaferin A using AutoDock Vina. The ligand demonstrated favourable binding affinities towards all three viral targets, indicating strong interaction potential at functionally relevant sites. Drug-likeness and pharmacokinetic properties predicted using SwissADME and pkCSM indicated acceptable ADMET profiles that comply with key drug-like criteria. To validate the stability of the docking, molecular dynamics simulations (GROMACS, 100 nanoseconds) were conducted. The protein-ligand complexes exhibited stable root mean square deviation (RMSD), root mean square fluctuation (RMSF), and consistent hydrogen bonding patterns throughout the simulation. The MM-GBSA binding free energy analysis further supported favorable binding energetics, predominantly driven by van der Waals and electrostatic interactions. Altogether, these findings demonstrate that Withaferin A exhibits promising multi-target inhibitory potential against key MARV proteins. This study provides molecular insights into ligand-protein interactions and supports further experimental validation of Withaferin A as a potential therapeutic candidate against Marburg virus.

Cisplatin is a cornerstone chemotherapeutic agent for a broad spectrum of solid malignancies, yet its clinical utility is substantially curtailed by dose-limiting organ toxicity, principally nephrotoxicity and hepatotoxicity, mediated through reactive oxygen species (ROS)-driven oxidative stress, glutathione depletion, and lipid peroxidation. Naringenin (NAR), a bioactive citrus flavanone, possesses potent free-radical scavenging, anti-inflammatory, and cytoprotective properties that make it a compelling candidate for chemoprotection. The present study investigated whether oral naringenin supplementation (50 mg/kg body weight/day for 30 days) could mitigate cisplatin-induced oxidative injury to the liver and kidney in male Swiss albino mice. Cisplatin was administered intraperitoneally at 2.3 mg/kg body weight in three cycles of five consecutive days followed by a five-day interval. Biochemical indices of oxidative stress, such as malondialdehyde (MDA), reduced glutathione (GSH), and glutathione S-transferase (GST) activity, were assayed in liver and kidney homogenates on day 45. Cisplatin administration significantly elevated hepatic and renal MDA levels, indicating pronounced lipid peroxidation, and markedly depleted the concentrations of GSH and the activity of GST in both organs. Compared with cisplatin alone, naringenin coadministration significantly attenuated the increase in the level of MDA, restored the level of GSH, and rescued the activity of GST in both tissues, with more pronounced effects in the kidney. Notably, compared with the control, naringenin alone did not alter any biochemical parameters, confirming its physiological safety at the administered dose. These findings demonstrate that naringenin has meaningful hepatoprotective and nephroprotective effects against cisplatin-induced oxidative toxicity, possibly through antioxidant augmentation, glutathione repletion, and membrane stabilization mechanisms. This study provides a rational preclinical basis for evaluating naringenin as a coadministered chemoprotectant in cisplatin-based chemotherapy regimens.

The repair of photo-induced DNA lesions through nucleotide excision repair machinery is still the source of important questions. It has been observed that the repair rate of the different cyclobutane pyrimidine dimers, i.e. the photoproducts induced by dimerization of two {pi}-stacked pyrimidines (T<>T, T<>C, C<>T, C<>C), depends on the nucleobases involved in the lesion. TT derivatives (T<>T) are removed more slowly than those containing cytosine, especially in 5. Using all-atom molecular dynamics simulations and free-energy calculations, we demonstrate that the variation of the repair rate observed in human skin and in cultured cutaneous cell is associated to the recognition of the four lesions by the DDB2 protein moiety, and more specifically by the differential structural deformation induced on the complementary strand. Indeed, while C<>C and C<>T induce a larger deviation on the groove parameters, T<>T and T<>C, instead, affect DNA structure to a lesser extent. less affected. These effects then hamper differentially the downstream recruitment of the repair complexes. The observed DNA deformation correlates with the experimental repair rate and provides a structural rationale for the different repair rates of CPD by nucleotide excision repair machinery. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=105 SRC="FIGDIR/small/724087v1_ufig1.gif" ALT="Figure 1"> View larger version (43K): org.highwire.dtl.DTLVardef@cf6b6dorg.highwire.dtl.DTLVardef@195e35forg.highwire.dtl.DTLVardef@1829296org.highwire.dtl.DTLVardef@165baba_HPS_FORMAT_FIGEXP M_FIG C_FIG

The M. tuberculosis ESX-1 secretion system EccA1 enzyme is involved in the secretion of virulence factors and is essential for virulence and bacterial survival within the phagosome. Development of the small molecular inhibitors abolishing EccA1 function can yield new antivirulence drugs. In this study, we modeled the full-length EccA1 (573 residues, Mw [~]62.4 kDa) structure, which contains N-terminal TPR domain and a C-terminal CbxX/CfqX type ATPase domain. We have identified five ZINC compounds having binding energy i. e. Z1 (ZINC000004513760, -43.45 kcal/mol), Z2 (ZINC000000001793, -49.56 kcal/mol), Z3 (ZINC000005390388, -55.83 kcal/mol), Z4 (ZINC000257294577, -52.33 kcal/mol), Z5 (ZINC000004824264, -44.44 kcal/mol) through virtual screening of the ZINC compounds targeting C-terminal ATPase pocket of EccA1. The Z1-Z5 compounds were compared with ADP substrate having binding energy (Adenosine diphosphate, -35.00 kcal/mol), p97 ATPase inhibitors i.e. NMS873 (3-[3-cyclopentylsulfanyl-5-[[3-methyl-4-(4 methylsulfonylphenyl)phenoxy]methyl]-1,2,4-triazol-4-yl]pyridine, -48.68 kcal/mol), and CB5083 (1-[4-(benzylamino)-5H,7H,8H-pyrano[4,3-d]pyrimidin-2-yl]-2-methyl-1H-indole-4-carboxamide, -50.88 kcal/mol) against EccA1. The Z1-Z5 compounds exhibited good Absorption, Distribution, Metabolism, and/or Excretion properties (ADMTE). Pharmacokinetic properties and Lipinskys rule of five for Z1-Z5 compounds showed drug-like properties. 100 ns dynamics simulation analysis on EccA1 complexed with (i) Z1-Z5 compounds (ii) ADP substrate and (iii) NMS873 and CB5083 inhibitors showed high stability and biologically relevant conformation during dynamics simulation. These data indicate that Z1-Z5 compounds may act as potential inhibitors against EccA1 and provide avenues for new antivirulence drug development after in vitro and in vivo clinical trials.

Predicting protein-ligand interactions in the modern drug discovery has revolved from the involvement of artificial intelligence and structural bioinformatics using Graph Neural Networks (GNNs). The limited explainability of GNN models presents an important encumbrance in biomedical research, but it has achieved a high degree of accuracy in determining and identifying binding affinity and active compounds, as evidenced by [1] [2] [3] [4]. Here this research focuses on the interpretation of protein-ligand interactions at a molecular level, a rapidly developing area within Graph Neural Networks (GNNs). Now days modern study handling techniques such as visualization techniques, attention mechanism and model-based feature ascription by model to boost, and make robust and decrease false predictions on binding. Along with some approaches include like graph pooling strategies, message-passing optimization, self-supervised learning, transfer learning and contrastive learning are rapidly utilized to enhance the representative learnings. Furthermore, integration of molecular docking simulations, hybrid deep learning architectures and protein language model gives more reliable & biological predictions of protein-ligand interactions. That focuses on given process that identifies key ligand atoms and binding residues, as well as physicochemical factors influencing affinity, through chemical thought processes. Here this research work identified the challenges of developing biologically significant explanations, transparency, and the corollary dataset biases on interpretability. The research work conducted an in-depth investigation into the consolidation of protein language models to establish more reliable pathways for future research, examining hybrid architectures, transparent and energy-efficient GNNs, and scientifically grounded AI models for drug discovery. My research work highlights that XGNNs establishes a connection between Deep Learning and Biochemical expertise with increased confidence, which will enhance the accuracy of predictive models and computational models.

Glioblastoma is a very aggressive brain tumor with few therapeutics options. Type I and II Interferons (IFNs) co-formulation HeberFERON has been used in cancer treatment, with promising results in high grade brain tumors. High throughput techniques in easy-to-handle models have been important to interrogate biomolecules changes, describe mechanisms and find pharmacodynamic biomarkers. This study aims to elucidate the effect of HeberFERON over the cell proteome in comparison to its individual IFNs components. Proteomic changes with HeberFERON in the glioblastoma-derived cell line U-87MG, in comparison with individual IFN-2b and IFN-{gamma}, were studied using a nanoLC instrument EasyLC coupled to Velos Pro mass spectrometer; Maxquant and Perseus were also used. Several enrichment tools, networking analysis and canSAR for drug targets were employed. Translation, RNA processing, mitotic cell cycle, cytoskeleton and chromosome organization, apoptosis, autophagy, DNA repair are enriched to limit cellular growing together with changes in immune response components, supporting HeberFERON as a multitarget treatment. This co-formulation is distinguished at modulating RNA splicing with SMN complex, cytoskeleton organization and microtubule-based movement, nuclear envelope breakdown, DNA conformational changes, and oxidative phosphorylation, with a better drawing of effects over a variety of systems inside the tumoral cell. Together with previous microarray experiment, informative genes and proteins as pharmacodynamic biomarkers for antiproliferative effects showed up (ex. STAT1/2, CENPE, ATRIP, MAP1B, LIMA1, VCP, several ribosomal, spliceosome and proteasomal complexes proteins). This study complements transcriptomic and phosphoproteomic previous experiments in this model and underscore HeberFERON as a glioblastoma therapeutic.

Prasugrel is a prodrug, widely used in antiplatelet strategy for secondary prevention after acute coronary syndrome. The metabolism of prasugrel leads to the formation of the Prasugrel Active Metabolite (PAM), an irreversible P2Y12 receptor antagonist. Its mode of binding has not yet been fully established, although it is known that it binds covalently to P2Y12 by forming a disulfide bridge with cysteines and its sulfur moiety. PAM is a molecule with two chiral centers, resulting in four stereoisomers which appear to be stereoselective upon binding. A combination of different molecular modeling methods, such as molecular dynamics, ensemble docking, and Density Functional Theory (DFT), were used to rationalize these differences in antagonism observed in vitro and to elucidate the mode of binding of PAM to P2Y12. PAM is found to bind to the closed P2Y12 conformation in a preferential way. Although the four stereoisomers have comparable affinity, the location of the RS stereoisomer makes the formation of a disulfide bond with cysteines more favorable, particularly with cysteine 175. Compared to the RR stereoisomer, the RS stereoisomer interacts less deeply with the P2Y12 receptor, interacting in particular with the second and third extracellular loops, explaining the competition observed with cangrelor and an intermediate metabolite of prasugrel. Furthermore, DFT calculations have shown that the formation of a disulfide bridge is energetically more favorable with the RS stereoisomer than with the RR stereoisomer. The physical interactions and chemical reaction between the RS stereoisomer and the P2Y12 receptor are key factors in explaining the stereoselective binding of PAM to P2Y12.

The emergence of multidrug-resistant (MDR) and extended-spectrum {beta}-lactamase (ESBL)-producing Escherichia coli poses a significant threat to global public health, necessitating the development of alternative antimicrobial strategies. In this study, biogenic silver nanoparticles (AgNPs) were synthesized using aqueous seed extract of Azadirachta indica as a green, eco-friendly reducing and stabilizing agent. Successful synthesis of nanoparticles was confirmed by a visible color change and characterized using UV-Visible spectroscopy, FTIR, XRD, DLS, SEM, EDAX, and TEM analyses. The synthesized AgNPs exhibited a strong surface plasmon resonance peak at 420 nm and were predominantly spherical with an average size of [~]38 nm and a zeta potential of -24.26 mV, indicating moderate stability. The antimicrobial efficacy of the synthesized AgNPs was evaluated against 88 clinical isolates of MDR and ESBL-producing E. coli. The nanoparticles demonstrated potent antibacterial activity with a minimum inhibitory concentration (MIC) ranging from 1.5625 to 3.125 {micro}g/mL and bactericidal effects at low concentrations, significantly outperforming the neem seed extract alone. Cytotoxicity assessment using HEK-293 cell lines revealed a relatively high IC50 value (297.01 {+/-} 10.04 {micro}g/mL), suggesting low toxicity at effective antimicrobial doses. Overall, the study highlights the potential of A. indica seed-mediated AgNPs as an effective and biocompatible antimicrobial agent against resistant bacterial strains, warranting further in vivo investigations for clinical applications.

Neurodegenerative diseases (NDDs) are currently raising their prevalences and new preclinical low-cost investigations of drug design are urging. NDDs encompass a wide range of disorders, including Alzheimers, Parkinsons, ALS and others, many of which share mitochondrial dysfunction as a common pathological feature. As such, targeting mitochondrial metabolism has emerged as a promising therapeutic strategy. However, while rodent models are widely used in NDD research, they are costly and time-consuming, raising the need to consider other alternatives to accelerate the search for novel therapies. In this line, zebrafish (Danio rerio) have gained outstanding popularity as a valuable option. This systematic review aims to provide an extensive overview about the current strategies that use zebrafish assays to investigate modulations of mitochondrial function as new therapies against NDDs. The review was performed following an electronic search of different databases (PubMed, Embase, Scopus and Web of Science) after the PRISMA procedure. Articles published in the English language were identified and screened based on the keywords used: mitochondrial metabolism, therapy, neurodegenerative diseases and zebrafish. Following 176 entries, exclusion criteria reduced the record to 34 final studies. Overall, we found that these studies investigate 37 compounds: 24 natural, 6 semisynthetic, 5 synthetic and 2 compounds of not-determined origin; to ameliorate 9 prevalent diseases: ARSACS, Alzheimers, Parkinsons, Huntingtons diseases, Leigh and Wolfram syndromes, Amyotrophic lateral sclerosis, Limb - girdle muscular dystrophy 2G and hyperglycemia-associated amnesia. Additionally, a meta-analysis of these compounds and their gene interactions provides insights into their mechanisms of action and advances our understanding of NDDs, and furnishes us with a powerful tool to predictive potential new drugs or to repurpose existing ones. To conclude, this systematic review suggests that zebrafish have become an effective model for screening potential drugs for NDDs with symptomatology difficult to replicate in rodent models. Moreover, the use of computational tools is also emphasized as a promising strategy to guide therapeutic discovery more efficiently, reducing both time and costs, in developing treatments for NDDs. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=102 SRC="FIGDIR/small/710294v1_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@18893a1org.highwire.dtl.DTLVardef@1943a12org.highwire.dtl.DTLVardef@709146org.highwire.dtl.DTLVardef@51a488_HPS_FORMAT_FIGEXP M_FIG C_FIG

Pseudomonas aeruginosa, a Gram-negative opportunistic pathogen is well known for life-threatening acute infections among the human population. The bacterium can withstand most antibiotics by using their high levels of inherent and acquired resistance mechanisms such as Biofilm-EPS, Persistence, and Quorum sensing (QS). Owing to the importance of adaptive antibiotic multi-drug resistance of P. aeruginosa, the current investigation is aimed to explore the phytochemicals derived from mangrove plants as potential agents to control biofilm and drug resistance mechanisms through a multi-mechanistic computational approach. For identifying potential compounds and target, In-silico drug repurposing technique is implemented by docking/virtual screening of 49 phytochemical compounds against 18 proteins involved in the Persister Cell formation, QS, and EPS synthesis in P. aeruginosa which resulted the proteins RelA and SpoT (persistence), PqsA, and PqSR (QS), and PelA and PelB (EPS synthesis) and compounds Taraxerone and Taraxerol to be potential. The results of docking were well corroborated with MD simulations. These targets and compounds explored through in-silico approach, are found to target potential antimicrobial pathways involving EPS synthesis, persistence genes, and QS, aiming to enhance antibiotic efficacy. Further, this study could be reference for in-vivo and in-vitro investigations to evaluate the further effectiveness of the compounds and potentiality of the proteins for MDR therapeutics of P. aeruginosa.

Glioblastoma (GBM) presents significant therapeutic challenges due to its aggressive nature, complex microenvironment and the limitations of conventional drug delivery systems. In this study, hybrid nanoparticles were developed by combining synthetic liposomes with macrophage-derived extracellular vesicles (EVs) to harness the strengths of both platforms. Two distinct liposomal formulations, DPPC:Chol:DSPE-mPEG2000 (F1) and DPPC:DPPS:Chol:DSPE-mPEG2000 (F2), were used as the basis for the synthesis. EVs derived from J774 macrophages were integrated with F1 and F2 to create hybrid nanoparticles (H-F1 and H-F2). Doxorubicin (DOX) was encapsulated using a pH gradient and a remote loading procedure. The mean particle size of H-F1-DOX and H-F2-DOX was 158.2 {+/-} 1 nm and 162.8 {+/-} 9 nm, respectively. The polydispersity index (PDI) was 0.130 {+/-} 0.012 and 0.084 {+/-} 0.033, while the zeta potential values were -14.9 {+/-} 0.7 mV and -26.7 {+/-} 3.1 mV, respectively. H-F2-DOX exhibited the highest encapsulation efficiency (EE%), reaching 76.5{+/-}3.4%. The encapsulated hybrids remained stable up to one week, at +5{degrees}C. The release of DOX from H-F2-DOX in DMEM supplemented with 10% serum showed pH sensitivity, with total DOX release of 64.9 {+/-} 5.3% at pH 7.4 and 90.7 {+/-} 6.5% at pH 5.5. The cell viability assay demonstrated that all formulations exhibited strong cytotoxic effects against GBM cells under normoxic conditions, with H-F2-DOX showing the most potent effect under hypoxia-mimetic conditions.

In light of the "silent" AMR pandemic, new avenues to combat pathogenic bacteria are needed. In this work, we screened a large molecule library (n=35 684 unique compounds) with the aim of identifying molecules being able to bind and block translation of the prfA-thermosensor transcript in the bacterial pathogen Listeria monocytogenes. Using a thiazole-orange displacement approach, 468 ([~]1.3% of all molecules) showed the ability to reduce fluorescence. After dose response testing, 32 compounds remained promising and eight of them showed sufficient purity and availability to be further validated. Interestingly, four compounds, being structurally very similar, showed specificity for prfA at a varying degree. All four compounds carried 3 aromatic rings with one connecting amine between two of the rings and an amide linking an aliphatic amine side chain. The most selective compounds, M5, showed a Kd of [~]0.8 {micro}M for the prfA RNA at 35{degrees}C. However, none of the eight most efficient compounds were able to inhibit prfA translation in vitro, suggesting that the molecules are able to bind but not affect the stability of the overall structure. Through this work, we have been able to identify a set of molecules, able to bind the prfA thermosensor RNA selectively, but without affecting translation. These molecules could constitute an important scaffold for further drug development.

BackgroundTriple-negative breast cancer (TNBC) is an aggressive subtype lacking well-defined molecular targets, leaving chemotherapy as the primary treatment despite drug resistance, systemic toxicity, and high recurrence rates. Therefore, the development of effective and less toxic therapeutic agents is essential. This study investigated the anti-cancer potential of gloriosine, a bioactive alkaloid with antiproliferative activity and low toxicity toward normal breast cells. MethodsPotential targets of gloriosine were predicted using SwissTargetPrediction, TargetNet, and PharmMapper, and overlapping genes related to TNBC and glutamine metabolism were selected. Protein-protein interaction networks, Gene Ontology, and KEGG pathway enrichment analyses were performed. Molecular docking evaluated binding affinity, followed by in vitro validation using cell viability, colony formation, and wound healing assays. ROS levels were measured by DCFDA and GSH assays, and ferroptosis was assessed by Western blot and FerroOrange staining in MDA{square}MB{square}231 cells. ResultsA total of 100 potential targets were identified, with 60 overlapping with TNBC and glutamine metabolism-related genes. Key targets included SRC, EGFR, mTOR, and HSP90AA1. Enrichment analyses indicated involvement in cancer progression, metabolic regulation, and resistance pathways, including central carbon metabolism, EGFR inhibitor resistance, and ErbB signaling. Gloriosine showed strong binding affinity toward hub targets. Experimental studies confirmed concentration-dependent inhibition of cell proliferation and migration. Mechanistically, gloriosine suppressed glutamine metabolism via GLS1 downregulation and induced ferroptosis, evidenced by increased ROS, glutathione depletion, GPX4 downregulation, and elevated intracellular iron levels. ConclusionsGloriosine exerts significant anti-cancer effects in TNBC through multi-target modulation and induction of ferroptosis, highlighting its potential as a promising therapeutic candidate. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=133 SRC="FIGDIR/small/725321v1_ufig1.gif" ALT="Figure 1"> View larger version (40K): org.highwire.dtl.DTLVardef@ce0ebcorg.highwire.dtl.DTLVardef@29603borg.highwire.dtl.DTLVardef@6d0025org.highwire.dtl.DTLVardef@249700_HPS_FORMAT_FIGEXP M_FIG C_FIG Flow chart of the network pharmacological and in vitro study of gloriosine

Parkinsons disease (PD) is the second most progressive degenerative disorder of the brain due to dopaminergic (DA) neuron degenerations and alpha-synuclein (-Syn) accumulations. At present, the disease has no effective treatment. Therefore, the current study objective is to identify a novel anti-PD formula (Zhi-Shi-Huang-Wu Formula, F-2) computed at 8:4:2:1 ratio from HSP 70 promoter activators Valeriana jatamansi (V), Acori talarinowii (A), Scutellaria baicalensis (S), Fructus Schisandrae (F). Traditionally, V is used to cure memory impairments, A treats mental disorders, and chronic mild stress, S for neuroprotection, and F showed multiple therapeutic actions to treat insomnia. This study investigated the neuroprotective potential of the V, A, S, F, formula F-2 and its underlying molecular mechanisms in transgenic Caenorhabditis elegans models. A, S, F, and F-2 successfully restored 6-hydroxydopamine intoxicated DA neuron degenerations, reduced food-sensing behavior disabilities, and attenuated -Syn aggregations. Moreover, activates the lipid deposition and proteasome expressions to confirm -Syn degradations at the cellular level. Reactive oxygen species (ROS) cause oxidative stress, and A, S, F, and F-2 repressed ROS and raised SOD-3 expressions. Overall, these data indicate that V, A, S, F combined into F-2 (22.3%) are more effective against PD progression-like symptom than individual drugs V (0.7%), A (11.4%), S (9.6%), and F (12.6%). These improved neuroprotective actions of F-2 possibly due to following the antioxidative pathway. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=144 SRC="FIGDIR/small/709540v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@1a6f1f7org.highwire.dtl.DTLVardef@157a270org.highwire.dtl.DTLVardef@69a238org.highwire.dtl.DTLVardef@1194b5e_HPS_FORMAT_FIGEXP M_FIG C_FIG