Bacterial infections caused by multidrug-resistant (MDR) strains pose a serious global health threat. This study aimed to evaluate the antibacterial efficacy of green-synthesized copper nanoparticles (G-CuNPs) against MDR strains of Acinetobacter baumannii, Pseudomonas aeruginosa, Klebsiella spp., and Escherichia coli. Emerging MDR pathogens necessitate the development of novel, eco-friendly alternatives. G-CuNPs synthesized using Citrus pseudolimon peel extract may offer a biocompatible and sustainable approach to combating MDR infections. Clinical bacterial isolates were obtained from diagnostic specimens including urine, pus, wound swabs, sputum, and blood collected from hospitalized patients at a tertiary care hospital. Bacterial identification and antimicrobial susceptibility testing were performed using the VITEK 2 automated system. Phenotypic detection of metallo-β-lactamase (MBL) and extended-spectrum β-lactamase (ESBL) production was conducted. G-CuNPs were synthesized and characterized for physicochemical properties. Antibacterial activity was assessed using a CFU-based time-kill assay. Mechanistic studies included evaluation of cell membrane integrity, reactive oxygen species (ROS) generation, and DNA degradation. Interaction with bacterial enzymes was analyzed via molecular docking. Hemolytic and cytotoxicity assays were performed to assess biocompatibility. G-CuNPs (10 mg/mL) displayed potent antibacterial activity by disrupting cell membranes, inducing ROS accumulation, and degrading bacterial DNA. Molecular docking confirmed strong binding affinities to key bacterial enzymes. Compared to chemically synthesized CuNPs, G-CuNPs (Indian Patent No. 202111048797) exhibited minimal hemolytic and cytotoxic effects. G-CuNPs demonstrate promising antibacterial potential and biocompatibility, highlighting their applicability in biomedical domains such as implant coatings and wound care. Further in vivo studies are warranted to validate their clinical utility.

Exploring Morin hydrate as a natural thymidine kinase inhibitor to combat Staphylococcus aureus infection.

Being a persistent and deadly infection, tuberculosis (TB) caused by Mycobacterium tuberculosis remains a global health challenge. Despite having a well-established 4-drug combination therapy for drug-sensitive TB, the emergence of drug-resistant TB has rendered the treatment less effective. Although fluoroquinolones (FQs) are among the prominent drugs in the second-line treatment for multidrug-resistant tuberculosis (MDR-TB), FQ resistance has readily emerged in cases of extensively drug-resistant tuberculosis (XDR-TB). Other than the mutations in DNA gyrase, a universally conserved bacterial enzyme targeted by FQs, several mechanisms contribute to the emergence of FQ resistance. Recently, post-translational modification of DNA gyrase is implicated as one of the mechanisms for FQ resistance. Here, we describe succinylation of GyrB by a promiscuous acetyltransferase, Eis of M. tuberculosis, as a new mechanism contributing to FQ resistance in mycobacteria. Lysine succinylation of GyrB results in a reduced interaction of FQs with the enzyme, thereby decreasing the DNA cleavage by DNA gyrase in the presence of FQs. Accordingly, Eis overexpressing mycobacterial strains exhibit increased minimum inhibitory concentration (MIC) to FQs. Thus, succinylation of DNA gyrase is yet another resistance mechanism against the FQs.

The global increase in antimicrobial resistance, along with the role of pathological angiogenesis in infection-associated inflammation, underscores the need for multifunctional therapeutic agents. In this study, a series of riparin-type benzamides were synthesized using efficient and accessible synthetic methods, yielding structurally diverse compounds, and their antimicrobial and antiangiogenic potential was evaluated. Among these, riparin III (3c) exhibited the most potent and broad-spectrum antimicrobial activity, effectively inhibiting Gram-positive and Gram-negative bacterial strains as well as Candida spp. Furthermore, riparin I (3a), riparin II (3b), and riparin III (3c) exhibited antiangiogenic activity, suggesting their potential therapeutic relevance in infection-associated inflammation. Toxicity assessments indicated low cytotoxicity at the effective concentrations, supporting the safety profile of the compounds. Molecular docking analysis corroborated the in vitro findings, revealing strong interactions between riparin I (3a) and fungal cell wall targets, as well as between riparin III (3c) and key bacterial enzymes involved in resistance and replication. Overall, these results highlight riparin I (3a), riparin II (3b), and riparin III (3c) as promising multifunctional candidates for the development of antimicrobial agents capable of addressing the current challenges in infection control, including antimicrobial resistance and inflammation-associatedpathologies.

The escalating prevalence of multidrug-resistant tuberculosis (MDR-TB) underscores the urgent need for new classes of antitubercular agents targeting novel pathways. Carbonic anhydrase, a ubiquitous metalloenzyme, catalyses the reversible hydration of carbon dioxide in the CO2 + H2O ⇋ HCO3 - + H+ reaction. Suppressing this enzymatic activity has recently been identified as a new pathway for the treatment of Mycobacterium tuberculosis. To address this, a series of isoxazole-sulphonamides was rationally designed, incorporating an isoxazole pharmacophore as the aromatic tail, amide as a linker, and sulphonamide as the zinc-binding group. These compounds were evaluated against Mycobacterium tuberculosis carbonic anhydrases (MtCA 1 and 3) and two human carbonic anhydrases (hCA I and II) to identify selective inhibitors of the bacterial enzymes. The findings indicated that molecules containing an isoxazole pharmacophore with amide-linked benzene-3-sulphonamide were significantly more selective for MtCA 3 than hCA I and II. Among these compounds, 12c, 12e, and 19b had the highest inhibition against the MtCA 3 with K i values between 0.08-0.09 μM compared to the standard acetazolamide with a K i value of 0.10 μM. Some of the best compounds exhibited potent and selective inhibition of MtCA 3 over hCA I and II, with the meta- and para-substituted derivatives demonstrating higher selectivity and stronger inhibition. Specifically, compound 19b proved to be 199 and 38 times more selective for MtCA 3 than hCA I and hCA II respectively, compared to the standard drug acetazolamide, which is a non-selective CA inhibitor. The potential of compound 19b as a promising antitubercular agent with a MIC value of 8 μg mL-1 against mc2 6230 was further strengthened by in silico ligand-target interaction studies. Thus, compound 19b is emphasised as a promising lead in the pursuit of new, selective agents targeting MtCA 3.

Background: Pulmonary hypertension (PH) is a fatal condition characterized by elevated pressure in pulmonary artery which could be partly caused by dysfunctional nitric oxide (NO) signaling. Previous studies identified that nitrate is actively concentrated in saliva and could be reduced to nitrite by bacterial nitrate reductase enzymes, which are rarely found in human genomes. Oral nitrate reduction (NR) capacity was previously reported to be predictive of blood pressure in systemic circulation; however, the role of the oral microbiome in oral NR and NO signaling in PH remains unclear. Hypothesis: We hypothesize that loss of nitrate reductase-expressing bacteria in oral microbiota leads to decreased NR capacity and is associated with worse PH phenotype. Method: We prospectively enrolled patients who underwent RHC at University of Pittsburgh Medical Center, following the ERS 2022 PH hemodynamic classification. Oral wash [OW] samples were collected&underwent 16S rRNA sequencing then analyzed with Qiime2 (v2024.10; SILVA). We predicted bacterial NR capacity in silico by Picrust2 (v2.6). Differential abundance analyses were done with ANCOMBC2. Plasma nitrate&nitrite levels were quantified. In a subset, NR capacity was verified by measuring ex vivo nitrite formation rate with NO analyzer (Sievers) after adding KNO 3 substrate to OW. Results: We enrolled n=184 patients who underwent RHC in 2014-2020. Of these, 144 (78%) had mPAP over 20mmHg (PH) with mPAP of 32.4 [10.2] mmHg (mean [SD]) and PVR of 3.6 [1.9] Wood Units. Between PH vs non-PH patients, alpha diversity, beta diversity and microbial load (16S qPCR) were similar in the oral microbiome (all p>0.05). Veillonella, Rothia, Neisseria etc are major NR capacity contributors, but their abundances did not differ between PH vs non-PH. NR index, defined as the ratio of sum abundance of NR contributors over non-contributors, was significantly associated with the predicted abundance of bacterial nitrate reductase genes, and also verified with the NR enzyme activity (p=0.016) measured in vitro . Increased NR index is associated with lower mPAP (R=-0.21, p=0.0129, adjusted for diastolic systematic blood pressure [DBP] and smoking history) and lower PVR (R= -0.19, adjusted p=0.019) among PH patients. Conclusions: We confirmed that predicted nitrate reduction capacity by the oral microbiome reflects functional nitrate reduction in vitro and is associated with PH severity. The oral microbiome represents a modifiable target in PH.

Metagenomic approaches have revolutionised the discovery of novel enzymes with ecological and biotechnological significance from different environments. Here, we report the comprehensive characterisation of a novel salicylaldehyde dehydrogenase (SALDAP) obtained from an alpine soil metagenome. Phylogenetic analysis revealed that SALDAP is the first experimentally characterised Alphaproteobacterial SALD, forming a distinct evolutionary clade among known bacterial enzymes. The recombinant enzyme exhibited strict specificity for NAD⁺ and exceptional catalytic efficiency toward aromatic aldehydes, with benzaldehyde as the preferred substrate. Kinetic analyses showed catalytic efficiencies exceeding 10⁶ M⁻¹ s⁻¹ for aromatics, whereas aliphatics were oxidised with much lower efficiency, consistent with ecological specialisation for aromatic catabolism in alpine soils enriched in lignin-derived compounds. SALDAP was most active under mildly alkaline conditions (optimum pH 8.0) and tolerated a range of chemical environments, though high concentrations of certain metals and solvents were inhibitory. Differential scanning fluorimetry demonstrated that the enzyme was stabilised by ligand binding, with maximal thermal stability observed when both substrate and cofactor were present. Structural alignment with Pseudomonas NahF and docking analyses revealed that SALDAP employs a distinctive catalytic configuration involving ASN-137, ARG-145, GLU-238, and CYS-272, highlighting a non-canonical role for ASN-137 in substrate binding and stabilisation. Based on these findings, we propose a mechanistic model for SALDAP that expands the catalytic diversity of the aldehyde dehydrogenase superfamily. This study establishes a new paradigm for aromatic aldehyde oxidation, underscores the ecological significance of SALDAP in alpine soil microbiomes, and provides a foundation for engineering novel biocatalysts for bioremediation and synthetic biology applications.

Advanced disinfection system using a 172nm excimer lamp and citric acid for Escherichia coli O157:H7.

Divergent acetyl-CoA binding modes mediate allosteric inhibition of bacterial hybrid-type malic enzymes.

Bioprocessing of Cressa cretica: From biopolymer extraction to pectinase-mediated juice treatment.

ABSTRACT In this study, a library of aryl‐substituted chroman‐4‐one ester derivatives 8(a–k) was synthesized through a sequential approach involving condensation–cyclization, Suzuki–Miyuara coupling, followed by nucleophilic acyl substitution reaction. The structures were finalized by 1 H and 13 C NMR, and mass spectral data. Pure titled compounds were assessed their antimicrobial activity against a panel of Gram‐positive and Gram‐negative bacteria in addition to fungal strains using standard in vitro methods. Of these, the Compound 8a exhibited the highest antibacterial activity, with minimum inhibitory concentration (MIC) values range of 0.98–0.12 µg/mL against across the tested bacterial strains, which matched the activity of ciprofloxacin. However, the antifungal activity of Compound 8a , MIC values for which were the lowest among all the compounds, with values of 0.98 µg/mL for Candida albicans and 3.90 µg/mL for Aspergillus niger and Aspergillus flavus , was equivalent to fluconazole. Attributable to the strong electron‐withdrawing –CF 3 group increases lipophilicity, enhancing π–π interactions and binding affinity with microbial targets. The combined biological and computational results highlight the potential of these novel chroman‐4‐one ester derivatives as promising candidates for antimicrobial drug development. Docking study revealed that the Compounds 8a–k bind effectively to the bacterial enzyme DNA gyrase B (PDB ID: 1KZN) with molecular interactions that are comparable to that of ciprofloxacin. These interactions provide valuable insights into their mechanism of enzyme inhibition.

The global rise of antimicrobial resistance underscores the need for novel inhibitors targeting essential bacterial enzymes such as UDP-N-acetylglucosamine enolpyruvyl transferase (MurA). This study evaluates the antibacterial potential of three natural polyphenols—Acertannin from African leaves and structurally modified with Thiophene-2-carbaldehyde (TC) to enhance MurA inhibition. A validated QSAR model, incorporating hydrophobic, electronic, and steric descriptors, predicted significantly lower EC₅₀ values for TC-modified compounds, with TC-acertannin showing the highest predicted potency (EC₅₀ = 0.382 µM). Molecular docking revealed strong binding affinity to MurA, with ΔG = −7.8 kcal/mol and Ki = 1.88 µM, involving key interactions such as hydrogen bonding, π-anion, and π-sulfur contacts with residues CYS115, ARG120, ASN23, ARG91, LYS22 and GLU188. PASS prediction further indicated enhanced antibacterial activity and membrane-related mechanisms, with TC-Acertannin showing a Pa of 0.923 for membrane integrity agonism. These results highlight TC-modified tannin as promising MurA-targeted antibacterial agents and support the rational design of natural product-based inhibitors to combat antibiotic-resistant bacteria

Infectious diseases and cancer remain leading global health challenges, with rising resistance to existing antibiotics and limited selectivity of many cytotoxic agents. Heterocyclic scaffolds, particularly thiazolidinones, offer a promising platform for the development of novel antimicrobial and anticancer compounds. The aim of the study. To evaluate the antimicrobial and cytotoxic properties of thiazolidinone-based compounds against a panel of pathogenic microorganisms and human cancer cell lines, and to identify the most promising derivatives with favorable safety, pharmacokinetic, and mechanistic profiles through molecular docking and dynamics studies. Materials and methods. A library of 5-enamine(hydrazine)-4-thiazolidinone derivatives was screened for antimicrobial activity against Gram-positive and Gram-negative bacteria and Candida albicans, and for cytotoxic activity against six human cancer cell lines. Minimum inhibitory concentrations (MIC) were determined, and IC₅₀ values were measured for selected compounds. Pharmacokinetic properties, including gastrointestinal absorption and lipophilicity, were assessed in silico. To investigate potential mechanisms of antibacterial action, molecular docking was performed against MurB (UDP-N-acetylenolpyruvylglucosamine reductase) and DNA gyrase subunit B (ATPase domain), followed by molecular dynamics (MD) simulations to evaluate the stability of the most promising complexes. Results. Thirty-two compounds exhibited antimicrobial activity (MIC ≤ 200 µM), and ten (6, 7, 10, 12, 13, 16, 19, 21, 22, and 29) were identified as the most active. Compound 29, an isatin–oxadiazole hybrid, demonstrated potent activity against Enterococcus faecalis and vancomycin-resistant E. faecium (MIC = 3.13 µM), outperforming vancomycin. Compound 21 was highly active against Staphylococcus epidermidis (MIC = 1.56 µM), while compound 6 showed efficacy against methicillin-susceptible and -resistant S. aureus (MIC = 6.25 µM). Moderate antifungal activity was observed for compound 27 (MIC = 100 µM), whereas Gram-negative bacteria were largely resistant. Cytotoxicity screening revealed selective anticancer activity of compounds 12 and 27, with high therapeutic indices against CCRF-CEM cells and minimal effects on normal fibroblasts. Compound 2 exhibited strong cytotoxicity (IC₅₀ = 1.1 µM), while compound 29 combined non-cytotoxicity with favorable pharmacokinetic characteristics. Molecular docking supported MurB as the primary antibacterial target, with the most active compounds (21 and 29) showing the most favorable binding energies. Compound 29 also exhibited strong affinity for GyrB, indicating a potential dual-target mechanism. Molecular dynamics confirmed that MurB–compound 29 complexes were particularly stable, correlating well with experimental antibacterial activity. Conclusions. Thiazolidinone-based hybrids demonstrated promising antimicrobial and anticancer properties. Compound 29 emerged as a particularly attractive dual-purpose candidate due to its potent activity, safety profile, favorable pharmacokinetics, and validated interaction with essential bacterial enzymes. Together, biological and computational results support the potential of thiazolidinone scaffolds as a basis for the development of selective or multitarget therapeutic agents

Antimicrobial resistance (AMR) poses a significant global health challenge, with methicillin-resistant Staphylococcus aureus (MRSA) being a major threat. The urgent need for novel therapeutic agents has driven research into natural bioactive compounds. Bactris guineensis, a native Colombian plant, is rich in polyphenols, which have demonstrated antimicrobial properties in prior studies. This research explores the potential of B. guineensis polyphenols as inhibitors of MRSA through molecular docking studies. Three-dimensional structures of 13 polyphenols from B. guineensis were retrieved from PubChem. Geometric optimization was performed using Gaussian 09 with Density Functional Theory (DFT) at the B3LYP/6-31G level. The optimized conformations served as ligands for molecular docking, which was conducted with AutoDock Vina, evaluating binding energy as the primary affinity metric. Seven S. aureus protein targets (PDB codes: 5M18, 6O9W, 4URO, 7JM2, 5TZJ, 3FRF, 6H5O) were obtained from the Protein Data Bank, prepared using SYBYL-X, and assigned Kollman charges with polar hydrogens in AutoDockTools. Validation was performed using crystallized endogenous ligands. The overall energy profile across all targets demonstrated favorable binding affinities, with values ranging from -5.9 to -10.3 kcal/mol. Isoorientin, a C-glucosylated flavone with known in vitro antimicrobial activity, exhibited the highest binding affinity (-10.3 kcal/mol) against the TarS enzyme (PDB: 5TZJ), which catalyzes β-OGlcNAcylation of teichoic acids in the MRSA cell wall. Isoorientin formed key interactions, including two hydrogen bonds with aspartate 178 and additional contacts with residues R206, E177, H210, Q179, and S175. Schaftoside showed strong affinity (-9.7 kcal/mol) for dihydrofolate reductase, interacting with critical residues (L28, V31, I50, L54) in the enzyme’s hydrophobic channel, mirroring known inhibitors. Polyphenols from B. guineensis, particularly isoorientin and schaftoside, demonstrate significant potential as anti-MRSA agents by targeting key bacterial enzymes. These findings highlight the value of characterizing bioactive metabolites from native Colombian plants for developing novel antimicrobial therapies.

Chondroitinase ABC (ChABC) is a bacterial enzyme that can potentially address the inhibitory effects of Chondroitin Sulfate Proteoglycans (CSPGs) in various neurological disorders and injuries. CSPGs are key components of the extracellular matrix that, when accumulated after Central Nervous System (CNS) injury or neurodegenerative diseases, inhibit axonal growth and tissue repair. This review explores the therapeutic potential of ChABC in Spinal Cord Injury (SCI), Traumatic Brain Injury (TBI), stroke, Parkinson's Disease (PD), Alzheimer's Disease (AD), and peripheral nerve regeneration. ChABC's mechanism of action involves the degradation of CSPGs, promoting neural plasticity, axonal regeneration, and functional recovery in SCI and other CNS injuries. In stroke and TBI, ChABC treatment has been shown to enhance neurogenesis, reduce glial scar formation, and support neuronal survival. In neurodegenerative conditions like PD and AD, ChABC's ability to modify the inhibitory extracellular environment offers novel strategies for promoting neuronal repair and cognitive function. Additionally, ChABC has been explored in cancer therapy, where its ability to degrade the tumor extracellular matrix facilitates improved drug delivery and tumor infiltration. While ChABC holds promise, challenges remain in its clinical application, particularly regarding stability, targeted delivery, and long-term effects. This review discusses the mechanism of action of ChABC and various delivery strategies, including viral vectors and localized infusion, and emphasizes the need for further research to optimize ChABC's potential. The future of ChABC in regenerative medicine depends on overcoming these barriers, improving delivery methods, and exploring synergistic treatments for enhanced recovery outcomes.

Abstract Diabetic foot ulcers (DFUs) often involve multidrug‐resistant (MDR) infections, complicating treatment. Plant extracts and green‐synthesized nanoparticles are promising alternatives for combating these pathogens. This study aimed to evaluate the antimicrobial and antibiofilm effects of Hibiscus sabdariffa extracts and magnesium oxide nanoparticles (MgO‐GNPs) against MDR bacteria isolated from DFUs, while exploring their mechanisms through in silico analysis. Qualitative phytochemical analysis was performed on Hibiscus extracts. The MgO‐NPs were characterized using Fourier transform infrared spectroscopy (FTIR), X‐ray diffraction (XRD), ultraviolet–visible spectroscopy (UV–vis), energy dispersive X‐ray analysis (EDX), and scanning electron microscope (SEM). Their antimicrobial effects by MIC against E. coli , P. aeruginosa , and S. aureus . Biofilm inhibition and disruption were assessed in mono‐ and mixed‐species cultures. Molecular docking and molecular dynamics simulations explored interactions between MgO‐GNPs and bacterial enzymes. MgO‐GNPs demonstrated superior antimicrobial activity, with MICs between 22 and 36 µg/mL. They significantly inhibited biofilm formation and disrupted existing biofilms more effectively than Hibiscus extracts. MgO‐GNPs increased ROS levels, damaged bacterial membranes, and suppressed exopolysaccharide and alginate production. In silico studies showed strong binding affinity of MgO‐GNPs to bacterial glycosyltransferases, supporting their mechanism of action. MgO‐GNPs show promising antimicrobial and antibiofilm activity against MDR pathogens. Their ability to target biofilms and bacterial mechanisms suggests they could serve as effective alternatives for managing drug‐resistant DFU infections.

Abstract A BF 3 ·OEt 2 catalyzed chemo‐ and regioselective Michael type addition of indoles with α,β‐unsaturated oxindole esters has been developed to synthesize new designed 3‐indolylmethyl oxindole scaffolds in excellent yields (61%–88%). Notably, the reaction exhibits high site‐selectivity, favoring C‐3 indole addition to the β‐position of the enone moiety while retaining full tolerance to the ester functionality. This metal‐free protocol offers the first selective access to biologically important NH‐free indolyl frameworks in a single step, relevant to naturally occurring indole alkaloids, including those isolated from the indole‐3‐carbinol‐induced culture of Daldinia eschscholzii . In silico studies of the synthesized compounds revealed strong binding affinities to key bacterial enzymes ( Staphylococcus aureus FtsZ, Sortase A, Thioredoxin Reductase) and cancer‐related human proteins (Bcl‐2, KSP, EGFR). Several compounds showed dual‐targeting potential, highlighting their promise as multifunctional therapeutic candidates.

Bioremediation effectiveness of crude oil-polluted soil can be monitored via bacterial enzyme activity. In this study, crude oil-polluted and non-polluted soil samples were used to prepare various treatment media, and petroleum hydrocarbon (PHC) degradation was assessed at different incubation periods and pH levels by measuring catalase activity. The stimulatory effects of kenaf-core combined with 95% rhamnolipid on in vitro PHC degradation were compared against bio-stimulation and bioaugmentation treatments. All treatments showed peak catalase activity on the 90th day of incubation. Among bio-augmented treatments, AZ1T6 (Azikoro-1) exhibited the highest catalase activity (20.19 mL KMnO4 g−1h−1), followed by OL1T6 (Ologbo-1) and OT1T6 (Otukpoti-1) (17.66 mL KMnO4 g−1h−1), with BN3T6 (Benin-3) showing the lowest (17.33 mL KMnO4 g−1h−1). In bio-stimulated treatments, media supplemented with kenaf-core and rhamnolipid (T4) demonstrated the highest catalase activity at day 90. All bio-stimulated treatments had significantly higher catalase activity compared to the negative control (T7) (P < 0.05), though their activity was statistically lower than in bio-augmented treatments (P < 0.05). Optimal degradation occurred at pH values between 6.0 and 8.0. The synergistic combination of kenaf-core and 95% rhamnolipid likely enhanced nutrient availability, stimulating indigenous microorganisms and catalase activity, thereby improving bioremediation efficiency beyond what either treatment achieved alone. The enhanced effectiveness observed when combining kenaf-core and rhamnolipid, relative to their separate use, provides evidence for a novel synergistic interaction.

Diabetic nephropathy is a kidney disease aggravated by the uremic toxin indoxyl sulfate, which is produced from indole by the gut microbiota. Targeting the bacterial enzyme tryptophan indole-lyase (TIL), which produces indole from L-tryptophan, could be a promising therapeutic strategy. This study investigates diketopiperazines, particularly cyclo-glycylproline [cyclo(Gly-Pro)], as potential TIL inhibitors. Cyclo(Gly-Pro) and other diketopiperazines moderately inhibited indole production from L-tryptophan in crude bacterial extracts. Cyclo(Gly-Pro) was not metabolized by the bacteria and did not affect their viability. Cyclo(Gly-Pro) inhibited the Escherichia coli TIL with a Ki​ value of 17μM through a mixed-type mechanism. Computational docking studies supported this finding, showing that cyclo(Gly-Pro) binds near the active site of TIL. Additionally, cyclo(Gly-Pro) significantly reduced indole production in bacterial cultures and human fecal samples. These findings suggest that cyclo(Gly-Pro) could be a promising dietary supplement or a lead compound for developing new therapeutics to prevent or treat diabetic nephropathy.

Background: Probiotics are increasingly used in aquaculture as eco-friendly alternatives to antibiotics for enhancing growth, feed efficiency, and fish health. Big Fish Probiotic, containing beneficial bacterial strains and digestive enzymes, has potential for improving carp farming productivity in Pakistan. Objective: This study aimed to evaluate the effects of Big Fish Probiotic on growth performance, feed efficiency, survival, and behavioral health of Cyprinus carpio (common carp) fry. Methods: A total of 45 fry were randomly assigned to three groups: (1) control group fed commercial feed, (2) 1% probiotic group with Big Fish Probiotic supplementation, and (3) 2% probiotic group with higher supplementation. The experiment was conducted for 15 days under controlled conditions with daily water exchange and aeration. Growth parameters, including weight gain (WG), specific growth rate (SGR), feed conversion ratio (FCR), condition factor (K), survival rate (SR%), and behavioral traits were measured. Results: The 1% probiotic group demonstrated the most favorable outcomes, recording the highest WG (4.33 g), best SGR (0.57%/day), lowest FCR (1.62), and 100% survival rate. Behavioral assessments also revealed greater vitality and feeding response in this group. The 2% probiotic group showed moderate improvements, while the control group had poor survival (60%) and weaker feed efficiency. Conclusion: Supplementation of Big Fish Probiotic at a 1% inclusion level significantly enhances growth, survival, and feed efficiency in Cyprinus carpio. The findings indicate that Big Fish Probiotic can serve as a sustainable feed additive to improve aquaculture productivity in Pakistan.