This study aimed to detect the co-occurrence of carbapenem resistance genes along with aminoglycoside-modifying enzyme (AME) genes in clinical Enterobacterales isolates to understand the distribution of multiple resistance genes among clinical isolates. This prospective study was conducted for six months (November 2024 to April 2025) in the department of microbiology of a tertiary care hospital. A total of 30 blood culture isolates were identified as resistant to both carbapenem and aminoglycoside antibiotics using the automated VITEK 2 compact system. The genes responsible for carbapenem resistance (blaNDM , blaOXA-48 , blaKPC , blaIMP , and blaVIM ) were detected by multiplex real-time PCR, and the aminoglycoside-modifying enzyme genes [APH(3')-Ia, APH(2")-Ib, AAC(3)-IIc, AAC(6')-Ib, and ANT(3")-I] were detected by the conventional polymerase chain reaction method. All the clinical data, patient demographics, and molecular findings were entered in an MS Excel spreadsheet version 14.0.4734.1000 and analyzed using GraphPad/PRISM software version 10.5.0. Of the 30 Enterobacterales isolates, Klebsiella pneumoniae was the most common isolate (66.7%). Molecular detection revealed blaNDM in 40% isolates and blaOXA48 in 10% isolates. The majority of the AME genes were in combination. The most common combination of the AME gene was AAC(6')-Ib+ AAC(3)-IIc+ ANT(3")-I + APH(3')-I detected in 4 (13.3%) isolates. The most common combination of carbapenem and aminoglycoside resistant genes was blaNDM + blaOXA48 + AAC(6')-Ib+ AAC(3)-IIc+ ANT(3")-I+ APH(3')-I (13.3%). The blaOXA-48 gene had a statistically significant association with AAC(6')-Ib, ANT(3")-I, and APH(3')-I (p <0.05). The Co-occurence of carbapenem resistance and aminoglycoside-modifying enzyme genes in clinical Enterobacterales isolates limits the therapeutic option.

Diabetes mellitus (DM), particularly type 2 DM (T2DM), is a chronic metabolic disorder requiring novel therapeutic approaches as the available therapies are not meeting the current challenges. This study investigates the anti-diabetic potential of Vigna unguiculata using a network pharmacology approach, supported by in vitro and in silico analyses. The plant was collected from Khyber Pakhtunkhwa, Pakistan, and subjected to hydroalcoholic extraction and fractionation. In vitro assays included α-amylase, α-glucosidase, and aldose reductase. Target prediction using STITCH and SwissTargetPrediction identified 88 common genes linked to T2DM. Protein-protein interaction (PPI) network analysis highlighted key genes like EGFR, PTGS2, and TLR4 as central nodes in diabetes-related pathways. Molecular docking was used to study the binding affinities of compounds. IC50 values were determined using IBM SPSS Statistics 21 software. The data underwent analysis using one-way ANOVA followed by Dunnett's multiple comparison test. Significance value was determined at *p 0.05, **p 0.01 and ***p 0.001. In-vitro assays demonstrated significant α-amylase, α-glucosidase, and aldose reductase inhibitory activities. Phytochemical screening identified several bioactive compounds. Functional annotation and KEGG pathway analysis confirmed these genes' roles in crucial metabolic pathways. Virtual screening revealed strong binding affinities of compounds like Stigmasterol, Luteoline, and Quercetin with GSK3B, PTGS2, and TLR4. The Molecular Dynamics (MD) simulation, binding free energy calculations (MM-PBSA and MM-GBSA), confirmed the results of Virtual screening. In short, these findings underscore V. unguiculata as a promising source for anti-diabetic agents, supporting further clinical trials for T2DM management.

Wound management presents significant challenges, requiring effective treatments. Herbal extracts have been traditionally used to support healing due to their anti-inflammatory, antimicrobial, and cell-regenerative properties. This study aimed to evaluate the therapeutic efficacy of Pau D'Arco (Tabebuia), Yarrow (Achillea millefolium), Gotu Kola (Centella asiatica), Figwort (Scrophularia nodosa), and Broadleaf (Plantago major) extracts, both individually and combined, on wound healing in vitro and in vivo in equine models. In vitro tests using human macrophages and keratinocyte cell lines to assess cellular responses such as cytokine secretion and phagocytic activity under simulated inflammatory conditions. Additionally, pilot case studies on equines with open wounds provided practical insights into the extracts' healing capabilities. MTT assay was used to assess cytotoxicity. The extracts did not significantly affect the viability of HaCaT or THP-1 cells. The herbal extracts reduced IL-8 levels and increased phagocytic activity in macrophages, indicating an ability to modulate inflammatory responses. In vivo, the extracts were well tolerated and associated with supported healing in equines. These effects were suggested to be attributed to the synergistic actions of the herbal components. These findings suggest that the herbal extracts may be useful for supporting wound healing. Their natural anti-inflammatory and healing properties could provide an additional option alongside traditional wound management approaches.

Anti-pathogenic potential of a polyherbal formulation Enteropan® was investigated against a multidrug-resistant strain of the bacterium Pseudomonas aeruginosa. Growth, pigment production, antibiotic susceptibility, etc., were assessed through appropriate in vitro assays. Virulence of the test pathogen was assessed employing the nematode worm Caenorhabditis elegans as a model host. Molecular mechanisms underlining the anti-pathogenic activity of the test formulation were elucidated through whole transcriptome analysis of the extract-exposed bacterial culture. Enteropan-pre-exposed P. aeruginosa displayed reduced (~70%↓) virulence towards the model host C. elegans. Enteropan affected various traits like biofilm formation, protein synthesis and secretion, quorum-modulated pigment production, antibiotic susceptibility, nitrogen metabolism, etc., in this pathogen. P. aeruginosa could not develop complete resistance to the virulence-attenuating activity of Enteropan even after repeated exposure to this polyherbal formulation. Whole transcriptome analysis showed 17% of P. aeruginosa genome to get differentially expressed under influence of Enteropan. Major mechanisms through which Enteropan exerted its anti-virulence activity were found to be generation of nitrosative stress, oxidative stress, envelop stress, quorum modulation, disturbance of protein homeostasis and metal homeostasis. Network analysis of the differently expressed genes resulted in identification of 10 proteins with high network centrality as potential targets from among the downregulated genes. Differential expression of genes coding for five (rpoA, tig, rpsB, rpsL, and rpsJ) of these targets was validated through real-time polymerase chain reaction too, and they can further be pursued as potential targets by various drug discovery programmes.

Immunotherapy has become an integral part of a comprehensive treatment approach to metastatic colorectal cancer (mCRC). Nivolumab (Opdivo) is a human immunoglobulin G4 monoclonal antibody that blocks the interaction between the programmed cell death 1 (PD-1) receptor and its ligands 1/2 (PD-L1/PD-L2), leading to inhibition of T-cell proliferation, cytokine secretion, and enhanced immune response. The US Food and Drug Administration (FDA) has approved this drug for use in high microsatellite instability (MSI-high)/deficiencies in mismatch repair (dMMR) advanced CRC patients. However, its efficacy is extremely limited in microsatellite stability (MSS)/mismatch repair proficient (pMMR) patients. We report a case of a 42-year-old man diagnosed with MSS/pMMR mCRC who has achieved a durable response to nivolumab after a progression under chemotherapy with antiangiogenic treatment. We observed for the first time an atypical response after 8 months of nivolumab treatment, with the regression of previous primary pulmonary lesions and the presence of new para-aortic lymph node lesions. This report demonstrates that a subset of pretreated mCRC patients with the MSS/pMMR phenotype may benefit from nivolumab and these patients need more attention.

Optimal treatment for metastatic non-small cell lung cancer (NSCLC) with mesenchymal epithelial transition gene (MET) exon 14 skipping mutation has not been established yet. MET inhibitors were demonstrated to be effective and tolerated in patients with this condition, while evidence on safety and efficacy of immunotherapy and/or chemotherapy in this population is limited. Here we report the case of an 86-year-old male with metastatic NSCLC harboring MET exon 14 skipping mutation and with high programmed cell death ligand 1 (PD-L1) expression (tumor proportion score ≥50%). The patient received the MET inhibitor tepotinib as first-line treatment, achieving a partial response, with G2 peripheral edema as adverse event that was successfully managed with temporary discontinuation, dose reduction, diuretics and physical therapy. After 31 months, the patient is still receiving tepotinib, with an ongoing response. Tepotinib is a valuable therapeutic option for first-line treatment of older patients with NSCLC harboring MET exon 14 skipping mutation, even in the presence of high PD-L1 expression.

Introduction: Antimicrobial resistance (AMR) is a serious global threat. Identification of novel antibacterial targets is urgently warranted to help antimicrobial drug discovery programs. This study attempted identification of potential targets in two important pathogens Pseudomonas aeruginosa and Staphylococcus aureus. Methods: Transcriptomes of P. aeruginosa and S. aureus exposed to two different quorum-modulatory polyherbal formulations were subjected to network analysis to identify the most highly networked differentially expressed genes (hubs) as potential anti-virulence targets. Results: Genes associated with denitrification and sulfur metabolism emerged as the most important targets in P. aeruginosa. Increased buildup of nitrite (NO2) in P. aeruginosa culture exposed to the polyherbal formulation Panchvalkal was confirmed through in vitro assay too. Generation of nitrosative stress and inducing sulfur starvation seemed to be effective anti-pathogenic strategies against this notorious gram-negative pathogen. Important targets identified in S. aureus were the transcriptional regulator sarA, immunoglobulin-binding protein Sbi, serine protease SplA, the saeR/S response regulator system, and gamma-hemolysin components hlgB and hlgC. Conclusion: Further validation of the potential targets identified in this study is warranted through appropriate in vitro and in vivo assays in model hosts. Such validated targets can prove vital to many antibacterial drug discovery programs globally.

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Objective:To review the pharmacology, pharmacokinetics, efficacy, safety, and place in therapy of a phenylalanine-metabolizing enzyme indicated to reduce blood phenylalanine concentrations, pegvaliase injection.Data Sources:Searches of MEDLINE (1946-September 1, 2018) were conducted using the terms pegvaliase and phenylalanine ammonia lyase (PAL). Additional data were obtained from the prescribing information, the product dossier obtained from the manufacturer, and Clinicaltrials.gov.Study Selection and Data Extraction:All English language articles related to pharmacology, pharmacokinetics, efficacy, or safety of the combination therapy in human subjects were reviewed.Data Synthesis:Pegvaliase is a pegylated PAL enzyme that converts phenylalanine to ammonia and trans-cinnamic acid. Blood phenylalanine levels were reduced by approximately 50% to 70% in patients receiving therapeutic doses of pegvaliase. However, most patients experienced adverse events.Conclusions and Relevance:The mainstay of therapy in phenylketonuria (PKU) has historically consisted of dietary restriction of phenylalanine. Pegvaliase injection is the first Food and Drug Administration (FDA)–approved enzyme substitution therapy for patients with PKU. The therapy may be a viable option for patients with documented blood phenylalanine >600 µmol/L who have failed existing management strategies.

This work explores the antiplasmodial potential of ethanol extract of Blighia sapida (Lin. Sapindaceae) in chloroquine (CQ)-resistant Plasmodium berghei (ANKA strain)–infected mice. Chloroquine-resistant (ANKA) strain of P berghei was inoculated intraperitoneally into Swiss albino mice. Mice were treated orally for 4 consecutive days, before and after inoculation (prophylactic, suppressive, and curative models) with graded doses of the plant extracts with Artemether-Lumefantrine (Coartem) as control. Prophylactically, the extract showed a remarkable activity in the chemosuppression of P berghei parasites (P < .01) ranging from 57% to 36.5% at doses of 200 to 800 mg/kg, respectively, whereas Coartem (10 mg/kg) produced 62.1% chemosuppression. No significant chemosuppression was observed in the curative and suppressive models. The plant extract appeared to be safe at the highest dose tested (5000 mg/kg) for acute toxicity, with no adverse effect on the different organs. The plant extract possesses prophylactic antimalarial activity, which supports its use in the prevention of malaria.