Mitochondrial diseases are common inherited neurometabolic disorders and frequently involve the nervous system, yet their multisystem nature often necessitates complex pharmacological management. Many commonly prescribed medications have off-target effects on mitochondrial function, and patients with mitochondrial disease may be particularly vulnerable to such effects due to impaired energy metabolism. However, systematic data on medication safety in this patient group remain scarce. In this retrospective, single-centre, cohort-based study at Turku University Hospital (Turku, Finland), we reviewed the medication data from all hospital stays and outpatient prescriptions of 44 mostly adult (20 women; mean age 50 years, range 12-83 years) patients with genetically and clinically confirmed mitochondrial disease for years 2010-2022. We used the Anatomical Therapeutic Chemical system for drug classification. Potential drug-drug interactions and potential adverse drug reactions were investigated. Special focus was on potential mitochondrial toxicity of drugs and clinically relevant drug-drug interactions. Altogether ~ 1000 individual medication entries were reviewed. We identified several common drugs with potentially adverse effects on mitochondria, including metformin, beta-blockers, statins, ciprofloxacin, fluoxetine, ibuprofen, and certain anti-seizure drugs. Medications generally considered contraindicated in mitochondrial disease were not observed. No high-risk drug interactions were detected. Additional finding of clinical relevance was the frequent use of analgesics. Further research regarding mitochondrial safety of several drug classes is needed for more evidence-based safety evaluations. Pain in the context of mitochondrial disease merits increased attention.
Adjunctive drug therapy is indispensable in periodontitis treatment. Currently, such pharmacotherapy is not ideal, so new formulations need to be explored to optimize prolonged in situ slow release of active ingredients, biocompatibility, and preparation costs. This study constructed an injectable dual drug delivery composite hydrogel system (Gel-Alg/MINO/MEL) to locally deliver minocycline hydrochloride (MINO) and melatonin (MEL) for antibacterial, anti-inflammatory, antioxidant effects and bone formation. We tested the release kinetics of Gel-Alg/MINO/MEL, evaluated its in vitro and in vivo biocompatibility, and detected its in vitro effects on multi-species biofilm, pro-inflammatory cytokines, related genes/proteins and reactive oxygen species, as well as in vivo effects on periodontal inflammation and tissue reconstruction. Release kinetics showed Gel-Alg/MINO/MEL continuously released MINO and MEL for over 10 days with good biocompatibility. In vitro, it inhibited biofilm formation, reduced pro-inflammatory cytokines, increased related genes/proteins expression and scavenged excessive reactive oxygen species; in vivo, it relieved periodontal inflammation and promoted tissue reconstruction and collagen fiber attachment. The Gel-Alg/MINO/MEL hydrogel has promising potential as a local drug delivery system for treating periodontitis, with favorable sustained release and biocompatibility. Regular placement of the Gel-Alg/MINO/MEL composite hydrogel in periodontal pockets, in conjunction with basic clinical periodontal treatment, might remodel the bone and soft tissues lost during periodontitis. Therefore, this is expected to be a more effective medication-assisted treatment modality for refractory periodontitis.
Due to their ecotoxicological potential toward aquatic biota, the escalating occurrence of pharmaceutical residues in aquatic ecosystems constitutes a critical global environmental issue. The research examined variations in the behavioural, haematological, biochemical, oxidative stress and morphometric indices of the African catfish, Clarias gariepinus juveniles, subjected to sub-lethal doses (3.80, 5.99, 11.38, and 22.60 mg/L) of pentoxifylline and dechlorinated carbon-filtered water (control) for 21 days, and a post-treatment duration of 7 days. Toxicity and stress- induced responses including alterations in behaviour, were documented on day 1, 7, 14, last day of exposure (day 21), and at the end of the 7 days outside the drug. Blood sample for the haematological and plasma biochemical indices, and liver for oxidative stress and morphometric indices, were also taken at the same intervals. Rapid opercula movement, excess mucus secretion, reduced feeding, and skin discoloration, increased with concentration, while corkscrew swimming, jumping and swimming rate, declined in the exposed fish. Behavioural toxicity index (BTI) increased significantly in a dose and duration dependent pattern from the fish exposure to pentoxifylline. There was a dose and duration-influenced significant fall in the packed cell volume, haemoglobin, red blood cell count, and mean corpuscular volume, while the opposite was registered for the white blood cells count, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration in the exposed fish. Of the differential leucocyte parameters sampled, values were only significant for the neutrophil and lymphocyte. The neutrophil increased, while the lymphocytes decreased significantly. A significant increase was registered in the aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase activities. Conversely, a reduction was registered in the protein and glucose levels in the exposed fish. A significant reduction in the oxidative stress biomarkers, liver catalase, superoxide dismutase, glutathione peroxidase, and induction in the glutathione and malondialdehyde levels, were reported in the exposed fish, in contrast to the control. The condition factor of the exposed fish, exhibited no significant difference from those of the control. On the contrary, the hepatosomatic index increased significantly, in a manner influenced by the dose of the drug and duration of the treatment, in comparison to the control. The effects of the drug abated after the 7 days- post treatment duration. The findings of this research imply that pentoxifylline is harmful to fish.
The review article is a critical examination of the growing global problem of multidrug-resistant (MDR) Salmonella and Shigella, focusing on their epidemiology, molecular resistance mechanisms, and clinical effects. It compiles current information on increasing resistance to major antibiotics, such as fluoroquinolones, third-generation cephalosporins, and sulfonamides, that is caused by a variety of factors, including ESBL production and plasmid-mediated gene transfer, especially in Asia, Africa, and the Middle East. The minireview discusses the new therapeutic approaches such as bacteriophage therapy, antimicrobial peptides, antimicrobials based on nanotechnology, and a combination of antibiotic regimens and their mechanisms, efficacy, and some limitations that exist in these approaches. In addition, it discusses the issues of diagnostic challenges, infection control practices, and antibiotic stewardship as part and parcel of resistance management. It will be a focused, evidence-based review to inform research, clinical practices, and policy to turn the trend of resistance around and enhance patient outcomes. The presented multidisciplinary review shows that MDR Salmonella and Shigella infections have become a threatening problem in the world, which requires urgent measures, including multilateral cooperation, improved molecular tracking, fast diagnostics, and new therapeutic options.
Continuous distillation is widely used in the commodity and petrochemical industry to purify fine chemicals on massive industrial scales; however, this key separation strategy is largely unused within the pharmaceutical industry. This article investigates the development of a pilot-scale continuous distillation system for in-line removal of an excess amine reactant (i.e., tert-butylamine) when incorporated within an advanced manufacturing technology (AMT) for continuous drug substance production. Initial design parameters were simulated in ASPEN + to assess potential process solvents, setpoint parameters, and azeotrope formation. Complementary analytical characterization was performed with high performance liquid chromatography (HPLC) and gas chromatography - flame ionization detection (GC-FID). Early continuous distillation prototypes and key engineering design decisions are highlighted. The final engineered system is comprised of a + 3-foot distillation column fitted with an Allihn condenser, a 250 mL three-necked round bottom flask, and an automated system for reflux ratio (RD) control. The column is readily integrated within an advanced manufacturing system to continuously remove amine reagent during continuous manufacturing of albuterol sulfate at a 3.0 mL/min process basis, corresponding to a theoretical throughput of approximately 1,800 doses per hour based on the demonstrated process flowrate, reaction stoichiometry, and operating conditions. The application of continuous distillation offers a unique strategy for impurity removal during advanced pharmaceutical manufacturing and can be scaled to meet target throughputs. Here we provide general criteria necessary for successful implementation of similar continuous in-line distillation systems.
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The clinical translation of niclosamide as an anticancer drug remains as a significant challenge mainly due to its poor aqueous solubility. To address this, a dual layered nanocarrier system, PLGA@Na-CMC-CaCO3 was developed in which the presence of PLGA enhanced the encapsulation of niclosamide, Na-CMC improved the bioavailability, and CaCO3 nanoparticles helped in pH-responsive drug release. The fabricated nanocarrier exhibited a uniform size with low polydispersity index when analyzed using DLS. The stepwise incorporation of all the elements along with the drug was confirmed using FTIR and XRD, and the thermal stability of the nanocarrier was assessed using TGA. The PLGA@Na-CMC-CaCO3 nanocarrier system demonstrated a high niclosamide encapsulation efficiency of ∼95.67%. Further a pH-dependent drug release profile was observed, with increased drug release under acidic pH mimicking the tumor microenvironment. The release kinetics followed the first-order and Higuchi models, confirming diffusion-dependent drug release. In vitro biological studies on MDA-MB-231 cells using the alamar blue assay demonstrated negligible cytotoxicity of PLGA@Na-CMC-CaCO3 nanocarriers alone, indicating their biocompatibility. In contrast, niclosamide-loaded PLGA@Na-CMC-CaCO3 nanocarriers showed enhanced dose-dependent cytotoxicity compared to niclosamide alone. The presence of apoptosis in niclosamide-loaded PLGA@Na-CMC-CaCO3 nanocarriers was confirmed by the JC-1 assay, which clearly demonstrated a dose-dependent decrease in mitochondrial membrane potential in the drug encapsulated nanocarrier system. Overall, this PLGA@Na-CMC-CaCO3 nanocarrier system has the potential to effectively address the key limitations of niclosamide delivery for improved cancer therapy.
To evaluate the impact of health literacy on medication adherence of hyperlipidemic outpatients, considering age, gender, perceived health, adverse drug reaction and lipid-lowering treatment. To examine the impact of a modifiable factor-health literacy (HL)-on medication adherence (MA), a cross-sectional observational study was conducted in outpatient lipid clinic (January to June 2025). The exposure of interest was HL, assessed using the Brief Health Literacy Screening Items scale. The primary outcome was MA, measured with the Morisky Medication Adherence Scale. 621completed questionnaires were analysed (age 61.2 ± 13.8 years; male 58.1%). Patients had 8 [2-15] years of lipid lowering therapy, their perceived well-being score was 75.8 ± 15.8 while 40.6% reported side effects due to hypolipidemic medications. Subjects with adequate adherence were older (p = 0.030) and showed a higher level of HL (p < 0.001) while patients with adequate literacy showed a higher level of medication adherence (p < 0.001), reported a greater perceived well-being (p = 0.003) and a lower incidence of side effects (p = 0.003). The network analysis revealed that unintentional MA is directly correlated with age (R = 0.141; p < 0.001) and intentional MA (R = 0.362; p < 0.001). Intentional MA is directly correlated with health numerical literacy (R = 0.227; p < 0.001) and inversely correlated with adverse drug reactions (rho = - 0.143; p < 0.001). The parallel mediation model was built showing the indirect effect of age on MA, mediated by HL (indirect effect1 = - 0.033, 95% CI - 0.060 to - 0.008). Combining the findings from both the network and mediation analyses revealed that HL mediates the relationship between patients' age and MA, with numerical literacy playing a central role. These insights could potentially inform the development of effective health policies.No part of the submitted work has been published or is under consideration for publication elsewhere.
Adaptive therapy is an evolution-based treatment paradigm that has been shown to delay resistance in prostate cancer through treatment breaks that control, rather than minimize, tumor burden. However, patient responses are highly heterogeneous, and there is a significant unmet clinical need for biomarkers to personalize treatment scheduling. To develop and retrospectively validate mathematical biomarkers that predict time to progression (TTP), mean daily dose, and overall survival (OS) under adaptive therapy from first-cycle prostate-specific antigen (PSA) dynamics. This retrospective modeling and validation study used longitudinal, nonrandomized clinical trial data from 2 independent cohorts: 40 patients with castrate-sensitive prostate cancer (CSPC) (June 1996 to September 2006) and 13 patients with metastatic castrate-resistant prostate cancer (mCRPC) (April 2015 to January 2022). A 2-population differential equation model was used to describe the overall tumor growth through the competing dynamics of drug-sensitive and drug-resistant cells. The statistical analysis was conducted from January 2025 to May 2026. Patients received either intermittent androgen deprivation therapy (for CSPC) or adaptive abiraterone acetate (for mCRPC). The initial treatment cycle served as the exposure period to extract longitudinal PSA kinetics. Mechanism-based mathematical biomarkers (adaptive therapy score, expected TTP, and expected mean daily dose) were derived from first-cycle PSA kinetics. Outcomes included in silico benchmarking experiments and retrospective validation against clinical TTP and OS. Performance was benchmarked against standard phenomenological PSA metrics (eg, PSA nadir, time to nadir, and doubling time). Overall, data from 53 patients across 2 clinical trials were included. In the CSPC cohort of 40 patients, the adaptive therapy score derived from first-cycle data was highly prognostic for prolonged clinical TTP (univariable hazard ratio [HR], 0.49; 95% CI, 0.31-0.76; P = .002). In the mCRPC cohort of 13 patients, the adaptive therapy score exhibited a strong rank correlation with clinical TTP (Spearman ρ = 0.76; P = .002) and was associated with prolonged TTP (HR, 0.41; 95% CI, 0.16-1.07; P = .07). Analysis of long-term survival data in the mCRPC cohort demonstrated that both the adaptive therapy score and expected TTP were significantly associated with prolonged OS, whereas standard empirical PSA metrics displayed no association with OS. In this modeling and validation study, mechanism-based mathematical biomarkers derived from the initial-cycle PSA dynamics accurately predicted patient-specific outcomes and survival, outperforming traditional phenomenological PSA monitoring. These accessible metrics could act as a mathematically informed decision support framework to stratify patients into personalized treatment protocols.
Staphylococcus pseudintermedius is a common opportunistic pathogen in dogs and an increasing concern in veterinary medicine due to rising antimicrobial resistance, particularly to methicillin. This study aimed to characterize resistance profiles and genetic mechanisms in isolates from healthy and diseased dogs in Montevideo, Uruguay. A total of 133 isolates was analyzed (83 from clinical infections and 50 from asymptomatic carriers). Antimicrobial susceptibility was assessed by disk diffusion following veterinary guidelines. Resistance genes and SCCmec types were detected by PCR. Ten representative isolates underwent whole genome sequencing. High resistance rates were observed for penicillin (81%), erythromycin (49.6%), and clindamycin (45%). Overall, 48.9% of isolates were multidrug-resistant. Phenotypic resistance to oxacillin was detected in 26% of isolates; however, 23% carried mecA gene and were therefore classified as genotypic MRSP. These isolates were more frequent among dogs with clinical infections. These strains showed higher resistance to all antimicrobials tested. SCCmec type V was the most prevalent, and greater genetic diversity was found among isolates from symptomatic dogs. Genomic analysis revealed circulating strains of unassigned sequence types (STs), a variety of resistance genes within specific lineages, the circulation of SCCmec XIV cassette carrying strains, and an Oxacillin-susceptible Methicillin-resistant Staphylococcus pseudintermedius (OS-MRSP) isolate. These findings demonstrate the clinical and epidemiological relevance of S. pseudintermedius in Uruguay and the role of asymptomatic dogs as reservoirs of resistant strains. The results emphasize the need for surveillance, prudent antimicrobial use, and integrated control strategies within a One Health framework.
Bruceantin (BCT), a natural product with promising anticancer properties, has limited clinical utility due to its hydrophobicity, low systemic bioavailability, and dose-limiting toxicity. Here, we report a redox-responsive polymeric nanoplatform based on a disulfide-linked diblock copolymer, poly(ethylene glycol)-block-poly(ε-caprolactone) (PEG-SS-PCL), functionalized with luteinizing hormone-releasing hormone (LHRH) peptides for the targeted delivery of BCT to pancreatic cancer cells overexpressing LHRH receptors. The nanoplatform exhibits favorable physicochemical characteristics, including optimal particle size (45.7 nm), zeta potential (+3.5 mV), and high drug encapsulation efficiency (74.8%). Notably, it demonstrates redox-responsive drug release under glutathione-rich conditions that mimic the tumor microenvironment. In vitro assays highlight the superior cytotoxicity of the LHRH-targeted BCT formulation (IC50, 224.1 μg mL-1) compared to the non-targeted formulation (IC50, 279.5 μg mL-1). Biodistribution studies in an orthotopic pancreatic cancer murine model revealed substantial accumulation of LHRH-targeted nanoparticles primarily in tumors following systemic administration, with minimal hepatic deposition. Furthermore, BCT-loaded nanoplatforms reduced tumor growth by ∼72.4% relative to controls, with no detectable adverse effects. Collectively, this work highlights the potential of LHRH-functionalized nanoplatforms as a promising strategy for the targeted treatment of pancreatic cancer with BCT, offering enhanced therapeutic efficacy with minimal systemic toxicity and paving the way for future clinical translation of this natural product.
Despite universal access to combined antiretroviral therapy in Brazil, substantial inequalities persist in the HIV/AIDS epidemic. This nationwide retrospective study analyzed 753,071 notifications from the Brazilian Notifiable Diseases Information System (SINAN) between 2014 and 2024 to characterize the epidemiological profile, temporal trends, spatial distribution, factors associated with AIDS-stage notification, and predictors of AIDS-related mortality. HIV infection notifications accounted for 491,976 records, while AIDS case notifications accounted for 261,095. HIV infection notifications were concentrated among young adults, particularly men aged 20-29 years, whereas AIDS case notifications were more frequent in older age groups. Spatial analysis showed significant positive autocorrelation of smoothed municipal notification rates (Moran's I = 0.2306; p < 0.001), with significant High-High clusters mainly in the South and Southeast regions. In the multivariable logistic regression model, older age was strongly associated with AIDS-stage notification, reaching the highest odds among individuals aged 60 years or older (AOR = 2.03; 95% CI: 1.96-2.09), while higher education was associated with lower odds. In the Cox model, age at diagnosis was the main predictor of mortality, with increased risk among individuals aged 30-49 years (AHR = 1.42; 95% CI: 1.36-1.49) and 50 years or older (AHR = 1.95; 95% CI: 1.86-2.05). Lower education and injecting drug use were also strongly associated with AIDS-related mortality. These findings indicate that, although AIDS case notifications have declined over time, AIDS-stage notification and mortality remain concentrated in socially and geographically vulnerable groups.
Human induced pluripotent stem cells (hiPSCs) are essential tools for disease modeling, drug testing, and regenerative medicine. These applications require differentiation of multiple cell lines in parallel, synchronized production of large numbers of hiPSCs and differentiated cells, and assessment of differentiation efficiency before proceeding to in-depth characterization through molecular and cellular assays. However, hiPSC culture protocols are often laborious manual processes, which affects reproducibility and makes high-throughput applications challenging. We describe high-throughput hiPSC maintenance and amplification protocols that are weekend-free, scalable, and can be performed manually or automated. These optimized protocols enable scale-up of hiPSC production and differentiation to multiple cell types, including endothelial cells, microglia, and retinal pigment epithelial cells. We characterized the differentiated cells using molecular and cellular assays and demonstrated that cells generated with this approach represent accurate cell identities. The approaches described here enable high-throughput hiPSC applications and improve reproducibility and scalability.If you want, I can also tighten this further to better fit a strict 350-word journal limit or adapt the heading style to a specific journal.
The management of carbapenem-resistant Acinetobacter baumannii (CRAB) infections remains a formidable clinical challenge. This study evaluated the in vitro antimicrobial activities of sulbactam-durlobactam (SUL-DUR) and eravacycline (ERV) against CRAB isolates and elucidated the genomic landscapes of resistance and virulence determinants in SUL-DUR-resistant strains to inform therapeutic decision-making. A total of 233 clinical CRAB isolates were collected and screened for susceptibility to SUL-DUR and ERV using the Kirby-Bauer (K-B) disk diffusion assay. Isolates exhibiting resistance to SUL-DUR were further characterized via metagenomic next-generation sequencing (mNGS) to identify key resistance and virulence factors. SUL-DUR and ERV demonstrated robust in vitro activity, with susceptibility rates of 92.3% and 91.4%, respectively. Notably, no isolates exhibited concurrent non-susceptibility to both agents. Genomic analysis of 14 SUL-DUR-resistant strains revealed a complex and heterogeneous distribution of genetic determinants. The presence of bla NDM-1 was identified as a critical driver of SUL-DUR resistance. Additionally, reduced susceptibility was potentially associated with specific mutations in bla OXA-23, bla OXA-66, and bla TEM-1, while hyperactive efflux systems and altered membrane permeability further synergized to enhance the resistance phenotype. Despite the extensive-drug-resistant (XDR) nature of current CRAB isolates, they maintain high sensitivity to SUL-DUR and ERV. Our findings underscore that SUL-DUR and ERV represent highly promising therapeutic options with significant development potential and broad clinical application prospects for the management of CRAB-related infections.
Pharmacist-led interventions significantly improved denosumab adherence in patients with osteoporosis at an academic medical center, reducing non-adherence from 20.8 to 4.9%. This quality improvement initiative included pharmacist-led medication counseling and laboratory monitoring, highlighting the effectiveness of pharmacist-physician collaboration in preventing rebound-associated vertebral fractures and enhancing patient outcomes. Interrupting or delaying denosumab beyond 7 months (210 days) from the last injection leads to transient increase in bone turnover known as rebound phenomenon, resulting in rapid loss of bone density and increased risk of vertebral fracture(s), now known as rebound-associated vertebral fractures (RAVF). Numerous studies suggest that clinical pharmacist participation in chronic disease state management leads to an increase in medication adherence and positive clinical outcomes. We evaluated the impact of a clinical pharmacist intervention on adherence to optimal denosumab timing for patients with osteoporosis. The model for improvement was used to design and evaluate this quality improvement project. This study analyzed 143 patients receiving denosumab therapy in an osteoporosis clinic every 6 months for the indication of osteoporosis. At baseline, 21% of patients receiving denosumab did not receive the medication at optimal time intervals for clinical effectiveness. An administration was considered correct or on time if denosumab was given within 210 days of the last injection received. If the injection was given outside this window, it was considered incorrect administration. The incidence of correct and incorrect administrations was gathered to calculate the non-adherence rate. Clinical pharmacist interventions included patient counseling and electronic dosing reminders and phone calls. The aim of this study was to reduce the nonadherence rate from 20.8% to less than 5.0% by July 31, 2023. With these interventions, the number of correct administrations between incorrect administrations improved from an average of 3.3 to 13.1. These effects became visible about 10 weeks after the second intervention and correspond to a reduction in nonadherence rate from 20.8 to 4.9% by July 31, 2023. Pharmacist-physician collaboration can reduce the nonadherence rate for drugs such as denosumab, interruption of which can result in serious consequences.
Cervical cancer remains a leading cause of cancer related mortality in women worldwide. Cisplatin (CP) is a cornerstone chemotherapeutic agent; however, its clinical utility is constrained by dose-limiting toxicity and the emergence of drug resistance, necessitating novel combinatorial strategies. This study investigated the therapeutic potential of combining CP with Aloe vera extract (AVE) against HeLa cervical cancer cells and the normal human skin fibroblast cell line CCD-1072Sk. Cytotoxicity assays performed using the xCELLigence Real-Time Cell Analysis (RTCA) system determined IC50 values of 18.45 µM for CP and 18.88 µg/mL for AVE in HeLa cells, compared with 53.31 µM and 19.09 µg/mL, respectively, in CCD-1072Sk cells. The CP + AVE combination exhibited markedly enhanced antiproliferative activity in HeLa cells compared with CP alone, while CCD-1072Sk cells exhibited continued but decelerated proliferation, suggesting a differential cellular response. Seahorse metabolic analysis revealed that AVE, both alone and in combination with CP, markedly increased mitochondrial oxygen consumption rate (OCR) and ATP-linked respiration in HeLa cells, indicating induction of sustained bioenergetic stress beyond the cells' adaptive capacity. Transcriptomic profiling of HeLa cells identified 453 differentially expressed genes (DEGs) upon combination treatment, with significant downregulation of oxidative phosphorylation (OXPHOS) components, mitochondrial electron transport chain subunits, and the fatty acid metabolism gene CPT1B, collectively indicating a severe disruption of cellular energy metabolism. DepMap database analysis confirmed that downregulated targets including CPT1B and CHN2 are functionally important for HeLa cell survival. Collectively, these data suggest that AVE drives HeLa cells into a hypermetabolic state that overloads mitochondrial capacity; in combination with CP, this converging bioenergetic stress produces enhanced cytotoxicity descriptively consistent with an additive interaction (a term used here in its dose-response sense, as no formal combination-index or Bliss/Loewe synergy analysis was performed) at the IC50 doses employed. This study highlights the potential of AVE as a metabolically active combinatorial agent capable of potentiating CP efficacy by targeting cancer cell energy metabolism, and provides a mechanistic basis for further investigation.
The KEYNOTE-689 trial demonstrated improved event-free survival with the addition of perioperative pembrolizumab to standard of care for patients with locally advanced head and neck squamous cell carcinoma (HNSCC), yet the cost-effectiveness remains unknown. To evaluate the cost-effectiveness of the KEYNOTE-689 perioperative pembrolizumab protocol. A partitioned survival model simulated the KEYNOTE-689 trial cohort with 3 mutually exclusive health states: event-free survival, disease progression, and death. Model inputs included Healthcare Cost and Utilization Project-derived and US Centers for Medicare & Medicaid Services-derived costs for drug acquisition (pembrolizumab and cisplatin), administration, surgery, radiation therapy, supportive care, management of serious adverse events, and disease progression. KEYNOTE-689 Kaplan-Meier survival estimates were digitized and data for the combined positive score (CPS) of 1 to 10 subgroup were derived from the reported CPS of 1 or greater and CPS of 10 or greater subgroups. Quality-adjusted life-year (QALY) values for different health states were sourced from published literature, with the US population preference-weighting algorithm applied to EuroQol 5-Dimension 3-Level health questionnaire data from the CheckMate 141 trial. The primary outcome was the incremental cost-effectiveness ratio (ICER) results stratified by programmed cell death 1 ligand 1 (PD-L1) combined positive score (CPS of 1 to 10 vs greater than 10). A probabilistic sensitivity analysis (PSA) was performed to evaluate parameter uncertainty. Subgroup analyses based on PD-L1 expression yielded divergent results. In the CPS greater than 10 cohort, perioperative pembrolizumab was cost-effective, producing approximately 1.35 additional QALYs at an incremental cost of $181 900, resulting in an ICER of $134 700. PSA showed a 57% probability that the pembrolizumab strategy was cost-effective at a $150 000 willingness-to-pay (WTP) threshold. In the CPS of 1 to 10 cohort, perioperative pembrolizumab resulted in approximately 0.08 additional QALYs at an incremental cost of $166 500, for an ICER of $2 179 400, with the PSA demonstrating an 11% probability of cost-effectiveness at a $150 000 WTP threshold. In this economic evaluation, addition of perioperative pembrolizumab to standard treatment for locally advanced HNSCC may represent a cost-effective intervention for patients with a PD-L1 CPS greater than 10 if KEYNOTE-689 data are replicable in clinical practice. Perioperative pembrolizumab is not a cost-effective strategy for patients with a CPS less than 10 based on current data. Biomarker stratification may inform the value-based integration of immunotherapy among patients with HNSCC.
Granulomatous amoebic encephalitis (GAE) is a rare but highly fatal central nervous system infection caused primarily by Acanthamoeba spp. and Balamuthia mandrillaris. Delayed diagnosis and the absence of standardized treatment protocols contribute to mortality exceeding 90%. This review summarizes current knowledge regarding GAE pathogenesis, diagnosis, therapeutic challenges, and emerging treatment strategies. The pathogenesis involves amoebic adhesion, secretion of proteases and phospholipases, host inflammatory responses, and blood-brain barrier disruption. Recent advances in molecular diagnostics improve early pathogen detection, while drug repurposing, nanotechnology-based delivery systems, and cyst-targeted approaches represent promising therapeutic strategies. Nevertheless, treatment remains challenging because of poor blood-brain barrier penetration, cyst-associated resistance, and limited clinical evidence. Continued integration of molecular diagnostics, mechanistic studies, translational research, and multicentre clinical investigations is essential for improving outcomes in this devastating disease.
Antibiotic resistance of Gram-positive bacteria poses a major clinical challenge. Daptomycin, a lipopeptide antibiotic, is a treatment option for drug-resistant organisms. However, there have been clinical reports of daptomycin resistance and in vitro reports of daptomycin tolerance that can result in treatment failure. Daptomycin possesses a fatty acid tail that inserts into the membrane, and membrane fatty acid content can influence its binding. Given that, we examined the potential of fatty acids to enhance daptomycin activity against enterococcal strains. We demonstrate that the saturated fatty acids myristic and palmitic acids increase bacterial susceptibility to the antibiotic, even for a daptomycin-resistant clinical isolate. These effects were highly specific with no impact on daptomycin susceptibility of Staphylococcus aureus. Growth of enterococci with myristic or palmitic acid reduced membrane fluidity, potentially promoting tighter daptomycin insertion. However, we noted no difference in membrane permeability for cells treated with fatty acids and daptomycin. While cells exposed to saturated fatty acids experience greater membrane depolarization, the lack of consistent, significant time-dependent changes upon daptomycin treatment suggests that depolarization alone does not fully explain the increased sensitivity. A strain that is unable to incorporate exogenous fatty acids into phospholipids was insensitive to the effects of saturated fatty acids on its susceptibility to daptomycin, highlighting the necessity of fatty acid incorporation onto lipid headgroups for potentiation. Overall, our data suggest that the incorporation of specific saturated fatty acids within the enterococcal membrane likely disrupts lipid composition, membrane organization and cellular envelope homeostasis, ultimately contributing to enhanced daptomycin susceptibility.
Pyogenic spondylitis is treated with high-dose antibiotics for at least six weeks. In this study, we aimed to develop a novel therapeutic agent that can enhance bacterial targeting. We used solid lipid nanoparticles (SLNs) containing the antibiotic ampicillin (AMP), which were functionalized with the positively charged peptide LL-37. AMP-loaded SLNs (AMP-SLNs) were prepared, and LL-37 was attached to the AMP-SLNs (LL-37@AMP-SLNs). The physicochemical properties of the nanoparticles were characterized by particle size, zeta potential, morphology, drug release behavior, and hemocompatibility analyses. Furthermore, antibacterial activity, bacterial-targeting capability, and therapeutic efficacy were evaluated using Enterococcus faecalis-infected mesenchymal stem cells and a rat model of pyogenic spondylitis. The antibacterial performance of LL-37@AMP-SLNs was compared with that of SLNs and AMP-SLNs. The average size of the SLNs was approximately 342.66 ± 48.04 nm, which increased to 528.58 ± 35.64 nm after AMP loading and 695.58 ± 75.58 nm following LL-37 modification. The LL-37@AMP-SLNs exhibited sustained AMP release over four days and demonstrated enhanced antibacterial activity and bacterial-targeting capability compared with unmodified SLNs and AMP-SLNs. LL-37@AMP-SLNs were shown to inhibit bacterial growth by targeting the cytoplasmic membrane and the cell walls of Gram-positive bacteria. Moreover, the LL-37@AMP-SLNs treatment led to significantly enhanced antibacterial effects. In vitro studies showed significantly greater inhibition of bacterial growth, while intravenous administration of LL-37@AMP-SLNs significantly reduced bacterial burden and improved therapeutic outcomes in the pyogenic spondylitis rat model compared with free AMP treatment. We suggest that LL-37@AMP-SLNs can be a useful therapeutic agent for targeting bacteria in pyogenic spondylitis.