Search Results (3,568)

Background: PEG-Asparaginase and Erwinia asparaginase are enzyme-based anticancer therapies used in the treatment of acute lymphoblastic leukaemia (ALL) and lymphoblastic lymphoma (LBL), where adequate plasma enzyme activity is required for therapeutic efficacy. In many study groups, therapeutic drug monitoring is routinely applied due to pharmacokinetic variability and the risk of hypersensitivity reactions followed by increased clearance and insufficient treatment. In clinical practice, samples may be exposed to prolonged transport and variable pre-analytical conditions. Knowledge on pre-analytical stability is important for correct interpretation of PEG-Asparaginase and Erwinia asparaginase activity in plasma. This study aimed to evaluate the in vitro stability of PEG-Asparaginase and Erwinia asparaginase under pre-analytical conditions. Methods: Three experimental stability studies were conducted at two activity levels. Enzyme stability in plasma was assessed during storage at 4 °C and 20 °C for up to 14 days and following three freeze–thaw cycles. Stability in whole blood prior to centrifugation was evaluated over 24 h. Enzyme activity was measured using a validated spectrophotometric assay, and stability was defined as a deviation within ±15% of baseline activity. Results: Both enzymes remained stable in plasma for up to 14 days at 4 °C and 20 °C, and no clinically relevant reduction in enzyme activity of freeze–thaw cycling was observed. In whole blood, Erwinia asparaginase and high-activity PEG-Asparaginase remained stable for 24 h at 20 °C, whereas low-activity PEG-Asparaginase showed a reduction in activity of approximately 22%, mainly within the first two hours. Conclusions: PEG-Asparaginase and Erwinia asparaginase are stable in plasma for up to 14 days at room temperature, enabling shipment of plasma samples by mail. However, prompt centrifugation is recommended for samples with low PEG-Asparaginase activity to ensure accurate therapeutic drug monitoring. Full article

Growth hormone (GH) and insulin-like growth factor 1 (IGF-1) play essential roles in the brain, influencing neuronal and dendritic growth, as well as neurotransmission. These effects persist throughout life. Numerous studies in animals and humans have demonstrated the beneficial effects of GH therapy on memory and cognitive function, as well as on the restoration of neuronal function following injury. All nerve cells, including neurons, glia, endothelial, epithelial, and perivascular cells, are affected by the actions of GH/IGF-1. IGF-1, in particular, has been associated with cognitive function. The GH-IGF-1 axis increases the proliferation of neuronal progenitor cells and the formation of new neurons, oligodendrocytes, and astrocytes. In this study, we searched databases such as PubMed, Google Scholar, and Embase for human clinical trials evaluating the effect of growth hormone (GH) therapy on dementia, Alzheimer’s disease (AD), post-traumatic brain injury (PTI), and stroke. The following search terms were used: “GH and dementia,” “GH and Alzheimer’s disease,” “GH and TBI,” and “GH and stroke.” Inclusion criteria were all randomized controlled trials and observational studies. Exclusion criteria included the lack of cognitive and memory assessments. We found 28 articles. Most studies show the beneficial effects of GH therapy on memory and recovery of brain function after traumatic injury and stroke; however, consistent data are still lacking. The limited number of clinical trials, the small number of patients, and the lack of data on plasma levels of sex hormones that clearly contribute to brain function are limiting factors. This is the case, for example, with androgens. Other critical factors are dosage and treatment duration. Prolonged administration and supraphysiological doses are more effective in inducing positive clinical changes. Growth hormone (GH) therapy is a very promising intervention for preventing and treating dementia and early-stage Alzheimer’s disease, and it contributes significantly to the recovery of brain function in patients after traumatic injury and stroke. Further studies with more robust methodologies are needed to confirm these results. Full article

This article discusses rheumatoid arthritis (RA) as a chronic, systemic autoimmune disease leading to progressive joint damage and multi-organ complications. The complex pathogenesis of the disease is presented, involving the interaction of environmental, genetic, and immunological factors, including the role of autoantibodies and proinflammatory cytokines. Particular attention is paid to leflunomide, a disease-modifying antirheumatic drug (DMARD), which primarily works by inhibiting the DHODH enzyme, leading to reduced T and B cell proliferation. The additional anti-inflammatory properties of the drug’s active metabolite, teriflunomide, and its impact on signaling pathways related to the immune response are also discussed. This article examines the variability in patient responses to leflunomide treatment in terms of both efficacy and toxicity, with particular emphasis on the potential role of pharmacogenetic factors. It was pointed out that polymorphisms in genes related to drug metabolism, transport, and mechanism of action may influence the pharmacokinetics and safety of the therapy. It was also emphasized that the available data are primarily derived from observational studies and small cohorts, and the results are often inconsistent. Although some genetic variants and plasma teriflunomide concentrations show potential as predictors of treatment response, the current level of evidence does not support the routine use of pharmacogenetic testing in clinical practice. The article emphasizes that the pharmacogenetics of leflunomide represents a promising, yet still exploratory, avenue of research in the context of personalized RA therapy. It emphasizes the need for larger, well-designed clinical trials and the development of standardized guidelines, which would be necessary before the potential implementation of such strategies in routine clinical practice. Full article

Viral infections of the central nervous system produce memory impairment through mechanisms that extend beyond acute neuronal injury. Herpes simplex virus type 1, human immunodeficiency virus, varicella zoster virus, cytomegalovirus, Epstein–Barr virus, influenza, SARS-CoV-2, West Nile virus, and Zika virus each enter or engage the brain through distinct routes, yet converge on four shared molecular pathways that selectively damage hippocampal circuits: mitochondria-associated membrane (MAM) dysfunction, chronic neuroinflammation, blood–brain barrier (BBB) disruption, and impaired CREB-BDNF signaling. These pathways specifically compromise the dentate gyrus, CA3, and CA1 subfields, producing predictable deficits in pattern separation, associative retrieval, and temporal memory binding. Antiretroviral and antiviral therapies suppress viral replication but fail to reverse organelle-level dysfunction, leaving most hippocampal injury unaddressed. Emerging plasma biomarkers, p-tau217, neurofilament light chain, and GFAP, combined with hippocampal subfield MRI, now enable mechanistic stratification before irreversible circuit loss occurs. This review proposes, as a unifying hypothesis, that virus-associated memory impairment represents a convergent hippocampal syndrome driven by shared downstream pathways, and that combination therapies targeting these pathways simultaneously offer greater therapeutic promise than pathogen-specific approaches alone. The evidentiary basis for this framework varies across pathogens and conditions; direct mechanistic evidence, mechanistic analogy, and preclinical data are distinguished throughout. Full article

Tumor cell heterogeneity and multicellular interactions critically influence drug resistance, recurrence, and prognosis. Here, CPcellsubpopulation, a computational framework integrating scRNA-seq, bulk RNA-seq, and clinical data was developed to identify cancer progression-associated cell subpopulations. Then, the integrated analyses of scRNA-seq and spatial transcriptomics were performed to predict potential interactions, identify critical transcription factors, and predict candidate anticancer drugs. Across nine cancers, we detected cancer progression-associated cell subpopulations significantly linked to prognosis, with consistent patterns across cancer types. In renal cell carcinoma (RCC), we identified conserved metabolic^high UBE2C+ cancer cells linked to poor outcomes, metabolic reprogramming and low differentiation, and PLK1+ NK cells, plasma cells, and CDC20+ macrophages associated with advanced stages and unfavorable prognosis. Spatial mapping revealed spatial association of RCC progression-associated cancer and immune cell subpopulations, suggesting the potential role of the VEGF, GDF, PTN and IL16 pathways in the remodeling of the tumor microenvironment. Gene regulatory network analysis highlighted RAD21 as a key regulator linking metabolism and therapy resistance. This study provides a systematic pipeline to delineate cancer progression-associated cell subpopulations, uncovers metabolic^high UBE2C+ cancer cells as progression-associated tumor cell population, and nominates critical regulators and compounds as therapeutic targets. Full article

Background/Objectives: Boron neutron capture therapy (BNCT) relies on the selective delivery of boron-10 to tumor cells. Although 4-[¹⁰B]borono-L-phenylalanine (BPA) is currently the only clinically approved BNCT agent, it is limited by poor L-type amino acid transporter 1 (LAT1)/LAT2 selectivity and aqueous solubility. We previously developed 3-borono-5-fluoro-α-methyl-L-phenylalanine (5F-αMe-3BPA), a novel BPA derivative designed to be a LAT1-targeted BNCT/positron emission tomography theranostic agent. This study comprehensively characterizes its pharmacological profile and explores its pharmacokinetic optimization by modulating renal organic anion transporter 1 (OAT1). Methods: Transport kinetics of BPA, related analogs, and 5F-αMe-3BPA were analyzed in HEK293 cells stably expressing LAT1 or LAT2 using Michaelis–Menten analysis. Time-dependent cellular uptake and intracellular retention of BPA and 5F-αMe-3BPA were evaluated in T3M-4 pancreatic cancer cells with or without the LAT1 inhibitor JPH203. In vivo biodistribution was examined in T3M-4 tumor-bearing mice after intravenous administration of 5F-αMe-3BPA or BPA, with assessment of probenecid pretreatment. Results: 5F-αMe-3BPA retained LAT1 affinity comparable to that of BPA while showing markedly reduced LAT2-mediated transport, indicating improved LAT1/LAT2 selectivity. In T3M-4 cells, 5F-αMe-3BPA showed stronger LAT1 dependence, higher steady-state accumulation, and better intracellular retention than BPA under amino acid-containing conditions. Although 5F-αMe-3BPA achieved favorable tumor-to-plasma and tumor-to-muscle ratios in vivo, it was rapidly cleared from circulation. Probenecid pretreatment increased plasma exposure, reduced early renal accumulation, and significantly enhanced tumor boron accumulation, reaching approximately twofold higher levels than control. Conclusions: These findings establish 5F-αMe-3BPA as a highly LAT1-selective BNCT candidate and identify probenecid pretreatment as a clinically translatable pharmacokinetic strategy for maximizing therapeutic boron delivery. Full article

Oxidative stress and inflammation are key drivers of kidney injury and disease progression. In this context, the role of sialic acids emerged as a critical regulatory layer linking redox imbalance, immune activation, and tissue damage. Sialic acids are terminal negatively charged residues that regulate complement activity, immune cell signaling, and the structural integrity of the glomerular filtration barrier. Alterations in sialylation, resulting from impaired biosynthesis or increased sialidase activity, disrupt immune homeostasis, enhance inflammatory responses, and promote complement-mediated injury. In the kidney, these mechanisms contribute to podocyte dysfunction, glomerular inflammation, and fibrosis and are implicated in glomerulopathies, transplantation, and plasma cell dyscrasias. Emerging evidence also highlights the therapeutic potential of targeting sialic acid metabolism through inhibition of desialylation or restoration of sialylation pathways. Overall, sialic acids represent dynamic modulators at the intersection of oxidative stress and immunity, offering novel opportunities for biomarker development and mechanism-based therapies in kidney disease. Full article

Polydeoxyribonucleotide (PDRN) activates the adenosine A2A receptor (A2AR), triggering anti-inflammatory signaling and providing essential nucleotides for the salvage pathway, thereby helping bypass metabolic bottlenecks and promoting tissue repair. Combining PDRN with biochemical agents and physical stimuli represents a significant shift in medical treatment, moving from monotherapy to an integrated, multi-target regenerative approach. These combinatorial strategies effectively address the limitations of PDRN, such as its rapid degradation and diffusion, by simultaneously meeting the structural, metabolic, and signaling needs of injured tissues. The mechanism of action for PDRN involves a synergistic effect with hyaluronic acid, amplification of growth factors (e.g., Platelet-Rich Plasma (PRP), Epidermal Growth Factor (EGF), Platelet-Derived Growth Factor (PDGF)), and enhancements from extracorporeal shockwave therapy (ESWT) and lasers. This results in a notable acceleration of the repair process for chronic wounds, musculoskeletal disorders, and neurological injuries. As intelligent delivery systems like responsive hydrogels and sustainable L-PDRN production continue to advance, these synergistic protocols are poised to redefine global standards of care in regenerative medicine and esthetic dermatology. Future clinical success will hinge on the standardization of sequence-specific protocols and large-scale validation to ensure long-term safety and efficacy. Full article

Background: Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-Kp) central nervous system (CNS) infections represent a major therapeutic challenge in neurosurgical patients. Intraventricular or intrathecal polymyxin B sulfate (PMB) is commonly used as salvage therapy but is limited by substantial neurotoxicity. Ceftazidime–avibactam (CZA) exhibits potent in vitro activity against KPC-Kp; however, prospective clinical and pharmacokinetic evidence supporting its use in CNS infections remains limited. Methods: In this prospective, single-centre observational study, adult neurosurgical patients with culture-confirmed KPC-Kp CNS infections admitted to the neurointensive care unit of Huashan Hospital were enrolled. Patients received either intravenous CZA (CZA group, n = 15) or intrathecal/intraventricular PMB-based therapy (PMB group, n = 10). Primary outcomes included clinical cure, microbiological eradication, 28-day mortality, and safety. Therapeutic drug monitoring was performed to determine steady-state plasma and cerebrospinal fluid (CSF) concentrations of ceftazidime, avibactam, and polymyxin B, enabling assessment of CSF penetration and exposure–toxicity relationships. Results: Overall clinical cure and microbiological eradication rates were 68.0% and 84.0%, respectively, with a 28-day mortality of 20.0%. Compared with PMB, CZA was associated with a significantly higher clinical cure rate (86.7% vs. 40.0%, p = 0.024) and a numerically higher eradication rate (93.3% vs. 70.0%). No neurological adverse events occurred in the CZA group, whereas neurological toxicity was observed in 60.0% of PMB-treated patients (p < 0.001). Functional outcomes favoured the CZA group, with lower modified Rankin Scale scores at discharge and at 6 months. Pharmacokinetic analyses demonstrated that steady-state CSF concentrations of ceftazidime and avibactam exceeded commonly accepted pharmacodynamic targets, while markedly elevated PMB CSF concentrations were observed in patients with neurological toxicity. Conclusions: While intravenous CZA showed potentially favourable efficacy and safety compared with local PMB in this cohort, these preliminary findings should be interpreted as hypothesis-generating given the small sample size and non-randomised design. These results provide a rationale for further validation in larger multicentre, randomised controlled trials. Full article

Background: Venous thromboembolism (VTE) remains a leading cause of global morbidity and mortality. The pathophysiological mechanisms leading to VTE are still not fully elucidated. Diagnostic biomarkers for VTE lack specificity, and biomarkers to guide the duration of anticoagulation therapy have not yet entered routine clinical practice. Proteomics has emerged as a high-throughput approach for novel biomarker discovery, offering insights into the complex biological processes across the VTE continuum. This review explores current VTE proteomic research and discusses the challenges and gaps hindering clinical translation. Methods: We performed a narrative review based on a search of Scopus and PubMed for original human proteomic studies published between 1995 and July 2025. Results were limited to whole-blood, plasma or serum-based studies. Results: A total of 1190 studies were retrieved, of which 27 studies were included in this review. Studies were mainly plasma-based. Studies compared VTE patients to non-VTE controls, different VTE subtypes, and various provoked VTE cohorts. Broad themes identified proteomic signatures involving dysregulated coagulation, complement activation, inflammation, and platelet activation. Conclusions: Current proteomic evidence supports VTE as a systemic immunothrombotic disorder that shows key differences even years before developing VTE. Proteomic research in VTE holds the promise to identify biomarkers that may aid in the diagnosis and guide management of VTE. However, most proteomic findings remain exploratory to date and methodologies are varied across studies. Future studies should prioritise the workflow standardisation and validate promising biomarker panels in large-scale, prospective, longitudinal cohorts. Full article

Background/Objectives: Acute myeloid leukemia (AML) is a hematological malignancy frequently driven by mutations in the FLT3 gene, particularly internal tandem duplications (FLT3-ITD), which contribute to aberrant cell proliferation and resistance to tyrosine kinase inhibitors (FLT3i). The limitations of current FLT3i therapies, including drug resistance, off-target effects, and poor selectivity, necessitate the development of novel therapeutic strategies. Proteolysis-targeting chimeras (PROTACs) represent a promising approach to achieving degradation of oncogenic proteins. Methods: We developed FLT3-targeting PROTACs based on the previously described compound MA49, with a focus on linker modifications to improve degradation efficiency and pharmacokinetic properties. Results: Among these, compounds MA190 and MA191, containing rigid cyclohexyl-piperidine/piperazine linkers, demonstrate superior degradation of FLT3-ITD in MV4-11 AML cells at nanomolar concentrations, achieving >95% reduction in FLT3-ITD levels, outperforming MA49. In addition to improved kinase selectivity, good solubility, and plasma stability, MA190 and MA191 also exhibit excellent metabolic stability, whereas the predecessor PROTAC MA49 was unstable in microsomal assays. In cellular assays, MA190 and MA191 induce potent apoptosis in FLT3-ITD⁺ AML cells but have minimal effects on cells with wild-type FLT3. Proteomics reveal that MA191 also degrades MAPK14 (p38α), a kinase upregulated in leukemia, in addition to FLT3. Conclusions: Dual targeting of FLT3-ITD and MAPK14 enhances proapoptotic signaling without any cytotoxic effect on normal human HEK293 cells. The co-inhibition using MA191 or a combination of doramapimod (a MAPK14 inhibitor) with a non-degrading FLT3 inhibitor result in greater caspase-3 activation than either treatment alone. This synergistic effect can be a therapeutic advantage, as several oncogenic drivers are switched off simultaneously by MA191. Full article

Background: Leptospirosis is a globally prevalent zoonosis with significant morbidity and mortality, especially in tropical regions like South Asia. In its severe form, the disease often leads to multiorgan dysfunction, with pulmonary haemorrhage being a major cause of death. Evidence supporting specific treatments for severe leptospirosis with pulmonary involvement remains limited. Recent studies suggest that immunomodulatory therapies, such as therapeutic plasma exchange (TPE), may offer survival benefits. This case series explores the application and outcomes of TPE in patients with severe leptospirosis at a tertiary care hospital in Sri Lanka. Methods: We studied a case series involving nine patients with confirmed severe leptospirosis and multiorgan involvement from September 2021 to October 2022. All patients received standard care, including intravenous antibiotics and methylprednisolone. TPE was initiated in all nine patients based on clinical severity, particularly in the presence of pulmonary haemorrhage. Clinical, laboratory, and radiological data were collected from patient records and follow-up. Leptospirosis diagnosis was confirmed through ELISA IgM testing. TPE decisions were made by a multidisciplinary team. Results: Of the nine patients who received TPE, seven survived (78%). Pulmonary haemorrhage was the primary indication for TPE in all cases. All patients had multiorgan involvement: renal failure (89%), hepatic dysfunction (55%), and myocarditis (67%). Mortality was associated with inotropic-dependent myocarditis and mechanical ventilation at TPE initiation. Patients requiring intubation had a 50% mortality rate, compared to 14% in those who were not intubated. Non-survivors also had significantly elevated lactate levels (>4 mmol/L) and worsening acid–base status. Four patients required dialysis: three survived. Conclusions: Early initiation of TPE may be safe and beneficial in severe leptospirosis, and future randomised controlled studies are necessary to examine its benefits further. These findings are hypothesis-generating for further research, particularly on patient selection for TPE. Full article

Chimeric antigen receptor (CAR) T-cell therapy represents a major advance in modern immunotherapy. This narrative review summarizes evidence from the past five years, including case reports, case series, and clinical trials, on its application beyond hematologic malignancies, focusing on autoimmune diseases such as systemic lupus erythematosus (SLE), systemic sclerosis (SSc), as well as solid tumors including melanoma and primary cutaneous lymphomas. CD19-directed CAR T-cells have demonstrated clinical benefits in SLE and SSc, with sustained immune reset, reduced autoreactive antibody levels, and clinical improvement. In melanoma, CAR T-cells targeting GD2, cMET, and CD20 have shown in vivo expansion and tumor infiltration; however, clinical efficacy remains limited, with transient stabilization or disease progression in most patients. In primary cutaneous lymphomas, early-phase studies with anti-CD70 and anti-CCR4.30 CAR T-cells indicate partial tumor regression and disease stabilization, often requiring additional therapy. Key challenges include limited durability of immune reset due to persistent plasma cells in autoimmune disorders, tumor heterogeneity, antigen loss or overlap, infiltration barriers, resistance mechanisms, and T-cell depletion in solid tumors, collectively reducing response durability and safety. The main toxicities include grade 1–2 cytokine release syndrome and rare hematologic complications, while immune effector cell-associated neurotoxicity syndrome is uncommon. Clinical translation remains limited and requires larger studies to improve efficacy and define safety profiles. Full article

Chronic obstructive pulmonary disease (COPD) is characterized by persistent airflow limitation, chronic airway inflammation, and immune dysregulation, and currently available therapies remain insufficient to effectively halt disease progression. In this study, we used an integrative, hypothesis-generating strategy to investigate the potential mechanisms of baicalein against COPD by combining multi-dataset transcriptomic analysis, single-cell transcriptomics, machine learning-based feature selection, Mendelian randomization (MR), molecular simulation, virtual knockout analysis, and in vitro validation. Putative targets of baicalein were predicted using CTD, SEA, and SwissTargetPrediction, and were intersected with COPD-related genes collected from GeneCards and OMIM. Four GEO datasets (GSE20257, GSE42057, GSE76925, and GSE130928) were integrated after batch-effect correction, yielding a combined cohort of 260 control samples and 250 COPD samples. Candidate genes were prioritized by intersecting the results of LASSO regression, random forest, and support vector machine. Immune-cell infiltration was estimated using CIBERSORT, and single-cell transcriptomic data were used to define the cellular localization of prioritized genes. Formal protein-level MR analysis was conducted for CD163 using deCODE plasma protein pQTL/GWAS summary statistics as the exposure dataset and the IEU OpenGWAS COPD dataset (ebi-a-GCST90018807) as the outcome dataset. Molecular docking, molecular dynamics simulation, and virtual knockout analysis were further used to provide structural and network-level supportive evidence. Finally, LPS-stimulated BEAS-2B cells were used as an epithelial inflammatory model to evaluate the effects of baicalein by CCK-8 assay, wound-healing assay, ELISA, and RT-qPCR. Five core genes were prioritized, namely ABCC1, CD163, CYP1B1, IKBKB, and PIK3CA. Immune infiltration and single-cell analyses suggested that macrophage-associated immune regulation may represent an important mechanistic direction. MR analysis provided supportive genetic evidence for prioritizing CD163 in COPD. Molecular simulation offered preliminary structural support for several target-compound interactions. In LPS-stimulated BEAS-2B cells, baicalein reduced inflammatory cytokine release and modulated the expression of IKBKB, PIK3CA, IL1B, IL6, and IL10, thereby providing epithelial-level support for the predicted network. Taken together, these findings suggest that baicalein may exert anti-inflammatory effects in COPD through a multi-target, immune-associated mechanism, with macrophage-related regulation and CD163 emerging as noteworthy candidate directions for further investigation. This study provides an integrative framework for target prioritization and mechanistic exploration, while the predicted macrophage-centered mechanisms still require dedicated validation in immune-cell and in vivo models. Full article

Background/Objectives: Cadherin-13 (CDH13), part of the cadherin family, is attached to the plasma membrane through glycosylphosphatidylinositol. CDH13 plays essential roles in the development of the neurological and vascular systems and is a risk factor for neural and cardiovascular diseases. CDH13 is expressed on the plasma membrane in both mature and uncleaved precursor forms with the prodomain. Although several anti-CDH13 monoclonal antibodies (mAbs) are available for basic research, there have been no reports of anti-CDH13 mAbs that can detect both the mature form and the uncleaved precursor in flow cytometry. Methods: We developed novel anti-human CDH13 mAbs (named Ca₁₃Mabs) using the mature form of CDH13-expressed cells as an antigen. Results: Among Ca₁₃Mabs, a clone, Ca₁₃Mab-17 (IgG_2b, κ) specifically recognized the mature and uncleaved precursor CDH13-overexpressed Chinese hamster ovary-K1 (CHO/CDH13) cells with no detectable cross-reactivity toward 21 other cadherins by flow cytometry. Ca₁₃Mab-17 also detected endogenous CDH13 in human glioblastoma (LN229 and U87MG) and lung mesothelioma (NCI-H2052) cell lines. The dissociation constant (K_D) value of Ca₁₃Mab-17 for LN229 was estimated at 4.1 × 10⁻⁸ M. Furthermore, Ca₁₃Mab-17 detected both the mature and uncleaved precursor CDH13 in Western blotting. It also identified new blood vessels and glioblastoma cells by immunohistochemistry. Conclusions: Ca₁₃Mab-17 is a versatile tool for detecting both mature and uncleaved precursor forms of CDH13 and has potential for tumor diagnosis and therapy. Full article