Search Results (404)

This study evaluated clinical characteristics, treatment outcomes, and CNS relapse patterns in patients with breast-involved diffuse large B-cell lymphoma (DLBCL), a rare extranodal presentation with limited real-world data. We conducted a retrospective study on 33 consecutive patients with newly diagnosed breast-involved DLBCL treated from 2019 to 2024. All patients received R-CHOP. Baseline CNS screening—including neurological examination, fundoscopy, brain magnetic resonance imaging (MRI), and cerebrospinal fluid (CSF) analysis—was routinely performed. High-dose methotrexate (HD-MTX) was offered as CNS prophylaxis after completion of systemic therapy based on multidisciplinary team evaluation and clinician–patient shared decision-making according to institutional treatment protocols. Median age was 52.6 years; 84.8% had ECOG 0. Non-GCB subtype predominated (84.8%), and Ki-67 >70% was present in 69.7%. The overall response rate was 90.9%, with 84.8% complete responses. At a median follow-up of 44 months, 5-year Overall Survival (OS) and Progression-Free Survival (PFS) were 84.8% and 66.7%. CNS relapse occurred in 4 of 6 patients (66.7%) without prophylaxis, all within 5–11 months after R-CHOP, whereas no CNS relapses were observed among prophylaxis recipients (p < 0.001), although this observation should be interpreted with extreme caution given the very small non-prophylaxis subgroup (n = 6), limited statistical power, and non-randomized treatment allocation. Exploratory analyses suggested that bulky disease was associated with inferior OS. R-CHOP achieved high response rates and favorable long-term outcomes in breast-involved DLBCL. The absence of CNS relapse among HD-MTX prophylaxis recipients, contrasted with a high relapse rate in those without prophylaxis, provides only a hypothesis-generating observation that requires confirmation in larger prospective studies; this warrants further investigation of the role of systemic CNS prophylaxis. Full article

Kaposi’s Sarcoma Herpesvirus (KSHV) is the etiological agent of Kaposi’s Sarcoma (KS), which induces metabolic stress in infected host cells. KSHV reprograms host metabolic pathways for efficient viral replication and infectious virion production. Here, we report a time-course global metabolomics study conducted in iSLK.BAC16 cells to compare latent and lytic KSHV infection. Our data show that amino acid, central carbon, and nucleotide metabolic pathways are highly dysregulated upon reactivation. During lytic KSHV infection, pathway enrichment analysis identifies purine and pyrimidine metabolism as the top two most significantly impacted and dysregulated pathways. Further experiments have shown that nucleotide metabolism is required during lytic KSHV infection to produce maximal infectious virus. Treatment with the FDA-approved drug, methotrexate (MTX), a folate antagonist that decreases nucleotide metabolism by reducing tetrahydrofolate cofactors, significantly reduced KSHV copy number and late lytic viral gene expression upon reactivation compared to controls. Additionally, titers of cell-free supernatants from MTX-treated lytic samples showed a significant reduction in infectious virion production. Furthermore, MTX significantly decreased the viral titer of murine herpesvirus 68 (MHV-68), a model virus to study gammaherpesvirus. Overall, our study demonstrates that metabolic inhibition during lytic gammaherpesvirus infection decreases productive infection and hence serves as a potential therapeutic antiviral target. Full article

Nanoparticle-laden contact lenses (CLs) represent a promising strategy for ocular drug delivery. Nanocarriers built from hyaluronic acid and chitosan offer a biomimetic alternative to synthetic polymers, by combining the lubricating property of hyaluronic acid with the mucoadhesive property of chitosan. However, achieving sustained drug release without compromising lens properties remains challenging, and the influence of lens material and replacement modality is unclear. This study evaluated methotrexate (MTX)-loaded chitosan/hyaluronic acid (CS/HA) nanoparticles incorporated into silicone hydrogel CLs with different replacement modalities, assessing their effect on drug release kinetics. Daily replacement lenses (Senofilcon A and Delefilcon A) released 26 ± 4% and 33 ± 5% of MTX after 24 h, respectively. The monthly lens Lehfilcon A showed slower diffusion-controlled release, with only 15 ± 2% released at 24 h and a 64 ± 3% cumulative release after 31 days (p < 0.01). Nanoparticle incorporation improved drug retention and reduced initial drug loss compared with lenses loaded with only MTX. Monthly lenses demonstrated sustained delivery potential, supporting prolonged ocular therapy, while daily lenses may be better suited for short-term treatment. These findings reinforce the value of bioinspired nanocarriers, mimicking natural retention mechanisms of the ocular surface, for next-generation therapeutic CLs. Full article

Liver involvement is frequent in rheumatoid arthritis (RA) and may progress from steatosis to fibrosis, cirrhosis, or hepatocellular carcinoma. Liver fibrosis (LF) in RA is multifactorial, but the available data are limited and the impact of methotrexate (MTX) remains controversial. In this cross-sectional study, 51 RA patients (46 females; mean age of 48.8 ± 8.2 years; and a median disease duration of 12 years) were enrolled. LF was assessed using non-invasive indices (the aspartate aminotransferase-to-platelet ratio index [APRI] and fibrosis-4 index [FIB-4]) and liver stiffness measurement (LSM) using shear wave elastography. Serum endothelin-1 (ET-1) and selected organokines (namely myostatin, resistin, and osteoprotegerin) were quantified by the ELISA. Associations of the APRI, FIB-4, and LSM with organokines and endothelin-1 were analyzed using univariable and multivariable linear regression, whereas correlations with the RA-specific cardiovascular (CV) risk score (ERS-RA) and echocardiographic parameters of left ventricular diastolic dysfunction (LVDD) were assessed using Spearman’s rank correlation. Myostatin and resistin showed a significant positive and significant negative association with LSM, respectively, while FIB-4 was negatively correlated with lateral and medial e’ velocities and positively with the ERS-RA. MTX use, mean weekly dose, cumulative dose, and endothelin-1 were not associated with the APRI, FIB-4, LSM, or LVDD. These exploratory findings support the potential role of myostatin and resistin in LF-related phenotypes and suggest that FIB-4 may capture aspects of both hepatic and cardiovascular risk in RA. RA patients with elevated FIB-4 values may require echocardiographic assessment and comprehensive CV risk evaluation. In this cohort, MTX exposure and endothelin-1 were not associated with LF or LVDD. Full article

Background and Objectives: Pulmonary fibrosis (PF) is a life-threatening respiratory disorder involving complex pathophysiological mechanisms such as inflammation, collagen deposition, and epithelial cell injury. Methotrexate (MTX), a chemotherapeutic and immunosuppressive agent, is frequently associated with pulmonary toxicity, representing a significant adverse effect with unpredictable outcomes. Thus, the current study investigates the possible protective effect of dulaglutide (DUL) against MTX-induced lung injury. Methods: Sixty male Wistar rats were allocated into six groups: group 1 served as a control group, group 2 received MTX (14 mg/kg/week, p.o) for 2 weeks, group 3 was treated with MTX treatment (14 mg/kg/week, p.o) prior to dexamethasone (DEXA) (0.5 mg/kg/week i.p), Groups 4–5 were treated concurrently with MTX (14 mg/kg/week, p.o) and DUL at doses of 0.05 and 0.1 mg/kg/week, s.c. and Group 6 received DUL (0.1 mg/kg/week, s.c.) only. Animals were sacrificed on day 15 for histopathological and biochemical assessments. Results: Our data demonstrated that MTX markedly increased some oxidative stress markers, inflammatory biomarkers and fibrotic and apoptotic indicators, alongside a significant decrease in Beclin-1 and Adenosine Monophosphate Activated Protein Kinase (AMPK) levels. DUL administration significantly ameliorated these alternations in a dose-dependent manner, with histopathological findings corroborating biochemical data through Hematoxylin & Eosin (H&E) and Mason’s Trichrome (MTC) staining. Conclusions: DUL confers significant protection against MTX-induced pulmonary fibrosis, likely through its antioxidant, anti-inflammatory, and anti-apoptotic properties, which are associated with the restoration of total AMPK and Beclin-1 levels. This highlights its therapeutic potential in preventing drug-induced lung toxicity. Full article

Background: B-cell acute lymphoblastic leukemia (B-ALL) is the most frequent malignancy of childhood; worldwide, 487,294 new cases and 305,405 deaths were reported in 2022, including more than 5000 new cases in Mexico. Chemotherapy, delivered in induction, consolidation and maintenance phases, remains the mainstay of treatment, yet 10–20% of patients relapse after induction because of chemoresistance, whose molecular basis is still incompletely understood. Methods: This retrospective study aimed to identify proteins associated with resistance to vincristine (VCR) and methotrexate (MTX) administered during the induction phase, using LC–MS/MS proteomics and bioinformatic analysis of treatment-naive bone marrow aspirates from responders and nonresponders. Results: Nonresponders showed a distinct proteomic profile, with deregulated processes converging on cytoskeletal structure and dynamics, nucleic acid metabolism, DNA repair and RNA processing. Within these processes, thymidine phosphorylase (TYMP) and gelsolin (GSN) emerged as differentially expressed proteins, both consistently overexpressed in nonresponders at the individual-patient level. Conclusions: We conclude that cytoskeletal remodeling and nucleotide metabolism are prominent features of intrinsic chemoresistance in pediatric B-ALL, and that TYMP and GSN represent candidate biomarkers of nonresponse to VCR- and MTX-based induction that warrant validation by orthogonal methods in independent, adequately powered cohorts. Full article

In this study, a pH-responsive chondroitin sulfate–methotrexate (MTX) polymeric prodrug was synthesized through Schiff base formation between oxidized chondroitin sulfate and MTX. The resulting amphiphilic conjugate exhibited a conjugation degree of 184 mg MTX per g conjugate and spontaneously self-assembled into stable nanoparticles (CSMXPs) with a mean diameter of 120 ± 10 nm, a polydispersity index of 0.24, and a critical aggregation concentration of 4.7 × 10⁻⁴ mg mL⁻¹. Drug release studies demonstrated a marked pH-dependent behavior, with complete MTX release after 24 h at pH 5.0 and a sustained release profile under physiological conditions. The release mechanism followed reversible first-order kinetics and was accelerated by acid-catalyzed hydrolysis of the imine linkage. Biological evaluation revealed enhanced therapeutic selectivity of CSMXPs compared with free MTX. At 36 μM MTX-equivalent concentration, CSMXPs reduced HeLa cell viability to 37%, while maintaining MCF-10A viability above 88%, whereas free MTX decreased viability in both cell lines (51% and 65%, respectively). Fluorescence confocal microscopy confirmed efficient nanoparticle uptake by cancer cells. These findings demonstrate that CSMXPs represent a promising self-assembling nanoprodrug platform for selective and targeted cancer therapy. Full article

Chronic inflammatory barrier diseases (CIBDs) affect many women of reproductive age, yet the safety of biologics during pregnancy remains inadequately investigated. This retrospective cohort study used the US Collaborative Network of TriNetX to evaluate the risk of adverse pregnancy outcomes (APOs) among women with CIBD receiving tumor necrosis factor inhibitors (TNFis), interleukin-23 inhibitors (IL-23is), or conventional synthetic disease-modifying anti-rheumatic drugs (csDMARDs), compared with untreated CIBD controls and the general pregnant population. Among 1842 pregnant women with CIBD receiving systemic therapy, 1188 were exposed to TNFis, 170 to IL-23is, 370 to azathioprine (csDMARD), and 114 to methotrexate (MTX) (csDMARD). The incidence of any APO ranged from 11% in untreated CIBD controls to 19% with ustekinumab (IL-12/23i). Among treatment groups, rates were 17% with TNFis, 18% with IL-23is, 13% with azathioprine, and 15% with MTX, compared with 17% in the general pregnant population. APO rates were comparable between the overall TNFi group and the TNFi group excluding certolizumab pegol (CZP) (TNFi). In the only non-exploratory comparison, TNFi exposure was associated with higher risks of (pre-)eclampsia and hypertension but a lower risk of abortion or intrauterine death versus CIBD controls, with no significant difference for overall APO. No treatment group showed a markedly increased overall APO risk, supporting tailored decision-making that balances the consequences of uncontrolled maternal disease against individual treatment-associated risks. Full article

In this work, we develop a coupled multiphysics model that integrates polymer carriers exhibiting time-dependent thixotropic structural recovery with Darcy flow, linear Biot poroelasticity and advection–diffusion transport in a spherically symmetric, isotropic and homogeneous tissue domain. The formulation explicitly links rheological evolution to pressure-driven flow, interstitial deformation and solute transport through a unified framework, enabling systematic prediction of post-injection behavior. Unlike conventional approaches that assume constant carrier properties, the present model incorporates a time-dependent viscosity evolution, capturing the transition from an initially shear-thinned state to a recovered, highly viscous structure. Numerical simulations using hydroxypropyl methylcellulose and methotrexate parameters as representative components demonstrate that rapid post-injection viscosity recovery suppresses pressure-driven transport and diffusion, thereby enhancing local drug retention near the injection site. A systematic sensitivity analysis identifies the equilibrium viscosity as the dominant parameter controlling spatial localization, whereas tissue mechanical properties exert a comparatively minor influence. An effectiveness metric based on the Kullback–Leibler divergence reveals a tumor-size-dependent trade-off between spatial coverage and retention. The proposed framework thus introduces a predictive tool for analyzing coupled rheological-transport interactions and for the rational design and optimization of thixotropy-enhanced local chemotherapy strategies. Full article

Dried droplet imaging has become an established approach for analyzing complex fluid systems as evaporation patterns preserve physicochemical and structural information. However, spatial variability of these patterns limits the effectiveness of conventional texture descriptors. This work presents a region-aware deep learning algorithm for the classification of dried droplets through spatial decomposition and convolutional learning. Dried droplet images were geometrically standardized prior to patch extraction. The Hough transform supported consistent crown–core localization and patch extraction across the dataset. Each image was divided into twenty localized regions. Twelve represented peripheral patches sampled at 30° intervals. The remaining eight patches were extracted from four central regions using two complementary angular orientations (30° and 90°) per region. This process allowed dataset augmentation and evaluation of regional contributions. Patches were processed using the VGG16 convolutional neural network, following repeated patch-level and droplet-level partitioning strategies designed to prevent data leakage. The proposed algorithm was evaluated on Methotrexate (MTX) droplets prepared with 40%, 60%, and 80% water dilution levels relative to a reference solution. Results show that the proposed strategy increases classification accuracy, outperforming traditional descriptors. These findings demonstrate that spatial decomposition combined with convolutional learning constitutes an effective approach for heterogeneous evaporative pattern classification. Full article

Methotrexate (MTX) is widely used in the treatment of malignancies and inflammatory disorders, but nephrotoxicity remains a major adverse effect. Naringenin (NAR), a natural flavonoid, has antioxidant, anti-inflammatory, and nephroprotective properties. This study investigated the potential protective effects of NAR against MTX-induced nephrotoxicity at biochemical, molecular, and histopathological levels. Forty-two adult male Wistar albino rats were assigned to seven groups (n = 6): Control, CMC (carboxymethyl cellulose), NAR100, MTX, and MTX combined with NAR (25, 50, or 100 mg/kg/day). NAR was administered for 7 days, with MTX given on day 3. Renal function, histopathology, and genes associated with oxidative stress, apoptosis, endoplasmic reticulum stress, and inflammation were evaluated. MTX administration caused marked renal damage, increased creatinine and BUN levels, elevated apoptosis-, inflammation-, and ER stress-related gene expression, and suppressed antioxidant defense-related genes. However, 50 and 100 mg/kg/day NAR attenuated these alterations, with greater effects at 100 mg/kg/day. Histopathological damage was attenuated by NAR treatment, although complete recovery was not observed. These findings suggest that NAR may protect against MTX-induced nephrotoxicity through the modulation of pathways associated with oxidative stress, inflammation, apoptosis, and ER stress. However, the persistence of certain histopathological alterations indicates that structural recovery of renal tissue may take a longer period compared with molecular changes. Full article

Background/Objectives: Rheumatoid arthritis (RA) has been associated with accelerated atherosclerosis and cerebrovascular alterations. Our 2017 study compared 60 RA patients to healthy controls, assessing vascular, neurological, and cognitive parameters. The present study is a follow-up of these RA patients to evaluate disease progression and vascular changes over time, using their 2017 results as baseline. Methods: In 2023, we reassessed 43 of the original 60 RA patients using laboratory testing, carotid ultrasound, functional transcranial Doppler (TCD) and brain magnetic resonance imaging (MRI) examinations. Changes over time were analyzed within the same individuals. Results: Inflammatory markers and lipid profiles showed a trend toward improvement, though changes were not statistically significant, except for a significant increase in vitamin D (p < 0.001) and a decrease in Disease Activity Score in 28 Joints (DAS28) scores (p < 0.001). Carotid ultrasound revealed a significant increase in plaque burden (p = 0.022 on the right side and p = 0.008 on the left), while carotid intima media thickness (cIMT) showed a non-significant rise. TCD measurements indicated significantly increased pulsatility (p < 0.001 on the right, p = 0.001 on the left side) and resistance (p = 0.001 on the right, p = 0.012 on the left side) indices and reduced flow velocities (p < 0.001 on the right and p = 0.001 on the left side) in bilateral middle cerebral arteries (MCAs). The cerebrovascular reserve capacity was significantly lower on the right side overall (p = 0.013), with further decline noted in the methotrexate (MTX)-treated subgroup on the left side (p = 0.043). MRI findings showed non-significant numerical trends toward worsening lacunar small-vessel disease (p = 0.405) and cerebral atrophy (p = 0.063), with higher but stable lacunar infarction scores among MTX users (p = 0.023). Conclusions: Despite improved inflammatory control, RA patients demonstrated progressive vascular and hemodynamic alterations over time, while MRI changes should be interpreted as trends. These findings support multimodal vascular monitoring in RA. Full article

The ICAM-1-derived cIBR peptide selectively binds to the I-domain of LFA-1, a receptor highly expressed on leukemia T cells; thus, the MTX-cIBR conjugate can be used to target methotrexate (MTX) to leukemic T cells and reduce its off-target toxicity. However, the uptake, biological mechanism, and selectivity of MTX-cIBR compared with unconjugated MTX remain unclear. Therefore, this study is aimed at evaluating the uptake, cytotoxicity, selectivity, apoptosis, cell cycle effects, and DHFR-related activity of MTX-cIBR in leukemia T cells compared with unconjugated MTX. MTX-cIBR exhibited cytotoxic activity comparable to MTX in LFA-1-expressing MOLT-4 cells but showed lower toxicity toward LFA-1-negative K562 cells, indicating improved selectivity. MTX uptake occurred through RFC and mFBP transport systems, whereas MTX-cIBR no longer depended on these pathways, suggesting altered cellular uptake after conjugation with cIBR by utilizing the LFA-1 receptor. Both compounds predominantly induced apoptosis with minimal necrotic cell populations. MTX induced S-phase arrest at lower concentrations and G2/M induced arrest at higher concentrations, whereas MTX-cIBR consistently promoted S-phase accumulation. In addition, MTX and MTX-cIBR downregulated the expression of DHFR, FPGS, and TYMS in MOLT-4 cells. Computational analyses further demonstrated that MTX exhibited lower binding free energy (ΔG) and greater binding stability toward DHFR than MTX-cIBR. These findings suggest that MTX-cIBR retains selective cytotoxic activity toward LFA-1-expressing leukemia T cells through altered cellular uptake and exhibits different interaction characteristics with DHFR compared with unconjugated MTX. Full article

Background: Methotrexate-induced oxidative stress is mechanistically linked not only to hepatocellular injury but also to DNA damage, indicating that oxidative stress, hepatotoxicity, and genotoxicity represent interconnected manifestations of the same antifolate-driven toxic cascade. Methotrexate (MTX)-induced hepatotoxicity is characterized not only by oxidative stress, but also by progressive fibrotic remodeling driven by activation of the TGF-β/SMAD signaling pathway. Objective: We aimed to examine the hepatoprotective effects of Loureirin B, with a particular focus on its anti-fibrotic potential and underlying molecular mechanisms in MTX-induced liver injury. Methods: Thirty female Wistar rats were assigned to normal control, MTX, and MTX + Loureirin B groups. Liver injury was induced with a single intraperitoneal MTX dose (20 mg/kg), followed by oral administration of Loureirin B (50 mg/kg/day) for 10 days. Biochemical, molecular, and histopathological analyses were performed, including ALT, AST, ALP, MDA, SIRT1, TGF-β, SMAD3, hydroxyproline, and VEGF levels, alongside the evaluation of necrosis, fibrosis, and inflammatory infiltration. Results: MTX induced significant hepatic injury characterized by elevated serum ALT, AST, and ALP levels, increased oxidative stress, suppression of SIRT1, and increased TGF-β and SMAD3 levels, accompanied by elevated collagen-associated markers. Loureirin B treatment significantly reduced the serum liver enzyme levels and oxidative stress, partially restored SIRT1 levels, and decreased fibrosis-associated markers, including hydroxyproline and VEGF. Although the TGF-β levels were significantly reduced following Loureirin B treatment, the reduction in SMAD3 levels did not remain statistically significant after correction for multiple comparisons. Histopathological findings further demonstrated attenuation of fibrosis-associated changes and partial improvement in hepatic architecture. Conclusions: Loureirin B may exert protective effects against methotrexate-associated liver injury through the modulation of oxidative stress, partial restoration of SIRT1 levels, attenuation of profibrotic alterations associated with the TGF-β/SMAD pathway, and modulation of VEGF-related responses. Full article

Background/Objectives: Superparamagnetic iron-oxide nanoparticles (SPIONs) bearing poly(methacrylate) brushes were synthesized via surface-initiated atom-transfer radical polymerization (SI-ATRP) as magnetically responsive nanoplatforms. Three brush architectures, poly(2-hydroxyethyl methacrylate) (PHEMA) and poly(poly(ethylene glycol) methacrylate) with six ethylene-oxide units (PPEGMA6) and ten units (PPEGMA10), were grown from a dopamine-anchored initiator and covalently loaded with methotrexate (MTX). Methods: Physicochemical characterization confirmed successful polymer grafting, tunable hydrodynamic size (185–1320 nm before MTX conjugation and 427–694 nm after), retained superparamagnetic properties (22–69 emu g⁻¹), and high drug payloads, with PPEGMA6 achieving 131 µg mg⁻¹. MTX conjugation induced partial compaction of the polymer shell yet maintained ζ-potentials conducive to colloidal stability. Results: In vitro assays showed negligible toxicity toward primary human fibroblasts, whereas MTX-decorated formulations induced a pronounced concentration-dependent cytotoxic effect in MCF-7 breast cancer cells, reaching 69% loss of viability—significantly higher than free MTX. Structure–activity analysis attributes the superior performance of PPEGMA6-MTX to its balanced brush density, high payload, and favorable surface charge. Conclusions: These findings demonstrate that precise modulation of polymer brush architecture via SI-ATRP yields SPION-based nanocarriers that integrate MRI visibility and the potential for magnetic guidance and targeted chemotherapy. The PPEGMA6-MTX construct is highlighted as a promising platform for future preclinical investigations. Full article