Search Results (708)

The autoantibodies against the NR1 subunit are well known in the pathomechanism of NMDAR encephalitis. The dysfunction of the NR2 subunit could be a critical factor in this neurological disorder due to its important role in the postsynaptic pathways that direct synaptic plasticity. We report a case of paraneoplastic anti-NMDAR encephalitis presented alongside very severe illness. Computed tomography (CT) of the brain, as well as FLAIR and T2-weighted MRI, was performed to rule out any other acute brain processes. A semi-quantitative method was applied to detect the presence of anti-NMDAR antibodies in the serum and CSF. A CT chest–abdomen–pelvis scan was performed that detected an ovarian teratoma. A histopathological examination was performed after a laparoscopic right-ovary cystectomy. Subsequent immunofluorescence immunohistochemical staining showed the expression of NMDA receptors of type NR2B. Treatment included first-line immunotherapy, second-line immunotherapy, tumor removal, and intrathecal injections with methotrexate and dexamethasone. The histological finding for our patient after tumor removal was ovarian teratoma. Hematoxylin–eosin (HE) staining revealed a characteristic spectrum of elements, including stratified squamous epithelium and fat tissue accompanied by neuroglial cells. Subsequent immunohistochemical staining showed an expression of NMDA receptors of type NR2B in different structures of the teratoma, including the neuroglial cells. The first-line immunotherapy following the tumor removal was insufficient in our patient. The paraneoplastic anti-NMDAR encephalitis with a coexpressed NR2B subunit on the neural cells of the ovarian teratoma may suggest a different inflammation process and could be the key factor in the pathomechanism and treatment of the refractory anti-NMDAR encephalitis. Full article

Methotrexate is an antimetabolic agent with proliferative and immunosuppressive activity. It has been demonstrated to be an effective treatment for acute lymphoblastic leukemia (ALL) in children. However, there is evidence of an association between methotrexate and toxicity risks, which influences the personalization of treatment, particularly in the case of childhood ALL. This article presents the development and implementation of an algorithm based on artificial neural networks to detect methotrexate toxicity in pediatric patients with acute lymphoblastic leukemia. The algorithm utilizes historical clinical and laboratory data, with an effectiveness of 99% in the tests performed with the patient dataset. The use of neural networks in medicine is often linked to disease diagnosis systems. However, neural networks are not only capable of recognizing examples but also hold very important information. For this reason, one of the main areas of application of neural networks is the interpretation of medical data. In this article, we diagnose, with the application of neural networks in medicine, a concrete example: detecting methotrexate in its early stages in pediatric patients. Full article

Bacterial and eukaryotic dihydrofolate reductase (DHFR) enzymes are essential for DNA synthesis and are differentially sensitive to the competitive inhibitors trimethoprim and methotrexate. Unexpectedly, trimethoprim did not reduce Wolbachia abundance, and the wStri DHFR homolog contained amino acid substitutions associated with trimethoprim resistance in E. coli. A phylogenetic tree showed good association of DHFR protein sequences with supergroup A and B assignments. In contrast, DHFR is not encoded by wFol (supergroup E) and wBm (supergroup D) or by genomes of the closely related genera Anaplasma, Ehrlichia, Neorickettsia, and possibly Orientia. In E. coli and humans, DHFR participates in a coupled reactions with the conventional thymidylate synthase (TS) encoded by thyA to produce the dTMP required for DNA synthesis. In contrast, Wolbachia and other Rickettsiales express the unconventional FAD-TS enzyme encoded by thyX, even when folA is present. The exclusive use of FAD-TS suggests that Wolbachia DHFR provides a supplementary rather than an essential function for de novo synthesis of dTMP, possibly reflecting the relative availability of, and competing demands for, FAD and NAD coenzymes in the diverse intracellular environments of its hosts. Whether encoded by thyA or thyX, TS produces dTMP by transferring a methyl group from methylene tetrahydrofolate to dUMP. In the Rickettsiales, serine hydroxymethyltransferase (SMHT), encoded by a conserved glyA gene, regenerates methylene tetrahydrofolate. Unlike thyA, thyX lacks a human counterpart and thus provides a potential target for the treatment of infections caused by pathogenic members of the Rickettsiales. Full article

Background: Lichen planus (LP) is a chronic inflammatory disease that can present with significant morbidity, particularly in children. Erosive lichen planus (ELP), its rare destructive subtype, can be particularly difficult to diagnose and manage. We present a rare pediatric case of ELP with multisite involvement and discuss the differential diagnosis. Case Presentation: A 12-year-old boy presented with painful erosions and ulcers on the lateral tongue and dystrophic nails. His six-year history of tongue and nail lesions prompted several comprehensive examinations. Laboratory tests did not reveal any abnormalities. Histopathological examination of the tongue lesions was representative of ELP. Line-field confocal optical coherence tomography (LC-OCT) examination of the tongue lesions showed features that strongly correlated with histopathology. The patient was later hospitalized due to dysphagia and esophageal food impaction, during which esophageal ELP was confirmed. The patient was initially managed with topical corticosteroids. He was later started on systemic therapy in the form of methotrexate and low-dose naltrexone to address his symptoms and disease presentation. Conclusions: This case highlights the complexities of diagnosis and management of ELP in pediatric patients. A multidisciplinary approach and regular follow-up are necessary to manage symptoms, prevent complications, and improve quality of life. Full article

Background/Objectives: Pteridine reductase 1 (PTR1) has been one of the prime targets for discovering novel antileishmanial therapeutics in the fight against Leishmaniasis. This enzyme catalyzes the NADPH-dependent reduction of pterins to their tetrahydro forms. While chemotherapy remains the primary treatment, its effectiveness is constrained by drug resistance, unfavorable side effects, and substantial associated costs. Methods: This study addresses the urgent need for novel, cost-effective drugs by employing in silico techniques to identify potential lead compounds targeting the PTR1 enzyme. A library of 1463 natural compounds from AfroDb and NANPDB, prefiltered based on Lipinski’s rules, was used to screen against the LmPTR1 target. The X-ray structure of LmPTR1 complexed with NADP and dihydrobiopterin (Protein Data Bank ID: 1E92) was identified to contain the critical residues Arg17, Leu18, Ser111, Phe113, Pro224, Gly225, Ser227, Leu229, and Val230 including the triad of residues Asp181-Tyr194-Lys198, which are critical for the catalytic process involving the reduction of dihydrofolate to tetrahydrofolate. Results: The docking yielded 155 compounds meeting the stringent criteria of −8.9 kcal/mol instead of the widely used −7.0 kcal/mol. These compounds demonstrated binding affinities comparable to the known inhibitors; methotrexate (−9.5 kcal/mol), jatrorrhizine (−9.0 kcal/mol), pyrimethamine (−7.3 kcal/mol), hardwickiic acid (−8.1 kcal/mol), and columbamine (−8.6 kcal/mol). Protein–ligand interactions and molecular dynamics (MD) simulation revealed favorable hydrophobic and hydrogen bonding with critical residues, such as Lys198, Arg17, Ser111, Tyr194, Asp181, and Gly225. Crucial to the drug development, the compounds were physiochemically and pharmacologically profiled, narrowing the selection to eight compounds, excluding those with potential toxicities. The five selected compounds ZINC000095486253, ZINC000095486221, ZINC000095486249, 8alpha-hydroxy-13-epi-pimar-16-en-6,18-olide, and pachycladin D were predicted to be antiprotozoal (Leishmania) with Pa values of 0.642, 0.297, 0.543, 0.431, and 0.350, respectively. Conclusions: This study identified five lead compounds that showed substantial binding affinity against LmPTR1 as well as critical residue interactions. A 100 ns MD combined with molecular mechanics Poisson–Boltzmann surface area (MM/PBSA) calculations confirmed the robust binding interactions and provided insights into the dynamics and stability of the protein–ligand complexes. Full article

Background: Incomptine A (IA) has been reported to have cytotoxic activity in non-Hodgkin lymphoma cancer cell lines and have effects on U-937 cells, including the induction of apoptosis, the production of reactive oxygen species, and the inhibition of glycolytic enzymes. Also, IA has cytotoxic activity in the triple-negative subtypes, HER2+, and luminal A of breast cancer cells, with its properties being associated with an effect on the antiapoptotic function of Hexokinase II (HKII). Objectives: In this research, we reviewed the altered levels of proteins present in the lymph nodes of male Balb/c mice inoculated with U-937 cells and treated with IA or methotrexate, as well as mice only inoculated with cancer cells. Methods: Five approaches, including Tandem Mass Tag (TMT), Gene ontology (GO), Reactome, KEGG pathway analysis, and molecular docking, were used. Results: TMT showed that 74 proteins were differentially expressed, out of which 12 presented overexpression (FC ≥ 1.5) and 62 were under expressed (FC ≤ 0.67). In general, the TMT approach showed that IA had a better effect on proteins than methotrexate. Gene ontology, Reactome, and KEGG pathway analysis showed that proteins with altered levels may be implicated in several processes, including gene silencing by RNA, oxidative phosphorylation, glycolysis/gluconeogenesis, cytoskeleton organization, and ATP metabolic and energetic processes. The molecular docking analysis, which used 23 altered proteins as targets, revealed that IA interacted with all the proteins used. Conclusions: The results obtained using the five bioinformatic approaches provide information and show that IA could be used to treat non-Hodgkin lymphoma induced with the U-937 cell line. Also, it could provide a basis for future research and the development of clinical trials. Full article

Background/Objectives: Psoriasis is a chronic dermatological condition with systemic implications, especially with metabolic syndrome (MS). This study evaluated the vicious cycle where obesity and MS exacerbate systemic inflammation that complicates the efficacy of psoriasis therapies by examining the PASI score over a one-year period. Patients were classified into two subgroups: those with psoriasis alone (PSO) and those with both psoriasis and metabolic syndrome (PSO-MS). Methods: A total of 150 patients, half of whom also concomitantly presented with metabolic syndrome, received biologic therapies comprising anti-IL-17, anti-IL-23, and anti-TNF-a, or methotrexate, with PASI scores assessed at baseline and at 3, 6, and 12 months. Results: All treatments showed significant reductions in PASI; however, patients with PSO showed more marked reductions in PASI score than those in the PSO-MS group. Anti-IL-17 treatments produced the greatest sustained long-term improvements, whereas anti-IL-23 produced prompt early improvements. Increases in BMI and leptin concentrations were associated with a modest rate of reduction in PASI score, underlining the impact of obesity and metabolic dysfunction on treatment efficacy. Conclusions: This study highlights the importance of managing comorbidities such as MS in the treatment of psoriasis, as the interplay between systemic inflammation and metabolic health further complicates therapeutic outcomes. Full article

Background/Objectives: Brain cancer is notoriously resistant to traditional treatments, including radiotherapy. Microbeam radiation therapy (MRT), arrays of ultra-fast synchrotron X-ray beams tens of micrometres wide (called peaks) and spaced hundreds of micrometres apart (valleys), is an effective alternative to conventional treatments. MRT’s advantage is that normal tissues can be spared from harm whilst maintaining tumour control. Combining MRT with targeted radiosensitisers, such as nanoparticles, chemotherapeutic drugs, and halogenated pyrimidine drugs, can further improve radiotherapy by enhancing radiation damage. However, the underlying mechanisms of MRT are still being understood, which is essential to ensuring the reliable and successful use of MRT. Methods: An in vitro study was performed using γH2AX imaging, and quantification was performed via confocal microscopy and a clonogenic cell survival assay. Results: We show that methotrexate chemotherapeutics and iododeoxyuridine enhance MRT cell-killing and thulium oxide nanoparticles (TmNPs) broaden MRT peaks, and using γH2AX immunofluorescent confocal microscopy to quantify DNA damage, we further our knowledge of MRT mechanisms. γH2AX images verify the biological responses of cells aligning with the physical collimation of MRT, and we can accurately measure MRT microbeam characteristics bio-dosimetrically. The peak-to-valley dose ratio (PVDR), the ratio of the peak dose to the valley dose that characterises an MRT field, was accurately measured biologically using γH2AX imaging, despite studies previously finding this challenging. Conclusions: The measurement of biological PVDR has been performed for the first time with high-Z radiosensitisers, including nanoparticles, and several novel radiosensitiser-enhanced MRT mechanisms were discovered. Our results deepen our understanding of MRT with radiosensitisers, and can contribute to its accurate and future successful use in treating cancer. Full article

In the present study, ultra-small, magnetic, oleyl amine-coated Fe₃O₄ nanoparticles were synthesized and stabilized with a cationic ligand, cetyltrimethylammonium bromide, and an anticancer drug, methotrexate, was incorporated into a micelle-like nanoparticle structure for glioblastoma treatment. Nanoparticles were further characterized for their physicochemical properties using spectroscopic methods. Drug incorporation efficiency, drug loading, and drug release profile of the nanoparticles were investigated. According to the results, max incorporation efficiency% of 89.5 was found for 25 µg/mL of methotrexate-loaded nanoparticles. The cumulative amount of methotrexate released reached 40% at physiological pH and 85% at a pH of 5.0 up to 12 h. The toxicity and anticancer efficacy of the nanoparticles were also studied on U87 cancer and L929 cells. IC₅₀ concentration of nanoparticles reduced cell viability to 49% in U87 and 72% in L929 cells. The cellular uptake of nanoparticles was found to be 1.92-fold higher in U87 than in L929 cells. The total apoptosis% in U87 cells was estimated to be ~10-fold higher than what was observed in the L929 cells. Nanoparticles also inhibited the cell motility and prevented the metastasis of U87 cell lines. Overall, designed nanoparticles are a promising controlled delivery system for methotrexate to the cancer cells to achieve better therapeutic outcomes. Full article

Sarcoidosis is a multisystem granulomatous inflammatory disorder, of unknown aetiology, which causes a wide spectrum of clinical phenotypes. It can present at any age, most commonly between 20 and 60 years, with a roughly equal sex distribution. Diagnosis is often delayed due to multiple diagnostic mimics, particularly joint disease. Common presenting features include pulmonary disease, with bilateral hilar lymphadenopathy and pulmonary infiltrates, cutaneous lesions, and ocular disease. Musculoskeletal manifestations are reported in 10–40% of patients with sarcoidosis and include bone lesions, acute arthritis, chronic arthritis, axial disease, dactylitis, and sarcoid myopathy, which are explored in detail in this review article. Diagnosis is confirmed through histological evidence of non-caseating granuloma on tissue biopsy. Newer imaging modalities, including ^18FFDG PET/CT, can help identify the extent of musculoskeletal involvement, and biomarkers can provide weight to a diagnosis, but there is no single biomarker with prognostic value for disease monitoring. The mainstay of treatment remains corticosteroids, followed by disease-modifying antirheumatic drugs such as methotrexate and antimalarials. More recently, biologic treatments have been used successfully in the treatment of sarcoidosis with rheumatic involvement. Full article

Background/Objectives: Standard chemotherapy is generally considered the best approach to treat many solid cancers, even accounting for severe side effects. Therefore, the development of a drug delivery system for chemotherapeutic administration could significantly improve standard chemotherapy by maintaining the cytotoxic effects of the drugs while decreasing the inherent side effects of the treatment. The aim of our study is the optimization of a loading strategy that conjugates the use of extracellular vesicles (EVs) as drug delivery carriers, by preserving their integrity, with the loading efficiency and activity maintenance of chemotherapeutics. Methods: We compared the EV loading of the chemotherapeutics epirubicin, mitomycin, methotrexate and mitoxantrone by co-incubation. Once loaded, the activity of drug-carrying EVs was tested on cancer cells and compared to that of free chemotherapeutics. Results: We defined a linear correlation between chemotherapeutics’ concentration and their absorbance at the drug-specific wavelength, which allowed the definition of a highly sensitive absorbance-based spectrophotometric quantification system, enabling the assessment of drug loading efficiency. Co-incubation of EVs and chemotherapeutics was sufficient to obtain quantifiable drug loading, and the efficacy of EV loading was drug-dependent. Epirubicin-loaded vesicles showed increased toxicity to bladder cancer cells with respect to the free chemotherapeutic. The cytotoxicity was maintained even upon 6-month storage at −80 °C of loaded EVs. Conclusion: We established an absorbance-based spectrophotometric quantification system that enables a straightforward measure of drug loading efficiency into EVs, and we demonstrated that chemotherapeutic-carrying EVs can be obtained by co-incubation, preserving and increasing drug cytotoxicity. Full article

In this paper, the application of NiO nanosheets (NiO NSs) for the detection of methotrexate (MTX) is described. The NiO NSs were synthesized using a hydrothermal method. The electrocatalytic activity of two modifiers, ionic liquid (IL) and NiO NSs, was examined on a carbon paste electrode (CPE) in relation to MTX, utilizing voltammetry methods such as cyclic voltammetry (CV), linear sweep voltammetry (LSV), differential pulse voltammetry (DPV), and chronoamperometry at 0.1 M phosphate buffer solution (PBS) pH = 7.0. The anodic peak currents for MTX on the NiO NSs/IL/CPE were approximately 3.5 times greater than those on unmodified CPE. Based on DPV measurements, the electrochemical sensor demonstrated a linear response in the concentration range (LDR: 0.01 µM to 160.0 µM), with a limit of detection (LOD: 0.003 µM). Moreover, the NiO NSs/IL/CPE sensor demonstrated good stability, repeatability, reproducibility, and selectivity, which were of importance in the electroanalysis of compounds. Lastly, the practicality of the NiO NSs/IL/CPE sensor was assessed by analyzing MTX levels in urine samples and pharmaceutical formulation, yielding satisfactory recovery rates of 97.1% to 103.3%. Full article

Objectives: To describe the frequency of neutropenia and Felty syndrome in patients with rheumatoid arthritis (RA) attended in routine clinical practice. Methods: We selected by randomization a sample of 270 RA patients attended from January 2014 to November 2022. Demographic, clinical, and neutropenia-related variables were collected from the electronic medical records. Neutropenia was defined as having an absolute neutrophil count (ANC) of less than 1500/mm³ once, and acute if it persisted for <3 months. Felty syndrome was defined as RA-related neutropenia, rheumatoid factor (RF) and/or anti citrullinated protein antibody (ACPA) positivity. Results: We found 50 patients who had at least one neutropenia episode, with an incidence of 18.5% (14.0–25.6%). Most were women, with age (mean, p25–p75) at the time of neutropenia of 61.5 (57.4–69.3) years, 85% RF+ and 76% ACPA+. The demographic and RA characteristics of patients with and without neutropenia were very similar, except for sex: most patients with neutropenia were women. The 50 patients had 99 episodes of neutropenia; 59% were acute. The lower ANC was 1240 (1000–1395) mm³, and most of the episodes were mild (74%). In 32% of cases, there was other cytopenia. The RA activity measured by DAS28 in patients with neutropenia was low, at 2.18 (1.75–2.97). A total of 82 of 99 neutropenia episodes were related to DMARDs, 60% to Anti-IL6 drugs in monotherapy, 13% to RA activity, 3% to infectious diseases and 1% to hematologic malignancy. There were five (1.8%) cases with Felty syndrome, but only one woman with the classic combination of RA, positivity of autoantibodies (RF and ACPA), neutropenia and splenomegaly. Conclusions: In the 21st century, neutropenia in RA patients is most commonly related to biologics, mostly IL6 inhibitors and methotrexate. Episodes are mild, acute, with low RA activity, and associated with severe infections in few cases. Felty syndrome is rare. Full article

Inflammatory bowel disease (IBD) is a complex, multisystemic disease and is associated with ocular pathology in 4–12% of patients. In general, ocular disease affects Crohn’s patients more frequently than those with ulcerative colitis. Episcleritis and uveitis are the most common presentations, with episcleritis often correlating with IBD flares, whereas uveitis presents independently of IBD activity and, in some cases, may even alert clinicians to a new diagnosis of IBD. Corneal EIMs encompass a range of pathologies, such as the common and benign keratoconjunctivitis sicca (dry eye disease), which nevertheless causes significant patient discomfort, and the rarer condition of peripheral ulcerative keratitis, which warrants urgent review due to the risk of corneal perforation. Alongside EIMs, clinicians should also be aware of the iatrogenic consequences to the eye following treatment of IBD. Corticosteroids may cause cataracts, glaucoma, and—indirectly via hyperglycaemia—diabetic retinopathy. Methotrexate is irritating to ocular tissues and may cause conjunctivitis and blepharitis. Biologic medications, such as anti-TNFα agents, overlap in their use as treatment of both IBD and uveitis, and yet in some patients may also increase the risk of acute uveitis flares, as well as opportunistic, sight-threatening infections. With integrated care between gastroenterology and ophthalmology, patient outcomes can be improved by facilitating earlier detection and management of ocular disease. This narrative review summarises the ocular extraintestinal manifestations of IBD, including pathophysiology, epidemiology, and current treatment strategies. Full article

Objectives: This study is a retrospective analysis of patients with plaque psoriasis treated with biological drugs at a single center in Poland. We sought to evaluate patient demographics, disease characteristics, comorbidity burden, and treatment patterns in this cohort. Methods: Data were collected from the medical records of patients with plaque psoriasis who received biological treatments. In total, data from 1 January 2013 to 2 August 2024 were analyzed, encompassing 159 patients. The variables analyzed included age, disease duration, affected areas, prior treatments, and treatment outcomes. Results: The mean age at the start of biological treatment was 48 years (range: 10–73 years), with an average psoriasis duration of 18.2 years (range: 1–51 years). Obesity was noted in 39% of patients. Psoriasis lesions commonly affected the scalp (74.66%) and nails (64.38%). Methotrexate was the most commonly used systemic therapy prior to biologics (86.30%). Risankizumab and adalimumab were the most frequently prescribed biologics. Secondary treatment failure led to the highest discontinuation rates with tildrakizumab, whereas bimekizumab, guselkumab, risankizumab, and secukinumab showed the lowest rates. Conclusions: Biological drugs play a pivotal role in managing plaque psoriasis, particularly for patients with comorbidities and in treating challenging areas such as the scalp and nails. Risankizumab and adalimumab were prominent in prescription patterns. Future research involving larger cohorts and prospective designs is needed to deepen understanding and optimize treatment strategies for plaque psoriasis in Poland. Full article