The standard alkaline comet assay is a straightforward and cost-effective genotoxicity test commonly used in genetic toxicology to detect strand breaks and alkali-labile sites in DNA. Furthermore, with certain modifications, other altered bases such as oxidised and alkylated bases and cross-links can also be detected. However, it cannot detect bulky DNA adducts, which are a significant DNA lesion formed by the attachment of a chemical to DNA. Given that bulky adducts are mainly repaired by nucleotide excision repair (NER), the modification of the comet assay by adding blockers of the repair synthesis stage of NER, lead to the accumulation of incision intermediates, i.e. DNA breaks. Although this modification has been proposed as a method for detecting bulky adducts, the literature suggests that it may also detect other DNA lesions and that it has not yet been validated. To study the DNA lesion specificity of the in vitro comet assay in combination with DNA repair inhibitors, we tested a set of seven genotoxic compounds with different mechanisms of action and two non-genotoxic compounds, in TK6 cells. The assays were performed in the absence or presence of the inhibitors, specifically the combination of hydroxyurea (HU) and cytosine arabinoside (Ara-C), or aphidicolin (APC). The panel of genotoxic compounds included a bulky DNA adducts inducer (BPDE), two alkylating agents (methyl methanesulfonate (MMS) and ethyl methanesulfonate (EMS)), two oxidising agents (KBrO₃ and H₂O₂), and two cross-linking agents (cisplatin (CisPt) and mitomycin C (MitC)). The MTS and comet assays were performed to determine cytotoxicity and DNA damage, respectively. Results showed that this approach increases the sensitivity of the comet assay for detecting DNA damage, but not specifically bulky DNA adducts.

Splanchnic venous thrombosis (SVT) is a potentially catastrophic manifestation of polycythaemia vera (PV). We report a woman in her 60s with masked PV who had been managed for 5 years with intermittent phlebotomy and aspirin without haematology referral, molecular testing or cytoreductive therapy. Chronic iron deficiency masked erythrocytosis, delaying diagnosis. She presented with extensive thrombosis involving the portal, splenic and superior mesenteric veins complicated by mesenteric ischaemia requiring exploratory laparotomy, jejunal resection and ileostomy. Bone marrow biopsy and JAK2 V617F testing confirmed PV. Therapeutic anticoagulation was initiated with enoxaparin and subsequently transitioned to apixaban, while hydroxyurea achieved sustained haematologic control. Interval imaging demonstrated thrombus regression with persistent cavernous transformation. Given ongoing portal hypertension, oesophageal varices, occasional mild ostomy bleeding and low body weight, anticoagulation was continued indefinitely at reduced intensity with apixaban 2.5 mg twice daily. At 21-month follow-up, the patient remained free of recurrent thrombosis and major bleeding.

Trypanosoma cruzi, the etiological agent of Chagas Disease (CD), represents a significant public health concern and serves as a valuable model for investigating the cell cycle in early-diverging eukaryotes. Its unique cellular features,including the absence of chromosome condensation and the coordination of nuclear division with specialized organelles, provide insights into non-canonical regulatory mechanisms, and underscore the pivotal role of the nucleus in maintaining genomic integrity and regulating fundamental processes such as DNA replication and gene expression. To investigate nuclear dynamics throughout the cell cycle, we synchronized T. cruzi epimastigotes at the G1/S transition using hydroxyurea (HU), followed their progression through S and G2/M phases, and quantitatively analyzed isolated nuclear proteins by LC-MS/MS. We identified 2,937 nuclear-enriched proteins, revealing distinct, phase-specific expression patterns. The G1/S phase was marked by increased levels of metabolic enzymes including those related to energy and nucleotide/nucleoside metabolisms. The S phase showed elevated abundance of canonical and variant histones, consistent with chromatin remodeling and DNA replication. The G2/M phase was enriched in proteins involved in protein synthesis and microtubule dynamics, essential for mitosis. Notably, metabolic enzymes represented approximately 6% of the proteins identified in the purified nuclear fraction, consistent with previously published proteomic analyses of T. cruzi nuclear-enriched fractions. Hexokinase, citrate synthase, and pyruvate kinase activities were detected in extracts from this fraction, while immunofluorescence supported the association of hexokinase and citrate synthase with the nuclear region. These results suggest that metabolic enzymes and/or their metabolites may modulate nuclear processes in T. cruzi, potentially influencing the epigenome and gene expression regulation, as proposed in other eukaryotic models. Altogether, our findings reveal dynamic remodeling of the nuclear proteome during the T. cruzi cell cycle and point to a previously underappreciated role for metabolic enzymes likely regulating nuclear functions in a cell cycle-dependent manner.

Sickle cell nephropathy (SCN) is one of the most consequential yet under-recognized complications of sickle cell disease (SCD), contributing significantly to long-term morbidity and premature mortality. Despite its clinical importance, SCN often escapes early detection, in part because standard measures of kidney function can mask disease that is already well established. This narrative review synthesizes current evidence on SCN, examining the diagnostic strategies and the evolving therapeutic landscape-with particular attention paid to hydroxyurea, erythropoiesis-stimulating agents, and renal replacement therapy-and identifies the research gaps that must be addressed to improve outcomes for this patient population.

Following the publication of the above article, a concerned reader drew to the authors' attention that, regarding the images of the mouse embryo fibroblast (MEF) cells with senescence‑associated β‑galactosidase (SA‑β‑gal) staining shown in Fig. 1B on p. 3254, the panels on the far left and the far right on the bottom row of data [showing treatments with 0 µM and 40 µM adjudin respectively in the presence of hydroxyurea (HU)] contained an overlapping section of data (albeit after turning the right‑hand panel through 90°), such that data which were intended to show the results from differently performed experiments had apparently been derived from the same original source. In addition, the reader also observed that one of the antibodies the authors had used in their study was apparently selected inappropriately: The antibody (cat. no. ab51243) that the authors reported to have used is specific for the protein known as p16‑ARC (or ARPC5), not the apoptosis‑associated protein p16‑INK4a, as was intended. After re‑examining their original data, the authors have realized that the data in Fig. 1B were assembled incorrectly. A revised version of Fig. 1 is shown on the next page, now featuring data for all the panels in Fig. 1B from one of the repeated sets of experiments. Concerning the potential selection of an inappropriate antibody to probe for p16 protein, the authors expressed their regret that they had not retained their original receipt for the purchase of this antibody, and were not in a position to prove whether or not the antibody had been chosen legitimately for the affected experiments; however, an increased abundance of p16‑INK4a was used as an indicator of cellular senescence in combination with the abundance of p21 and the fraction of cells that contained a SA‑β‑gal signal. In view of this, the authors considered that removing the p16 data from the paper would not compromise the conclusions drawn in this study. After considering this appeal, the Editor has approved the authors' request that the p16 data be omitted from this study. Therefore, all references to 'p16' in this study (as were featured in the Abstract, and the Materials and methods, Results and Discussion sections) should be considered to be eliminated from this paper; the revised version of Fig. 1 that has been provided to take account of the new data in Fig. 1B also no longer includes the p16 data, and a new version of Fig. 3 has also been provided (on the third page of this corrigendum), again with the p16 data excluded. Note that the revisions made to these figures, and the omission of the data relating to p16, do not affect the overall conclusions reported in the paper. The authors express their gratitude to the Editor of International Journal of Molecular Medicine for allowing them the opportunity to publish this corrigendum, and apologize to the readership for any inconvenience caused. [International Journal of Molecular Medicine 42: 3522‑3529, 2018; DOI: 10.3892/ijmm.2018.3917].

This article presents checklists designed to guide clinicians in the comprehensive management of pregnant individuals with sickle cell disease, including prepregnancy, antepartum, delivery, and postpartum care, as well as the specific management of vaso-occlusive crises. The checklists aim to standardize care, enhance documentation and communication, and improve maternal and neonatal outcomes. Implementation strategies are discussed, emphasizing the need for integration of checklists into clinical workflows and continuous evaluation of quality indicators to address disparities and optimize patient care.

To compare clinical and laboratory characteristics of transfusion-dependent and non-transfusion-dependent beta thalassaemia patients with leg ulcers, and to identify contributing and protective factors We retrospectively reviewed 12 patients with beta thalassaemia and leg ulcers (seven transfusion-dependent and five non-transfusion-dependent) at a tertiary centre. We analysed demographics, haemoglobin levels, ferritin, foetal haemoglobin, transfusion status, comorbidities, and ulcer characteristics. Transfusion-dependent patients were predominantly younger males (mean age: 31.9 years), while non-transfusion-dependent patients were older (mean age: 44.0 years). Ulcers were larger in transfusion-dependent patients (mean area: 3.1 cm[2]) but more frequent in non-transfusion-dependent patients (mean episodes: 2.2). Splenectomy was performed in all transfusion-dependent patients and most non-transfusion-dependent patients. Transfusion-dependent patients had higher ferritin levels, while non-transfusion-dependent patients had higher foetal haemoglobin and slightly better ulcer resolution rates (80% vs. 71%). Major exacerbating factors included chronic anaemia, iron overload, trauma, and hypercoagulability. Protective factors included higher foetal haemoglobin levels, hydroxyurea use, occasional transfusions, and early conservative wound care. Although more commonly reported in non-transfusion-dependent patients, leg ulcers were also clinically significant in transfusion-dependent patients, where they were associated with larger ulcer size, higher iron burden, and incomplete healing despite regular transfusion and chelation. These findings highlight the need for heightened recognition and tailored preventive strategies for leg ulcers in transfusion-dependent thalassaemia.

Hydroxyurea (HU) is an established anticancer drug used to treat chronic myelogenous leukemia, but its therapeutic efficacy is limited by rapid clearance, variable bioavailability, and dose-related toxicity. This study aimed to improve HU delivery by developing hydroxyurea-loaded nanostructured lipid carriers (NLCs) using different surfactant combinations. Three formulations, including NX-ST (Tween 80-SDS), NX-CT (Cremophor EL-Tween 80), and NX-CS (Cremophor EL-SDS), were prepared and evaluated for particle size, polydispersity index (PDI), zeta potential, encapsulation efficiency (EE), and cytotoxic activity. The formulations exhibited nanoscale particle sizes (67.377 to 110 nm), acceptable PDI (<0.35), high zeta potential (-30.033 to -46.433 mV), and good encapsulation efficiency (78.189 to 86.626%), indicating stable formulations. The release profile exhibited an approximately linear increase over the 0.5-6 h period, followed by a plateau. Cytotoxicity studies showed enhanced anticancer activity compared with free HU (IC50 = 722.57 µg/mL). The formulation NX-ST demonstrated the highest cytotoxicity (IC50 = 225 ± 21.47 µg/mL), consistent with enhanced drug delivery; the contribution of improved cellular uptake is inferred and was not directly measured in this study. The NLC formulations studied, particularly NX-ST, effectively enhanced HU's therapeutic performance by improving stability, delivery efficiency, and anticancer activity.

Nitric oxide (NO)-based therapy represents a promising strategy against multidrug-resistant infections owing to its broad-spectrum antibacterial activity and minimal propensity to induce resistance. Hydroxyurea (HyU), a clinically approved drug for sickle cell disease, has recently emerged as a hydrogen peroxide (H2O2)-responsive NO precursor. However, the hydroxamic acid moiety responsible for NO generation is highly susceptible to hydrolytic degradation in aqueous environments, severely limiting its therapeutic utility. Herein, we report a coordination-driven stabilization strategy that incorporates HyU as a coordinating component within a Cu(II)-based metal-organic framework (CuPH). Coordination with Cu(II) preserves the chemical integrity of HyU for at least 14 days under physiological conditions, enabling its use as a stable NO precursor within a glucose-responsive nanoreactor. Using this stabilized framework, glucose oxidase (GOx) is co-confined within CuPH to construct a nanoreactor (GOx@CuPHcs). Under hyperglycemic wound conditions, GOx catalyzes in situ H2O2 generation from glucose, which subsequently activates Cu-mediated oxidation of coordinated HyU to achieve glucose-gated and on-demand NO release. The generated NO effectively eliminates methicillin-resistant Staphylococcus aureus (MRSA) and disrupts mature biofilms. GOx@CuPHcs was also associated with macrophage phenotype remodeling, enhanced fibroblast migration, and accelerated wound healing in a diabetic murine model with negligible systemic toxicity. This work demonstrates that coordination interactions can improve the stability of hydrolytically labile therapeutic molecules while enabling their integration into stimulus-responsive nanoreactors, highlighting the potential of coordination-based strategies for developing advanced therapeutic systems.