Candida albicans is an opportunistic pathogen whose virulence factors, such as polymorphism and biofilm formation, facilitate infection. Due to rising antifungal resistance, essential oils (EOs) from Citrus limon and Citrus sinensis are being investigated for their antimicrobial potential. This study analyzes the chemical composition, antifungal activity against Candida albicans in planktonic and biofilm forms, cytotoxicity on human erythrocytes, and pharmacokinetic properties of Citrus limon and Citrus sinensis essential oils. The essential oils were extracted from the leaves by hydrodistillation, and their composition was analyzed by GC-MS. Antifungal activity was assessed using broth microdilution and time-kill kinetics. Combined effects were assessed using checkerboard assays, while antibiofilm tests were evaluated using polystyrene plate assays and microscopy. Hemolytic activity tests were performed using percentage hemolysis. C. limon essential oil had limonene 40.8%, β-pinene 16.5%, and citronellal 12.3%, while C. sinensis had limonene 36.7%, β-pinene 14.6%, and eucalyptol 10.5% as its main constituents. The MIC was 0.312 - 1.25 mg/mL, with kill kinetics of up to 10 h. It showed additive effects for C. limon/fluconazole and synergistic effects for C. sinensis/fluconazole. It exhibited anti-adhesion and anti-polymorphism effects, inhibiting biofilm formation. It also exhibited low cytotoxicity, with an LC50 of 14 000 µg/mL for C. limon and 13 000 µg/mL for C. sinensis. ADMET and BOILED-egg pharmacokinetic predictions indicated safety for systemic use, with caution for topical application of limonene, citronellal, and eucalyptol. Both EOs exhibited strong anti-Candida activity with minimal cytotoxicity. Its additive/synergistic effects with fluconazole highlight its potential as an alternative or adjuvant therapy against C. albicans infections.

Bioprospecting of the Amazonian mapati fruit (Pourouma cecropiifolia Martius): Antiproliferative and antimalarial potential of peel and seed hydroethanolic extracts.

Computational epitope discovery and in silico design of a bispecific antibody model (COMED hook) to block Plasmodium invasion into erythrocytes.

Quercetin mitigates size-dependent oxidative and metabolic toxicity of citrate-coated silver nanoparticles in human erythrocytes.

Abstract Human erythrocyte band 3 is an anion exchanger and comprised of 14 transmembrane segments (TMs). Its function is inhibited by the modification of Lys-539 on TM 5 using 4,4’-diisothiocyanatodihydrostilbene 2,2’-disulfonate (H2DIDS). Upon exposure of band 3 to alkaline pH, another isothiocyanate group of H2DIDS can react with Lys-851 on TM 13. Thus, Lys-539 on TM 5 and Lys-851 on TM13 are cross-linked by H2DIDS. Such cross-linking was facilitated by a pressure of 100 MPa and suppressed the 200 MPa-induced hemolysis of erythrocytes pre-exposed to alkaline pH. Thus, the cross-linkage between TMs by H2DIDS stabilizes bilayer-cytoskeleton interactions and results in the reduction in the volume of band 3.

Uranium, being chemo-radiotoxic, requires confinement within a safe environment to prevent severe health consequences. Chelation therapy, utilizing multidentate ligands, is a widely accepted approach for developing effective uranium decorporation treatments. In this study, a systematic investigation of uranium complexation by pyrazine-2-amidoxime (PAM) was undertaken for the first time, employing chemical speciation, in vitro studies, and computational calculations. Potentiometric and spectrophotometric titrations, corroborated by electrospray ionization mass spectrometry (ESI-MS), revealed that PAM forms 1:1 (ML) and 1:2 (ML2) complexes with the uranyl ion (UO22+). PAM acts as a bidentate chelator, coordinating via the oxygen and nitrogen atoms of amidoxime, with high stability constants (log β) of 8.54 ± 0.04 for ML and 16.33 ± 0.07 for ML2 complexes. Density functional theory (DFT) calculations further elucidated the preferred coordination mode, donor-site contributions, and electronic factors governing the stability of the PAM-uranyl complexes. Ex vivo experiments with human erythrocytes (RBCs) confirmed that PAM is cytocompatible and significantly enhanced uranium decorporation from human RBCs, facilitating removal of 25-40% of uranium at 100-200 μM compared to uranium-treated controls. Additionally, PAM effectively reduced uranium content by 10-20% from physiologically relevant proteins such as human serum albumin and from native human blood plasma.

Efficacy of phosphonium amphiphilic salts against Acanthamoeba genotype T4

Perfluorooctane sulfonate (PFOS) causes dysfunction of erythrocytes by calcium dysregulation, and a diabetic condition exacerbates PFOS-induced prothrombotic activation.

Excipient toxicity and tolerability in self-emulsifying drug delivery systems: insights from cell-based assays.

A liquid chromatography-mass spectrometry method for profiling global phospholipid methylation via Headgroup analysis revealing cell-type specificity.

A novel model recently proposed by Doktorova etal. challenges current concepts for the molecular organization of the plasma membrane bilayer. It is at odds with previously published research. The model posits that there are far fewer phospholipid molecules in the outer leaflet of the bilayer than in the inner leaflet and that the resulting area deficit is filled by cholesterol. This conclusion is based on the incomplete hydrolysis of the phosphatidylcholine in intact erythrocytes by phospholipase A2, leading to the inference that the undigested fraction is endofacial. But the incomplete digestion can be explained by product inhibition. Furthermore, ultrastructural analysis has shown that almost all of the phosphatidylcholine in the erythrocyte bilayer resides in the outer leaflet. Finally, the high concentration of cholesterol predicted for the outer leaflet of resting human erythrocytes is not detectable by probes. The conflict of the new model with the literature makes it insecure.

This study sought to comprehensively explore the thrombolytic and membrane-stabilizing activities of allyl isothiocyanate (AITC) through both in vitro experimental assays and computational analysis. To investigate its thrombolytic activity, we conducted clot-lysis assays that mimic physiological clot formation and breakdown. In parallel, the membrane-stabilizing activity of AITC was assessed via human erythrocyte hemolysis assays, designed to cellular membrane damage that occurs during inflammation and oxidative stress. These assays allowed us to investigate the therapeutic relevance of AITC in conditions involving thrombosis and cell lysis. Additionally, molecular docking studies made us understand the potential interactions of AITC with two key molecular targets: plasminogen, which contributes significantly to the breakdown of fibrin clots, and cyclooxygenase-1 (COX-1), a major enzyme involved in the inflammatory responses. The in vitro results showed that AITC exerted concentration-dependent effects. At 100 μg/ml concentration, AITC achieved 82.18 ± 0.01% clot dissolution and 92.10 ± 0.01% membrane protection. The IC₅₀ values were observed to be 41.70 ± 1.47 μg/ml for clot-lysis and 36.01 ± 1.71 μg/ml for membrane-stabilization, indicating potent bioactivity. Docking analysis demonstrated favorable interaction of AITC with plasminogen (−4.2 kcal/mol) and COX-1 (-4.5 kcal/mol), suggesting possible mechanisms underlying its observed effects. Taken together, these results suggest that AITC may function through dual fibrinolytic and anti-inflammatory pathways. Its ability to interact with key proteins involved in clot resolution and inflammation suggests it as a promising natural therapeutic candidate. However, additional clinical and pre-clinical investigations are required to establish its therapeutic safety and efficacy profile. Bangladesh Pharmaceutical Journal 29(1): 53-67, 2026 (January)

When liquid medications are administered intravenously, the first cellular defense encountered by the drug is erythrocytes. Catalase is the main antioxidant system in erythrocytes. Drug-related catalase inhibition can cause adverse effects. Although catalase is a well-known enzyme, studies investigating drug-catalase interactions are scarce in the literature. Therefore, we investigated the impact of liquid cardiac drugs on human erythrocyte catalase activity in vitro. Catalase activity was determined by a spectrophotometric method using a procedure developed by Aebi. Liquid cardiac drugs were incubated with human blood in vitro. IC50 values were compared among the drugs. The most potent inhibitors were noradrenaline (IC50: 4.61 µM), adrenaline (IC50: 32.58 µM), and amiodarone hydrochloride (IC50: 41.86). Dopamine hydrochloride (IC50: 429.15 µM) and lidocaine hydrochloride (IC50: 453.1 µM) showed less inhibitory effects on catalase activity compared with adenosine (IC50: 58.49 µM), atropine sulfate (IC50: 68.75 µM), dobutamine hydrochloride (IC50: 80.79 µM), glyceryl trinitrate (IC50: 86.66 µM), and heparin sodium (IC50: 92.4 µM). Noradrenaline, adrenaline, and amiodarone hydrochloride have strong inhibitory effects on catalase activity. Catalase inhibition may be responsible for the side effects of these drugs. Therefore, when these drugs are used in treatment, their dosages and duration of administration should be carefully controlled to prevent adverse effects due to catalase enzyme inhibition.

Antiplatelet drugs are commonly used to prevent cardiovascular diseases (CVD). However, they are associated with side effects, highlighting the need for safer and more effective alternatives. The purpose of the present work was to evaluate the antiplatelet potential of a series of Morita-Baylis-Hillman (MBH) adducts and to test their safety. The antiplatelet activity was assessed using microplate aggregometry and confirmed by the turbidimetric method, utilizing platelet-rich plasma (PRP) from healthy volunteers. Next, we examined the effect of the selected compounds on platelet secretion, GPIIb/IIIa complex activation, and blood coagulation. The safety of the antiplatelet agents was evaluated by testing their cytotoxicity on platelets and erythrocytes. Among the tested compounds, Al4, OAc1, OAc2, OAc3, R12, R22, R32, and R52 demonstrated potent inhibition of Adenosine 5'-diphosphate (ADP)-induced platelet aggregation, with IC50 values ranging from 0.042 to 0.4 mM. The selected compounds also inhibited platelet aggregation induced by arachidonic acid (AA) and collagen. Notably, they significantly inhibited P-selectin expression and GPIIb/IIIa activation, without affecting coagulation parameters. Toxicological evaluation showed that these compounds did not induce hemolytic or cytotoxic effects on human erythrocytes or platelets. Collectively, these results identify MBH adducts as promising scaffolds for the development of novel selective antiplatelet agents.

The rise of Multidrug-Resistant (MDR) bacterial infections and the limited efficacy of conventional antibiotics have underscored the urgent need for innovative antimicrobial therapies. Antimicrobial Peptides (AMPs) represent a promising class of agents due to their broad-spectrum activity; however, their clinical application is often hindered by cytotoxicity and poor stability. This study aimed to develop and evaluate a novel ultrashort AMP-antibiotic conjugate, UP5-C-Levo, with improved antimicrobial efficacy and safety. UP5-C-Levo was synthesized by covalently linking the ultrashort peptide UP5 to the antibiotic levofloxacin. The antimicrobial activity of the conjugate was assessed against MDR Gram-positive and Gram-negative bacteria using Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. Antibiofilm efficacy was tested on Staphylococcus aureus biofilms. Cytotoxicity was evaluated via hemolysis of human erythrocytes and MTT assays on MDCK cells. Scanning Electron Microscopy (SEM) was employed to investigate bacterial membrane disruption. UP5-C-Levo exhibited potent antimicrobial activity, with MIC values ranging from 12.5 to 25 μM across all tested MDR strains. It significantly reduced biofilm biomass, achieving nearcomplete eradication of S. aureus biofilms. Hemolysis and cytotoxicity assays indicated minimal toxicity to human cells. SEM imaging revealed extensive bacterial membrane disruption, suggesting a dual mechanism of action. The conjugation of UP5 with levofloxacin resulted in a synergistic antimicrobial agent with enhanced efficacy and low cytotoxicity. The ability of UP5-C-Levo to disrupt bacterial membranes and eradicate biofilms addresses two major challenges in infection treatment. These findings align with current strategies aimed at overcoming antibiotic resistance through AMPbased drug design. UP5-C-Levo is a promising therapeutic candidate for the treatment of MDR and biofilm-associated infections. Its strong antibacterial and antibiofilm activity, combined with a favorable safety profile, supports further preclinical development and potential clinical translation.

ABSTRACT Lysimachia arvensis (L.) U. Manns & Anderb. is a medicinal plant traditionally used for various therapeutic purposes; however, its bioactive properties and potential toxicological effects remain insufficiently characterized. This study aimed to investigate the biological activity, cytotoxic potential, phytochemical composition, and molecular interaction mechanisms attributed to exposure to methanolic extract of L. arvensis using integrated in vitro and in silico approaches. The methanolic extract exhibited antioxidant activity with a DPPH radical scavenging IC50 value of 88.59 ± 23.29 mg/ml, and demonstrated strong 71% xanthine oxidase inhibition. Total flavonol, flavonoid, phenolic, and tannin contents were determined to be 25.33 mg QE/g, 16.55 mg QE/g, 23.18 mg GAE/g, and 0.91 mg GAE/g extract, respectively. GC–MS analysis identified 17 compounds, with guanosine, quinic acid, methyl linolenate, and myo-inositol as predominant constituents. Cytotoxicity assays demonstrated marked toxicity of the extract toward HEK293 cells with an IC50 value of 17.47 μg/mL, indicating significantly higher cytotoxicity compared to methanol alone. Molecular docking identified guanosine as the strongest binder to human erythrocyte catalase (PDB ID: 1DGF) with a binding energy of −8.6 kcal/mol, followed by neophytadiene and a pyran derivative. Molecular dynamics simulations over 100 ns confirmed stability of protein–ligand complexes while revealing ligand-specific mobility patterns. DFT and NCI analyses noted strong hydrogen-bonding interactions and distinct electronic properties among key compounds. Overall, L. arvensis exhibits notable bioactivity associated with significant cytotoxicity indicating the need for careful safety risk assessment.

Human erythrocytes (red blood cells; RBCs) undergo spontaneous disassembly after several hours of exposure to n-butyl acetate (nBA). Images of the morphological changes were captured in time-lapse sequences using differential interference contrast (DIC) light microscopy. On exposure to less than 10mM nBA dramatic events did not take place, but with ∼60mM aqueous solutions of nBA, discocytes became spherical with a single contiguous 'geographical' indentation patch. Over the next ∼2h the cells became echinocyte-like with rounded projections; and several hours later they discharged filaments that writhed in Brownian motion. In parallel with these changes, vesicles budded from the cells, followed by their alignment on the filaments, like balloons on a string. The vesicles then serially fused, finally giving rise to a single large vesicle that was ∼0.1-0.2 times the diameter of the spherical parent cell; it then fused with the parent cell that a short while later ruptured and became a 'ghost'. Owing to the striking nature of this phenomenon that was evocative of party decor, the term coined for it wasFeierzeit (German: celebration time). The background to this serendipitous discovery, and the deductive odyssey that identified the causative agent, nBA, are presented. Broader implications for understanding the assembly of the RBC cytoskeleton-plasma membrane complexes, and their disassembly under normal, pathological, and forensic conditions are discussed.

Protective effect of crocin against glycated LDL-induced cytotoxicity and oxidative stress in isolated human erythrocytes.

Electronic computer-based neural networks can recover high-resolution quantitative phase images from inline holograms to reveal the fine structure of samples, which may face issues such as intensive computing, energy consumption, and time delay. In contrast, optical diffraction computing not only has super-fast parallel processing capabilities and extremely low energy consumption but also is naturally compatible with the physical process of holographic imaging. However, since existing optical diffraction computing models are not able to preserve the phase detail in phase reconstruction, they focus on intensity recovery of holograms. To address this problem, this paper proposes an all-optical residual diffraction neural network (RDNN) architecture to obtain a high-precision phase distribution of inline holograms. In this method, residual optical structures and cascaded diffraction layer designs are combined to effectively reduce the loss of phase information during reconstruction, thereby ensuring a high degree of consistency between the reconstructed phase and the original phase. A series of numerical simulation results shows that the proposed architecture can be generalized to the phase reconstruction task of three-dimensional biological samples. In phase reconstruction of human erythrocytes, the average SSIM and PSNR reach 0.82 and 28, respectively. Additionally, we assess the network’s performance with noisy holograms, demonstrating that, unlike fully connected diffraction neural networks, this approach significantly enhances phase reconstruction quality, even under random Gaussian noise. These results indicate that the proposed RDNN has the potential to replace the electronic neural network to achieve accurate phase reconstruction of inline holograms and provide an efficient solution to observe the living cell dynamics.

Erythrocytes play a crucial role in human life and are a good model for single-cell ageing research. Therefore, the mechanism of erythrocyte ageing has been a serious concern for researchers. Here, we investigate the ageing mechanism of human erythrocytes in circulation with a new approach. We first try to discover all the factors that affect erythrocyte ageing from the existing findings about the interactions of erythrocytes with the environment during circulation and the resulting alterations in the cells during their ageing process. Then, we define the main events in erythrocyte ageing according to the incidence and strength of the interactions and the magnitude of the biophysical and biochemical alterations induced by these interactions. With the main events as a guide for tracking the ageing process and analysing the causes and consequences of the events taking place during erythrocyte ageing, we depict the pathways and framework of the ageing mechanism. We hope that understanding the ageing mechanism of erythrocytes will help people to have a deep insight into the biophysical and biochemical processes of cell ageing and their molecular basis. Additionally, it may offer solutions to different fundamental and practical problems related to erythrocyte ageing, storage lesions, transfusion and diseases.