Search Results (54)

Preeclampsia (PE) is associated with systemic oxidative stress and vascular dysfunction, yet its effects on red blood cell (RBC) stability and mechanics remain incompletely understood. Here, we investigate the structural and nanomechanical alterations of RBCs in third-trimester pregnancies complicated by non-severe and severe PE, compared with normotensive controls. RBCs are analyzed using differential scanning calorimetry (DSC) to assess protein thermal stability and atomic force microscopy (AFM) to determine membrane elasticity (Young’s modulus) during in vitro aging. Linear mixed-effects models are applied to evaluate the effects of disease severity, storage time, and their (group × storage time) interaction. DSC reveals that Band 3 and hemoglobin exhibited pronounced destabilization in PE, with severe cases showing earlier and larger reductions in transition temperatures and heat capacities, indicative of disrupted membrane–cytoskeletal interactions. AFM confirms that these molecular changes translate into functional consequences: control and non-severe PE RBCs show physiological softening over time, whereas severe PE RBCs undergo pathological stiffening. Statistical modeling demonstrates strong time, group, and interaction effects for both thermodynamic and mechanical parameters. Together, these findings identify the Band 3–hemoglobin macrocomplex as a primary target of PE-induced RBC alterations and suggest that combined thermodynamic–nanomechanical profiling can serve as a sensitive approach to detect early subclinical RBC damage not detectable by routine hematological tests. Full article

Yeast vitality during storage is essential for maintaining consistent fermentation performance. This study compares the physiological responses of top- and bottom-cropped Saccharomyces cerevisiae stored at 4 °C for 20 days and evaluates Optical Nanomotion Detection (ONMD) as a rapid, label-free vitality assessment tool. Classical assays (FUN-1, methylene blue, propidium iodide, glucose acidification power, glycogen content, and ethanol tolerance) were used to monitor metabolic activity, membrane integrity, and stress resilience. Bottom-cropped yeast retained metabolic activity, membrane stability, and energy reserves longer than top-cropped cells. ONMD revealed distinct single-cell nanomotion signatures and detected mechanically active subpopulations even when traditional vitality indicators declined. Analysis of nanomotion slopes showed an increasingly negative trend in the decline over storage time in top-cropped cells, indicating reduced temporal stability of nanomechanical activity during the 180 min recordings. Ethanol-challenge experiments confirmed the vitality dependence and stress-sensitivity of the ONMD signal. Together, these findings demonstrate that ONMD resolves cold-storage-induced changes in yeast nanomechanical vitality and provides complementary information beyond conventional vitality and viability assays. ONMD offers a fast, reagent-free method for monitoring brewing yeast physiology and represents a promising basis for future development toward brewery quality-control applications. Full article

Postharvest fungal decay is a primary cause of losses in blueberries, motivating the development of sustainable alternatives to conventional fungicides. This study aimed to develop and evaluate antifungal active films based on polylactic acid (PLA) enriched with citronella essential oil to control phytopathogenic fungi associated with blueberry spoilage. PLA films containing 7.5, 10, and 12.5% (w/w) citronella essential oil were produced by solvent casting and characterized for water vapor transmission rate and nanomechanical properties. The antifungal effect was tested in vitro against Epicoccum nigrum, Alternaria alternata, and Cladosporium herbarum. Active films exhibited concentration-dependent antifungal activity, with C. herbarum being the most sensitive fungus. The incorporation of citronella essential oil did not significantly alter the water vapor barrier properties of PLA, while mechanical analysis revealed a reduction in elastic modulus only at the highest concentration. The antifungal mechanism was elucidated using scanning electron microscopy, fatty acid profiling, absorbance at 260 nm, and conductivity measurements. The results indicate that the released citronella essential oil induced membrane disruption and morphological damage in fungal hyphae, with species-specific responses. Overall, PLA–citronella essential oil films represent a promising biodegradable packaging solution to control postharvest blueberry losses. Full article

Background/Objectives: Cardiopulmonary bypass (CPB) facilitates complex cardiac surgery but can damage erythrocyte membranes, impairing microcirculation and oxygen transport. Standard rheological tests assess overall blood properties but fail to define specific cellular mechanisms. In this study, atomic force microscopy (AFM) was employed to characterize morphological, nanostructural, and mechanical changes in erythrocytes following CPB and CPB combined with hypothermic circulatory arrest (HCA). Methods: The study included 14 patients who underwent cardiac surgery with CPB. Patients were divided into two groups. Group 1 underwent heart valve surgery with normothermic CPB (n = 7), and Group 2 underwent aortic arch surgery with CPB combined with HCA and moderate hypothermia (28 °C) (n = 7). Arterial blood samples were collected before the induction of anesthesia and immediately after CPB. The morphology and surface roughness (Rtm) of the erythrocyte membrane were evaluated on air-dried blood smears. Young’s modulus (E) was estimated from force-distance curves on living cells; measurements were performed at 24 °C in PBS. Results: Following CPB, both groups exhibited a decrease in the proportion of discocytes and an increase in echinocytes. In the CPB+HCA group, discocytes were absent after surgery. The mean Rtm increased 1.4-fold in Group 1 and 1.6-fold in Group 2, indicating greater nanostructural membrane damage in the latter. In Group 1, Young’s modulus increased by an average of 1.6 times, indicating increased cell stiffness. In Group 2, the increase was smaller (mean: 1.1 times) and was not statistically significant in some patients. Conclusions: Normothermic CPB primarily affects the nanomechanical properties of erythrocytes, whereas CPB+HCA induces more severe morphological and membrane surface damage while relatively preserving cytoskeletal elasticity. AFM-derived parameters of membrane roughness and cell elasticity may serve as sensitive indicators of erythrocyte biophysical integrity. Full article

Erythrocyte aging is a fundamental physiological phenomenon that involves significant structural and nanomechanical alterations of the cells’ structure and function. Coupling optical, fluorescence, and Atomic Force Microscopy (AFM), we analyzed morphology, membrane roughness and nanomechanical properties of the very same RBCs arising from favism subjects, measured at different stages of their aging in vitro. We also investigated the evolution and abundance of vesicles arising from the cells over their senescence pathway. This approach combines high-resolution fluorescence imaging with the correlation of membrane topology and biomechanics. This explores the differences between investigation based on statistical morphometric parameters, such as membrane roughness, and those based on the measure of point-dependent nanomechanical properties. Our ultra-morphological study evidences the existence of clear differences in the aging of normal and favism erythrocytes that results in a larger number of cells with abnormal shapes and in a hyper-production of vesicles along the senescence pathway of favism cells. In explaining these differences, we focused on the roles played by the hemoglobin evolution and by the morpho-mechanical properties that are responsible for the skeletal alterations. In particular, our data reported evidence that the two corresponding degradative pathways are coupled and play an important enhancement role in promoting the progression of cell senescence. Full article

Chronic lymphocytic leukemia (CLL) has systemic effects that extend beyond malignant lymphocytes, potentially altering the structure and function of circulating red blood cells (RBCs). In this study, atomic force microscopy (AFM) was combined with complementary calorimetric analysis to investigate the membrane ultrastructure, nanomechanical characteristics, and thermodynamic behavior of RBCs from untreated CLL patients and those receiving targeted therapies (Obinutuzumab/Venetoclax or Ibrutinib). RBCs from untreated patients exhibited pronounced reduction in membrane roughness, increased stiffness and adhesion forces, and altered thermal unfolding of cytoskeletal and membrane proteins, indicative of impaired structural flexibility and stability. Treatment with Obinutuzumab/Venetoclax partially restored surface topography, but stiffness and adhesion forces remained elevated, suggesting persistent cytoskeletal rigidity. The obscured spectrin and Band 2–4 thermal transitions and the elevated total enthalpy change revealed by differential scanning calorimetry indicated a modified conformation or binding state of membrane proteins. In contrast, Ibrutinib therapy produced near-normal nanomechanical and thermal characteristics, reflecting a more comprehensive restoration of RBC integrity. These findings demonstrate that CLL and its therapies distinctly influence erythrocyte morphology and mechanics, underscoring the systemic impact of the disease. The strong correspondence between AFM and calorimetric data highlights the potential of integrated biophysical approaches to detect subtle RBC alterations and to serve as complementary indicators for therapeutic monitoring. Full article

Background/Objectives: The accumulation of specific amyloid proteins and peptides in the human brain is a hallmark of neurodegenerative disorders such as Alzheimer’s disease (AD) and Parkinson’s disease (PD). Beyond the central nervous system, circulating peripheral blood cells are also exposed to these pathological proteins, which may contribute to the systemic disease manifestation. Human platelets (PLTs) were used as an in vitro model to investigate the impacts of amyloid Aβ1-42 peptide oligomers (Aβ42) and on-pathway α-synuclein (α-syn), two key amyloids implicated in AD and PD, on platelet biophysical properties. Methods: Using atomic force microscopy, imaging and force–distance modes, we analyzed changes in surface nanostructure, morphometric and nanomechanical signatures of PLTs, derived from healthy donors, following exposure to increasing concentrations of Aβ42 and α-syn. Results: Our findings show that platelet activation progresses with increasing amyloid concentration, characterized by cytoskeletal remodeling (filopodia-to-pseudopodia and lamellipodia transformation). While Aβ42 causes progressive decrease in the platelet membrane roughness, α-syn exhibits a biphasic effect—initial smoothing followed by a pronounced increase in the roughness at high concentrations. Both amyloids induce substantial increase in membrane stiffness (Young’s modulus). Conclusions: The changes in PLTs’ biophysical properties closely resemble the previously observed modification in PLTs derived from AD and PD patients, suggesting that amyloid proteins’ interactions with PLTs may contribute to their dysfunction. Our results underscore the potential of platelets as peripheral indicators of neurodegeneration and point to their role in the systemic pathology of amyloid-associated diseases. Full article

Background/Objectives: Leukoreduced low-titer group O whole blood (LTOWB) is a promising option for early transfusion in massive hemorrhages, by providing red blood cells (RBCs), plasma, and platelets in a single unit. This study aimed to assess the effects of pathogen reduction (PR) on the LTOWB quality and its suitability for emergency transfusions, including its post-thaw RBC usability. Methods: Whole blood from 24 donors was processed and analyzed over seven days, including post-thaw assessments. Hematological and coagulation parameters, along with atomic force microscopy, were used to assess the RBCs’ morphology, cytoskeletal integrity, and nanomechanical properties. Results: The PR reduced the leukocyte content while preserving platelet counts at the level of the control group. Although certain clotting factors, such as fibrinogen (p < 0.001) and factor VIII (p < 0.001), were diminished after the PR, thromboelastometry results remained within reference ranges, indicating a maintained hemostatic potential. Morphological changes in RBCs were observed, but the membrane elasticity remained stable throughout storage and after thawing, indicating preserved macromechanical properties essential for hemostasis. Conclusions: These findings demonstrate that LTOWB treated with partial a leukoreduction and PR maintains functional and structural integrity, supporting its potential utility as a safe and effective transfusion product for managing acute blood loss. Full article

Two proline-rich antimicrobial peptides (PrAMPs), named P1 and P2, purified from hemolymph of the greater wax moth Galleria mellonella, were studied for their effects on Gram-negative (Escherichia coli) and Gram-positive (Micrococcus luteus) bacteria. Both peptides decreased the M. luteus bacterial survival rate and caused E. coli bacterial membrane permeabilization. However, in both cases, the P2 peptide was approximately three times more effective than the P1 peptide. Fluorescence microscopy imaging demonstrated binding of both FITC-labeled peptides to E. coli and M. luteus cells. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) imaging of peptide-treated bacteria revealed considerable changes in cell morphology, cell surface topography, and nanomechanical properties. The interactions of the PrAMPs with bacterial cells were also analyzed by FTIR spectroscopy. The P1 peptide action toward E. coli led to partial aggregation of proteins, whereas treatment with P2 resulted in reduced protein aggregation, reflecting differences between both G. mellonella PrAMPs antibacterial action. Moreover, both PrAMPs caused a decrease and an increase in the protein content in relation to lipids on the E. coli and M. luteus cell surface, respectively. The obtained results reflect not only differences between the G. mellonella P1 and P2 peptides but also differences in the cell surface between Gram-negative and Gram-positive bacteria. Both characterized G. mellonella PrAMPs are further representatives of proline-rich peptides with a membrane-permeabilizing antimicrobial mode of action. Full article

Erythrocyte (RBC) aging involves significant structural and nanomechanical alterations crucial to their function. This study aims to bridge the gap between analyses based on statistical morphometric parameters, e.g., membrane roughness, and those based on point-dependent nanomechanical properties, e.g., stiffness or Young’s modulus. Using Atomic Force Microscopy, we investigated morphology, membrane roughness, and nanomechanical properties on the very same RBCs under dehydrated (air) and hydrated (physiological buffer) conditions. The cells were studied at different stages of in vitro aging: one, seven, and 12 days. Our results quantitatively show that across dehydration, as well as along the aging pathway, RBCs become progressively more rigid while their membrane roughness decreases, a trend observed in both environments. Notably, the differences between the hydrated and dehydrated states were large in young cells but diminished when erythrocytes aged. Despite these parallel trends, high-resolution mapping on the nanoscale revealed that roughness and Young’s modulus do not correlate, indicating that these parameters are linked to different properties. In conclusion, this work provides a comprehensive protocol for a biophysical description of RBC aging and establishes that the simultaneous measurement of membrane roughness and nanomechanical properties offers a complementary approach, yielding a more complete characterization of cellular properties. Full article

Tumor-Treating Fields (TTFields), a novel therapeutic avenue, is approved for therapy in Glioblastoma multiforme, malignant pleural mesothelioma, and metastatic non-small cell lung cancer (NSCLC). In pancreatic ductal adenocarcinoma (PDAC), several clinical trials are underway to improve outcomes, yet a significant knowledge gap prevails involving the cell-extracellular matrix (ECM) crosstalk. Herein, we hypothesized that treatment with TTFields influence this crosstalk, which is reflected by the dynamic alteration in nanomechanical properties (NMPs) of cells and the ECM in a co-culture system. We employed an ECM gel comprising collagen, fibronectin, and laminin mixed in 100:1:1 stoichiometry to co-culture of Panc1 and AsPC1 individually. This ECM mixture mimics the in vivo tumor microenvironment closely when compared to the individual ECM components studied before. A comprehensive frequency-dependent study revealed the optimal TTFields frequency to be 150 kHz. We also observed that irrespective of the ECM’s presence, TTFields increase cell membrane stiffness and decrease deformation several-folds in both Panc1 and AsPC1 cells at both 48 h and 72 h. Although adhesion for AsPC1 decreased at 48 h, at 72 h it was observed to increase irrespective of ECM’s presence. Moreover, it significantly alters the NMPs of ECM gels when co-cultured with PDAC cell lines. However, AsPC1 cells were observed to be more detrimental to these changes. Lastly, we attribute the stiffness changes in Panc1 cells to the membrane F-actin reorganization in the presence of TTFields. This study paves a path to study complex PDAC TME as well as the effect of various chemotherapeutic agents on such TME with TTFields in the future. Full article

Although commonly appreciated for their anti-oxidative and neuroprotective properties, flavonoids can also exhibit pro-oxidative activity, potentially reducing cell survival, particularly in the presence of metal ions. Disrupted copper homeostasis is a known contributor to neuronal dysfunction through oxidative stress induction. This study investigated the effects of myricitrin (1–20 μg/mL) on copper-induced toxicity (0.5 mM CuSO₄) in the neuroblastoma SH-SY5Y cell line. At non-toxic concentrations, myricitrin exacerbated copper’s toxic effects. The myricitrin-induced decrease in survival was accompanied with increased reactive oxygen species (ROS) production, reduced superoxide dismutase activity, and a lower GSH/GSSG ratio. In combination with copper, myricitrin also activated caspase-3/7, promoted nuclear chromatin changes, and compromised membrane integrity. At the protein level, myricitrin upregulated p53 and PUMA expression. The toxic effects of myricitrin were alleviated by the p38 inhibitor SB203580, the intracellular calcium chelator BAPTA-AM, and the NMDA receptor blocker MK-801, highlighting the significant role of the ROS/p53/p38 axis in cell death and the critical involvement of calcium ions in apoptosis induction. The atomic force microscopy was used to assess the surface morphology and nanomechanical properties of SH-SY5Y cells, revealing changes following myricitrin treatment. This research highlights the toxic potential of myricitrin and emphasizes the need for caution when considering flavonoid supplementation in conditions with elevated copper levels. Full article

Functional amyloids (protein nanofibrils, PNF) synthesized from plant sources exhibit unique physicochemical and nanomechanical properties that could improve food texture. While environmental factors affecting PNFs are well-known, scientific evidence on how cells (focus on the oral cavity) respond to them under physiological conditions is lacking. Self-assembled PNFs synthesized from fava bean whole protein isolate show a strong pH- and solvent-dependent morphology and elasticity modification measured by atomic force microscopy (AFM). After incubation of PNFs with an oral mechanosensitive model cell line at pH 7.3, difference in cell-surface roughness without significant changes in the overall cell elasticity were measured. The role of cell membrane composition on supported lipid bilayers was also tested, showing an increase in membrane elasticity with increasing fibril concentration and the possible impact of annular phospholipids in binding. Genetic responses of membrane proteins involved in texture and fat perception were detected at the mRNA level by RT-qPCR assay and both mechano- and chemosensing proteins displayed responses highlighting an interface dependent interaction. The outcomes of this study provide a basis for understanding the changing physicochemical properties of PNFs and their effect on flavor perception by altering mouthfeel and fat properties. This knowledge is important in the development of plant-based texture enhancers for sensory-appealing foods that require consumer acceptance and further promote healthy diets. Full article

The glycocalyx is a brush-like layer that covers the surfaces of the membranes of most cell types. It consists of a mixture of carbohydrates, mainly glycoproteins and proteoglycans. Due to its structure and sensitivity to environmental conditions, it represents a complicated object to investigate. Here, we review studies of the glycocalyx conducted using scanning probe microscopy approaches. This includes imaging techniques as well as the measurement of nanomechanical properties. The nanomechanics of the glycocalyx is particularly important since it is widely present on the surfaces of mechanosensitive cells such as endothelial cells. An overview of problems with the interpretation of indirect data via the use of analytical models is presented. Special insight is given into changes in glycocalyx properties during pathological processes. The biological background and alternative research methods are briefly covered. Full article

The mechanical properties of yeast play an important role in many biological processes, such as cell division and growth, maintenance of internal pressure, and biofilm formation. In addition, the mechanical properties of cells can indicate the degree of damage caused by antifungal drugs, as the mechanical parameters of healthy and damaged cells are different. Over the past decades, atomic force microscopy (AFM) and micromanipulation have become the most widely used methods for evaluating the mechanical characteristics of microorganisms. In this case, the reliability of such an estimate depends on the choice of mathematical model. This review presents various analytical models developed in recent years for studying the mechanical properties of both cells and their individual structures. The main provisions of the applied approaches are described along with their limitations and advantages. Attention is paid to the innovative method of low-invasive nanomechanical mapping with scanning ion-conductance microscopy (SICM), which is currently starting to be successfully used in the discovery of novel drugs acting on the yeast cell wall and plasma membrane. Full article