Search Results (130)

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

The white snook (Centropomus viridis) is an emerging aquaculture species with high market acceptance, exhibiting catadromous and protandric hermaphroditic characteristics in adulthood. This study aimed to preliminarily characterize certain hematological and biochemical parameters, as well as blood cell morphology, for identifying possible variations between sexes maintained under aquaculture recirculating system (RAS) conditions. The white snook broodstock was anesthetized with clove oil, and biometric values, as well as sex classification, were measured. Then, blood samples were collected from 14 females (7132 ± 1610 g) and 20 males (2200 ± 0.963 g) via caudal vessel puncture to analyze selected hematological parameters, blood biochemistry, and cellular morphology. Fulton’s condition factor (K) showed no differences between sexes, indicating a healthy fish status. Females showed significantly higher serum cholesterol, glucose, and triglyceride levels than males. Also, hematocrit (HCT) and mean corpuscular volume (MCV) were elevated in females. No sex-related differences were observed in red or white cell counts or in blood cell dimensions. Morphological characterization identified erythrocytes, thrombocytes, and three types of leukocytes: lymphocytes (small and large lymphocytes), neutrophils, and monocytes, with no eosinophils or basophils detected in either sex. These findings provide fundamental reference values for the hematological and biochemical profiles of C. viridis broodstock in captivity and highlight sex-specific differences relevant for reproductive and health monitoring. However, it should be considered that the sample size used to establish reference ranges for the species is small, so it is recommended to implement a monitoring plan for this and other broodstocks of this emerging species. Full article

Background and Objectives: Malaria is a disease that can result in a variety of complications. Diagnosis is carried out by an optical microscope and depends on operator experience. The use of artificial intelligence to identify morphological patterns in erythrocytes would improve our diagnostic capability. The object of this study was therefore to establish computer viewing models able to classify blood cells infected with Plasmodium spp. to support malaria diagnosis by optical microscope. Materials and Methods: A total of 27,558 images of human blood sample extensions were obtained from a public data bank for analysis; half were of parasite-infected red cells (n = 13,779), and the other half were of uninfected erythrocytes (n = 13,779). Six models (five machine learning algorithms and one pre-trained for a convolutional neural network) were assessed, and the performance of each was measured using metrics like accuracy (A), precision (P), recall, F1 score, and area under the curve (AUC). Results: The model with the best performance was VGG-19, with an AUC of 98%, accuracy of 93%, precision of 92%, recall of 94%, and F1 score of 93%. Conclusions: Based on the results, we propose a convolutional neural network model (VGG-19) for malaria diagnosis that can be applied in low-complexity laboratories thanks to its ease of implementation and high predictive performance. Full article

Background: Physical activity in elderly adults reduces the risk of numerous diseases, enhances their quality of life, and improves their physical performance and overall strength. This study aimed to evaluate the effects of smovey^® exercise on morphological and rheological blood parameters in a group of women over 60 years of age. Methods: The study included 30 women aged 71.08 ± 6.41 years, with a control group of 15 age-matched women. Participants in the experimental group underwent a three-month smovey^® training program, consisting of two 45 min sessions per week. The control group did not engage in any regular physical activity. Blood samples were collected from the experimental group before and after the training period, while in the control group, blood was collected once. Results: After three months of smovey^® training, a statistically significant increase in red blood cell, hemoglobin, and platelet counts was observed. Smovey^® training also enhanced erythrocyte deformability at the lowest shear force. Conclusions: Smovey^® training leads to changes in the morphological properties of blood in elderly women, leading to increases in RBC, HGB, PLT, and MCHC indices. Additionally, smovey^® training improves erythrocyte deformability at a shear stress of 0.30 [Pa], reducing the half-time of complete aggregation. Full article

Background: In vitro blood circulation loop systems are utilized to assess the hemocompatibility and performance of medical devices that come into contact with blood, in accordance with the international standards ASTM F1830 and ASTM F1841. However, a method for evaluating the specific type of anticoagulant and the blood characteristics of each animal species is necessary to ensure consistent and reliable results. Methods: Blood was collected from healthy rabbits, pigs, rhesus monkeys, and cynomolgus monkeys to evaluate whole blood preserved in anticoagulants (ACD-A, CPDA-1, and heparin). For each sample, red blood cells were monitored over time, and their morphological characteristics were documented. Results: The morphological grade of erythrocytes gradually decreased over time. Significant differences were observed based on the type of anticoagulant used in the experiment, and variations were noted among different animal species. Conclusions: The hemocompatibility of in vitro blood circulation loop systems may vary depending on the animal species. Observing erythrocyte morphology can serve as a control measure to ensure reproducible results. Full article

Analyses of the hematological statuses of animals inhabiting areas of anthropogenic pollution may provide valuable insights into the extent of disturbance of their living conditions and the mechanisms of adaptation to various environmental stressors. The current work compares the erythrogram and erythrocyte-metric parameters of marsh frogs (Pelophylax ridibundus) inhabiting the polluted sedimentation lake of Brikel TPP in southern Bulgaria to those in frogs inhabiting a relatively clean habitat (reference population). The study includes a total of 120 individuals (30 females and 30 males from each site). For all of them, total erythrocyte count, hemoglobin concentration, hematocrit, MCV, MCH, and MCHC were evaluated via standard laboratory techniques. All erythrocyte metrics were determined microscopically in two blood smears from each animal. Our study reveals alterations in the erythrogram parameters and the erythrocyte sizes in marsh frogs living in conditions of chronic pollution with industrial wastewater compared to the animals from the reference site. The mean values for all erythrocyte morphology parameters are significantly lower in both female and male individuals inhabiting the polluted area compared to those originating from the reference one. Conversely, three erythrogram parameters—erythrocyte count, hemoglobin, and hematocrit—appeared significantly higher in females and males from the polluted site. The observed changes in the erythrogram parameters and erythrocyte sizes result from the deteriorated water quality of the sedimentation lake. Full article

Erythrocytes, the most abundant blood cells, are a prevalent cell model for the analysis of the membrane-damaging effects of different molecules, including drugs. In response to stimuli, erythrocytes can change their morphology, e.g., shape or volume, which in turns influences their main function to transport oxygen. Membrane active molecules can induce hemolysis, i.e., release of hemoglobin into the blood plasma. Free hemoglobin in the blood circulation is toxic causing serious health problems including vasoconstriction, high blood pressure and kidney damage. Therefore, early recognition of the risk of massive hemolysis is highly important. Here, we investigated surfactant induced hemolysis applying UV–vis spectrophotometry. Saponin, sodium dodecyl sulfate and Triton X-100, detergents known to provoke hemolysis at different concentrations and by different mechanisms, were applied to initiate the process. Whole absorption spectra of erythrocyte suspensions in the range 300–750 nm were recorded every 15 s for following the process in real-time. The hemolysis process, with respect to morphological changes in the erythrocytes and their influence on the oxygenation state of hemoglobin, was characterized by the absorbance at 700 nm, the height relative to the background and the wavelength of the Soret peak. The results suggest that these UV–vis spectrophotometry parameters provide reliable information in real-time; not only about the process of hemolysis itself, but also about pre-hemolytic changes in the erythrocytes, even at sub-hemolytic surfactant concentrations. Full article

Antimicrobial peptides (AMPs) are biocompatible and biodegradable, making them an attractive alternative to traditional antimicrobial agents and chemical preservatives. Here, a novel α-helix amphiphilic anionic AMP Lc149 was screened from a large yellow croaker (Larimichthys crocea) using a Bacillus subtilis expression system. Lc149 is a hypothesized protein fragment not annotated in the genome of a large yellow croaker. Both extracellular protein and recombinant Lc149 (rLc149) exhibited significant killing effects against Gram-negative Escherichia coli and Vibrio harveyi. Scanning and transmission electron microscopy revealed that rLc149 had the ability to disrupt bacterial cell membranes, causing irregular cell morphology, severe cell membrane damage, cytoplasm agglutination, and intracellular content leakage. Confocal laser scanning microscopy and flow cytometry further confirmed bacterial cell destruction and mortality rates of over 80%. Gel retardation assays and SDS-PAGE electrophoresis showed that rLc149 was unable to bind to bacterial DNA, but did reduce bacterial protein contents. Additionally, rLc149 maintained antibacterial activity against E. coli and V. harveyi upon exposure to temperatures of 25–100 °C, UV radiation time of 0–60 min, pH levels of 3–12, and different proteases. Biosafety assays revealed low hemolytic toxicity to erythrocytes of large yellow croaker, rabbit, and shrimp, and low cytotoxicity to large yellow croaker kidney cells and HEK 293T cells. More deeply, rLc149 also possessed significant killing activity against parasites. Therefore, rLc149 can be considered an antibacterial and antiparasitic drug in fisheries. Full article

Morphological changes in erythrocytes during disease, aging, or reactions to external agents are significant as they can influence disease progression. However, the exact mechanisms behind these temporary alterations and their potential to cause dysfunction remain unclear. Using a saponin-induced erythrocyte shape transition (EST) model, we studied the gradual shift of erythrocytes towards echino-stomato-spherocytic forms and its link to hemolysis and thrombosis. We observed that different saponin concentrations elicited varying shape transformations. At low concentrations, erythrocytes transition from discocytic shapes to echinocytic, echino-stomatocytic, and ultimately stomatocytic forms. As the concentration moderately increases, the morphology evolves into stomato-spherocytic forms. At higher saponin concentrations, the erythrocytes completely transform into spherocytic forms. Regardless of the transformation degree, all forms showed increased phosphatidylserine exposure (PS) and microvesicle (MV) production, primarily due to increased scramblase and decreased flippase activity, which were influenced by elevated calcium levels and caspase 3 activity, effectively managing PS distribution and influencing cell membrane expansion and invagination. These alterations increased thrombin production, erythrocyte adhesion, and aggregation, promoting thrombosis in rats. Altogether, our findings indicate that the shift towards echino-stomato-spherocytic forms fosters a hypercoagulable state through PS externalization, heightening thrombotic risk. Full article

Dogs have multiple blood type antigens, among which DEA 1, DEA 4, and Dal can induce severe acute hemolytic transfusion reactions. Various antigen modulation techniques have been developed to reduce immunogenicity and transfusion reactions. Recently, trypsin has been suggested as a potential tool for modulating the antigenicity of DEA 1 in veterinary medicine. Following this rationale, this study aims to evaluate the effects of trypsin on the antigenicity of these three antigens. A 50% RBC suspension treated with 1 mg/mL trypsin was incubated at 37 °C for 120 min. The antigenicity of DEA 1, DEA 4, and Dal was assessed using blood typing assays before and after trypsin treatment. As a result, trypsin did not reduce the antigenicity of DEA 1 and DEA 4; instead, trypsin significantly increased their antigenicity (p = 0.008) and promoted agglutination, whereas Dal exhibited a significant reduction in antigenicity (p = 0.008). Quantitative morphological parameters obtained from an automated hematology analyzer revealed no significant differences between trypsin-treated and negative control groups. However, morphological scoring under an optical microscope showed significantly fewer echinocytes in the trypsin-treated group (p = 0.008). Consequently, broad-spectrum proteases like trypsin are unsuitable for universal blood production due to their variable effects on erythrocyte surface antigens. Full article

An important area of health is health promotion. A healthy lifestyle supports health improvement and early prevention of chronic diseases. Stimulation of the body by cold water swimming and swimming in a swimming pool can lead to adaptive changes beneficial for the human cardiovascular system. Within the winter swimming season of 2023/2024, for a period of 5 months, from November to March, once a week, study participants (n = 30; n = 15 females and n = 15 males) from the Krakow Society of Winter Swimmers ‘Kaloryfer’ in Krakow (Poland) practiced winter swimming in cold water (4–5 °C) and swam in the sports pool of the University of Physical Culture in Krakow in water at a temperature of 28 °C. After a full season of winter swimming and swimming pool sessions, both males and females exhibited a tendency towards lower erythrocyte (p = 0.002), leukocyte (p < 0.001), and platelet counts (p < 0.001), as well as an increase in blood plasma viscosity (within normal limits) (p = 0.001), without any changes in blood aggregation or fibrinogen indicators. The remaining morphological indicators and the elongation index demonstrated only limited variation. Winter swimming induces positive changes in blood morphology and rheology. Full article

Baru nut oil (Dipteryx alata Vogel) is a lipidic extract from a species endemic to the Cerrado biome, renowned for its antioxidant potential. This study aimed to develop a nanoemulsion containing baru nut oil (BNON) using lecithin and polysorbate 80, and to evaluate its antioxidant activity and cytotoxicity. The physicochemical properties of BNON were characterized, and its cytotoxicity was assessed in human erythrocytes and keratinocytes. Antioxidant activity was evaluated using the DPPH method and inhibition of AAPH-induced hemolysis. BNON exhibited a droplet size of 530.1 ± 20.48 nm, a polydispersity index of 0.496 ± 0.057, and a zeta potential of −35.7 ± 2.19 mV. Free baru nut oil showed no cytotoxicity to keratinocytes or erythrocytes within the concentration ranges tested (1.0–0.031 mg/mL and 0.8–0.006 mg/mL, respectively). In contrast, BNON displayed cytotoxic effects on keratinocytes and erythrocytes only at the highest tested concentration. Atomic force microscopy analysis of erythrocytes from the hemolysis assay revealed normal morphology for cells treated with free oil at 0.8 mg/mL, whereas cells treated with BNON at the same concentration exhibited a slightly widened concave center. Free oil at 0.8 mg/mL significantly protected erythrocytes from AAPH-induced hemolysis, while BNON did not. However, BNON (5 mg/mL) demonstrated free radical scavenging activity, quantified at 0.0074 mg Trolox equivalents/mg via the DPPH assay. These findings suggest that baru nut oil has potential as an antioxidant product, although further optimization of the nanoformulation is required. Full article

Background/Objectives: In recent years the global incidence of cancer during pregnancy is rising, occurring in 1 out of every 1000 pregnancies. In this regard, the most used chemotherapy drugs to treat cancer are alkylating agents such as cyclophosphamide (Cp). Despite its great efficacy, has been associated with the production of oxidative stress and DNA damage, leading to embryotoxicity, genotoxicity, and teratogenicity in the developing conceptus. Therefore, this study aimed to investigate the protective role of phycobiliproteins (PBP) derived from Arthrospira maxima (spirulina) in reducing Cp-induced embryotoxicity and genotoxicity in pregnant CD1 mice. Methods: Pregnant CD1 mice were divided into five groups: control, Cp 20 mg/kg, and three doses of PBP (50, 100, and 200 mg/kg) + Cp co-treatment. PBP were administered orally from day 6 to 10.5 dpc, followed by a single intraperitoneal dose of Cp on 10.5 dpc. Embryos were collected at 12.5 dpc to assess morphological development and vascular alterations, while maternal DNA damage was evaluated using micronucleus assays and antioxidant enzyme activity in maternal plasma. Results: PBP exhibited a dose-dependent protective effect against Cp-induced damage. The 200 mg/kg PBP dose significantly reduced developmental abnormalities, micronucleated polychromatic erythrocytes, and oxidative stress, (as evidenced by increased SOD and GPx activity). Conclusions: Phycobiliproteins from Arthrospira maxima (spirulina) effectively reduced Cp-induced morphological and vascular alterations in embryos and genotoxicity in pregnant mice. These findings highlight their potential as a complementary therapy to mitigate teratogenic risks during chemotherapy. Further research is needed to optimize dosing and explore clinical applications. Full article

Studying the blood cell morphology of marine mammals provides an opportunity to elucidate the physiological mechanisms of adaptive changes associated with the aquatic habitat that occur at the cellular level, as well as adaptations to changing environmental conditions and under various physiological and pathological processes. The Baikal seal [Pusa sibirica (family Phocidae)] is endemic to the freshwater Lake Baikal, but comprehensive hematology data are not available. We studied the morphological features of blood cells of twelve clinically normal, adult Baikal seals (n = 6 males, n = 6 females) from two oceanariums under professional care for eight years. The morphology of mature and immature erythrocytes and inclusions are described. The blood of Baikal seals is characterized by the presence of erythrocytes with a size of 8.2 ± 0.6 µm; Howell–Jolly bodies were rarely observed, the number of reticulocytes ranged from 4.1 to 93.1 × 10⁹/L, and nucleated erythrocytes were absent. The morphological features of neutrophils, eosinophils, basophils, and platelets were described. Inter-individual and sex differences in the counts of basophils, platelets, red blood cells, and levels of hemoglobin, the mean corpuscular volume, and the mean corpuscular hemoglobin concentration were statistically observed. The results could be useful for Baikal seal veterinary care, immune response research, and comparative studies with other pinnipeds. Full article

Oxidative stress is widely recognized as a key mechanism in the development of hypertension. Under pathological conditions, such as in hypertension, oxidative stress leads to irreversible posttranslational modifications of proteins, which result in loss of protein function and cellular damage. We have previously documented physiological and morphological changes across various blood and bone marrow cell lineages, all of which exhibit elevated oxidative stress. While cytoskeletal changes in erythrocytes have been well characterized in hereditary diseases, this is the first study, to our knowledge, to investigate cytoskeletal reorganization in erythrocytes from hypertensive patients. To this end, we compared the expression patterns and subcellular distribution of key cytoskeletal proteins in erythrocytes from hypertensive individuals with those from normotensive subjects using Western blot, flow cytometry, and confocal microscopy. Our results revealed the presence of three erythrocyte subpopulations with differential expression of glycophorin A. The persistent oxidative environment in hypertensive patients causes dysregulation in the expression of glycophorin A, Band 3 protein, protein 4.1, and ankyrin, as well as the reorganization of spectrin. These alterations in protein expression and distribution suggest that oxidative stress in hypertensive individuals may induce structural modifications, ultimately impairing erythrocyte membrane elasticity and function. Full article