Search Results (171)

In familial Alzheimer’s disease (FAD), presenilin 1 (PSEN1) E280A cholinergic-like neurons (ChLNs) induce aberrant secretion of extracellular amyloid beta (eAβ). How PSEN1 E280A ChLNs-eAβ affects microglial activity is still unknown. We obtained induced microglia-like cells (iMG) from human peripheral blood cells (hPBCs) in a 15-day differentiation process to investigate the effect of bolus addition of Aβ42, PSEN1 E280A cholinergic-like neuron (ChLN)-derived culture supernatants, and PSEN1 E280A ChLNs on wild type (WT) iMG, PSEN1 E280A iMG, and sporadic Alzheimer’s disease (SAD) iMG. We found that WT iMG cells, when challenged with non-cellular (e.g., lipopolysaccharide, LPS) or cellular (e.g., Aβ42, PSEN1 E280A ChLN-derived culture supernatants) microenvironments, closely resemble primary human microglia in terms of morphology (resembling an “amoeboid-like phenotype”), expression of surface markers (Ionized calcium-binding adapter molecule 1, IBA-1; transmembrane protein 119, TMEM119), phagocytic ability (high pHrodo™ Red E. coli BioParticles™ phagocytic activity), immune metabolism (i.e., high generation of reactive oxygen species, ROS), increase in mitochondrial membrane potential (ΔΨm), response to ATP-induced transient intracellular Ca²⁺ influx, cell polarization (cluster of differentiation 68 (CD68)/CD206 ratio: M1 phenotype), cell migration activity according to the scratch wound assay, and especially in their inflammatory response (secretion of cytokine interleukin-6, IL-6; Tumor necrosis factor alpha, TNF-α). We also found that PSEN1 E280A and SAD iMG are physiologically unresponsive to ATP-induced Ca²⁺ influx, have reduced phagocytic activity, and diminished expression of Triggering Receptor Expressed on Myeloid Cells 2 (TREM2) protein, but when co-cultured with PSEN1 E280A ChLNs, iMG shows an increase in pro-inflammatory phenotype (M1) and secretes high levels of cytokines IL-6 and TNF-α. As a result, PSEN1 E280A and SAD iMG induce apoptosis in PSEN1 E280A ChLNs as evidenced by abnormal phosphorylation of protein TAU at residue T205 and cleaved caspase 3 (CC3). Taken together, these results suggest that PSEN1 E280A ChLNs initiate a vicious cycle between damaged neurons and M1 phenotype microglia, resulting in excessive ChLN death. Our findings provide a suitable platform for the exploration of novel therapeutic approaches for the fight against FAD. Full article

In the context of critical challenges in curcumin-modified polyurethane synthesis—including limited curcumin bioavailability and suboptimal biodegradability/biocompatibility—a novel polyurethane material (Cur-PU) with good mechanical, shape memory, pH-responsive, and biocompatibility was synthesized via a one-pot, two-step synthetic protocol in which HO-PCL-OH served as the soft segment and curcumin was employed as the chain extender. The experimental results demonstrate that with the increase in Cur units, the crystallinity of the Cur-PU material decreases from 32.6% to 5.3% and that the intensities of the diffraction peaks at 2θ = 21.36°, 21.97°, and 23.72° in the XRD pattern gradually diminish. Concomitantly, tensile strength decreased from 35.5 MPa to 19.3 MPa, and Shore A hardness declined from 88 HA to 65 HA. These observations indicate that the sterically hindered benzene ring structure of Cur imposes restrictions on HO-PCL-OH crystallization, leading to lower crystallinity and retarded crystallization kinetics in Cur-PU. As a consequence, the material’s tensile strength and hardness are diminished. Except for the Cur-PU-3 sample, all other variants exhibited exceptional shape-memory functionality, with R_f and R_r exceeding 95%, as determined by three-point bending method. Analogous to pure curcumin solutions, Cur-PU solutions demonstrated pH-responsive chromatic transitions: upon addition of hydroxide ion (OH⁻) solutions at increasing concentrations, the solutions shifted from yellow-green to dark green and finally to orange-yellow, enabling sensitive pH detection across alkaline gradients. Hydrolytic degradation studies conducted over 15 weeks in air, UPW, and pH 6.0/8.0 phosphate buffer solutions revealed mass loss <2% for Cur-PU films. Surface morphological analysis showed progressive etching with the formation of micro-to-nano-scale pores, indicative of a surface-erosion degradation mechanism consistent with pure PCL. Biocompatibility assessments via L929 mouse fibroblast co-culture experiments demonstrated ≥90% cell viability after 72 h, while relative red blood cell hemolysis rates remained below 5%. Collectively, these findings establish Cur-PU as a biocompatible material with tunable mechanical properties, and pH responsiveness, underscoring its translational potential for biomedical applications such as drug delivery systems and tissue engineering scaffolds. 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 Clinical Significance: Kaposi sarcoma (KS) is a rare angio-proliferative mesenchymal tumor that predominantly affects the skin and mucous membranes but may involve lymph nodes and visceral organs. Clinically, it manifests as red-purple-brown papules, nodules, or plaques, either painless or painful, often with disfiguring potential. The diagnosis is traditionally based on clinical and histopathological evaluation, although non-invasive imaging techniques are increasingly used to support diagnosis and treatment monitoring. We report a case of HHV-8-negative Kaposi sarcoma evaluated with multiple non-invasive imaging modalities to highlight their diagnostic utility. Case Presentation: An 83-year-old man presented with multiple painful, violaceous papulo-nodular lesions, some ulcerated, on the lateral aspect of his left foot. Dermoscopy revealed the characteristic rainbow pattern. Dynamic Optical Coherence Tomography (D-OCT) allowed real-time visualization of microvascular abnormalities, identifying large serpentine and branching vessels with clearly delineated capsules. Line-field Optical Coherence Tomography (LC-OCT) showed irregular dermal collagen, vascular lacunae, and the presence of spindle cells and slit-like vessels. Histological analysis confirmed the diagnosis of Kaposi sarcoma, revealing a proliferation of spindle-shaped endothelial cells forming angulated vascular spaces, with red blood cell extravasation and a mixed inflammatory infiltrate. Conclusions: Non-invasive imaging tools, including dermoscopy, D-OCT, and LC-OCT, have emerged as valuable adjuncts in the diagnosis and monitoring of KS. These techniques enable in vivo assessment of vascular architecture and tissue morphology, enhancing clinical decision-making while reducing the need for immediate biopsy. Dermoscopy reveals polychromatic vascular features, such as the rainbow pattern, while D-OCT and LC-OCT provide high-resolution insights into vascular proliferation, tissue heterogeneity, and cellular morphology. Dermoscopy, dynamic OCT, and LC-OCT represent promising non-invasive diagnostic tools for the assessment of Kaposi sarcoma. These technologies provide detailed morphological and vascular information, enabling earlier diagnosis and more personalized management. While histopathology remains the gold standard, non-invasive imaging offers a valuable complementary approach for diagnosis and follow-up, particularly in complex or atypical presentations. Ongoing research and technological refinement are essential to improve accessibility and clinical applicability. Full article

Label-free, high-throughput, and 3D morphological analysis of blood cells remains a major challenge in biomedical optics. In this study, we investigate this issue using flow cytometry with digital holographic microscopy (DHM) to enable real-time, label-free imaging of red blood cells (RBCs) and blood coagulation structures (BCSs) without the need for staining or chemical pretreatment. We demonstrate an approach for the automated quantification and statistical characterization of these cells using quantitative phase information reconstructed from digital holograms. Although established image processing techniques such as phase unwrapping and segmentation are used, this study presents, to the best of our knowledge, the first statistical characterization of the 3D morphological features of BCSs. This is particularly useful in analyzing the heterogeneous and complex 3D structures of BCSs, which are difficult to assess using conventional microscopy. The results suggest that this DHM-based flow cytometry system provides a promising platform for non-invasive, real-time morphological evaluation of blood samples and has potential applications in hematological diagnostics and research related to blood coagulation. Full article

The spleen plays a critical role in the immune and hematopoietic systems of teleost fish, functioning as a major secondary lymphoid organ. This study provides a detailed morphological and ultrastructural assessment of the spleen in goldfish (Carassius auratus), focusing on its immunological organization and cellular diversity. Through light and transmission electron microscopy, we examined red and white pulps, identifying key features such as melanomacrophage centers (MMCs), ellipsoids, and various immune cell types. The red pulp was rich in sinusoidal capillaries and splenic cords, whereas the white pulp housed lymphocytes, dendritic cells, macrophages, and telocytes, all contributing to immune regulation. Notably, ellipsoids were surrounded by reticular and macrophage sheaths, forming a filtration barrier against pathogens. Ultrastructural analysis revealed diverse immune cells with active morphological traits, including macrophages with pseudopodia and pigment granules, dendritic cells with dendrite-like extensions, and epithelial reticular cells involved in forming the blood–spleen barrier. These findings highlight the complex immunological microarchitecture of the goldfish spleen and its functional relevance in teleost immune responses. 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

Fish red blood cells (RBCs) are nucleated, transcriptionally active, and key players in both gas transport and immune responses. They are the primary targets of Orthoreovirus piscis (PRV), the etiological agent of heart and skeletal muscle inflammation (HSMI), which includes three genotypes (PRV-1, PRV-2, and PRV-3), linked to circulatory disorders in farmed salmon. In Chile, PRV-3 affects the coho salmon (Oncorhynchus kisutch), but host–pathogen interactions remain poorly characterized. This study compared the interactions of PRV-3 in coho salmon and PRV-1 in Atlantic salmon (Salmo salar) using RBC infection models. RBCs were isolated from healthy juvenile salmon (n = 3) inoculated with either PRV-1 (Ct = 18.87) or PRV-3 (Ct = 21.86). Poly I:C (50 µg/mL) was used as a positive control for the antiviral response. Cells were monitored for up to 14 days post-infection (dpi). PRV-3 infection in coho salmon RBCs caused significant metabolic disruption, apoptosis from 7 dpi, and correlated with increasing viral loads. In contrast, PRV-1 infection in Atlantic salmon RBCs showed limited apoptosis and maintained cell viability. Coho salmon RBCs upregulated rig-i, mx, and pkr transcripts, indicating activation of the type I interferon pathway, whereas Atlantic salmon RBCs exhibited a more attenuated response. PRV-3 induced notable morphological changes in coho salmon RBCs, although neither PRV-3 nor PRV-1 caused hemolysis. These findings highlight species-specific differences in RBC responses to PRV infection and provide new insights into the pathogenesis of PRV-3 and PRV-1. Full article

Background/Objectives: The prevalence of obesity worldwide has rapidly increased. Numerous studies showed a beneficial effect of probiotics in obese individuals, and changes in hematological parameters are observed in obesity. Therefore, the aim of this study was to investigate the effect of a novel probiotic approach on the red blood cells (RBCs) and platelets. Methods: Twenty-five obese women participated in a randomized, placebo-controlled study and were divided into the experimental group (one capsule daily containing Lactiplantibacillus plantarum 299v (DSM9843), Saccharomyces cerevisiae var. boulardii, and 40 mg octacosanol; n = 13) and the placebo group (n = 12). Blood samples were collected for light microscopic examination, morphometric analysis, and an automated hematology analyzer. A possible relationship between hematological parameters and body mass index (BMI), a common indicator of obesity, was investigated using Spearman correlation. The plasma concentration of soluble P-selectin and fibrinogen were determined using an ELISA assay. All measurements were performed before (T0) and after 12 weeks of supplementation (T1). Results: The three-month supplementation of probiotics improved hemoglobin levels, chromic status, and red blood cell morphology. The mean platelet volume (MPV), a measure of platelet size, was restored to normal levels, platelet morphology was improved, and the number of activated platelets was significantly reduced (p < 0.05). A strong negative correlation (r = −0.5904, p < 0.05) was found between BMI and platelet distribution width (PDW), a measure of variation in platelet size and shape. Conclusions: The results show that the probiotic approach improves morphology and normalizes the values of disturbed hematological parameters of RBCs and platelets in obese women. 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: Splenic B-cell lymphomas and leukaemias include hairy cell leukaemia, splenic marginal zone lymphoma, splenic diffuse red pulp small B-cell lymphoma, and splenic B-cell lymphoma/leukaemia with prominent nucleoli. The main diagnostic challenge is to differentiate between splenic B-cell lymphomas and leukaemias due to highly overlapping clinical, morphologic, and phenotypic features in the absence of splenectomies for diagnostic purposes. Case presentation: We describe a case of a 78-year-old woman who presented with weight loss and was subsequently found to have pancytopenia, lymphocytosis, and splenomegaly. Peripheral blood smear showed a homogenous population of small- to medium-sized lymphocytes having oval nuclei, condensed chromatin, and villous cytoplasmic projections. Bone marrow showed B-cell infiltrate in a predominantly intrasinusoidal pattern. Except for cyclin D1 and BRAF, the immunophenotype was similar to that of hairy cell leukaemia. This was further supported by the lack of BRAF p.V600E mutation by polymerase chain reaction. A diagnosis of splenic diffuse red pulp small B-cell lymphoma was thus favored based on the lack of cyclin D1 expression and pattern of infiltration in the bone marrow biopsy. Conclusions: Awareness of this infrequent clinical presentation and immunophenotype of splenic diffuse red pulp small B-cell lymphoma is crucial for diagnosis and devising appropriate therapeutic strategies for the patient. 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

Background: Capillary tortuosity is a morphological variant of microcirculation. However, the mechanisms by which tortuous vessels meet metabolic requirements in health and disease remain unknown. We recently reported that capillary tortuosity score (CTS) is significantly higher in patients with septic shock than in steady-state individuals, and that CTS is significantly associated with alveolar-to-arterial oxygen (A-a O₂) gradient and oxygen debt in septic shock patients. Objective: We aimed to investigate the characteristics of the magnetic fields in the sublingual microcirculation of individuals with normal physiology and patients with septic shock. Methods: Systemic hemodynamics were recorded, and sublingual microcirculation was monitored using sidestream dark field (SDF+) imaging. The number of capillary red blood cells (N_RBC), the intensity of the magnetic field of a red blood cell (H_RBC), the intensity of the magnetic field of each capillary (H_CAP), and the intensity with which the magnetic field of a capillary acts on an RBC (F_CAP) were calculated. Results: Significant differences in macro- and microhemodynamic variables were observed between the two groups. Although N_RBC was significantly higher in individuals with steady-state physiology [87.4 (87.12) vs. 12.23 (6.9)], H_RBC was significantly stronger in patients with septic shock [5.9 × 10⁻¹⁶ (6.9 × 10⁻¹⁶) A m⁻¹ vs. 1.6 × 10⁻¹⁵ (1.4 × 10⁻¹⁵) A m⁻¹]. No significant difference was observed in H_CAP [2.16 × 10⁻¹⁴ (2.17 × 10⁻¹⁴) A m⁻¹ vs. 1.34 × 10⁻¹⁴ (1.23 × 10⁻¹⁴) A m⁻¹] and F_CAP [1.66 × 10⁻²⁴ (3.36 × 10⁻²⁴) A m⁻¹ vs. 6.44 × 10⁻²⁵ (1.1 × 10⁻²⁴) A m⁻¹] between the two groups. In patients with septic shock, H_RBC was associated with De Backer score (rho = −0.608) and venous–arterial carbon dioxide difference (rho = 0.569). In the same group, H_CAP was associated with convective oxygen flow (rho = 0.790) and oxygen extraction ratio (rho = −0.596). Also, F_CAP was significantly associated with base deficit (rho = 0.701), A-a O₂ gradient (rho = 0.658), and oxygen debt (rho = −0.769). Conclusions: Despite the microcirculatory impairment in patients with septic shock, H_RBC was significantly stronger in that group than in steady-state individuals. Also, H_CAP and F_CAP were comparable between the two groups. Tortuous vessels may function as biomagnetic coils that amplify RBC-induced magnetic fields, enhancing perfusion and oxygenation of adjacent tissues. Full article

Silver nanoparticles (AgNPs), synthesised using Chlorella vulgaris algal extract and silver nitrate, are studied in medicine for their antibacterial properties in poultry. This study assessed the effect of AgNPs on bacterial inhibition and early development and blood parameters in Ross 308 chicken embryos. AgNPs were characterised using transmission electron microscopy, scanning electron microscopy with a focused ion beam, UV–Vis spectroscopy, and a zetasizer. The antibacterial properties of the AgNP colloid against S. enterica were assessed using minimal inhibitory concentration, minimal bacterial concentration, and PrestoBlue assays. AgNP colloid (2 mg/L) was injected into egg albumen on day 0. Chicken embryos were incubated for 3 and 16 d. The effect of AgNPs on 3 d old embryos was evaluated based on mortality and somite count using the Hamburger–Hamilton classification. For older embryos, mortality, dimensions, anatomical changes, organ mass, plasma liver enzymes and antioxidants, and red blood cell morphology were determined. Blood samples from the control group embryos were assessed for the impact of AgNPs on hemolysis. AgNPs inhibited S. enterica growth at concentrations >6.75 mg/L. A 3 d exposure to AgNPs caused an insignificant decrease in the number of somites without affecting embryo mortality. However, a 16 d exposure to AgNPs reduced live embryos and plasma antioxidants, changed the levels of ALT, AST, and GGT, altered red blood cell morphology, and caused hemolysis. Toxicity of AgNPs was model-dependent, whereby the chicken embryo was more sensitive to AgNPs than the bacterium. Full article

Tungsten disulfide (WS₂) nanoparticles have emerged in the biomedical field as potential theranostic agents due to their unique properties, including biocompatibility. However, their impact on the immune response remains unexplored. This study aimed to evaluate the effects of inorganic fullerene-like WS₂ (IF-WS₂) nanostructures on human peripheral blood mononuclear cells (PBMCs) in vitro. The study investigated several parameters to evaluate the effects of IF-WS₂ nanoparticles. Cytotoxicity was assessed by measuring cell viability, apoptosis, and necrosis. Internalization of IF-WS₂ by PBMCs was analyzed using morphological and flow cytometric techniques. Proliferation was studied in CellTrace Far Red-prestained total PBMCs stimulated with phytohemagglutinin (PHA) and in isolated T cell cultures stimulated with CD3/CD28-coated beads. Additionally, the production of cytokines and chemokines was measured in culture supernatants of total PBMCs and T cells. IF-WS₂ nanoparticles were non-cytotoxic up to a concentration of 200 µg/mL. Concentrations ≥25 µg/mL inhibited PHA-stimulated PBMC proliferation but did not affect T cell proliferation. Morphological and flow cytometric analysis demonstrated dose- and time-dependent internalization of IF-WS₂ by macrophages. Additionally, IF-WS₂ significantly reduced the production of pro-inflammatory cytokines (IL-1β, TNF-α, IL-8, MCP-1, and GRO-α) in PHA-stimulated PBMCs. Th1, Th17, and Th21 cytokines were downregulated, while Th2, Th9, and T regulatory cytokines were upregulated. In conclusion, this study demonstrated for the first time that pristine IF-WS₂ nanoparticles, at non-cytotoxic concentrations, exhibit notable anti-inflammatory and immunomodulatory properties on activated PBMCs in vitro. Full article