Treatment of gingival fibroblasts with PDL extracellular vesicles results in promotion of Wnt signalling pathway and osteogenic differentiation. PDL secretome shows selective wound healing and matrix remodelling which can have implications for future periodontal regenerative strategies.

• MeSH
• Cell Differentiation MeSH
• Extracellular Vesicles * physiology   MeSH
• Fibroblasts physiology   MeSH
• Gingiva cytology   MeSH
• Wound Healing physiology   MeSH
• Humans MeSH
• Osteogenesis physiology   MeSH
• Periodontal Ligament * cytology   physiology   MeSH
• Regeneration * physiology   MeSH
• Wnt Signaling Pathway physiology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

BACKGROUND: Despite secondary prevention with aspirin, patients with stable cardiovascular disease (CVD) remain at elevated long-term risk of major adverse cardiovascular events. The Cardiovascular Outcomes in People Using Anticoagulant Strategies (COMPASS) double-blind, randomized clinical trial demonstrated that aspirin plus low-dose rivaroxaban (COMPASS regime) significantly decreased the incidence of major adverse cardiovascular events by 24% compared with aspirin alone. However, the mechanisms underlying these potential synergistic/nonantithrombotic effects remain elusive. Extracellular vesicles (EVs) are crucial messengers regulating a myriad of biological/pathological processes and are highly implicated in CVD. OBJECTIVES: We hypothesized that circulating EV profiles reflect the cardioprotective properties of the COMPASS regime. METHODS: A cohort of stable CVD patients (N = 40) who participated in the COMPASS trial and were previously randomized to receive aspirin were prospectively recruited and assigned a revised regimen of open-label aspirin plus rivaroxaban. Blood samples were obtained at baseline (aspirin only) and 6-month follow-up. Plasma EV concentration, size, and origin were analyzed by nanoparticle tracking analysis and flow cytometry. EVs were enriched by ultracentrifugation for proteomic analysis. RESULTS: The COMPASS regime fundamentally altered small (<200 nm) and large (200-1000 nm) EV concentration and size compared with aspirin alone. Crucially, levels of platelet-derived and myeloperoxidase-positive EVs became significantly decreased at follow-up. Comparative proteomic characterization further revealed a significant decrease in highly proinflammatory protein expression at follow-up. CONCLUSION: The observed changes in EV subpopulations, together with the differential protein expression profiles, suggest amelioration of an underlying proinflammatory and prothrombotic state upon dual therapy, which may be of clinical relevance toward understanding the fundamental mechanism underlying the reported superior cardiovascular outcomes associated with this antithrombotic regimen.

• MeSH
• Aspirin * administration & dosage   therapeutic use   adverse effects   MeSH
• Double-Blind Method MeSH
• Extracellular Vesicles * metabolism   drug effects   MeSH
• Platelet Aggregation Inhibitors * administration & dosage   adverse effects   therapeutic use   MeSH
• Factor Xa Inhibitors * administration & dosage   adverse effects   therapeutic use   MeSH
• Cardiovascular Diseases * blood   prevention & control   drug therapy   MeSH
• Drug Therapy, Combination * MeSH
• Middle Aged MeSH
• Humans MeSH
• Inflammation Mediators blood   MeSH
• Prospective Studies MeSH
• Proteomics methods   MeSH
• Rivaroxaban * administration & dosage   MeSH
• Aged MeSH
• Thrombosis blood   prevention & control   drug therapy   MeSH
• Treatment Outcome MeSH
• Inflammation blood   MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Randomized Controlled Trial MeSH

There is growing interest in the role of extracellular vesicles (EVs) in neonatal pathology. This study aimed to characterise circulating EVs following preterm birth. This single-centre prospective observational study included cord and postnatal plasma from preterm (n = 101) and full-term infants (n = 66). EVs were analysed using nanoparticle tracking analysis, flow cytometry, proteomics and procoagulant activity assay. We found changes in the concentration, size, cellular origin and proteomic content of circulating EVs in preterm infants during perinatal adaptation. To understand if these changes were related to prematurity or normal adaptation to extrauterine life, they were also investigated in term infants. There was a dramatic increase in the concentration of small and large EVs on Day 3 in the preterm group; specific subsets of platelet (CD42b+ and CD62P+), endothelial (VEGFR2) and tissue factor EVs were elevated. Differentially expressed proteins relating to haemostasis, pulmonary physiology and immunity were identified between Day 1 and 3 in preterm infants. These changes have never previously been described in a large cohort of preterm infants and differ from healthy term infants. These findings have major implications for future neonatal EV studies, particularly the timing of sample collection. Further work is required to understand the clinical implications of this unique EV profile following preterm birth.

• MeSH
• Extracellular Vesicles * metabolism   MeSH
• Adaptation, Physiological * MeSH
• Humans MeSH
• Infant, Premature * blood   MeSH
• Infant, Newborn MeSH
• Prospective Studies MeSH
• Proteomics methods   MeSH
• Pregnancy MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Infant, Newborn MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Observational Study MeSH

The use of microfluidic sperm sorting (MFSS) systems in infertility treatment is increasing due to their practicality and ease of use. While often presented as highly effective, their efficacy in patients with varying sperm analysis results remains uncertain. In this study, we evaluated the effectiveness of MFSS compared with the swim-up (SU) technique in terms of oxygen radical levels and spermiogram parameters. Samples from each patient were processed using both methods, followed by assessments of sperm concentration, motility, morphology, DNA integrity, acrosomal status, and mitochondrial membrane potential. Participants were selected based on sperm analysis and categorized as normozoospermic (n = 40) or non-normozoospermic (n = 28). An analysis of separation techniques revealed no significant differences, except for a lower percentage of DNA-fragmented sperm in the MFSS group compared with SU within the non-normozoospermic cohort (SU: 10.0% vs. MFSS: 5.69%, p = 0.027). No differences were observed between SU and MFSS in normozoospermic men. The MFSS method is a simple technique, frequently used in laboratories, that yields good results but does not offer a substantial advantage over SU. The primary benefit of MFSS appears to be a significant reduction in the proportion of sperm with DNA fragmentation compared with SU in patients with abnormal sperm analysis results.

• MeSH
• Semen Analysis methods   MeSH
• Adult MeSH
• DNA Fragmentation MeSH
• Sperm Injections, Intracytoplasmic * methods   MeSH
• Humans MeSH
• Membrane Potential, Mitochondrial MeSH
• Microfluidics * methods   MeSH
• Sperm Motility * MeSH
• Infertility, Male therapy   MeSH
• Cell Separation * methods   MeSH
• Spermatozoa * cytology   metabolism   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH

Mužská neplodnost je komplexní stav s mnoha etiologickými faktory, včetně hormonálních, anatomických, genetických a vlivů životního stylu. Přestože se mužský faktor podílí až na 40 % případů neplodnosti, přesná příčina zůstává často neznámá (tzv. idiopatická neplodnost). Současné diagnostické metody zahrnují detailní klinické vyšetření, analýzu ejakulátu, hormonální profil, genetické testování a zobrazovací techniky. Výrazný pokrok byl zaznamenán v oblasti testování fragmentace DNA spermií a měření oxidačního stresu, jež poskytují širší pohled na kvalitu spermií. Terapeutické postupy se liší podle příčiny neplodnosti – od farmakologické léčby a chirurgických intervencí až po asistovanou reprodukci. Rychle se rozvíjející výzkum v oblasti regenerativní a genové terapie slibuje nové možnosti léčby. Nedílnou součástí prevence i léčby je rovněž zdravý životní styl a management rizikových faktorů.

Male infertility is a multifactorial condition influenced by hormonal, anatomical, genetic, and lifestyle factors. Although the male factor contributes to up to 40% of infertility cases, the exact etiology often remains unknown (so-called idiopathic infertility). Current diagnostic approaches include detailed clinical evaluation, semen analysis, hormonal assessment, genetic testing, and imaging techniques. Significant progress has been achieved in sperm DNA fragmentation testing and oxidative stress evaluation, offering a broader insight into sperm quality. Treatment strategies vary depending on the underlying cause, ranging from pharmacological therapy and surgical interventions to assisted reproductive technologies. Rapid advances in regenerative medicine and gene therapy show promise for novel therapeutic options. A healthy lifestyle and risk factor management are integral to both prevention and treatment.

• MeSH
• Semen Analysis MeSH
• Andrology trends   MeSH
• Reproductive Techniques, Assisted MeSH
• Diagnostic Techniques and Procedures classification   MeSH
• DNA Fragmentation MeSH
• Humans MeSH
• Infertility, Male * diagnosis   etiology   therapy   MeSH
• Oxidative Stress MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Publication type
• Review MeSH

INTRODUCTION: The immunosuppressive roles of galectin-3 (Gal-3) in carcinogenesis make this lectin an attractive target for pharmacological inhibition in immunotherapy. Although current clinical immunotherapies appear promising in the treatment of solid tumors, their efficacy is significantly weakened by the hostile immunosuppressive tumor microenvironment (TME). Gal-3, a prominent TME modulator, efficiently subverts the elimination of cancer, either directly by inducing apoptosis of immune cells or indirectly by binding essential effector molecules, such as interferon-gamma (IFNγ). METHODS: N-(2-Hydroxypropyl)methacrylamide (HPMA)-based glycopolymers bearing poly-N-acetyllactosamine-derived tetrasaccharide ligands of Gal-3 were designed, synthesized, and characterized using high-performance liquid chromatography, dynamic light scattering, UV-Vis spectrophotometry, gel permeation chromatography, nuclear magnetic resonance, high-resolution mass spectrometry and CCK-8 assay for evaluation of glycopolymer non-toxicity. Pro-immunogenic effects of purified glycopolymers were tested by apoptotic assay using flow cytometry, competitive ELISA, and in vitro cell-free INFγ-based assay. RESULTS: All tested glycopolymers completely inhibited Gal-3-induced apoptosis of monocytes/macrophages, of which the M1 subtype is responsible for eliminating cancer cells during immunotherapy. Moreover, the glycopolymers suppressed Gal-3-induced capture of glycosylated IFNγ by competitive inhibition to Gal-3 carbohydrate recognition domain (CRD), which enables further inherent biological activities of this effector, such as differentiation of monocytes into M1 macrophages and repolarization of M2-macrophages to the M1 state. CONCLUSION: The prepared glycopolymers are promising inhibitors of Gal-3 and may serve as important supportive anti-cancer nanosystems enabling the infiltration of proinflammatory macrophages and the reprogramming of unwanted M2 macrophages into the M1 subtype.

• MeSH
• Acrylamides chemistry   pharmacology   MeSH
• Apoptosis drug effects   MeSH
• Galectin 3 * antagonists & inhibitors   MeSH
• Galectins MeSH
• Interferon-gamma * metabolism   MeSH
• Blood Proteins MeSH
• Humans MeSH
• Macrophages drug effects   MeSH
• Monocytes * drug effects   MeSH
• Tumor Microenvironment drug effects   MeSH
• Polymers * chemistry   pharmacology   MeSH
• Antineoplastic Agents * pharmacology   chemistry   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

The sodium/calcium exchanger (NCX) type 1 has been well described in various cancers, but little is known about the other two NCX types (NCX2 and NCX3). In this study, we used the selective blocker of NCX3 - YM-244769 to investigate changes in apoptosis induction, migration, proliferation, intracellular calcium and ATP in four cancer cell lines - DLD1, HeLa, MDA-MB-231 and JIMT1. In all four cell lines we observed a concentration-dependent increase in the number of apoptotic cells, as well as reduced migration and proliferation. Induction of hypoxic conditions did not alter the response of these cells to YM-244769 in any of the above-mentioned parameters. These results indicate the role of NCX3 in cancer cell migration, proliferation and apoptosis, as inhibition of NCX1 by the specific blocker SEA0400 had no significant effect on these parameters. However, we verified the effect of NCX3 inhibition by using CRISPR/Cas9 to generate clones in which the SLC8A3 (NCX3) gene was deleted, and we obtained the same results. In addition, mitochondrial respiration was impaired in the clones with NCX3 knocked-out, suggesting that NCX3 also play a role in bioenergetics. In conclusion, we have clearly shown that NCX3 plays an important anti-apoptotic, pro-migratory and proliferative role in the cancer cells by affecting mitochondrial bioenergetics, thus supporting their survival and fate.

• MeSH
• Apoptosis drug effects   MeSH
• Humans MeSH
• Mitochondria metabolism   drug effects   MeSH
• Cell Line, Tumor MeSH
• Neoplasms * metabolism   pathology   genetics   MeSH
• Cell Movement drug effects   MeSH
• Cell Proliferation drug effects   MeSH
• Sodium-Calcium Exchanger * metabolism   genetics   antagonists & inhibitors   MeSH
• Calcium metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Extracellular vesicles can play an important role in the processes occurring after stem cell transplantation, preventing cell apoptosis, stimulating immunological processes, and promoting the synthesis of extracellular matrix. Human follicular fluid (FF) can be a source of a subpopulation of cells with mesenchymal stem cells (MSCs) properties. Moreover these subpopulations of FF cells can differentiate into osteoblasts. In presented studies flow cytometry of ovarian FF cells confirmed positive expression of MSCs markers such as: CD44, CD90, CD105, CD73 and negative expression of a hematopoietic marker: CD45. The CD90+, CD105+, CD45- cell subpopulation has been obtained during magnetic separation using appropriate antibodies conjugated with microbeads. The extracellular vesicles (EVs) secreted by the cells during osteodifferentiation process differed from those secreted by cells culture in the basal medium. Based on the previous and current electron microscopy research, changes in size, number, and shape would support the notion that released EVs could be crucial to the ovarian FF cell subpopulation differentiation process. Osteogenic differentiation has been confirmed via Alizarin red staining. Therefore, follicular fluid (FF) can be a new source of a cell subpopulation with MSC properties, with the cells capable of differentiating into the osteogenic lineage. EVs could play a key role as mediators in tissue regeneration, especially bone tissue regeneration.

• MeSH
• Cell Differentiation * MeSH
• Extracellular Vesicles * ultrastructure   metabolism   MeSH
• Follicular Fluid * cytology   metabolism   MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Mesenchymal Stem Cells * cytology   metabolism   MeSH
• Osteoblasts cytology   metabolism   MeSH
• Osteogenesis * MeSH
• Flow Cytometry MeSH
• Check Tag
• Humans MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

The parasitic protozoan Entamoeba histolytica secretes extracellular vesicles (EVs), but so far little is known about their function in the interaction with the host immune system. Infection with E. histolytica trophozoites can lead to formation of amebic liver abscesses (ALAs), in which pro-inflammatory immune responses of Ly6Chi monocytes contribute to liver damage. Men exhibit a more severe pathology as the result of higher monocyte recruitment and a stronger immune response. To investigate the role of EVs and pathogenicity in the host immune response, we studied the effect of EVs secreted by low pathogenic EhA1 and highly pathogenic EhB2 amebae on monocytes. Size and quantity of isolated EVs from both clones were similar. However, they differed in their proteome and miRNA cargo, providing insight into factors potentially involved in amebic pathogenicity. In addition, EVs were enriched in proteins with signaling peptides compared with the total protein content of trophozoites. Exposure to EVs from both clones induced monocyte activation and a pro-inflammatory immune response as evidenced by increased surface presentation of the activation marker CD38 and upregulated gene expression of key signaling pathways (including NF-κB, IL-17 and TNF signaling). The release of pro-inflammatory cytokines was increased in EV-stimulated monocytes and more so in male- than in female-derived cells. While EhA1 EV stimulation caused elevated myeloperoxidase (MPO) release by both monocytes and neutrophils, EhB2 EV stimulation did not, indicating the protective role of MPO during amebiasis. Collectively, our results suggest that parasite-released EVs contribute to the male-biased immunopathology mediated by pro-inflammatory monocytes during ALA formation.

• MeSH
• Liver Abscess, Amebic immunology   parasitology   MeSH
• Cytokines metabolism   MeSH
• Entamoebiasis immunology   parasitology   MeSH
• Entamoeba histolytica * immunology   pathogenicity   genetics   MeSH
• Extracellular Vesicles * immunology   metabolism   MeSH
• Humans MeSH
• Monocytes * immunology   parasitology   MeSH
• Signal Transduction * MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

The critical role of the immune system in brain function and dysfunction is well recognized, yet development of immune therapies for psychiatric diseases has been slow due to concerns about iatrogenic immune deficiencies. These concerns are emphasized by the lack of objective diagnostic tools in psychiatry. A promise to resolve this conundrum lies in the exploitation of extracellular vesicles (EVs) that are physiologically produced or can be synthetized. EVs regulate recipient cell functions and offer potential for EVs-based therapies. Intranasal EVs administration enables the targeting of specific brain regions and functions, thereby facilitating the design of precise treatments for psychiatric diseases. The development of such therapies requires navigating four dynamically interacting networks: neuronal, glial, immune, and EVs. These networks are profoundly influenced by brain fluid distribution. They are crucial for homeostasis, cellular functions, and intercellular communication. Fluid abnormalities, like edema or altered cerebrospinal fluid (CSF) dynamics, disrupt these networks, thereby negatively impacting brain health. A deeper understanding of the above-mentioned four dynamically interacting networks is vital for creating diagnostic biomarker panels to identify distinct patient subsets with similar neuro-behavioral symptoms. Testing the functional pathways of these biomarkers could lead to new therapeutic tools. Regulatory approval will depend on robust preclinical data reflecting progress in these interdisciplinary areas, which could pave the way for the design of innovative and precise treatments. Highly collaborative interdisciplinary teams will be needed to achieve these ambitious goals.

• MeSH
• Biomarkers MeSH
• Mental Disorders * therapy   immunology   metabolism   MeSH
• Extracellular Vesicles * immunology   metabolism   transplantation   MeSH
• Precision Medicine * methods   MeSH
• Humans MeSH
• Cell Communication MeSH
• Brain immunology   metabolism   MeSH
• Neuroglia * metabolism   immunology   MeSH
• Neurons * metabolism   immunology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH