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Invasive fungal infections (IFIs) are responsible for elevated rates of morbidity and mortality, causing around of 1.5 million deaths annually worldwide. One of the main causative agents of IFIs is Candida albicans, and non-albicans Candida species have emerged as a spreading global public health concernment. Furthermore, COVID-19 has contributed to a boost in the incidence of IFIs, such as mucormycosis, in which Rhizopus oryzae is the most prevalent causative agent. The effector host immune response against IFIs depends on the activity of T cells, which are susceptible to the regulatory effects triggered by fungal virulence factors. The fungal cell wall plays a crucial role as a virulence factor, and its remodeling compromises the development of a specific T-cell response. The redirection of Jurkat T cells to target Candida spp. by recognizing targets expressed on the fungal cell wall can be facilitated using chimeric antigen receptor (CAR) technology. This study generated an M-CAR that contains an scFv with specificity to α-1,6 mannose backbone of fungal mannan, and the expression of M-CAR on the surface of modified Jurkat cells triggered a strong activation against Candida albicans (hyphae form), Candida tropicalis (hyphae form), Candida parapsilosis (pseudohyphal form), and Candida glabrata (yeast form). Moreover, M-CAR Jurkat cells recognized Rhizopus oryzae spores, which induced high expression of cell activation markers. Thus, a novel Mannan-specific CAR enabled strong signal transduction in modified Jurkat cells in the presence of Candida spp. or R. oryzae.

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Bone regeneration remains a critical challenge in regenerative medicine, particularly in dentistry, where conditions such as periodontal disease and trauma can lead to significant bone defects. Traditional treatment methods, such as autogenous bone grafting, face limitations, including donor site morbidity and postoperative complications. Recent advancements in biomaterials, particularly silk fibroin-based scaffolds, have shown promise due to their excellent biocompatibility and tunable mechanical properties. Incorporating bioactive glass and metal ions, such as cobalt, into these scaffolds can enhance osteogenic properties and antibacterial effects, creating an optimal environment for bone regeneration. The primary objective of this study was to develop and characterize SilkMA/silicated-chlorinated cobalt-doped bioactive glass composites with the potential for bone regeneration applications. Utilizing the sol-gel method, we synthesized cobalt-doped bioglass, enhancing its bioactivity and antibacterial properties. Mechanical testing, swelling assessments, degradation analysis, and in vitro evaluations using alveolar bone-derived mesenchymal stem cells (aBMSCs) demonstrated the scaffolds' cytocompatibility and favorable physical properties. The structural integrity of the electrospun fibers was confirmed through Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR), and Raman Spectroscopy analyses. Incorporating bioglass reduced swelling ratios, while in vitro assays showed that cobalt ions effectively inhibited the biofilm formation of Porphyromonas gingivalis. In vivo analysis using hematoxylin-eosin and von Kossa (vK) staining demonstrated that the SilkMA + 20% BGCo scaffold elicited a minimal inflammatory response, confirming its biocompatibility. However, the absence of positively stained structures in the vK analysis indicated its lack of mineralization potential. In sum, SilkMA/BGCo scaffolds showed promising in vitro potential for bone tissue regeneration and excellent biocompatibility in vivo despite lacking calcium deposition. Further studies with alternative in vivo models are needed to confirm their efficacy.

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This study analyzes the morphology of melanomacrophage centers (MMCs) in the kidney, liver, and spleen of Gymnotus carapo across two different environmental settings with varying degrees of anthropogenic impact, aiming to link habitat conditions to fish health and validate MMCs as bioindicators for environmental monitoring. A total of 28 specimens, captured and humanely sacrificed under anesthesia, were processed using conventional histological techniques. Morphometric measurements were recorded, and tissue samples were examined on an Olympus BX 41 microscope, selecting 10 random 10x fields per organ to count MMCs. Macroscopic and histological examination of the liver, kidney, and spleen highlighted MMCs distribution patterns and notable differences across sex and age groups. Adult and juvenile males showed major MMCs counts in the kidney and spleen compared to females, while liver samples from both sexes contained only pigmented cells without significant MMCs formation. Differences in MMCs quantity and structure were also observed between environments: natural habitats displayed lower MMCs counts in both sexes compared to samples from locations with higher anthropogenic impact, where MMCs frequency and pigmentation intensity were notably elevated. These findings suggest that MMCs variations in G. carapo may reflect environmental conditions and stressors, supporting the potential application of MMCs as health indicators for aquatic organisms and ecosystem monitoring. This work provides comprehensive morphometric and histological data on MMCs distribution in G. carapo, emphasizing its potential as a bioindicator species, particularly valuable in regions with varying pollution levels.

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Glial cells play a pivotal role in the Central Nervous System (CNS), constituting most brain cells. Gliogenesis, crucial in CNS development, occurs after neurogenesis. In the hypothalamus, glial progenitors first generate oligodendrocytes and later astrocytes. However, the precise molecular mechanisms governing the emergence of glial lineages in the developing hypothalamus remain incompletely understood. This study reveals the pivotal role of the transcription factor KLF10 in regulating the emergence of both astrocyte and oligodendrocyte lineages during embryonic hypothalamic development. Through transcriptomic and bioinformatic analyses, we identified novel KLF10 putative target genes, which play important roles in the differentiation of neurons, astrocytes, and oligodendrocytes. Notably, in the absence of KLF10, there is an increase in the oligodendrocyte population, while the astrocyte population decreases in the embryonic hypothalamus. Strikingly, this decline in the number of astrocytes persists into adulthood, indicating that the absence of KLF10 leads to an extended period of oligodendrocyte emergence while delaying the appearance of astrocytes. Our findings also unveil a novel signaling pathway for Klf10 gene expression regulation. We demonstrate that Klf10 is a target of CREB and that its expression is upregulated via the BDNF-p38-CREB pathway. Thus, we postulate that KLF10 is an integral part of the hypothalamic developmental program that ensures the correct timing for glial phenotypes' generation. Importantly, we propose that the Klf10-/- mouse model represents a valuable tool for investigating the impact of reduced astrocyte and microglia populations in the homeostasis of the adult hypothalamus.

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Bone related disorders are highly prevalent, and many of these pathologies still do not have curative and definitive treatment methods. This is due to a complex interplay of multiple factors, such as the crosstalk between different tissues and cellular components, all of which are affected by microenvironmental factors. Moreover, these bone pathologies are specific, and current treatment results vary from patient to patient owing to their intrinsic biological variability. Current approaches in drug development to deliver new drug candidates against common bone disorders, such as standard two-dimensional (2D) cell culture and animal-based studies, are now being replaced by more relevant diseases modelling, such as three-dimension (3D) cell culture and primary cells under human-focused microphysiological systems (MPS) that can resemble human physiology by mimicking 3D tissue organization and cell microenvironmental cues. In this review, various technological advancements for in vitro bone modeling are discussed, highlighting the progress in biomaterials used as extracellular matrices, stem cell biology, and primary cell culture techniques. With emphasis on examples of modeling healthy and disease-associated bone tissues, this tutorial review aims to survey current approaches of up-to-date bone-on-chips through MPS technology, with special emphasis on the scaffold and chip capabilities for mimicking the bone extracellular matrix as this is the key environment generated for cell crosstalk and interaction. The relevant bone models are studied with critical analysis of the methods employed, aiming to serve as a tool for designing new and translational approaches. Additionally, the features reported in these state-of-the-art studies will be useful for modeling bone pathophysiology, guiding future improvements in personalized bone models that can accelerate drug discovery and clinical translation.

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INTRODUCTION: The Transverse Rectus Abdominis Myocutaneous (TRAM) flap is used for breast reconstruction, but involves the risk of necrosis. Adipose tissue-derived mesenchymal Stem Cells (ADSCs) can be used to stimulate neovascularization and reduce the risk of TRAM flap necrosis. AIM: Determine the effect of ADSCs on TRAM flap viability in rats. METHODS: Twenty-four Wistar-EPM rats were distributed into three groups (n = 8). A right caudal pedicled TRAM flap was performed in all the animals and was the only procedure performed in Group TRAM. The additional procedures of intradermal injection of α-MEM culture medium and intradermal injection of α-MEM containing ADSCs labeled with a fluorescent marker were performed in Groups α-MEM and α-MEM-SC, respectively. The percentage of flap necrosis was determined, and the level of neovascularization and distribution of stem cells in the TRAM flap was assessed using immunohistochemical analysis and fluorescence microscopy, respectively. RESULTS: The percentage of necrosis observed in Group α-MEM-SC was lower than that observed in Groups TRAM and α-MEM, namely 23.36 % vs. 50.42 % and 53.57 %, respectively (p < 0.05). In Zone IV of the flap, the number of vessels was greater in Group α-MEM-SC than in the other groups (p < 0.05). Multiple stem cells were observed in the four zones of the flap in Group α-MEM-SC. No stem cells were observed in Groups TRAM or α-MEM. CONCLUSION: ADSCs increased TRAM flap viability and the number of vessels in Zone IV of the flap in rats.

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Retinal ganglion cells (RGCs) are the projection neurons of the retina, and their death promotes an irreversible blindness. Several factors were described to control their genesis during retinal development. These include Atoh7, a major orchestrator of the RGC program, and downstream targets of this transcription factor, including Pou4f factors, that in turn regulate key aspects of terminal differentiation. The absence of POU4F family genes results in defects in RGC differentiation, aberrant axonal elaboration and, ultimately, RGC death. This confirms the requirement of POU4F factors for RGC development and survival, with a crucial role in regulating RGC axon outgrowth and pathfinding. Here, we have investigated in vivo whether ectopic Pou4f2 expression in late retinal progenitor cells (late RPCs) is sufficient to induce the generation of cells with RGC properties, including long-range axon projections. We show that Pou4f2 overexpression generates RGC-like cells that share morphological and transcriptional features with RGCs that are normally generated during early development and extend axonal projections up to the brain. In conclusion, these results show that POU4F2 alone is sufficient to promote the crucial properties of projection neurons that arise from retinal progenitors outside their developmental window.

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OBJECTIVES: This study aimed to evaluate the effect of different irrigation protocols on the viability and metabolism of stem cells from the apical papilla (SCAP) using a new in vitro protocol that simulates the clinical situation. MATERIALS AND METHODS: Forty-eight bovine dentin cylinders were obtained and prepared to simulate teeth with incomplete rhizogenesis, positioned under a three-dimensional (3D) culture of SCAPs to mimic the apical papilla. The cylinders were divided into four groups (n = 8) according to the irrigating solution: Control; NaOCl (Sodium hypochlorite 1%); EDTA (17% ethylenediaminetetraacetic acid); and NaOCl + EDTA. Subsequently, the viability (Live/Dead n = 2) and metabolism (Alamar Blue n = 6) of the cells were assessed (ISO 10993). Data were analyzed using Kruskal-Wallis and Dunn tests (p < 0.05). RESULTS: In the 1 to 3 days period, Control and EDTA had significantly higher increases in metabolism compared to NaOCl and NaOCl + EDTA (p < 0.05). In the 3- to 7-day period, metabolism significantly decreased in NaOCl + EDTA compared to EDTA (p < 0.05) but was similar to Control and NaOCl. Additionally, significant differences were observed within groups Control, EDTA, and NaOCl + EDTA across the two periods (p < 0.05). CONCLUSIONS: The tested in vitro model allows for the analysis of the response of SCAPs to different irrigating solutions, simulating the clinical situation. Sodium hypochlorite 1% demonstrated high cytotoxicity to SCAPs, whose effects were partially reversed by 17% EDTA. CLINICAL RELEVANE: The methodology developed provides a tool for future investigations, allowing for the assessment of new irrigants and techniques that may optimize tissue regeneration.

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Photobiomodulation therapy (PBMT) is gaining recognition as an effective method for repairing damaged tissues, particularly in skin and mucosa injuries. Despite its growing use, there remains a significant gap in understanding the interaction of ectomesenchymal components during light therapy. This study explored various near-infrared light parameters on stressed skin keratinocytes (KC) cultured alone or in association with oral fibroblasts (FB) using a transwell system. Reactive oxygen species (ROS) levels were quantified, alongside observations of cell viability/proliferation, migration, and pro-resolutive gene expression in KC cultivated conventionally (at 1, 5, and 50 J/cm2; 1.4, 7 and, 70 s; 0.03, 0.14, and 1.4 J, respectively; 0.71 W/cm2; 20 mW) and co-cultured with FB (at 5 J/cm2). Murine tongue fragments were also treated at 5 J/cm2 for three days. The results revealed increased ROS levels in stressed cells. PBMT enhanced cell viability/proliferation and migration of KC in vitro, while pro-inflammatory cytokines were predominantly suppressed following irradiation. All energy densities and culture models showed upregulation of amphiregulin. In the co-culture system, PBMT notably upregulated interleukin 1 - receptor antagonist and vascular endothelial growth factor genes while downregulating transforming growth factor-ß1 expression. In ex vivo tongue fragments, PBMT significantly increased lamina propria thickness and spindle-shaped cell counts compared to non-irradiated samples. Overall, near-infrared PBMT demonstrated pro-regenerative effects by modulating inflammation. The activation of pro-resolutive molecules and the dampening of cytokines may favor early tissue remodeling in the dermis counterpart.

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Piscirickettsia salmonis is the pathogen that has most affected the Chilean salmon industry for over 30 years. Considering the problems of excessive use of antibiotics, it is necessary to find new strategies to control this pathogen. Antivirulence therapy is an alternative to reduce the virulence of pathogens without affecting their growth. Polyphenolic compounds have been studied for their antiviral capacity. In this study, the capacity of quercetin and silybin to reduce the intracellular replication of P. salmonis in SHK-1 cells was evaluated. For this, three different infection protocols in Salmon Head Kidney-1(SHK-1) cells were used: co-incubation for 24 h, pre-incubation for 24 h prior to infection, and post-incubation for 24 h after infection. In addition, the effect of co-incubation in rainbow trout intestinal epithelial cells (RTgutGC) and the effect on the phagocytic capacity of SHK-1 cells were evaluated. The results obtained showed that quercetin and silybin decreased the intracellular replication of P. salmonis in SHK-1 cells when they were co-incubated for 24 h; however, they did not have the same effect in RTgutGC cells. On the other hand, both compounds decreased the phagocytosis of SHK-1 cells during co-incubation. These results are promising for the study of new treatments against P. salmonis.

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Umbilical cord blood (UCB) is an alternative therapeutic resource for treating both hematological and non-hematological diseases, especially for pediatric patients. However, UCB transplantation faces challenges, including delayed engraftment, increased risk of graft failure, and slower immune recovery. To maximize its clinical potential, it is essential to understand the variability and functionality of its nucleated cells. This study focused on characterizing UCB cellular populations, viability, and functionality at three key processing stages: freshly collected, post-volume reduction, and post-thawing. Using EuroFlow-based flow cytometry, significant changes were observed in granulocyte and T-cell populations during processing. Additionally, integrating EuroFlow data with hematology counts revealed variability that could affect the yield of specific cell populations, potentially influencing therapeutic decisions. An in vitro migration assay, designed to mimic the vascular niche, was employed to study donor variability in cellular migratory patterns. Notably, thawed UCB cells displayed two distinct migration profiles, distinguishing lymphocyte-like cells from monocyte-like cells. These findings underscore the importance of reproducible cellular quality control measures, such as immunophenotypic and functional donor characterization, to ensure the integrity of UCB composition. A better understanding of these parameters could improve the consistency and reliability of UCB as a starting material for the development of advanced therapies.

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κ-Casein glycomacropeptide (GMP) exerts anti-inflammatory and immune modulatory effects. A bovine GMP concentrate and its in vitro digestion product were obtained. GMP was also microencapsulated with phycocolloids and further digested. These products were tested in three-dimensional (3D) and open monolayer two-dimensional (2D) mouse jejunal organoids. Almost no effect was observed on the 2D organoids. In 3D organoids, GMP induced intestinal proliferation (Axin2, Pcna) and differentiation (Vil1, Alpl) genes together with Muc3, antibacterial genes (Lyz1, Pla2g2a), and Cxcl1. GMP also induced interferon I defense genes (Ifnb1, Ifr3, Oas2, Oas3, Rnasel) under basal conditions and in TNF-stimulated organoids. In vitro digestion abrogated the effects of GMP and induced new genes (Lgr5, Olfm4, and Lct). In TNF-stimulated organoids, digested GMP repressed multiple genes. Encapsulation largely preserved the GMP effects. In conclusion, GMP showed differential effects in 3D and 2D organoids. The effects of digestion peptides were also different, suggesting distinct potential as functional foods.

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The activity of matrix metalloproteinases (MMPs) plays a crucial role in the aging of the resin-dentin interface. The in situ action of MMP-2 and MMP-9 has been confirmed in the process of dentin-collagen degradation. However, the involvement of dental pulp cells in MMP secretion as a response to oxidative stress induced by contact with resin monomers has not been fully elucidated. Myricetin (MYR), like proanthocyanidin (PAC), has antioxidant properties and may help prevent extracellular matrix degradation. The objective was to evaluate the effect of MYR on the MMP expression and activity in response to reactive oxygen species (ROS) increase induced by triethylene glycol dimethacrylate (TEGDMA) in human odontoblast-like cells (hOLCs). hOLCs differentiated from dental pulp mesenchymal stem cells were exposed to TEGDMA released from dentin blocks using a barrier model with transwell inserts for 18, 24, and 36 h. Intracellular oxidation was evaluated using the 2',7'-dichlorofluorescein probe. The effect of 600 µM MYR on the MMP-2 and MMP-9 expression was determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The extracellular MMP levels were quantified using enzyme-linked immunosorbent assay, and their activation by means of a proteolytic fluorometric assay. The results were analyzed by one-way analysis of variance and Tukey's post hoc test, p ≤ 0.05. TEGDMA exposure increased intracellular ROS and upregulated MMP-2 and MMP-9 mRNA in hOLCs (p < 0.001). The levels of MMPs increased significantly 24 h after TEGDMA exposure (p = 0.013). These secreted proteases exhibited high activation ability. MYR reduced ROS production and downregulated MMP expression and activity at both mRNA and protein levels, similar to the effect found for PAC, which was used as a control. A relationship was observed between MMP-2 and MMP-9 expression, secretion, and early activation with ROS increase due to TEGDMA exposure. MYR showed potential as a therapeutic strategy to control MMP expression and modulate redox imbalance, offering a protective effect on cellular response.

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Arthropods are diverse, abundant, successful animals that exploit all available ecological niches. They sense the environment, move, interact with prey/predators/conspecifics, learn, and so forth using small brains with five orders of magnitude less neurons than mammals. Hence, these brains need to be efficient in information processing. One distinct aspect is the presence of large, easily identifiable single neurons that act as functional units for information processing integrating a high volume of information from different sources to guide behavior. To understand the synaptic organization behind these high-integration nodes research on suitable neurons is needed. The lobula giant neurons (LGs) found in the third optic neuropil, the lobula, of semiterrestrial crabs Neohelice granulata respond to moving stimuli, integrate information from both eyes, and show short- and long-term plasticity. They are thought to be key elements in the visuomotor transformation guiding escape responses to approaching objects. One subgroup, the MLG1 (monostratified LG type 1), is composed of 16 elements that have very wide main branches and a regular arrangement in a deep layer of the lobula which allows their identification even in unstained preparations. Here, we describe the types and abundance of synaptic contacts involving MLG1 profiles using transmission electron microscopy (TEM). We found an unexpected diversity of synaptic motifs and an apparent compartmentalization of the dendritic arbor in two domains where MLG1s act predominantly as presynaptic or postsynaptic, respectively. We propose that the variety of contact types found in the dendritic arbor of the MLG1s reflects the multiple circuits in which these cells are involved. Regarding the detection of approaching objects, the distinctive input contact motifs shared by lobula giant neurons in crabs and locusts suggest a similar organization of the collision-detecting pathways in both species.

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Mesenchymal stem cells (MSCs) are emerging as promising therapeutic agents due to their immunomodulatory effects, primarily mediated via paracrine signaling. Similarly, anthocyanins, such as cyanidin-3-glucoside (C3G), have demonstrated significant anti-inflammatory properties. In this context, this study investigated the immunomodulatory potential of C3G on MSCs, and subsequent effects on macrophage and lymphocyte responses. Cytotoxicity assays identified 50 µM as the highest nontoxic C3G concentration for MSCs. Flow cytometry confirmed that C3G treatment did not affect MSC viability or cell cycle distribution, even under LPS stimulation. Cytokine production by MSCs was evaluated after treatment with C3G and LPS. While no significant changes were observed in IL-6, IL-10, TGF-ß, or PGE2 levels, IL-1ß production was significantly reduced in LPS-stimulated MSCs treated with C3G. Protein expression analysis revealed decreased NFκB phosphorylation in LPS-stimulated MSCs treated with C3G, with no changes detected in STAT-3 or PCNA expression. The immunomodulatory effects of MSC-derived conditioned media on macrophages and lymphocytes were also assessed. In LPS-stimulated macrophages, conditioned media from MSCs reduced the production of IL-1ß, IL-6, and IL-12. Interestingly, conditioned media from C3G-treated MSCs specifically decreased TNF-α levels, enhanced IL-10 secretion, and further inhibited NFκB phosphorylation. In LPS-stimulated lymphocytes, conditioned media from C3G-treated MSCs suppressed IL-2 production while increasing IL-10 levels. In summary, these findings demonstrate that conditioned media from C3G-treated MSCs modulates immune cell responses more effectively than C3G alone. C3G influences the paracrine activity of MSCs, resulting in a shift in the secretory profile and subsequent effects on immune cell behavior.

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Introducción. La encefalopatía hipóxico-isquémica (EHI) causada por la falta de oxígeno y/o perfusión al cerebro puede provocar daño neurológico agudo en el recién nacido. La hipotermia terapéutica (HT) es el tratamiento más eficaz y seguro. Sin embargo, la mortalidad sigue siendo alta y con numerosas secuelas a largo plazo. Las terapias celulares y, en particular, la sangre de cordón umbilical (SCU) se estudian como terapias alternativas. El objetivo de este trabajo es evaluar la factibilidad y seguridad de la infusión de células autólogas de la sangre de cordón junto con la hipotermia moderada. Población y métodos. Se incorporaron 12 bebés de 36 semanas o más de gestación con diagnóstico de EHI moderada o grave y que contaban con colecta de SCU. La SCU se redujo en volumen y se obtuvieron hasta 4 dosis, que fueron infundidas en las primeras 72 horas posnatales. Se evaluó el tiempo a la primera infusión y posibles reacciones adversas a la infusión. Resultados. Entre los años 2014 y 2019, 12 bebés fueron incluidos en el protocolo (HT + SCU), 9 con diagnóstico de EHI moderada y 3 con EHI grave. En todos los casos, se obtuvo al menos 1 dosis de SCU para infundir. En todos los casos, fue posible infundir la primera dosis antes de las 24 horas y no hubo reacciones adversas atribuibles a la infusión. Conclusiones. La colecta, preparación e infusión de sangre de cordón umbilical autóloga fresca para su uso en recién nacidos con EHI es factible y segura en nuestras condiciones

Introduction. Hypoxic-ischemic encephalopathy (HIE) caused by lack of oxygen and perfusion to the brain can lead to acute neurological damage in newborns. Therapeutic hypothermia (TH) is the most effective and safest treatment. However, mortality remains high with numerous long-term sequelae. Cellular therapies, particularly umbilical cord blood (UCB), are being studied as alternative therapies. The aim of this study is to assess the feasibility and safety of combining autologous cord blood cell infusion with moderate hypothermia. Population and methods. Twelve infants of 36 weeks gestational age or older, diagnosed with moderate or severe HIE and with umbilical cord blood (UCB) collected were included. UCB was volume-reduced, and up to four doses were obtained. These doses were infused within the first 72 postnatal hours. Time to the first infusion and possible adverse reactions to the infusion were evaluated. Results. Between 2014 and 2019, 12 infants were included in the protocol (TH + UCB), 9 with a diagnosis of moderate HIE and 3 with severe HIE. In all cases, at least one dose of UCB was obtained for infusion. In all cases, the first dose was infused within 24 hours in every case, and no adverse reactions attributable to the infusion were observed. Conclusions. The collection, processing, and infusion of fresh autologous umbilical cord blood for use in newborns with HIE are feasible and safe under our conditions.

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Our knowledge about the consumption of cannabinoids during pregnancy lacks consistent evidence to determine whether it compromises neurodevelopment. Addressing this task is challenging and complex since pregnant women display multiple confounding factors that make it difficult to identify the real effect of cannabinoids' consumption. Recent studies shed light on this issue by using pluripotent stem cells of human origin, which can recapitulate human neurodevelopment. These revolutionary platforms allow studying how exogenous cannabinoids could alter human neurodevelopment without ethical concerns and confounding factors. Here, we review the information to date on the clinical studies about the impact of exogenous cannabinoid consumption on human brain development and how exogenous cannabinoids alter nervous system development in humans using cultured pluripotent stem cells as 2D and 3D platforms to recapitulate brain development.

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BACKGROUND: Despite many years of investigation into mesenchymal stem cells (MSCs) and their potential for treating inflammatory conditions such as COVID-19, clinical outcomes remain variable due to factors like donor variability, different tissue sources, and diversity within MSC populations. Variations in MSCs' secretory and proliferation profiles, and their proteomic and transcriptional characteristics significantly influence their therapeutic potency, highlighting the need for enhanced characterization methods to better predict their efficacy. This study aimed to evaluate the biological characteristics of MSCs from different tissue origins, selecting the most promising line for further validation in a K18-hACE2 mouse model of SARS-CoV-2 infection. METHODS: We studied nine MSC lines sourced from either bone marrow (hBMMSC), dental pulp (hDPMSC), or umbilical cord tissue (hUCMSC). The cells were assessed for their proliferative capacity, immunophenotype, trilineage differentiation, proteomic profile, and in vitro immunomodulatory potential by co-culture with activated lymphocytes. The most promising MSC line was selected for further experimental validation using the K18-hACE2 mouse model of SARS-CoV-2 infection. RESULTS: The analyzed cells met the minimum criteria for defining MSCs, including the expression of surface molecules and differentiation capacity, showing genetic stability and proliferative potential. Proteomic analysis revealed distinct protein profiles that correlate with the tissue origin of MSCs. The immunomodulatory response exhibited variability, lacking a discernible pattern associated with their origin. In co-culture assays with lymphocytes activated with anti-CD3/CD28 beads, all MSC lines demonstrated the ability to inhibit TNF-α, to induce TGF-ß and Indoleamine 2,3-dioxygenase (IDO), with varying degrees of inhibition observed for IFN-γ and IL-6, or induction of IL-10 expression. A module of proteins was found to statistically correlate with the potency of IL-6 modulation, leading to the selection of one of the hUCMSCs as the most promising line. Administration of hUCMSC to SARS-CoV-2-infected K18 mice expressing hACE2 was effective in improving lung histology and modulating of a panel of cytokines. CONCLUSIONS: Our study assessed MSCs derived from various tissues, uncovering significant variability in their characteristics and immunomodulatory capacities. Particularly, hUCMSCs demonstrated potential in mitigating lung pathology in a SARS-CoV-2 infection model, suggesting their promising therapeutic efficacy.

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BACKGROUND: Cavernous nerve injury-induced erectile dysfunction (CNI-ED) is a common complication following radical prostatectomy and severely affects patients' quality of life. The mitochondrial impairment in corpus cavernosum smooth muscle cells (CCSMCs) may be an important pathological mechanism of CNI-ED. Previous studies have shown that transplantation of human adipose derived stem cells (ADSC) can alleviate CNI-ED in a rat model. However, little is known about the effect of human umbilical cord mesenchymal stem cells (hUC-MSC) on CNI-ED. It remains unclear whether hUC-MSC can ameliorate mitochondrial damage in CCSMCs. In this study, we aimed to investigate the impacts of hUC-MSC on the mitochondrial mass and function of CCSMCs, as well as elucidate its underlying molecular mechanism. METHODS: The CNI-ED rat model was established by bilaterally crushing cavernous nerves. Subsequently, hUC-MSC were transplanted into the cavernosum and ADSC were injected as a positive control group. Erectile function evaluation and histological detection were performed 4 weeks after cell transplantation. In vitro, CCSMCs underwent hypoxia and were then co-cultured with ADSC or hUC-MSC using a transwell system. The mitochondrial mass and function, as well as signaling pathways, were investigated. To explore the role of the SIRT1/PGC-1α/TFAM pathway in regulating mitochondrial biogenesis of CCSMCs, we knocked down SIRT1 by siRNA. RESULTS: The administration of hUC-MSC significantly improved erectile function of CNI-ED rats and reduced the ratio of collagen to smooth muscle. Specifically, hUC-MSC treatment restored mitochondrial mass and function in CCSMCs injured by CNI or hypoxia, and inhibited the apoptosis of CCSMCs. Mechanistically, the application of hUC-MSC activated SIRT1/PGC-1α/TFAM pathway both in rat penile tissues and CCSMCs. In addition, knockdown of SIRT1 in CCSMCs abolished the protective effects of hUC-MSC on mitochondrial mass and function, while leading to an increase in cellular apoptosis. CONCLUSIONS: hUC-MSC contribute to the recovery of erectile function in CNI-ED rats by restoring mitochondrial mass and function of CCSMCs through the SIRT1/PGC-1α/TFAM pathway. Our present study offers new insights into the role and molecular mechanisms of hUC-MSC in regulating mitochondrial homeostasis, thereby facilitating the restoration of the erectile function in CNI-ED.

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STUDY DESIGN: Experimental study utilizing a standardized Balb C mouse model. OBJECTIVES: Evaluate histological changes and motor function recovery in the acute and subacute phases of Spinal Cord Injury (SCI) in mice using human Umbilical cord blood-derived mononuclear cells. METHODS: Forty mice were divided into five groups, with two receiving human Umbilical cord blood-derived mononuclear cells immediately after SCI and after 7 days, and three control groups. Motor assessment utilized BMS, MFS, and horizontal plane scales over six weeks. Necropsy evaluated macroscopic and histological spinal cord changes. RESULTS: Histologically, Umbilical cord blood-derived mononuclear cells-treated groups exhibited significant reductions in necrosis, hemorrhage, and degeneration compared to controls. Motor recovery showed partial improvement across all groups, with no statistically significant differences in scales between intervention and control groups. CONCLUSIONS: In the acute phases of SCI, Umbilical cord blood-derived mononuclear cells applied directly to Balb C mice lesions demonstrated histological improvement but played a limited role in functional enhancement. The study highlights distinctions in the treatment's efficacy, potentially related to these cells' diverse differentiation capacities and intrinsic properties.