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Self-driving laboratories (SDLs) promise an accelerated application of the scientific method. Through the automation of experimental workflows, along with autonomous experimental planning, SDLs hold the potential to greatly accelerate research in chemistry and materials discovery. This review provides an in-depth analysis of the state-of-the-art in SDL technology, its applications across various scientific disciplines, and the potential implications for research and industry. This review additionally provides an overview of the enabling technologies for SDLs, including their hardware, software, and integration with laboratory infrastructure. Most importantly, this review explores the diverse range of scientific domains where SDLs have made significant contributions, from drug discovery and materials science to genomics and chemistry. We provide a comprehensive review of existing real-world examples of SDLs, their different levels of automation, and the challenges and limitations associated with each domain.
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Obesity is a major risk factor for cancer. Conventional thought suggests that elevated adiposity predisposes to heightened inflammatory stress and potentiates tumor growth, yet underlying mechanisms remain ill-defined. Here, we show that tumors from patients with a body mass index >35 carry a high burden of senescent cells. In mouse syngeneic tumor models, we correlated a pronounced accretion of senescent cancer cells with poorly immunogenic tumors when mice were subjected to diet-induced obesity (DIO). Highly immunogenic tumors showed lesser senescence burden suggesting immune-mediated elimination of senescent cancer cells, likely targeted as a consequence of their senescence-associated secretory phenotype. Treatment with the senolytic BH3 mimetic small molecule inhibitor ABT-263 selectively stalled tumor growth in mice with DIO to rates comparable to regular diet-fed mice. Thus, consideration of body adiposity in the selection of cancer therapy may be a critical determinant for disease outcome in poorly immunogenic malignancies.
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Senescência Celular , Neoplasias , Camundongos , Animais , Obesidade/complicaçõesRESUMO
Long-term synaptic plasticity is typically associated with morphological changes in synaptic connections. However, the molecular mechanisms coupling functional and structural aspects of synaptic plasticity are still poorly defined. The catalytic activity of type I phosphoinositide-3-kinase (PI3K) is required for specific forms of synaptic plasticity, such as NMDA receptor-dependent long-term potentiation (LTP) and mGluR-dependent long-term depression (LTD). On the other hand, PI3K signaling has been linked to neuronal growth and synapse formation. Consequently, PI3Ks are promising candidates to coordinate changes in synaptic strength with structural remodeling of synapses. To investigate this issue, we targeted individual regulatory subunits of type I PI3Ks in hippocampal neurons and employed a combination of electrophysiological, biochemical and imaging techniques to assess their role in synaptic plasticity. We found that a particular regulatory isoform, p85α, is selectively required for LTP. This specificity is based on its BH domain, which engages the small GTPases Rac1 and Cdc42, critical regulators of the actin cytoskeleton. Moreover, cofilin, a key regulator of actin dynamics that accumulates in dendritic spines after LTP induction, failed to do so in the absence of p85α or when its BH domain was overexpressed as a dominant negative construct. Finally, in agreement with this convergence on actin regulatory mechanisms, the presence of p85α in the PI3K complex determined the extent of actin polymerization in dendritic spines during LTP. Therefore, this study reveals a molecular mechanism linking structural and functional synaptic plasticity through the coordinate action of PI3K catalytic activity and a specific isoform of the regulatory subunits.
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Fatores de Despolimerização de Actina , Actinas , Espinhas Dendríticas , Hipocampo , Potenciação de Longa Duração , Animais , Espinhas Dendríticas/metabolismo , Potenciação de Longa Duração/fisiologia , Actinas/metabolismo , Hipocampo/metabolismo , Hipocampo/citologia , Fatores de Despolimerização de Actina/metabolismo , Ratos , Proteínas rac1 de Ligação ao GTP/metabolismo , Sinapses/metabolismo , Polimerização , Proteína cdc42 de Ligação ao GTP/metabolismo , Plasticidade Neuronal/fisiologia , Fosfatidilinositol 3-Quinases/metabolismo , Classe Ia de Fosfatidilinositol 3-Quinase/metabolismo , Classe Ia de Fosfatidilinositol 3-Quinase/genética , Neurônios/metabolismo , Transdução de Sinais , Camundongos , Células CultivadasRESUMO
Age-related macular degeneration (AMD) is invariably associated with the chronic accumulation of activated mononuclear phagocytes in the subretinal space. The mononuclear phagocytes are composed of microglial cells but also of monocyte-derived cells, which promote photoreceptor degeneration and choroidal neovascularization. Infiltrating blood monocytes can originate directly from bone marrow, but also from a splenic reservoir, where bone marrow monocytes develop into angiotensin II receptor (ATR1)+ splenic monocytes. The involvement of splenic monocytes in neurodegenerative diseases such as AMD is not well understood. Using acute inflammatory and well-phenotyped AMD models, we demonstrate that angiotensin II mobilizes ATR1+ splenic monocytes, which we show are defined by a transcriptional signature using single-cell RNA sequencing and differ functionally from bone marrow monocytes. Splenic monocytes participate in the chorio-retinal infiltration and their inhibition by ATR1 antagonist and splenectomy reduces the subretinal mononuclear phagocyte accumulation and pathological choroidal neovascularization formation. In aged AMD-risk ApoE2-expressing mice, a chronic AMD model, ATR1 antagonist and splenectomy also inhibit the chronic retinal inflammation and associated cone degeneration that characterizes these mice. Our observation of elevated levels of plasma angiotensin II in AMD patients, suggests that similar events take place in clinical disease and argue for the therapeutic potential of ATR1 antagonists to inhibit splenic monocytes for the treatment of blinding AMD.
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Neovascularização de Coroide , Degeneração Macular , Humanos , Camundongos , Animais , Idoso , Monócitos/patologia , Angiotensina II , Degeneração Macular/genética , Inflamação/genéticaRESUMO
BACKGROUND: Around 10% of people with HIV (PWH) exhibit a low-level viremia (LLV) under antiretroviral therapy (ART). However, its origin and clinical significance are largely unknown, particularly at viremias between 50 and 200 copies/mL and under modern ART based on integrase strand transfer inhibitors (INSTIs). Our aim was to characterize their poor immune response against HIV in comparison to individuals with suppressed viremia (SV) and non-HIV controls (NHC). METHODS: Transversal observational study in 81 matched participants: 27 PWH with LLV, 27 PWH with SV, and 27 NHC. Activation (CD25, HLA-DR, and CD38) and senescence [CD57, PD1, and HAVCR2 (TIM3)] were characterized in peripheral T-cell subsets by spectral flow cytometry. 45 soluble biomarkers of systemic inflammation were evaluated by immunoassays. Differences in cell frequencies and plasma biomarkers among groups were evaluated by a generalized additive model for location, scale, and shape (GAMLSS) and generalized linear model (GLM) respectively, adjusted by age, sex at birth, and ART regimen. RESULTS: The median age was 53 years and 77.8% were male. Compared to NHC, PWH showed a lower CD4+/CD8+ ratio and increased activation, senescence, and inflammation, highlighting IL-13 in LLV. In addition, LLV showed a downtrend in the frequency of CD8+ naive and effector memory (EM) type 1 compared to SV, along with higher activation and senescence in CD4+ and CD8+ EM and terminally differentiated effector memory RA+ (TEMRA) subpopulations. No significant differences in systemic inflammation were observed between PWH groups. CONCLUSION: LLV between 50 and 200 copies/mL leads to reduced cytotoxic activity and T-cell dysfunction that could affect cytokine production, being unable to control and eliminate infected cells. The increase in senescence markers suggests a progressive loss of immunological memory and a reduction in the proliferative capacity of immune cells. This accelerated immune aging could lead to an increased risk of developing future comorbidities. These findings strongly advocate for heightened surveillance of these PWH to promptly identify potential future complications.
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Senescência Celular , Infecções por HIV , HIV-1 , Ativação Linfocitária , Viremia , Humanos , Masculino , Pessoa de Meia-Idade , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , Infecções por HIV/virologia , Feminino , Ativação Linfocitária/efeitos dos fármacos , Viremia/imunologia , Viremia/tratamento farmacológico , Senescência Celular/efeitos dos fármacos , Idoso , Adulto , Linfócitos T/imunologiaRESUMO
Although it is known that increasing age is associated with increased morbidity and mortality in allogeneic transplantation (allo-HSCT), individualization of the process may allow to perform it in progressively older patients.This study analyzed the outcome of 97 patients older than 60 years with a first allo-HSCT performed at our institution between 2011 and 2019.Median age was 66 years (range 60-79) and 15.4% were older than 70 years. The most frequent diagnosis was acute leukemia (50.5%), and 58.8% received a myeloablative conditioning. With a median follow-up of 33.9 months (range 7.9-111.5), at 3-years overall survival (OS) was 50%; progression-free survival (PFS), 46%; cumulative incidence of relapse, 22%; and non-relapse mortality (NRM), 32%. There were no significant differences in OS (p = 0.415), PFS (p = 0.691), cumulative incidence of relapse (p = 0.357) or NRM (p = 0.658) between patients of 60-64 years (n = 37), 65-69 (n = 45) and ≥ 70 years (n = 15). No differences were observed either depending on the intensity of the conditioning regimen in terms of OS (p = 0.858), PFS (p = 0.729), cumulative incidence of relapse (p = 0.416) or NRM (p = 0.270).In conclusion, older adults can safely and effectively undergo allo-HSCT with proper patient selection and individualized transplantation procedures.
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Doença Enxerto-Hospedeiro , Transplante de Células-Tronco Hematopoéticas , Leucemia Mieloide Aguda , Humanos , Idoso , Pessoa de Meia-Idade , Estudos de Viabilidade , Estudos Retrospectivos , Leucemia Mieloide Aguda/complicações , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Recidiva , Condicionamento Pré-Transplante/métodos , Doença Enxerto-Hospedeiro/etiologiaRESUMO
Reactive molecular dynamics simulations are used to model the covalent functionalization of amorphous silica with aromatic hydrocarbons. Simulations show that the surface density of silanol-terminated phenyl, naphthyl, and anthracenyl molecules is lower than the maximum value calculated based on molecule geometry, and the simulation densities decrease faster with the number of aromatic rings than the geometric densities. The trends are analyzed in terms of the surface-silanol bonding configurations, tilt angles, local conformational ordering, and aggregation of surface-bound molecules under steady-state conditions. Results show that the surface density is affected by both the size and symmetry of the aromatic hydrocarbons. The correlations among bonding, orientation, and surface density identified here may guide the selection or design of molecules for functionalized surfaces.
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PURPOSE: In an era characterized by rapid progression in neurosurgical technologies, traditional tools such as the non-navigated two-dimensional intraoperative ultrasound (nn-2D-IOUS) risk being overshadowed. Against this backdrop, this study endeavors to provide a comprehensive assessment of the clinical efficacy and surgical relevance of nn-2D-IOUS, specifically in the context of glioma resections. METHODS: This retrospective study undertaken at a single center evaluated 99 consecutive, non-selected patients diagnosed with both high-grade and low-grade gliomas. The primary objective was to assess the proficiency of nn-2D-IOUS in generating satisfactory image quality, identifying residual tumor tissue, and its influence on the extent of resection. To validate these results, early postoperative MRI data served as the reference standard. RESULTS: The nn-2D-IOUS exhibited a high level of effectiveness, successfully generating good quality images in 79% of the patients evaluated. With a sensitivity rate of 68% and a perfect specificity of 100%, nn-2D-IOUS unequivocally demonstrated its utility in intraoperative residual tumor detection. Notably, when total tumor removal was the surgical objective, a resection exceeding 95% of the initial tumor volume was achieved in 86% of patients. Additionally, patients in whom residual tumor was not detected by nn-2D-IOUS, the mean volume of undetected tumor tissue was remarkably minimal, averaging at 0.29 cm3. CONCLUSION: Our study supports nn-2D-IOUS's invaluable role in glioma surgery. The results highlight the utility of traditional technologies for enhanced surgical outcomes, even when compared to advanced alternatives. This is particularly relevant for resource-constrained settings and emphasizes optimizing existing tools for efficient patient care. NCT05873946 - 24/05/2023 - Retrospectively registered.
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Neoplasias Encefálicas , Glioma , Padrão de Cuidado , Humanos , Glioma/cirurgia , Glioma/diagnóstico por imagem , Glioma/patologia , Estudos Retrospectivos , Neoplasias Encefálicas/cirurgia , Neoplasias Encefálicas/diagnóstico por imagem , Neoplasias Encefálicas/patologia , Masculino , Feminino , Pessoa de Meia-Idade , Adulto , Idoso , Procedimentos Neurocirúrgicos/métodos , Procedimentos Neurocirúrgicos/normas , Monitorização Intraoperatória/métodos , Monitorização Intraoperatória/normas , Ultrassonografia/métodos , Ultrassonografia/normas , Adulto Jovem , Neoplasia Residual/diagnóstico por imagem , Neoplasia Residual/cirurgia , Imageamento por Ressonância Magnética/métodos , Imageamento por Ressonância Magnética/normasRESUMO
In this work, we describe the unexpected 2-fold Csp3-Csp3 bond cleavage suffered by cyclobutanols in the presence of a catalytic amount of Pd(OAc)2 and promoted by the bulky biaryl JohnPhos ligand. Overall, the sequential cleavage of a strained and an unstrained Csp3-Csp3 bond leads to the formal [2 + 2]-retrocyclization products, namely, styrene and acetophenone derivatives. This procedure might enable the use of cyclobutanols as masked acetyl groups, resisting harsh conditions in organic synthesis.
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The role of extracellular nucleotides as modulators of inflammation and cell stress is well established. One of the main actions of these molecules is mediated by the activation of purinergic receptors (P2) of the plasma membrane. P2 receptors can be classified according to two different structural families: P2X ionotropic ion channel receptors, and P2Y metabotropic G protein-coupled receptors. During inflammation, damaged cells release nucleotides and purinergic signaling occurs along the temporal pattern of the synthesis of pro-inflammatory and pro-resolving mediators by myeloid and lymphoid cells. In macrophages under pro-inflammatory conditions, the expression and activity of cyclooxygenase 2 significantly increases and enhances the circulating levels of prostaglandin E2 (PGE2), which exerts its effects both through specific plasma membrane receptors (EP1-EP4) and by activation of intracellular targets. Here we review the mechanisms involved in the crosstalk between PGE2 and P2Y receptors on macrophages, which is dependent on several isoforms of protein kinase C and protein kinase D1. Due to this crosstalk, a P2Y-dependent increase in calcium is blunted by PGE2 whereas, under these conditions, macrophages exhibit reduced migratory capacity along with enhanced phagocytosis, which contributes to the modulation of the inflammatory response and tissue repair.
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Inflamação , Prostaglandina-Endoperóxido Sintases , Humanos , Prostaglandina-Endoperóxido Sintases/metabolismo , Inflamação/metabolismo , Nucleotídeos/metabolismo , Macrófagos/metabolismo , Receptores Purinérgicos/metabolismoRESUMO
This research introduces an Artificial Intelligence (AI) based model designed to concurrently optimize energy supply management, biocide dosing, and maintenance scheduling for heat exchangers. This optimization considers energetic, technical, economic, and environmental considerations. The impact of biofilm on heat exchangers is assessed, revealing a 41% reduction in thermal efficiency and a 113% increase in flow frictional resistance of the fluid compared to the initial state. Consequently, the pump's power consumption, required to maintain hydraulic conditions, rises by 9%. The newly developed AI model detects the point at which the heat exchanger's performance begins to decline due to accumulating dirt, marking day 44 of experimentation as the threshold to commence the antifouling biocide dosing. Leveraging this AI model to monitor heat exchanger efficiency represents an innovative approach to optimizing antifouling biocide dosing and reduce the environmental impact stemming from industrial plants.
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Inteligência Artificial , Biofilmes , Incrustação Biológica , Desinfetantes , Água do Mar , Água do Mar/química , Biofilmes/efeitos dos fármacos , Incrustação Biológica/prevenção & controle , Modelos TeóricosRESUMO
INTRODUCTION: The aim of this study was to compare the biological effects of four chemical caries removal materials and to assess their cytotoxicity using human dental pulp stem cells (hDPSCs). METHODS: The products evaluated are: 1 - papain-based product (BRIX 3000®); 2 - papain/chloramine based products (NATURAL-CARE and Papacárie Duo®); and 3 - chloramine based product (Cariesolut). The following in vitro experiments were carried out: IC50 measurement, cell metabolic activity (MTT) assay, cell migration, immunofluorescence experiment, cell apoptosis analysis, and reactive oxygen species (ROS) production analysis. Statistical analyses were performed using one-way ANOVA followed by Tukey's post hoc test (p < 0.05). RESULTS: The IC50 values were: Brix 3000: 0.596%; Papacárie Duo: 0.052%; NATURAL CARE: 1.034%; and Cariesolut: 0.020%. The MTT assays showed non-adequate cell viability of all chemical-mechanical caries removal tested at 2% at 24, 48, and 72 h (p < 0.001). The same behaviour was observed at 0.1% in the Papacárie Duo and Cariesolut groups. In contrast, 0.1% of Brix 3000 at all times and NATURAL CARE at 24 h treated cells showed cell viability rates similar to the control group. At 0.01% only Brix 3000 did not show statistically significant differences at any time. Delayed cell migration was observed in all hDPSCs treated with Papacárie Duo and Cariesolut (p < 0.01 and p < 0.001). Phalloidin staining images showed a high confluence of cells in the presence of NATURAL CARE, similar to the control group. On the contrary, no cells were observed in Brix 3000 and Cariesolut at 2% and 0.1% concentrations. Papacárie Duo showed cells at all concentrations, but hDPSCs treated at 0.01% concentration exhibited better proliferation and spreading than those in the control group. Apoptosis essay showed that Brix 3000 at both 0.1% and 0.01% had a percentage of live cells higher than 99%, with 68.4% live cells at 2%, 3.69% early apoptotic cells, and 27.9% late apoptotic cells. Conversely, the rest of the materials showed an abundance of apoptotic cells, even at low concentrations. 0.1% and 0.01% of BRIX 3000 did not affect the ROS production levels, while 2% of BRIX 3000 counterpart very significantly increased the percentage of CM-H2DCFDA positive cells. Again, all concentrations of Cariesolut showed significantly higher levels of ROS production than those observed in control cells. CONCLUSION: Our results suggest that Brix 3000 would be the most suitable material for chemical caries removal, with Papacárie Duo and NATURAL CARE also being good options, and discourage the use of Cariesolut due to its low cytocompatibility on dental pulp stem cells.
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Controlled studies have extensively documented that concentrate supplements typically increase enteric CH4 emissions and milk yield and reduce emissions per unit of milk produced and dry matter intake. However, no studies have been conducted to determine the effect of concentrate on predicted greenhouse gas emissions from dairy farms representing the Australian pasture-based farming system. Thus, this study sought to determine how dietary concentrate supplementation affects enteric and manure CH4, and N2O of Australian pasture-based dairy farms. The Australian Dairy Carbon Calculator was used, which incorporates emission factors and methodologies used in the National Greenhouse Gas Inventory as reported to the Intergovernmental Panel on Climate Change. Primary data were collected and analyzed from 120 commercial farms in Australia's major dairy regions. Then the farms were divided into 4 groups based on their dietary concentrate supplementation: ≤1 (low; 15 farms), 1 to 2 (moderate; 35 farms), 2 to 3 (high; 35 farms), and ≥3 (very high; 35 farms) t of concentrate dry matter (tDM) per cow per year. Sources of greenhouse gas emissions were CO2 from concentrate production, enteric CH4, and manure CH4 and N2O. Total dry matter intake, milk yield, and daily enteric CH4 production (g/d) quadratically increased with concentrate level, whereas greenhouse gas emission intensity of milk production (kg of CO2 equivalent per kg of fat- and protein-corrected milk) decreased by 14% for farms supplementing with ≥3 tDM/cow per year compared with those supplementing with ≤1 tDM/cow per year of dietary concentrate. The N2O and CH4 emissions from manure increased quadratically and linearly, respectively, with the increasing supplementation of concentrate. Farms supplementing 2 to 3 tDM/cow per year showed substantial increases in gross income, gross margin, earnings before interest and tax, and net income ($/cow per year) compared with those supplementing of ≤1, 1 to 2, and ≥3 tDM/cow per year. Overall, increasing dietary concentrate supplementation for dairy cows resulted in increased milk production per cow, reduced greenhouse gas emissions per unit of milk produced, and increased income and profit. However, a comprehensive life cycle assessment study is needed to account for carbon sequestration by other farm components, such as pastures and trees, which were not considered in the present study. In addition, the present study was based on modeling and did not gather ground truth information for DMI, digestibility, crude protein, and urinary and fecal N excretion. Therefore, data should be interpreted with caution, and studies gathering such information are encouraged.
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Dieta , Suplementos Nutricionais , Gases de Efeito Estufa , Lactação , Leite , Animais , Leite/química , Bovinos , Gases de Efeito Estufa/análise , Feminino , Dieta/veterinária , Indústria de Laticínios , Metano/biossíntese , Metano/análise , Austrália , Ração Animal/análise , Fazendas , Esterco/análise , Efeito EstufaRESUMO
INTRODUCTION: Epilepsy is a disease characterized by an excessive discharge in neurons generally provoked without any external stimulus, known as convulsions. About 2 million people are diagnosed each year in the world. This process is carried out by a neurological doctor using an electroencephalogram (EEG), which is lengthy. METHOD: To optimize these processes and make them more efficient, we have resorted to innovative artificial intelligence methods essential in classifying EEG signals. For this, comparing traditional models, such as machine learning or deep learning, with cutting-edge models, in this case, using Capsule-Net architectures and Transformer Encoder, has a crucial role in finding the most accurate model and helping the doctor to have a faster diagnosis. RESULT: In this paper, a comparison was made between different models for binary and multiclass classification of the epileptic seizure detection database, achieving a binary accuracy of 99.92% with the Capsule-Net model and a multiclass accuracy with the Transformer Encoder model of 87.30%. CONCLUSION: Artificial intelligence is essential in diagnosing pathology. The comparison between models is helpful as it helps to discard those that are not efficient. State-of-the-art models overshadow conventional models, but data processing also plays an essential role in evaluating the higher accuracy of the models.
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Inteligência Artificial , Epilepsia , Humanos , Epilepsia/diagnóstico , Convulsões/diagnóstico , Algoritmos , Aprendizado de Máquina , EletroencefalografiaRESUMO
BACKGROUND: The recruitment of effector cells is one of the novel functions described for extracellular vesicles (EVs) that needs further study. For instance, cell recruitment by mesenchymal stromal cell derived-EVs (MSC-EVs) is one of the features by which MSC-EVs may induce regeneration and ameliorate tissue injury. On the other hand, increasing evidence suggests that cancer EVs play an important role in the preparation of the pre-metastatic niche (PMN) by recruiting their primary tumour cells. Understanding and measuring the potential of MSC-EVs or cancer-EVs to induce cell migration and recruitment is essential for cell-free therapeutic approaches and/or for a better knowledge of cancer metastasis, respectively. In this context, classical in vitro migration assays do not completely mimic the potential situation by which EVs exert their chemotactic capacity. RESULTS: We adapted an agarose spot migration assay as an in vitro system to evaluate the cell recruitment capacity of locally delivered or localized EVs. Cell migration was tracked for 12 h or 48 h, respectively. Thereafter, endpoint migration images and time-lapse videos were analysed to quantify several parameters aiming to determine the migration of cells to either MSC-EV or pro-metastatic EV. The number of cells contained inside the agarose spots, the migration distance, the area occupied by cells, the directionality of the cell movement, and the Euclidean distance were measured. This multi-parametric evaluation revealed the potential of different MSC-EV preparations to recruit endothelial cells and to detect an enhanced recruitment capacity of highly metastatic PC3-derived EVs (PC3-EVs) compared to low-metastatic LNCaP-EVs in a tumour cell-specific manner. CONCLUSIONS: Overall, this agarose spot migration assay may offer a diversity of measurements and migration settings not provided by classical migration assays and reveal its potential use in the EV field in two different contexts with recruitment in common: regeneration and cancer metastasis.
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Vesículas Extracelulares , Neoplasias , Humanos , Medicina Regenerativa , Sefarose , Fatores Quimiotáticos , Células Endoteliais , Neoplasias/terapiaRESUMO
The flowers of Yucca aloifolia ("flor de izote") are considered a millenary food in the Northeastern Highlands of Puebla, Mexico. The present investigation reports on the chemical and biological activities of the hydroalcoholic extract (YAHF) obtained from this edible source. HPLC-MS profiling revealed twenty bioactive phenolic compounds with chlorogenic acid (16.5â mg g-1 DW), quercetin (9.5â mg g-1 FW), and their glycosides (rutin and quercitrin), as well as caffeic acid (8.4â mg g-1 DW) and ferulic acid (7.9â mg g-1 DW) as major compounds dissolved in YAHF. Six metabolites had potent anti-lipase (IC50<100â µg mL-1) and anti-ornithine decarboxylase activity (IC50<100â µg mL-1), whereas thirteen exerted strong anti-alpha-glucosidase properties (IC50<100â µg mL-1). The evaluation of YAHF in mice subjected to standard oral glucose tolerance tests and prolonged administration of hypercaloric/atherogenic diet (30â days), unraveled their ability to improve glucose and lipid profiles. YAHF and six phenolic compounds significantly reduced DLD-1 cell viability (IC50, 117.9â µg mL-1) and avoided polyamine accumulation linked to anti-ornithine decarboxylase activity. YAHF and its twenty constituents exerted low toxicity in probiotics (>1000â µg mL-1) and 3T3 fibroblasts (>2.5â mg-mL-1), sustaining their safeness for human consumption.
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Herein we describe the inâ situ inhibitory activity of three hybrid films (FL1, FL2, and FL3) against two wild strains of Colletotrichum gloeosporioides and Penicillium digitatum as causal agents of rot in Persian limes. The films FL2 and FL3 contained 1.0 and 1.3 % weight/volume Litsea glaucescens essential oil (LgEO) and significantly (p<0.05) delayed rot emergence in Persian limes caused by both pathogens up to 10â days. The physicochemical properties of LgEO and hybrid films were obtained, whereas detailed HPLC profiling revealed that fruit covered with these films significantly (p<0.01) preserved reducing sugars (sucrose, fructose, and glucose), organic acids (citric acid, ascorbic acid, malic acid, and oxalic acid), and flavonoids with nutraceutical activity (hesperidin, eriocitrin, naruritin, neohesperidin, diosmin, vitexin, rutin, and quercetin). This evidence sustains that the composites generated in this investigation improve the shelf life of Persian limes and conserve their nutraceutical content.
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Citrus , Litsea , Óleos Voláteis , Antifúngicos/farmacologia , Antifúngicos/análise , Óleos Voláteis/análise , Frutas/químicaRESUMO
OBJECTIVES: To evaluate the cytocompatibility, bioactivity, and anti-inflammatory potential of the new pre-mixed calcium silicate cement-based sealers Bioroot Flow (BrF) and AH Plus Bioceramic Sealer (AHPbcs) on human periodontal ligament stem cells (hPDLSCs) compared to the epoxy resin-based sealer AH Plus (AHP). MATERIALS AND METHODS: Standardized discs and 1:1, 1:2, and 1:4 eluates of BrF, AHPbcs and AHP after setting were prepared. The following assays were performed: cell attachment and morphology via SEM, cell viability via a MTT assay, cell migration/proliferation via a wound-healing assay, cytoskeleton organization via immunofluorescence staining; cytokine release via ELISA; osteo/cemento/odontogenic marker expression via RT-qPCR, and cell mineralized nodule formation via Alizarin Red S staining. HPDLSCs were isolated from extracted third molars from healthy patients. Comparisons were made with hPDLSCs cultured in unconditioned (negative control) or osteogenic (positive control) culture media. Statistical significance was established at p < 0.05. RESULTS: Both BrF and AHPbcs showed significantly positive results in the cytocompatibility assays (cell metabolic activity, migration, attachment, morphology, and cytoskeleton organization) compared with a negative control group, while AHP showed significant negative results. BrF exhibited an upregulation of at least one osteo/cementogenic marker compared to the negative and positive control groups. BrF showed a significantly higher calcified nodule formation than AHPbcs, the negative and positive control groups, while AHPbcs was higher than the negative control group. Both were also significantly higher than AHP group. CONCLUSION: BrF and AHPbcs exhibit adequate and comparable cytocompatibility on hPDLSCs. BrF also promoted the osteo/cementogenic differentiation of hPDLSCs. Both calcium silicate-based sealers favored the downregulation of the inflammatory cytokine IL-6 and the calcified nodule formation from hPDLSCs. BrF exerted a significantly higher influence on cell mineralization than AHPbcs. CLINICAL RELEVANCE: This is the first study to elucidate the biological properties and immunomodulatory potential of Bioroot Flow and AH Plus Bioceramic Sealer. The results act as supporting evidence for their use in root canal treatment.
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Alopecia/congênito , Compostos de Cálcio , Resinas Epóxi , Ligamento Periodontal , Materiais Restauradores do Canal Radicular , Silicatos , Humanos , CitocinasRESUMO
OBJECTIVES: To assess the biocompatibility, bioactivity, and immunomodulatory properties of three new calcium silicate cement-based sealers: Ceraseal (CS), Totalfill BC Sealer (TFbc) and WellRoot ST (WR-ST) on human periodontal ligament stem cells (hPDLSCs). MATERIALS AND METHODS: HPDLSCs were isolated from extracted third molars from healthy patients. Eluates (1:1, 1:2, and 1:4 ratio) and sample discs of CS, TFbc and WR-ST after setting were prepared. A series of assays were performed: cell characterization, cell metabolic activity (MTT assay) cell attachment and morphology (SEM assay), cell migration (wound-healing assay), cytoskeleton organization (phaloidin-based assay); IL-6 and IL-8 release (ELISA); differentiation marker expression (RT-qPCR assay), and cell mineralization (Alizarin Red S staining). HPDLSCs cultured in unconditioned (negative control) or osteogenic (positive control) culture media were used as a comparison. Statistical significance was established at p < 0.05. RESULTS: All the tested sealers exhibited similar results in the cytocompatibility assays (cell metabolic activity, migration, attachment, morphology, and cytoskeleton organization) compared with a negative control group. CS and TFbc exhibited an upregulation of at least one osteo/cementogenic marker compared to the negative and positive control groups. CS and TFbc also showed a significantly higher calcified nodule formation than the negative and positive control groups. Both the marker expression and calcified nodule formation were significantly higher in CS-treated cells than TFbc treated cells. WR-ST exhibited similar results to the control group. CS and TFbc-treated cells exhibited a significant downregulation of IL-6 after 72 h of culture compared to the negative control group (p < 0.05). CONCLUSION: All the tested sealers exhibited an adequate cytocompatibility. CS significantly enhances cell differentiation by upregulating the expression of key genes associated with bone and cementum formation. Additionally, CS was observed to facilitate the mineralization of the extracellular matrix effectively. In contrast, the effects of TFbc and WR-ST on these processes were less pronounced compared to CS. Furthermore, both CS and TFbc exhibited an anti-inflammatory potential, contributing to their potential therapeutic benefits in regenerative endodontics. CLINICAL RELEVANCE: This is the first study to compare the biological properties and immunomodulatory potential of Ceraseal, Totalfill BC Sealer, and WellRoot ST. The results act as supporting evidence for their use in root canal treatment.
Assuntos
Materiais Biocompatíveis , Compostos de Cálcio , Teste de Materiais , Ligamento Periodontal , Silicatos , Compostos de Cálcio/farmacologia , Silicatos/farmacologia , Humanos , Ligamento Periodontal/citologia , Ligamento Periodontal/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Técnicas In Vitro , Células Cultivadas , Células-Tronco/efeitos dos fármacos , Materiais Restauradores do Canal Radicular/farmacologia , Diferenciação Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Adesão Celular/efeitos dos fármacos , Dente SerotinoRESUMO
Children with beta-thalassemia (BT) present with an increase in carotid intima-medial thickness, an early sign suggestive of premature atherosclerosis. However, it is unknown if there is a direct relationship between BT and atherosclerotic disease. To evaluate this, wild-type (WT, littermates) and BT (Hbbth3/+) mice, both male and female, were placed on a 3-mo high-fat diet with low-density lipoprotein receptor suppression via overexpression of proprotein convertase subtilisin/kexin type 9 (PCSK9) gain-of-function mutation (D377Y). Mechanistically, we hypothesize that heme-mediated oxidative stress creates a proatherogenic environment in BT because BT is a hemolytic anemia that has increased free heme and exhausted hemopexin, heme's endogenous scavenger, in the vasculature. We evaluated the effect of hemopexin (HPX) therapy, mediated via an adeno-associated virus, to the progression of atherosclerosis in BT and a phenylhydrazine-induced model of intravascular hemolysis. In addition, we evaluated the effect of deferiprone (DFP)-mediated iron chelation in the progression of atherosclerosis in BT mice. Aortic en face and aortic root lesion area analysis revealed elevated plaque accumulation in both male and female BT mice compared with WT mice. Hemopexin therapy was able to decrease plaque accumulation in both BT mice and mice on our phenylhydrazine (PHZ)-induced model of hemolysis. DFP decreased atherosclerosis in BT mice but did not provide an additive benefit to HPX therapy. Our data demonstrate for the first time that the underlying pathophysiology of BT leads to accelerated atherosclerosis and shows that heme contributes to atherosclerotic plaque development in BT.NEW & NOTEWORTHY This work definitively shows for the first time that beta-thalassemia leads to accelerated atherosclerosis. We demonstrated that intravascular hemolysis is a prominent feature in beta-thalassemia and the resulting increases in free heme are mechanistically relevant. Adeno-associated virus (AAV)-hemopexin therapy led to decreased free heme and atherosclerotic plaque area in both beta-thalassemia and phenylhydrazine-treated mice. Deferiprone-mediated iron chelation led to deceased plaque accumulation in beta-thalassemia mice but provided no additive benefit to hemopexin therapy.