RESUMO
Ferroptosis is an iron-dependent cell death mechanism involving the accumulation of lipid peroxides. As a critical regulator, glutathione peroxidase 4 (GPX4) has been demonstrated to be downregulated in epilepsy. However, the mechanism of ferroptosis in epilepsy remains unclear. In this study, bioinformatics analysis, analysis of epilepsy patient blood samples and cell and mouse experiments revealed strong associations among epilepsy, ferroptosis, microRNA-211-5p and purinergic receptor P2X 7 (P2RX7). P2RX7 is a nonselective ligand-gated homotrimeric cation channel, and its activation mainly increases neuronal activity during epileptic seizures. In our study, the upregulation of P2RX7 in epilepsy was attributed to the downregulation of microRNA (miR)-211-5p. Furthermore, P2RX7 has been found to regulate GPX4/HO-1 by alleviating lipid peroxidation induced by suppression of the MAPK/ERK signaling pathway in murine models. The dynamic decrease in miR-211-5p expression induces hypersynchronization and both nonconvulsive and convulsive seizures, and forebrain miR-211-5p suppression exacerbates long-lasting pentylenetetrazole-induced seizures. Additionally, in this study, induction of miR-211-5p expression or genetic-silencing of P2RX7 significantly reduced the seizure score and duration in murine models through the abovementioned pathways. These results suggest that the miR-211-5p/P2RX7 axis is a novel target for suppressing both ferroptosis and epilepsy.
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Epilepsia , Ferroptose , MicroRNAs , Humanos , Animais , Camundongos , Epilepsia/genética , Estresse Oxidativo , Convulsões , MicroRNAs/genética , Receptores Purinérgicos P2X7/genéticaRESUMO
The pathogenesis of Parkinson's disease (PD) is strongly associated with neuroinflammation, and type I interferons (IFN-I) play a crucial role in regulating immune and inflammatory responses. However, the specific features of IFN in different cell types and the underlying mechanisms of PD have yet to be fully described. In this study, we analyzed the GSE157783 dataset, which includes 39,024 single-cell RNA sequencing results for five PD patients and six healthy controls from the Gene Expression Omnibus database. After cell type annotation, we intersected differentially expressed genes in each cell subcluster with genes collected in The Interferome database to generate an IFN-I-stimulated gene set (ISGs). Based on this gene set, we used the R package AUCell to score each cell, representing the IFN-I activity. Additionally, we performed monocle trajectory analysis, and single-cell regulatory network inference and clustering (SCENIC) to uncover the underlying mechanisms. In silico gene perturbation and subsequent experiments confirm NFATc2 regulation of type I interferon response and neuroinflammation. Our analysis revealed that microglia, endothelial cells, and pericytes exhibited the highest activity of IFN-I. Furthermore, single-cell trajectory detection demonstrated that microglia in the midbrain of PD patients were in a pro-inflammatory activation state, which was validated in the 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mouse model as well. We identified transcription factors NFATc2, which was significantly up-regulated and involved in the expression of ISGs and activation of microglia in PD. In the 1-Methyl-4-phenylpyridinium (MPP+)-induced BV2 cell model, the suppression of NFATc2 resulted in a reduction in IFN-ß levels, impeding the phosphorylation of STAT1, and attenuating the activation of the NF-κB pathway. Furthermore, the downregulation of NFATc2 mitigated the detrimental effects on SH-SY5Y cells co-cultured in conditioned medium. Our study highlights the critical role of microglia in type I interferon responses in PD. Additionally, we identified transcription factors NFATc2 as key regulators of aberrant type I interferon responses and microglial pro-inflammatory activation in PD. These findings provide new insights into the pathogenesis of PD and may have implications for the development of novel therapeutic strategies.
Assuntos
Interferon Tipo I , Neuroblastoma , Doença de Parkinson , Camundongos , Animais , Humanos , Doença de Parkinson/genética , Doença de Parkinson/metabolismo , Doença de Parkinson/patologia , Doenças Neuroinflamatórias , Células Endoteliais/metabolismo , NF-kappa B/metabolismo , Análise de Célula Única , Camundongos Endogâmicos C57BLRESUMO
PURPOSE: The objective was to determine the accuracy of 3D-printed dental models subjected to different storage conditions using six different material and printer combinations. MATERIALS AND METHODS: Three completely dentate models were designed using dental CAD software (3Shape Dental System). A horseshoe-shaped solid base with a posterior horizontal bar was used. The models were printed in a horizontal direction against the building platform without support. The models were printed using six printers with the corresponding recommended resin material: Carbon M2 (DPR10), HeyGears A2D4K (Model HP UV2.0), Stratasys J5 (MED610), Stratasys Origin One (DM200), Envision One (E-Model LightDLP), and Asiga Pro4K (VeriModel) with a standard layer thickness of 50 µm. All printed models underwent scanning using a laboratory scanner (Sirona inEOS X5) after printing. Subsequently, the models were randomly assigned into three groups of storage conditions, LT: cold environment (4 ± 1°C), HT: hot and dry environment (50 ± 2°C), and RT: room temperature (25 ± 2°C) serving as the control. Each group was kept under the designated condition and was scanned at 1, 2, 3, 4, and 8 weeks. The total number of models (N) was 72, with 6 printers producing 12 models per printer for 3 storage conditions, resulting in 4 models for each storage condition and each printer. The generated STL files were imported into a 3D inspection software for comparison with the original STL files. In-tolerance percentage, the deviation RMS, trueness, and precision were obtained and analyzed with least square mean linear regression using JMP Pro 15 to identify the significant effects (α = 0.05). RESULTS: The in-tolerance percentage as-printed was significantly different among different printers. Significant dimension deviations were observed after the first week of storage at HT and with subsequent weeks of storage. RT and LT did not show significant dimensional changes. Models printed with Carbon M2 showed the highest in-tolerance percentages compared to the other printers. CONCLUSIONS: The model deviations were affected by storage conditions and the printer used, with high-temperature storage showed least stability compared to low and room temperatures. No significant difference was observed between low and room temperature storage conditions. The Carbon M2 printer showed the highest accuracy among all printers tested. The region had a significant effect on the deviation measured, with the abutment body showing the least deviation. Among the 3D printers evaluated, A2D4K by HeyGears and Carbon M2 printers demonstrated the highest accuracy in terms of both precision and trueness.
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OBJECTIVES: To investigate the risk factors for bronchopulmonary dysplasia (BPD) in twin preterm infants with a gestational age of <34 weeks, and to provide a basis for early identification of BPD in twin preterm infants in clinical practice. METHODS: A retrospective analysis was performed for the twin preterm infants with a gestational age of <34 weeks who were admitted to 22 hospitals nationwide from January 2018 to December 2020. According to their conditions, they were divided into group A (both twins had BPD), group B (only one twin had BPD), and group C (neither twin had BPD). The risk factors for BPD in twin preterm infants were analyzed. Further analysis was conducted on group B to investigate the postnatal risk factors for BPD within twins. RESULTS: A total of 904 pairs of twins with a gestational age of <34 weeks were included in this study. The multivariate logistic regression analysis showed that compared with group C, birth weight discordance of >25% between the twins was an independent risk factor for BPD in one of the twins (OR=3.370, 95%CI: 1.500-7.568, P<0.05), and high gestational age at birth was a protective factor against BPD (P<0.05). The conditional logistic regression analysis of group B showed that small-for-gestational-age (SGA) birth was an independent risk factor for BPD in individual twins (OR=5.017, 95%CI: 1.040-24.190, P<0.05). CONCLUSIONS: The development of BPD in twin preterm infants is associated with gestational age, birth weight discordance between the twins, and SGA birth.
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Displasia Broncopulmonar , Recém-Nascido Prematuro , Gêmeos , Humanos , Displasia Broncopulmonar/etiologia , Displasia Broncopulmonar/epidemiologia , Fatores de Risco , Recém-Nascido , Feminino , Estudos Retrospectivos , Masculino , Idade Gestacional , Peso ao Nascer , Modelos LogísticosRESUMO
Microalgae have great potential as a future source to meet the increasing global demand for foods. Several microalgae are permitted as safety sources in different countries and regions, and processed as commercial products. However, edible safety, economic feasibility, and acceptable taste are the main challenges for microalgal application in the food industry. Overcome such challenges by developing technology accelerates transition of microalgae into sustainable and nutritious diets. In this review, edible safety of Spirulina, Chlamydomonas reinhardtii, Chlorella, Haematococcus pluvialis, Dunaliella salina, Schizochytrium and Nannochloropsis is introduced, and health benefits of microalgae-derived carotenoids, amino acids, and fatty acids are discussed. Technologies of adaptive laboratory evolution, kinetic model, bioreactor design and genetic engineering are proposed to improve the organoleptic traits and economic feasibility of microalgae. Then, current technologies of decoloration and de-fishy are summarized to provide options for processing. Novel technologies of extrusion cooking, delivery systems, and 3D bioprinting are suggested to improve food quality. The production costs, biomass values, and markets of microalgal products are analyzed to reveal the economic feasibility of microalgal production. Finally, challenges and future perspectives are proposed. Social acceptance is the major limitation of microalgae-derived foods, and further efforts are required toward the improvement of processing technology.
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Nasopharyngeal carcinoma (NPC) is one of the most lethal head and neck cancers, threatening the health of people across the globe, especially in East and Southeast Asia, the Arctic, the Middle East and North Africa. Long non-coding RNAs (lncRNA) have been reported to regulate multiple cancers, including NPC. However, the role of LINC01140 in NPC remains to be covered. In this study, we found that LINC01140 is downregulated in NPC cells. It was uncovered from functional assays that LINC01140 inhibits the proliferation and improves the apoptosis and radiosensitivity of NPC cells. The downstream mechanism by which LINC01140 exerted its functions was explored in subsequent. As proven by mechanism experiments, cytoplasmic LINC01140 positively regulated the expression of ZNF621 through competitively binding to miR-452-5p. ZNF621 can also enhance the radiosensitivity of NPC cells. To summarize, LINC01140 regulates the radiosensitivity of NPC cells through the competing endogenous RNA (ceRNA) mode. Our study aims to identify novel biomarkers for regulating the radiosensitivity of NPC.
Assuntos
MicroRNAs , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas , RNA Longo não Codificante , Tolerância a Radiação , Linhagem Celular Tumoral , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Carcinoma Nasofaríngeo/genética , Carcinoma Nasofaríngeo/radioterapia , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/radioterapia , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Tolerância a Radiação/genética , Dedos de Zinco/genética , Dedos de Zinco/fisiologiaRESUMO
Adaptive laboratory evolution (ALE) has been widely utilized as a tool for developing new biological and phenotypic functions to explore strain improvement for microalgal production. Specifically, ALE has been utilized to evolve strains to better adapt to defined conditions. It has become a new solution to improve the performance of strains in microalgae biotechnology. This review mainly summarizes the key results from recent microalgal ALE studies in industrial production. ALE designed for improving cell growth rate, product yield, environmental tolerance and wastewater treatment is discussed to exploit microalgae in various applications. Further development of ALE is proposed, to provide theoretical support for producing the high value-added products from microalgal production.
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Microalgas/crescimento & desenvolvimento , Animais , Aquicultura , Organismos Aquáticos , BiotecnologiaRESUMO
OBJECTIVE: Measure the size and shape of talc particles in talcum powder and compare this data to the size and shape of talc particles found in surgically resected tissues from patients with ovarian carcinoma. METHODS: Using polarized light microscopy (PLM) and scanning electron microscopy (SEM), we measured the size and shape of talc particles in samples of talc-containing baby powder (TCBP) and surgically resected pelvic tissues (hysterectomies) from talc-exposed patients with ovarian carcinoma. RESULTS: The most frequent class of particles in TCBP can be unequivocally identified as talc, using both polarized light microscopy and scanning electron microscopy with energy dispersive X-ray analysis (SEM/EDX). The talc particles found in resected tissues from ovarian carcinoma patients are similar in size and shape to the most abundant morphological class of particles in TCBP. CONCLUSIONS: This finding, combined with previous epidemiological literature and tissue-based analytical studies, provides further evidence that the small, isodiametric particles that dominate TCBP can migrate from the perineum and become lodged in distal structures in the female reproductive tract, where they may lead to an increased risk of developing ovarian carcinoma.
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Linfonodos/química , Omento/química , Ovário/química , Talco/análise , Adulto , Idoso , Carcinoma Epitelial do Ovário/patologia , Feminino , Humanos , Linfonodos/ultraestrutura , Microscopia Eletrônica de Varredura , Pessoa de Meia-Idade , Omento/ultraestrutura , Neoplasias Ovarianas/patologia , Ovário/ultraestrutura , Talco/efeitos adversos , Talco/farmacocinéticaRESUMO
BACKGROUND: Talc, a hydrous magnesium silicate, often used for genital hygiene purposes, is associated with ovarian carcinoma in case-control studies. Its potential to cause inflammation, injury, and functional changes in cells has been described. A complication of such studies is that talc preparations may be contaminated with other materials. A previous study by (Beck et al. Toxicol Appl Pharmacol 87:222-34, 1987) used a hamster model to study talc and granite dust exposure effects on various biochemical and cellular inflammatory markers. Our current study accessed key materials used in that 1987 study; we re-analyzed the original talc dust with contemporary scanning electron microscopy and energy dispersive x-ray analysis (SEM/EDX) for contaminants. We also examined the original bronchoalveolar lavage (BAL) cells with polarized light microscopy to quantify cell-associated birefringent particles to gain insight into the talc used. RESULTS: SEM/EDX analyses showed that asbestos fibers, quartz, and toxic metal particulates were below the limits of detection in the original talc powder. However, fibers with aspect ratios ≥3:1 accounted for 22% of instilled material, mostly as fibrous talc. Talc (based on Mg/Si atomic weight % ratio) was the most abundant chemical signature, and magnesium silicates with various other elements made up the remainder. BAL cell counts confirmed the presence of acute inflammation, which followed intratracheal instillation. Measurements of cell associated birefringent particles phagocytosis revealed significant differences among talc, granite, and control exposures with high initial uptake of talc compared to granite, but over the 14-day experiment, talc phagocytosis by lavaged cells was significantly less than that of granite. Phagocytosis of talc fibers by macrophages was observed, and birefringent particles were found in macrophages, neutrophils, and multinucleate giant cells in lavaged cells from talc-exposed animals. CONCLUSION: Our data support the contention that talc, even without asbestos and other known toxic contaminants, may elicit inflammation and contribute to lung disease. Our findings support the conclusions of (Beck et al. Toxicol Appl Pharmacol 87:222-34, 1987) study. By analyzing particulate exposures with polarized light microscopy and SEM/EDX, fibrous talc was identified and a distinctive pattern of impaired particulate ingestion was demonstrated.
Assuntos
Exposição por Inalação/efeitos adversos , Pulmão/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Silicatos de Magnésio/toxicidade , Neutrófilos/efeitos dos fármacos , Talco/toxicidade , Animais , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Células Cultivadas , Cricetinae , Poeira , Exposição por Inalação/análise , Pulmão/metabolismo , Pulmão/patologia , Macrófagos/metabolismo , Macrófagos/ultraestrutura , Silicatos de Magnésio/química , Silicatos de Magnésio/farmacocinética , Masculino , Microscopia Eletrônica de Varredura , Neutrófilos/metabolismo , Neutrófilos/ultraestrutura , Tamanho da Partícula , Quartzo/química , Quartzo/farmacocinética , Quartzo/toxicidade , Dióxido de Silício/química , Dióxido de Silício/farmacocinética , Dióxido de Silício/toxicidade , Espectrometria por Raios X , Propriedades de Superfície , Talco/química , Talco/farmacocinéticaRESUMO
Talc may lodge in human tissues through various routes, and has been associated with the development of ovarian carcinoma in case control epidemiologic studies. Talc is detected in tissues with scanning electron microscopy and energy dispersive X-ray analysis (SEM/EDS), with expected magnesium (Mg) and silicon (Si) peaks. The theoretical atomic weight % Mg/Si ratio for talc is 0.649, and for diagnostic purposes, a range of ± 5% (~0.617 to 0.681) is often used as a standard. Our goal was to establish empirically the quantitative range for talc identification by SEM/EDS using two source materials: a Johnson's Baby PowderTM (cosmetic-grade) consumer sample, and talc from Sigma-Aldrich (particle-grade material intended for scientific use). We examined 401 Mg-Si particles with SEM/EDS across the two samples, using two different SEM microscopes. Overall, we found a mean Mg/Si atomic weight % ratio of 0.645, with minimal differences between study subsets. The standard deviation was 0.025; (± 1σ = 0.620-0.670). The currently used ± 5% diagnostic range (Mg/Si 0.617-0.681) is thus reasonably close to this study's ± 1σ range, and well within a ± 2 σ confidence interval span (Mg/Si 0.595-0.695). The ± 5% range is thus an appropriately conservative standard whose continued use seems justified.
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Microscopia Eletrônica de Varredura/normas , Espectrometria por Raios X/normas , Talco/análise , HumanosRESUMO
Perineal talc use is associated with ovarian carcinoma in many case-control studies. Such talc may migrate to pelvic organs and regional lymph nodes, with both clinical and legal significance. Our goal was to differentiate talc in pelvic lymph nodes due to exposure, versus contamination with talc in the laboratory. We studied 22 lymph nodes from ovarian tumor patients, some of which had documented talc exposure, to quantify talc using digestion of tissue taken from paraffin blocks and scanning electron microscopy/energy dispersive X-ray analysis (SEM/EDX). Talc particles correlated significantly with surface contamination assessments using polarized light microscopy. After adjusting for surface contamination, talc burdens in nodes correlated strongly with perineal talc use. In a separate group of lymph nodes, birefringent particles within the same plane of focus as the tissues in histological sections were highly correlated with talc particles within the tissue by in situ SEM/EDX (r = 0.80; p < 0.0001). We conclude that since talc can be a surface contaminant from tissue collection/preparation, digestion measurements may be influenced by contamination. Instead, because they preserve anatomic landmarks and permit identification of particles in cells and tissues, polarized light microscopy and in situ SEM/EDX are recommended to assess talc in lymph nodes.
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Linfonodos/patologia , Microscopia Eletrônica de Varredura , Neoplasias Ovarianas/patologia , Estudos de Casos e Controles , Feminino , Humanos , Microscopia Eletrônica de Varredura/métodos , Microscopia de Polarização/métodos , Pelve/patologiaRESUMO
STATEMENT OF PROBLEM: Estimating the width of the maxillary anterior teeth when creating an esthetic smile can be challenging. Valid metrics to assist in this process are needed. PURPOSE: The purpose of this systematic review was to evaluate the validity of interalar distance and inner canthal distance with the golden proportion, golden mean, and recurring esthetic dental proportion in predicting intercanine distance and the combined width of central incisors to potentially provide a guide for tooth restoration. MATERIAL AND METHODS: A literature search was conducted using PubMed, Medline, Google Scholar, EMBASE, CNKI, Web of Science, and the Cochrane Collaboration, identifying English- and non-English-language articles reporting on interalar width, inner canthal width, and maxillary anterior tooth width. Additional studies were identified by searching reference lists of the articles identified. Only studies that fulfilled inclusion criteria were included. Two examiners independently performed the literature search and data extraction. Using a meta-analysis software program, data extracted from each selected study were statistically combined using the random-effects model. Weighted mean differences, 95% confidence intervals, and heterogeneity were calculated for each measurement. RESULTS: The search strategy resulted in a total of 282 articles, but only 41 articles fulfilling the inclusion criteria were included in the meta-analysis. The interalar distance was found to be significantly larger than intercanine distance, and the inner canthal distance was found to be substantially smaller than the intercanine distance. When predicting the central incisors combined width by interalar distance, both the golden proportion and golden mean predicted value were larger than the combined width of the central incisors. Only the recurring esthetic dental proportion (70%) predicted value showed no significant difference from the combined width of central incisors. When predicting the central incisors combined width by inner canthal distance, the golden proportion predicted value was larger than the combined width of central incisors, whereas both the golden mean and recurring esthetic dental proportion (70%) predicted value were found to be significantly smaller than the combined width of central incisors. CONCLUSIONS: By analyzing the data from the literature, only the recurring esthetic dental proportion (70%) with interalar distance could be an accurate method for predicting the combined width of central incisors. Neither interalar distance nor inner canthal distance could directly be used to predict the intercanine distance.
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Dente Canino , Incisivo , Estética Dentária , Maxila , OdontometriaRESUMO
The goal of this study was to precisely and unambiguously identify foreign particles in human tissues using a combination of polarized light microscopy and Raman microscopy, which provides chemical composition and microstructural characterization of complex materials with submicrometer spatial resolution. This identification for patient care and research has been traditionally studied using polarized light microscopy, electron microscopy with X-ray analysis, and electron diffraction, all with some limitations. We designed a model system of stained and unstained cells that contained birefringent talc particles and systematically investigated the influence of slide and coverslip materials, laser wavelengths, and mounting media on the Raman spectra obtained. Hematoxylin and eosin stained slides did not produce useful results because of fluorescence interference from the stains. Unstained cell samples prepared with standard slides and coverslips produce high quality Raman spectra when excited at 532 nm; the spectra are uniquely assigned to talc. We also obtain high quality Raman spectra specific for talc in unstained tissue samples (pleural tissue following talc pleurodesis and ovarian tissue following long-term perineal talc exposure). Raman microscopy is sufficiently sensitive and compositionally selective to identify particles as small as one micrometer in diameter. Raman spectra have been catalogued for thousands of substances, which suggests that this approach is likely to be successful in identifying other particles of interest in tissues, potentially making Raman microscopy a powerful new tool in pathology.
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Macrófagos/ultraestrutura , Microscopia de Polarização/métodos , Microscopia Óptica não Linear/métodos , Ovário/ultraestrutura , Pleura/ultraestrutura , Talco/análise , Animais , Feminino , Humanos , Camundongos , Modelos Moleculares , Tamanho da Partícula , Pleurodese , Células RAW 264.7RESUMO
STATEMENT OF PROBLEM: Data on the retention of individually formed fiber-reinforced composite posts (everStick) cemented with self-adhesive cement are lacking. PURPOSE: The purpose of this in vitro study was to investigate the pull-out strength of 2 different fiber-reinforced composite posts (prefabricated and individually formed) cemented into extracted teeth with self-adhesive resin cement. MATERIAL AND METHODS: Thirty extracted single-rooted human teeth were decoronated, endodontically treated, and prepared with post spaces of equal length. Prepared specimens were divided into 2 groups (n=15 each) based on the type of post: commercially prefabricated fiber posts (GC) and individually formed resin posts (using GC reinforcing fibers). Self-adhesive resin cement (G-CEM LinkAce; GC) was used to cement all posts. Each post was held with moderate pressure, and root surfaces were light polymerized for 20 seconds (650 mW/cm2). After cementation, the specimens were stored in saline solution for 30 days. Treated teeth were kept in water for 24 hours before pull-out testing parallel to the longitudinal axis of the posts. Data were analyzed using the Student t test (α=.05) and the coefficient of variance as the ratio of the standard deviation to the mean. RESULTS: The mean pull-out retention strength was 185.7 ±61.2 N for the prefabricated fiber posts and 98.9 ±56.5 N for the individually formed fiber posts (P=.026). CONCLUSIONS: The prefabricated fiber posts exhibited significantly higher retention forces than the individually formed posts.
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Resinas Compostas/uso terapêutico , Técnica para Retentor Intrarradicular , Análise do Estresse Dentário , Humanos , Técnicas In Vitro , Projetos Piloto , Técnica para Retentor Intrarradicular/instrumentação , Cimentos de Resina/uso terapêuticoRESUMO
AIMS: Resin infiltration is an emerging technique for management of noncavitated lesions. This study evaluated the in vitro hydrolytic and color stability of the ICON® resin infiltration system (IC) in 42 extracted human teeth. MATERIALS AND METHODS: ICON® resin infiltration system was compared with dental adhesive (DA) and dental sealant (DS). The products were applied according to manufacturer's instructions. The baseline weight and color of the samples were recorded. Color was recorded by spectoral colorimeter. The samples were subjected to four experimental conditions: (1) group 1: Stored in lactic acid solution (pH 4.9) for 24 hours; (2) group 2: Thermocycled for 100 cycles (temperatures: 5°C, 55°C, and dwell time of 15 seconds); (3) group 3: Stored in 0.1 N sodium hydroxide solution (pH 12.48) for 14 days at 60°C; (4) group 4: Stored in phosphate-buffered saline solution (pH 7.2) at 37°C for 4 months. The weight and color were recorded again after removal of the samples from the experimental conditions. Two-factor analysis of variance models and Tukey's Honestly Significant Difference were performed to assess statistical differences among the groups. Scanning electron microscopy imaging was performed for samples from groups 1, 3, and 4. RESULTS: All the samples showed loss of material and change in color. In the demineralizing solution, IC showed significantly greater weight loss (p = 0.032) and color change (p = 0.038) compared with DA. Dental Sealant showed significantly greater weight loss than IC (p = 0.027) after thermocycling. Teeth in group 3 exhibited the greatest weight loss (p < 0.001). Teeth in group 2 exhibited the greatest color change (p < 0.001). CONCLUSION: All tested materials showed loss of retention and color change in the experimental conditions. Infiltration system exhibited greatest weight loss and color change in demineralizing solution. Dental sealant exhibited greatest weight loss upon thermocycling. CLINICAL SIGNIFICANCE: Clinicians should be cautious about the limitations of retention and color stability when considering resin infiltration for incipient lesions.
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Condicionamento Ácido do Dente , Selantes de Fossas e Fissuras , Cimentos de Resina , Condicionamento Ácido do Dente/métodos , Cor , Colorimetria , Cárie Dentária , Cimentos Dentários , Esmalte Dentário , Teste de MateriaisRESUMO
Streptococcus mutans, a major etiological agent of human dental caries, lives primarily on the tooth surface in biofilms. Limited information is available concerning the extracellular DNA (eDNA) as a scaffolding matrix in S. mutans biofilms. This study demonstrates that S. mutans produces eDNA by multiple avenues, including lysis-independent membrane vesicles. Unlike eDNAs from cell lysis that were abundant and mainly concentrated around broken cells or cell debris with floating open ends, eDNAs produced via the lysis-independent pathway appeared scattered but in a structured network under scanning electron microscopy. Compared to eDNA production of planktonic cultures, eDNA production in 5- and 24-h biofilms was increased by >3- and >1.6-fold, respectively. The addition of DNase I to growth medium significantly reduced biofilm formation. In an in vitro adherence assay, added chromosomal DNA alone had a limited effect on S. mutans adherence to saliva-coated hydroxylapatite beads, but in conjunction with glucans synthesized using purified glucosyltransferase B, the adherence was significantly enhanced. Deletion of sortase A, the transpeptidase that covalently couples multiple surface-associated proteins to the cell wall peptidoglycan, significantly reduced eDNA in both planktonic and biofilm cultures. Sortase A deficiency did not have a significant effect on membrane vesicle production; however, the protein profile of the mutant membrane vesicles was significantly altered, including reduction of adhesin P1 and glucan-binding proteins B and C. Relative to the wild type, deficiency of protein secretion and membrane protein insertion machinery components, including Ffh, YidC1, and YidC2, also caused significant reductions in eDNA.
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Biofilmes/crescimento & desenvolvimento , Membrana Celular/fisiologia , DNA Bacteriano/metabolismo , Regulação Bacteriana da Expressão Gênica/fisiologia , Biossíntese de Proteínas/fisiologia , Streptococcus mutans/metabolismo , DNA Bacteriano/genética , Streptococcus mutans/genética , Streptococcus mutans/fisiologia , Streptococcus mutans/ultraestrutura , Regulação para CimaRESUMO
PURPOSE: The aim of this study was to determine and compare color differences of pressed lithium disilicate ceramic specimens after repeated firing cycles. An additional objective was to determine and evaluate correlation of CIEDE2000 values analyzed by X-Rite Color i5 Spectrophotometer, VITA EasyShade Advance 4.0 and Adobe Photoshop. MATERIAL AND METHODS: Tile specimens (N=36) with 8 x 10 x 1.5mm dimensions were prepared by IPS e.max Press lithium disilicate MT Monochromatic ingots and IPS e.max Multi Press lithium disilicate Multichromatic ingots. Specimens were exposed to 7 repeated firing cycles. Color analysis was performed after the 1st, 2nd, 3rd, 5th, and 7th firing cycles. CIE L*a*b* values were measured by X-Rite Color i5 Spectrophotometer, VITA EasyShade Advance 4.0 and Adobe Photoshop. CIE DE*2000 (ΔE*00) was calculated to estimate color differences. RESULTS: Linear regression and multiple comparison analysis (Tukey's HSD test) showed a statistically significant (p<.001) color difference ΔE*00 after multiple firing cycles. Statistically significant differences (p<.05) were also noted in different shade groups and between different instruments used for shade evaluation. Moreover, significant differences (p<.05) were found in interactive effects between different shades tested by different instrument, different shades tested after multiple firing cycles and different instruments after multiple firing cycles. CONCLUSIONS: Lithium disilicate material shows significant color differences after repeated firing cycles tested by three color analysis instruments. Measuring instruments used to evaluate CIE L*a*b* color values showed significant differences in color values analysis, which may lead to altered level of interpretation, particularly to determine perceptibility and clinical acceptability thresholds.
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Streptococcus mutans, the primary causative agent of dental caries, contains two paralogues of the LytR-CpsA-Psr family proteins encoded by brpA and psr, respectively. Previous studies have shown that BrpA plays an important role in cell envelope biogenesis/homeostasis and affects stress responses and biofilm formation by Strep. mutans, traits critical to cariogenicity of this bacterium. In this study, a Psr-deficient mutant, TW251, was constructed. Characterization of TW251 showed that deficiency of Psr did not have any major impact on growth rate. However, when subjected to acid killing at pH 2.8, the survival rate of TW251 was decreased dramatically compared with the parent strain UA159. In addition, TW251 also displayed major defects in biofilm formation, especially during growth with sucrose. When compared to UA159, the biofilms of TW251 were mainly planar and devoid of extracellular glucans. Real-time-PCR and Western blot analyses revealed that deficiency of Psr significantly decreased the expression of glucosyltransferase C, a protein known to play a major role in biofilm formation by Strep. mutans. Transmission electron microscopy analysis showed that deficiency of BrpA caused alterations in cell envelope and cell division, and the most significant defects were observed in TW314, a Psr-deficient and BrpA-down mutant. No such effects were observed with Psr mutant TW251 under similar conditions. These results suggest that while there are similarities in functions between BrpA and Psr, distinctive differences also exist between these two paralogues. Like Bacillus subtilis but different from Staphylococcus aureus, a functional BrpA or Psr is required for viability in Strep. mutans.
Assuntos
Proteínas de Bactérias/metabolismo , Glucanos/metabolismo , Proteínas Repressoras/metabolismo , Streptococcus mutans/genética , Streptococcus mutans/metabolismo , Ácidos/toxicidade , Proteínas de Bactérias/genética , Biofilmes/crescimento & desenvolvimento , Western Blotting , Parede Celular/ultraestrutura , Deleção de Genes , Perfilação da Expressão Gênica , Viabilidade Microbiana/efeitos dos fármacos , Microscopia Eletrônica de Transmissão , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Repressoras/genética , Streptococcus mutans/efeitos dos fármacos , Streptococcus mutans/fisiologiaRESUMO
Bacterial vaginosis (BV) is an infection of the vagina associated with thriving anaerobes, such as Gardnerella vaginitis and other associated pathogens. These pathogens form a biofilm responsible for the recurrence of infection after antibiotic therapy. The aim of this study was to develop a novel mucoadhesive polyvinyl alcohol and polycaprolactone electrospun nanofibrous scaffolds for vaginal delivery, incorporating metronidazole, a tenside, and Lactobacilli. This approach to drug delivery sought to combine an antibiotic for bacterial clearance, a tenside biofilm disruptor, and a lactic acid producer to restore healthy vaginal flora and prevent the recurrence of bacterial vaginosis. F7 and F8 had the least ductility at 29.25% and 28.39%, respectively, and this could be attributed to the clustering of particles that prevented the mobility of the crazes. F2 had the highest at 93.83% due to the addition of a surfactant that increased the affinity of the components. The scaffolds exhibited mucoadhesion between 31.54 ± 0.83% and 57.86 ± 0.95%, where an increased sodium cocoamphoacetate concentration led to increased mucoadhesion. F6 showed the highest mucoadhesion at 57.86 ± 0.95%, as compared to 42.67 ± 1.22% and 50.89 ± 1.01% for the F8 and F7 scaffolds, respectively. The release of metronidazole via a non-Fickian diffusion-release mechanism indicated both swelling and diffusion. The anomalous transport within the drug-release profile pointed to a drug-discharge mechanism that combined both diffusion and erosion. The viability studies showed a growth of Lactobacilli fermentum in both the polymer blend and the nanofiber formulation that was retained post-storage at 25 °C for 30 days. The developed electrospun scaffolds for the intravaginal delivery of Lactobacilli spp., along with a tenside and metronidazole for the management of bacterial vaginosis, provide a novel tool for the treatment and management of recurrent vaginal infection.
RESUMO
Bronchopulmonary dysplasia (BPD) is characterized by abnormal development of the blood vessels and alveoli in lungs, which largely occurs in premature infants. Exosomes (EXO) from very preterm infants (VPI) with BPD (BPD-EXO) impair angiogenic activities of human umbilical vein endothelial cells (HUVECs) via EXO-miRNAs cargo. This study aimed to determine whether and how BPD-EXO affect the development of BPD in a mouse model. We showed that treating BPD mice with BPD-EXO chronically and irreversibly aggravated lung injury. BPD-EXO up-regulated 139 and down-regulated 735 genes in the mouse lung tissue. These differentially expressed genes were enriched to the MAPK pathway (e.g., Fgf9 and Cacna2d3), which is critical to angiogenesis and vascular remodeling. BPD-EXO suppressed expression of Fgf9 and Cacna2d3 in HUVECs and inhibited migration, tube formation, and increased cell apoptosis in HUVECs. These data demonstrate that BPD-EXO aggravate lung injury in BPD mice and impair lung angiogenesis, plausibly leading to adverse outcomes of VPI with BPD. These data also suggest that BPD-EXO could serve as promising targets for predicting and treating BPD.