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BACKGROUND: Occupational exposures to ultrafine particles in the plume generated during laser hair removal procedures, the most commonly performed light based cosmetic procedure, have not been thoroughly characterized. Acute and chronic exposures to ambient ultrafine particles have been associated with a number of negative respiratory and cardiovascular health effects. Thus, the aim of this study was to measure airborne concentrations of particles in a diameter size range of 10 nm to 1 µm in procedure rooms during laser hair removal procedures. METHODS: TSI Model 3007 Condensation Particle Counters were used to quantify the particle count concentrations in the waiting and procedure rooms of a dermatology office. Particle concentrations were sampled before, during, and after laser hair removal procedures, and characteristics of each procedure were noted by the performing dermatologist. RESULTS: Twelve procedures were sampled over 4 days. Mean ultrafine particle concentrations in the waiting and procedure rooms were 14,957.4 particles/cm3 and 22,916.8 particles/cm3 (p < 0.0001), respectively. Compared to background ultrafine particle concentrations before the procedure, the mean concentration in the procedure room was 2.89 times greater during the procedure (p = 0.009) and 2.09 times greater after the procedure (p = 0.007). Duration of procedure (p = 0.006), body part (p = 0.013), and the use of pre-laser lotion/type of laser (p = 0.039), were the most important predictors of ultrafine particle concentrations. Use of a smoke evacuator (a recommended form of local exhaust ventilation) positioned at 30.5 cm from the source, as opposed to the recommended 1-2 in., lowered particle concentrations, but was not a statistically significant predictor (p = 0.49). CONCLUSIONS: Laser hair removal procedures can generate high exposures to ultrafine particles for dermatologists and other individuals performing laser hair removal, with exposure varying based on multiple determinants.
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Poluentes Atmosféricos/análise , Remoção de Cabelo , Exposição Ocupacional/análise , Material Particulado/análise , Monitoramento Ambiental , Humanos , Lasers , Tamanho da PartículaRESUMO
A good understanding of the mechanism of interaction between quantum dots (QDs) and heavy metal ions is essential for the design of more effective sensor systems. In this work, CE was introduced to explore how l-cysteine-capped-CdTe QDs (l-cys-CdTe QDs) interacts with Hg(2+) . The change in electrophoretic mobility can synchronously reflect the change in the composition and property of QDs. The effects of the free and capping ligands on the system are discussed in detail. ESI-MS, dynamic light scattering (DLS), zeta potential, and fluorescence (FL) were also applied as cooperative tools to study the interaction mechanism. Furthermore, the interaction mechanism, which principally depended on the concentration of Hg(2+) , was proposed reasonably. At the low concentration of Hg(2+) , the formation of a static complex between Hg(2+) and the carboxyl and amino groups of l-cys-CdTe QDs surface was responsible for the FL quenching. With the increase of Hg(2+) concentration, the capping l-cys was stripped from the surface of l-cys-CdTe QDs due to the high affinity of Hg(2+) to the thiol group of l-cys. Our study demonstrates that CE can reveal the mechanism of the interaction between QDs and heavy metal ions, such as FL quenching.
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Compostos de Cádmio/química , Cisteína/química , Eletroforese Capilar/métodos , Mercúrio/química , Pontos Quânticos/química , Telúrio/química , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
Multiple myeloma (MM) induced bone lesion is one of the most crippling characteristics, and the MM secreted Dickkopf-1 (DKK1) has been reported to play important role in this pathologic process. However, the underlying regulation mechanisms involved in DKK1 expression are still unclear. In this study, we validated the expression patterns of microRNA (miR) 15a, 34a, 152, and 223 in MM cells and identified that miR-152 was significantly downregulated in the MM group compared with the non-MM group, and that miR-152 level was negatively correlated with the expression of DKK1 in the MM cells. Mechanistic studies showed that manipulating miR-152 artificially in MM cells led to changes in DKK-1 expression, and miR-152 blocked DKK1 transcriptional activity by binding to the 3'UTR of DKK1 mRNA. Importantly, we revealed that MM cells stably expressing miR-152 improved the chemotherapy sensitivity, and counteracted the bone disruption in an intrabone-MM mouse model. Our study contributes better understanding of the regulation mechanism of DKK-1 in MM, and opens up the potential for developing newer therapeutic strategies in the MM treatment.
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Regulação para Baixo/genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , MicroRNAs/genética , Mieloma Múltiplo/genética , Regiões 3' não Traduzidas/genética , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Sequência de Bases , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/patologia , Linhagem Celular Tumoral , Regulação para Baixo/efeitos dos fármacos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Melfalan/farmacologia , Camundongos SCID , MicroRNAs/metabolismo , Dados de Sequência Molecular , Ligação Proteica/efeitos dos fármacos , Ligação Proteica/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
This study investigates the nexus between carbon dioxide emissions, economic development, and development finance in seeking an empirical answer to the conundrum at the intersection of development and environmental economics. Employing a theoretical framework that incorporates three dimensions of endowments, the real economy, and the financial sector, our empirical model accounts for the bi-directional causality of environmental degradation and economic growth in the Global South by adopting the simultaneous equations model. Our results confirm an inverted N-shaped environment Kuznets curve which is statistically significant and robust, and consistent with the conceptualized theoretical framework. The results provide insights to enhance the effectiveness of future development finance and policy design by promoting sustainable growth and green transformation through facilitating renewable energy adoption, investing in human capital, and preserving renewable natural capital.
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The reuse of activated sludge as a solid waste is severely underutilized due to the limitations of traditional treatment and disposal methods. Given that, the sulfur-containing activated sludge catalyst doped with cobalt (SK-Co(1.0)) was successfully prepared by one-step pyrolysis and calcinated at 850 â. The generation of CoSx was successfully characterized by X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS), indicating that the sulfur inside the sludge was the anchoring site for the externally doped cobalt. Cobalt (â ¡) (Co2+), as the main adsorption site for peroxymonosulfate(PMS), formed a complex (SK-Co(1.0)-PMS* ) and created the conditions for the generation of surface radicals. The SK-Co(1.0)/PMS system showed high degradation efficiency and apparent rate constants for Sulfamethoxazole (SMX) (91.56% and 0.187 min-1) and Sulfadiazine (SDZ) (90.73% and 0.047 min-1) within 10 min and 30 min, respectively. Three sites of generation of 1O2, which played a dominant role in the degradation of SMX and SDZ in the SK-Co(1.0)/PMS system, were summarized asï¼sulfur vacancies (SVs), the Co3+/Co2+ cycles promoted by sulfur(S) species, oxygen-containing functional groups (C-O). The degradation mechanisms and pathways had been thoroughly investigated using DFT calculations. In view of this, a new idea for the resource utilization of activated sludge solid waste was provided and a new strategy for wastewater remediation was proposed.
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Antibacterianos , Carvão Vegetal , Esgotos , Cobalto , Resíduos Sólidos , Sulfonamidas , Sulfametoxazol , Sulfanilamida , Peróxidos/química , Sulfadiazina , CatáliseRESUMO
Ovarian cancer is one of the most common gynecological malignant tumors with insidious onset, strong invasiveness, and poor prognosis. Metabolic alteration, particularly aerobic glycolysis, which is tightly regulated by transcription factors, is associated with the malignant behavior of OC. We screened FOXK2 in this study as a key transcription factor that regulates glycolysis in OC. FOXK2 is overly expressed in OC, and poor prognosis is predicted by overexpression. FOXK2 promotes OC cell proliferation both in vitro and in vivo and cell migration in vitro. Further studies showed that PDK2 directly binds to the forkhead-associated (FHA) domain of FOXK2 to phosphorylate FOXK2 at Thr13 and Ser30, thereby enhancing the transcriptional activity of FOXK2. FOXK2 transcriptionally regulates the expression of PDK2, thus forming positive feedback to sustain glycolysis in OC cells.
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Proliferação de Células , Fatores de Transcrição Forkhead , Glicólise , Neoplasias Ovarianas , Piruvato Desidrogenase Quinase de Transferência de Acetil , Humanos , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/genética , Feminino , Glicólise/genética , Piruvato Desidrogenase Quinase de Transferência de Acetil/metabolismo , Piruvato Desidrogenase Quinase de Transferência de Acetil/genética , Fatores de Transcrição Forkhead/metabolismo , Fatores de Transcrição Forkhead/genética , Linhagem Celular Tumoral , Fosforilação , Animais , Proliferação de Células/genética , Camundongos , Movimento Celular/genética , Regulação Neoplásica da Expressão Gênica , Retroalimentação Fisiológica , Camundongos Nus , PrognósticoRESUMO
Accurate genotyping is important for genetic testing. Sanger sequencing-based typing is the gold standard for genotyping, but it has been underused, due to its high cost and low throughput. In contrast, short-read sequencing provides inexpensive and high-throughput sequencing, holding great promise for reaching the goal of cost-effective and high-throughput genotyping. However, the short-read length and the paucity of appropriate genotyping methods, pose a major challenge. Here, we present RCHSBT-reliable, cost-effective and high-throughput sequence based typing pipeline-which takes short sequence reads as input, but uses a unique variant calling, haploid sequence assembling algorithm, can accurately genotype with greater effective length per amplicon than even Sanger sequencing reads. The RCHSBT method was tested for the human MHC loci HLA-A, HLA-B, HLA-C, HLA-DQB1, and HLA-DRB1, upon 96 samples using Illumina PE 150 reads. Amplicons as long as 950 bp were readily genotyped, achieving 100% typing concordance between RCHSBT-called genotypes and genotypes previously called by Sanger sequence. Genotyping throughput was increased over 10 times, and cost was reduced over five times, for RCHSBT as compared with Sanger sequence genotyping. We thus demonstrate RCHSBT to be a genotyping method comparable to Sanger sequencing-based typing in quality, while being more cost-effective, and higher throughput.
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Técnicas de Genotipagem , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Reação em Cadeia da Polimerase Multiplex , Análise Custo-Benefício , Testes Genéticos/métodos , Antígenos HLA/genética , Sequenciamento de Nucleotídeos em Larga Escala/economia , Sequenciamento de Nucleotídeos em Larga Escala/normas , Humanos , Reprodutibilidade dos TestesRESUMO
Because of the unusual properties of the structure, the metal organic frameworks (MOFs) have received great interest in separation science. However, the most existing methods for the applications of MOFs in separation science require an off-line procedure to prepare the materials. Here, we report an in situ, layer-by-layer self-assembly approach to fabricate MIL-100(Fe) coated open tubular (OT) capillary columns for capillary electrochromatography. By a controllable manner, the OT capillary columns with a tailored MIL-100(Fe) coating have been successfully synthesized. The results of SEM, XRD, FT-IR, and ICP-AES indicated that MIL-100(Fe) was successfully grafted on the inner wall of the capillary. Some neutral, acidic and basic analytes were used to evaluate the performance of the MIL-100(Fe) coating OT capillary column. Because of the size selectivity of lattice aperture and hydrophobicity of the organic ligands, three types of analytes were well separated with this novel MIL-100(Fe) coating OT capillary column. For three consecutive runs, the intraday relative standard deviations (RSDs) of migration time and peak areas were 0.4-4.6% and 1.2-6.6%, respectively. The interday RSDs of migration time and peak areas were 0.6-8.0% and 2.2-9.5%, respectively. The column-to-column reproducibility of retention time was in range of 0.6-9.2%. Additionally, the 10 cycles OT capillary column (10-LC) could be used for more than 150 runs with no observable changes on the separation efficiency.
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Easily developed chemoresistance is a major characteristic of multiple myeloma (MM) and the main obstacle in curing MM in the clinic, but the key regulators have not been fully identified. In the current study, we find that PPFIA Binding Protein 1 (PPFIBP1) is highly expressed in the plasma cells from MM patients, and higher PPFIBP1 expression predicts poorer outcomes. PPFPIBP1 enhances chemoresistance of MM cells to the treatment of bortezomib (BTZ), a proteasome inhibitor, and manipulation of PPFPIBP1 can alter chemosensitivity of MM cells to BTZ. Mechanistic studies reveal that PPFPIBP1 directly binds and stabilizes RelA, promotes the cyto-nuclear translocation of RelA, and activates NF-κB signaling pathway. Targeting PPFPIBP1 in a xenograft mouse model of MM prohibits tumor growth and prolongs overall survival of mice. Taken together, our findings suggest that PPFIBP1 is a crucial regulator of chemoresistance to PIs in MM cells, and shed light on developing therapeutic strategies to overcome chemoresistance by targeting PPFIBP1.
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Metal organic frameworks-Covalent organic frameworks (MOFs-COFs) nanocomposites could improve the catalytic performance. Herein, a novel nanocomposite catalyst (CC@Co3O4) derived from MOFs-COFs (COF@ZIF-67) was prepared on peroxymonosulfate (PMS) activation for bisphenol A (BPA) and rhodamine B (RhB) degradation. Owing to the Co species, oxygen vacancy (OV), surface hydroxyl (-OH), graphite N and ketone groups (C=O), the CC@Co3O4 exhibited higher catalytic degradation performance and total organic carbon (TOC) for BPA (93.8% and 22.3%) and RhB (98.2% and 82.5%) with a small quantity of catalyst (0.10 g/L) and low concentration of PMS (0.20 g/L) even without pH adjustment. Sulfate radicals (â¢SO4-), hydroxyl radicals (â¢OH), single oxygen (1O2), superoxide radicals (â¢O2-) and electron transfer process were all involved in the degradation of BPA and RhB. Among them, the degradation of BPA and RhB mainly depended on â¢O2- and 1O2, respectively. Meanwhile, the degradation pathways of BPA and RhB were proposed, and the biotoxicity of the degradation products was evaluated by freshwater chlorella. The results illustrated that the degradation products were environmentally friendly to organisms. In addition, the role of COF in the nanocomposites was also studied. The addition of COF remarkably improved the catalytic performance of CC@Co3O4 due to the faster electron transfer, more graphite N and C=O. Overall, this work may open the door to the development of COF-based catalysts in the field of water pollutant remediation.
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Chlorella , Poluentes Ambientais , Grafite , Estruturas Metalorgânicas , Nanocompostos , Peróxidos/química , Nanocompostos/química , OxigênioRESUMO
Ischemic stroke, the third leading cause of disability globally, imposes a notable economic burden. Tetrandrine (Tet), which has been widely used clinically, exhibits potential protective effects against stroke. However, there has been little preclinical research to evaluate the therapeutic effects of Tet on stroke. The present study investigated the beneficial effect of Tet on ischemiareperfusion (I/R) injury and its underlying mechanism in rats. Rats were subjected to occlusion of the middle cerebral artery, then treated with Tet (30 mg/kg/day, intraperitoneal) in the subacute phase for 7 days. In order to detect the effects of Tet on the behavior of rats, modified neurological severity score and longa behavior, grasping capability and inclined plane tests were conducted on days 1, 3 and 7 following cerebral ischemia. In addition, neuronal apoptosis in the cortex and hippocampus following ischemia was assessed by Nissl staining and TUNEL assay. Finally, oxidative stress was evaluated by measurement of free radicals and immunofluorescence staining of LC3 was used to assess autophagy. Tet improved neurological function and decreased infarct volume in I/R injury rats. Tet also prevented neuronal apoptosis in the cortex and hippocampus region. In addition, Tet protected against oxidative damage following ischemia, which was reflected by decreased levels of nitric oxide and malondialdehyde and increased levels of glutathione (GSH) and GSH peroxidase. In addition, the expression levels of the autophagy marker LC3 decreased in the Tet treatment group. In conclusion, Tet attenuated I/Rinduced neuronal damage in the subacute phase by decreasing oxidative stress, apoptosis and autophagy.
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Apoptose/efeitos dos fármacos , Benzilisoquinolinas/farmacologia , Neurônios/efeitos dos fármacos , Traumatismo por Reperfusão/prevenção & controle , Acidente Vascular Cerebral/prevenção & controle , Animais , Autofagia/efeitos dos fármacos , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Córtex Cerebral/patologia , Modelos Animais de Doenças , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Hipocampo/patologia , Humanos , Masculino , Malondialdeído/metabolismo , Neurônios/patologia , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/complicações , Acidente Vascular Cerebral/etiologiaRESUMO
Excessive monocyte activation with the development of excessive or uncontrolled release of proinflammatory cytokines often results in host tissue injury and even death in patients with pneumonia caused by the 2019 novel coronavirus. However, the changes of cytokine profiles of coronavirus disease 2019 (COVID-19) patients, as well as the underlying mechanisms that are involved, remain unknown. Using a cytokine array containing 174 inflammation-related cytokines, we found significantly altered cytokine profiles in severe COVID-19 patients compared with those in mild patients or healthy controls, and identified leptin, CXCL-10, IL-6, IL-10, IL-12, and TNF-α as the top differentially expressed cytokines. Notably, leptin showed high consistency with CXCL-10 and TNF-α in predicting disease severity, and correlated with body mass index, decreased lymphocyte counts, and disease progression. Further analysis demonstrated that monocytes in severe patients with higher leptin levels were inclined toward M1 polarization. Mechanistic studies revealed that leptin synergistically up-regulated expression levels of inflammatory cytokines and surface markers with IL-6 in monocytes through STAT3 and NF-κB signaling pathways. Collectively, our results suggest that overweight COVID-19 patients were prone to have higher leptin levels, which further activated monocytes, resulting in amplified or dysregulated immune responses. Taken together, our findings argue that leptin correlates severity of COVID-19 and may indicate a possible mechanism by which overweight patients have a greater tendency to develop severe conditions.
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COVID-19/patologia , Leptina/metabolismo , Monócitos/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , COVID-19/sangue , COVID-19/virologia , Polaridade Celular , Criança , Citocinas/sangue , Citocinas/metabolismo , Progressão da Doença , Feminino , Humanos , Mediadores da Inflamação/metabolismo , Leptina/sangue , Masculino , Pessoa de Meia-Idade , NF-kappa B/metabolismo , SARS-CoV-2/fisiologia , Fator de Transcrição STAT3 , Índice de Gravidade de Doença , Transdução de Sinais , Adulto JovemRESUMO
Myeloma cells produce excessive levels of dickkopf-1 (DKK1), which mediates the inhibition of Wnt signaling in osteoblasts, leading to multiple myeloma (MM) bone disease. Nevertheless, the precise mechanisms underlying DKK1 overexpression in myeloma remain incompletely understood. Herein, we provide evidence that hypoxia promotes DKK1 expression in myeloma cells. Under hypoxic conditions, p38 kinase phosphorylated cAMP-responsive element-binding protein (CREB) and drove its nuclear import to activate DKK1 transcription. In addition, high levels of DKK1 were associated with the presence of focal bone lesions in patients with t(4;14) MM, overexpressing the histone methyltransferase MMSET, which was identified as a downstream target gene of hypoxia-inducible factor (HIF)-1α. Furthermore, we found that CREB could recruit MMSET, leading to the stabilization of HIF-1α protein and the increased dimethylation of histone H3 at lysine 36 on the DKK1 promoter. Knockdown of CREB in myeloma cells alleviated the suppression of osteoblastogenesis by myeloma-secreted DKK1 in vitro. Combined treatment with a CREB inhibitor and the hypoxia-activated prodrug TH-302 (evofosfamide) significantly reduced MM-induced bone destruction in vivo. Taken together, our findings reveal that hypoxia and a cytogenetic abnormality regulate DKK1 expression in myeloma cells, and provide an additional rationale for the development of therapeutic strategies that interrupt DKK1 to cure MM.
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Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Histona-Lisina N-Metiltransferase/genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , Mieloma Múltiplo/tratamento farmacológico , Proteínas Repressoras/genética , Cromatina/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Mieloma Múltiplo/genética , Mieloma Múltiplo/patologia , Nitroimidazóis/farmacologia , Osteoblastos/efeitos dos fármacos , Osteólise/genética , Mostardas de Fosforamida/farmacologia , Hipóxia Tumoral/efeitos dos fármacos , Via de Sinalização Wnt/efeitos dos fármacosRESUMO
Long noncoding RNAs (lncRNAs) may serve as potential molecular diagnostic markers to improve the capacity of earlier and more accurate diagnosis of dilated cardiomyopathy (DCM). We integrated five independent transcriptomic datasets (n = 504) from Gene Expression Omnibus for systematic identification of lncRNA-based diagnostic biomarkers in DCM. The multivariate logistic regression model based on the six lncRNAs (AC016722.3, AL589986.2, AC006007.1, AC092687.3, GS1-124K5.4, and AC007126.1) in the ceRNA networks showed high sensitivity and specificity (area under curves >0.8, p < 0.0001) of DCM diagnosis in the training and validation datasets. Functional analysis revealed that the autophagy, protein acetyltransferase, and DNA polymerase activity were associated with high levels of the six-lncRNA signature, while the collagen trimer, extracellular matrix structural constituent, and MHC protein complex were associated with low levels of the signature. Pathway analysis showed that high levels of the six-lncRNA signature were associated with upregulated selective autophagy, interleukin 17 signalings, and extracellular matrix interactions, while were associated with downregulated extracellular matrix organization and collagen formation. The identified six-lncRNA signature, with high performance in molecular diagnosis of DCM, might be applied in future clinical practices combined with traditional markers.
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We report acute antibody responses to SARS-CoV-2 in 285 patients with COVID-19. Within 19 days after symptom onset, 100% of patients tested positive for antiviral immunoglobulin-G (IgG). Seroconversion for IgG and IgM occurred simultaneously or sequentially. Both IgG and IgM titers plateaued within 6 days after seroconversion. Serological testing may be helpful for the diagnosis of suspected patients with negative RT-PCR results and for the identification of asymptomatic infections.
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Anticorpos Antivirais/sangue , Formação de Anticorpos/efeitos dos fármacos , Betacoronavirus/patogenicidade , Infecções por Coronavirus/tratamento farmacológico , Pneumonia Viral/tratamento farmacológico , Adulto , Idoso , Formação de Anticorpos/imunologia , Antivirais/uso terapêutico , Betacoronavirus/genética , COVID-19 , Infecções por Coronavirus/sangue , Infecções por Coronavirus/imunologia , Infecções por Coronavirus/virologia , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Pandemias/prevenção & controle , Pneumonia Viral/sangue , Pneumonia Viral/imunologia , Pneumonia Viral/virologia , SARS-CoV-2RESUMO
OBJECTIVE: This study was aimed to explore effect of baicalin on IKK/IKB/NF-kB signaling pathway and apoptosis-related proteins in rats with ulcerative colitis (UC). METHODS: Histopathological observation and scores of colon tissue were performed in the UC rat model. IKK/IKB/NF-kB signaling pathway and apoptosis-related proteins were measured by Western blotting. RESULTS: Baicalin significantly increased the activity of SOD, CAT and GSH-Px in colon tissue of rats with UC, but significantly decreased the content of MDA, IL-1ß, MPO, PEG2 and TNF-α in colon tissue of rats with UC. In the molecular mechanism, baicalin significantly decreased the expression of cleaved-caspase3, cleaved-caspase9, Bcl-2/Bax, cyt-c, NF-kB p-65, p-IKKß/IKKß and p-IKBα/IKBα. Baicalin could significantly inhibit p-IKBα/IKBα content change, but had no significant effect on p-IKKß/IKKß. CONCLUSION: Baicalin may have a regulating effect on IKK/IKB/NF-kB signaling pathway and apoptosis-related proteins in UC rats.
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Colite Ulcerativa/metabolismo , Flavonoides/farmacologia , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Colite Ulcerativa/patologia , Colo/efeitos dos fármacos , Colo/metabolismo , Colo/patologia , Quinase I-kappa B/metabolismo , Interleucina-1beta/metabolismo , Masculino , Camundongos , NF-kappa B/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Peroxidase/metabolismo , Células RAW 264.7 , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The effector function of natural killer, lymphokine--activated killer cells and T lymphocytes is commonly evaluated by radioactive chromiumrelease cytotoxicity assays. In addition to this indirect method, fluorescence assays have been described for the assessment of in vitro cellmediated cytotoxicity. In the present study, target cells were stained with 5(and6)carboxyfluorescein diacetate succinimidyl ester (CFSE), which is a stable integrated fluorescent probe that allows target and effector cells to be distinguished from one another. Staining of target THP1 cells with 8 µM CFSE revealed high and stable loading of fluorescence and no effect of the viability of cells. After 4 h of in vitro coculture between γδ T cells and CFSElabeled infected or uninfected THP1 cells, staining with propidium iodide (PI) was performed to distinguish between vital and dead cells. During sample acquisition, target cells were gated on the CFSE positivity and examined for cell death based on the uptake of PI. CFSE and PI double positive cells were recognized as the dead target cells. The percentage of cytotoxicity in the CFSEgated cell population was calculated by subtracting the value obtained for nonspecific PIpositive target cells, which was measured in a control group that did not contain effector cells. The present study describes a simple and convenient assay that is based on the direct quantitative and qualitative analysis of cell damage at a single cell level utilizing a twocolor flow cytometric assay. In conclusion, the flow cytometricbased assay described in the current study is a simple, sensitive and reliable tool to determine the cytolytic activity of γδ T lymphocytes against mycobacteria. Therefore, the present study may provide valuable information concerning the methods employed to investigate the function of γδ T cells and potentially other lymphocyte subsets.
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Citotoxicidade Imunológica/fisiologia , Linfócitos Intraepiteliais/citologia , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Técnicas de Cocultura , Citometria de Fluxo , Corantes Fluorescentes/química , Humanos , Linfócitos Intraepiteliais/imunologia , Linfócitos Intraepiteliais/metabolismo , Receptores de Lipopolissacarídeos/metabolismo , Microscopia de Fluorescência , Mycobacterium tuberculosis/fisiologia , Fagocitose , Acetato de Tetradecanoilforbol/análogos & derivados , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
The popularity of novel nanoparticles coated capillary column has aroused widespread attention of researchers. Metal organic frameworks (MOFs) with special structure and chemical properties have received great interest in separation sciences. This work presents the investigation of HKUST-1 (Hong Kong University of Science and Technology-1, called Cu3(BTC)2 or MOF-199) nanoparticles as a new type of coating material for capillary electrochromatography. For the first time, three layers coating (3-LC), five layers coating (5-LC), ten layers coating (10-LC), fifteen layers coating (15-LC), twenty layers coating(20-LC) and twenty-five layers coating (25-LC) capillary columns coated with HKUST-1 nanoparticles were synthesized by covalent bond with in situ, layer-by-layer self-assembly approach. The results of scanning electron microscopy (SEM), X-ray diffraction (XRD) and plasma atomic emission spectrometry (ICP-AES) indicated that HKUST-1 was successfully grafted on the inner wall of the capillary. The separating performances of 3-LC, 5-LC, 10-LC, 15-LC, 20-LC and 25-LC open tubular (OT) capillary columns were studied with some neutral small organic molecules. The results indicated that the neutral small organic molecules were separated successfully with 10-LC, 15-LC and 20-LC OT capillary columns because of the size selectivity of lattice aperture and hydrophobicity of organic ligands. In addition, 10-LC and 15-LC OT capillary columns showed better performance for the separation of certain phenolic compounds. Furthermore, 10-LC, 15-LC and 20-LC OT capillary columns exhibited good intra-day repeatability with the relative standard deviations (RSDs; %) of migration time and peak areas lying in the range of 0.3-1.2% and 0.5-4.2%, respectively. For inter-day reproducibility, the RSDs of the three OT capillary columns were found to be lying in the range of 0.3-5.5% and 0.3-4.5% for migration time and peak area, respectively. The RSDs of retention times for column-to-column for three batches of 10-LC, 15-LC and 20-LC OT capillary columns were in the range from 2.3% to 7.2%. Moreover, the fabricated 10-LC, 15-LC and 20-LC OT capillary columns exhibited good repeatability and stability for separation, which could be used successively for more than 120 runs with no observable changes on the separation efficiency.
Assuntos
Eletrocromatografia Capilar/métodos , Compostos Orgânicos/isolamento & purificação , Compostos Organometálicos/química , Soluções Tampão , Etanol/análise , Concentração de Íons de Hidrogênio , Estruturas Metalorgânicas , Microscopia Eletrônica de Varredura , Nanopartículas/química , Nanopartículas/ultraestrutura , Fenóis/análise , Reprodutibilidade dos Testes , Soluções , Difração de Raios XRESUMO
Covalent organic frameworks (COFs) have attracted much attention because of their permanent nanoscale porosity and higher surface area compared to zeolites as well as robustness. COFs have great potential in several fields such as hydrogen storage, gas separation, and catalysis. However, COFs have not yet been applied in capillary electrochromatography. Herein, covalent organic frameworks-LZU1 (COF-LZU1) was used as the stationary phase in open-tubular capillary electrochromatography for the first time. Compared to the monoliths used in electrochromatography, the preparation technique of a COF-LZU1-coated capillary was simple and practical. The baseline separation of model analytes including alkylbenzenes, polyaromatic hydrocarbons, and anilines by the COF-LZU1-coated capillary was achieved based on the size selectivity of COF-LZU1 porous structure and hydrophobic interactions between the model analytes and organic ligands of COF-LZU1. The load capacity of the COF-LZU1-coated capillary for naphthalene was 0.6mg/mL. For three consecutive runs, the intraday relative standard deviations (RSDs) were 1.4-2.6% for the migration time and 2.7-8.7% for the peak area. The interday RSDs were 1.3-3.9% for the migration time and 3.7-9.7% for the peak area. The column-to-column reproducibility of migration time was in the range 1.0-3.9%. Moreover, the coated capillary was used for >300 runs with no changes in the separation efficiency. Thus, COFs have great potential in capillary electrochromatography and may provide a new method for chromatographic separation.