Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 42
Filtrar
Mais filtros

Base de dados
País/Região como assunto
Tipo de documento
Intervalo de ano de publicação
1.
Cerebrovasc Dis ; 2024 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-38527440

RESUMO

INTRODUCTION: Although statin therapy reduces cardiovascular events, statin use is associated with the risk of new-onset diabetes mellitus (NODM). Using a linked dataset, we evaluated the effect of statin treatment on vascular outcomes and NODM development in patients with ischemic stroke. METHODS: From the dataset, we identified 20,250 patients with acute ischemic stroke who had neither a prior history of DM nor a previous history of statin use before the index stroke. Patients were divided into statin users and non-users. The outcomes were NODM and vascular outcomes, including recurrent ischemic stroke and acute myocardial infarction (AMI). RESULTS: Of the 20,250 patients, 13,706 (67.7%) received statin treatment after the index stroke. For the risk of NODM, a time-response relationship was observed between the use of statins and NODM; a longer post-stroke follow-up duration substantially increased the risk of NODM. Among those with ischemic stroke exceeding 3 years, statin users had an approximately 1.7-fold greater risk of NODM than statin non-users. Statin therapy significantly reduced the risk of recurrent ischemic stroke by 54% (HR 0.46, 95% CI, 0.43-0.50, P < 0.001) across all stroke subtypes. CONCLUSION: Statin therapy following ischemic stroke increased the occurrence of NODM in patients over a period of 3 years. Despite the increased risk of NODM, statin therapy shows a beneficial effect in reducing major cardiovascular events such as recurrent ischemic stroke and AMI in patients with ischemic stroke.

2.
J Korean Med Sci ; 39(14): e127, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38622936

RESUMO

BACKGROUND: To overcome the limitations of relying on data from a single institution, many researchers have studied data linkage methodologies. Data linkage includes errors owing to legal issues surrounding personal information and technical issues related to data processing. Linkage errors affect selection bias, and external and internal validity. Therefore, quality verification for each connection method with adherence to personal information protection is an important issue. This study evaluated the linkage quality of linked data and analyzed the potential bias resulting from linkage errors. METHODS: This study analyzed claims data submitted to the Health Insurance Review and Assessment Service (HIRA DATA). The linkage errors of the two deterministic linkage methods were evaluated based on the use of the match key. The first deterministic linkage uses a unique identification number, and the second deterministic linkage uses the name, gender, and date of birth as a set of partial identifiers. The linkage error included in this deterministic linkage method was compared with the absolute standardized difference (ASD) of Cohen's according to the baseline characteristics, and the linkage quality was evaluated through the following indicators: linked rate, false match rate, missed match rate, positive predictive value, sensitivity, specificity, and F1-score. RESULTS: For the deterministic linkage method that used the name, gender, and date of birth as a set of partial identifiers, the true match rate was 83.5 and the missed match rate was 16.5. Although there was bias in some characteristics of the data, most of the ASD values were less than 0.1, with no case greater than 0.5. Therefore, it is difficult to determine whether linked data constructed with deterministic linkages have substantial differences. CONCLUSION: This study confirms the possibility of building health and medical data at the national level as the first data linkage quality verification study using big data from the HIRA. Analyzing the quality of linkages is crucial for comprehending linkage errors and generating reliable analytical outcomes. Linkers should increase the reliability of linked data by providing linkage error-related information to researchers. The results of this study will serve as reference data to increase the reliability of multicenter data linkage studies.


Assuntos
Armazenamento e Recuperação da Informação , Registro Médico Coordenado , Humanos , Reprodutibilidade dos Testes , Registro Médico Coordenado/métodos , Valor Preditivo dos Testes , Serviços de Saúde
3.
FASEB J ; 34(7): 9141-9155, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32421247

RESUMO

Human-induced pluripotent stem cells (hiPSCs) are invaluable sources for drug screening and toxicity tests because of their differentiation potential and proliferative capacity. Recently, the CRISPR-Cas9-mediated homologous recombination system has enabled reporter knock-ins at desired loci in hiPSCs, and here, we generated a hiPSC reporter line expressing mCherry-tagged cytochrome P450 1A1 (CYP1A1), which can be utilized to screen for the modulators of aryl hydrocarbon receptor (AHR) in live cells. CYP1A1-mCherry hiPSCs exhibited typical characteristics of pluripotent stem cells such as marker expression, differentiation potential, and normal karyotype. After differentiation into hepatocyte-like cells (HLCs), CYP1A1-mCherry fusion protein was expressed and localized at the endoplasmic reticulum, and induced by AHR agonists. We obtained 23 hits modulating CYP1A1 expression from high-content screening with 241 hepatotoxicity chemicals and nuclear receptor ligands, and identified three upregulating chemicals and two downregulating compounds. Responses of hiPSC-HLCs against an AHR agonist were more similar to human primary hepatocytes than of HepG2 hepatocellular carcinoma cells. This platform has the advantages of live-cell screening without sacrificing cells (unlike previously available CYP1A1 reporter cell lines), as well as an indefinite supply of cells, and can be utilized in a wide range of screening related to AHR- and CYP1A1-associated diseases in desired cell types.


Assuntos
Citocromo P-450 CYP1A1/química , Fluorescência , Hepatócitos/citologia , Células-Tronco Pluripotentes Induzidas/citologia , Preparações Farmacêuticas/metabolismo , Bibliotecas de Moléculas Pequenas/farmacologia , Diferenciação Celular , Citocromo P-450 CYP1A1/metabolismo , Células Hep G2 , Hepatócitos/efeitos dos fármacos , Ensaios de Triagem em Larga Escala , Humanos , Células-Tronco Pluripotentes Induzidas/efeitos dos fármacos , Células-Tronco Pluripotentes Induzidas/enzimologia , Transdução de Sinais
4.
J Korean Med Sci ; 35(21): e135, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32476299

RESUMO

BACKGROUND: Neurocritical care by dedicated neurointensivists may improve outcomes of critically ill patients with severe brain injury. In this study, we aimed to validate whether neurointensive care could improve the outcome in patients with critically ill acute ischemic stroke using the linked big dataset on stroke in Korea. METHODS: We included 1,405 acute ischemic stroke patients with mechanical ventilator support in the intensive care unit after an index stroke. Patients were retrieved from linking the Clinical Research Center for Stroke Registry and the Health Insurance Review and Assessment Service data from the period between January 2007 and December 2014. The outcomes were mortality at discharge and at 3 months after an index stroke. The main outcomes were compared between the centers with and without dedicated neurointensivists. RESULTS: Among the included patients, 303 (21.6%) were admitted to the centers with dedicated neurointensivists. The patients treated by dedicated neurointensivists had significantly lower in-hospital mortality (18.3% vs. 26.8%, P = 0.002) as well as lower mortality at 3-month (38.0% vs. 49.1%, P < 0.001) than those who were treated without neurointensivists. After adjusting for confounders, a treatment without neurointensivists was independently associated with higher in-hospital mortality (odds ratio [OR], 1.59; 95% confidence intervals [CIs], 1.13-2.25; P = 0.008) and 3-month mortality (OR, 1.48; 95% CIs, 1.12-1.95; P = 0.005). CONCLUSION: Treatment by dedicated neurointensivists is associated with lower in-hospital and 3-month mortality using the linked big datasets for stroke in Korea. This finding stresses the importance of neurointensivists in treating patients with severe ischemic stroke.


Assuntos
AVC Isquêmico/patologia , Papel do Médico , Idoso , Idoso de 80 Anos ou mais , Comorbidade , Bases de Dados Factuais , Feminino , Mortalidade Hospitalar , Humanos , Unidades de Terapia Intensiva , AVC Isquêmico/epidemiologia , AVC Isquêmico/mortalidade , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Alta do Paciente , Sistema de Registros , República da Coreia/epidemiologia , Fatores de Risco , Análise de Sobrevida , Resultado do Tratamento
5.
Biotechnol Lett ; 40(5): 755-763, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29464570

RESUMO

OBJECTIVES: The aim of the study is to generate a spherical three-dimensional (3D) aggregate of hepatocyte-like cells (HLCs) differentiated from human embryonic stem cells and to investigate the effect of the 3D environment on hepatic maturation and drug metabolism. RESULTS: Quantitative real-time PCR analysis indicated that gene expression of mature hepatocyte markers, drug-metabolizing enzymes, and hepatic transporters was significantly higher in HLCs cultured in the 3D system than in those cultured in a two-dimensional system (p < 0.001). Moreover, hepatocyte-specific functions, including albumin secretion and bile canaliculi formation, were increased in HLCs cultured in the 3D system. In particular, 3D spheroidal culture increased expression of CES1 and BCHE, which encode hepatic esterases (p < 0.001). The enhanced activities of these hepatic esterases were confirmed by the cholinesterase activity assay and the increased susceptibility of HLCs to oseltamivir, which is metabolized by CES1. CONCLUSIONS: 3D spheroidal culture enhances the maturation and drug metabolism of stem cell-derived HLCs, and this may help to optimize hepatic differentiation protocols for hepatotoxicity testing.


Assuntos
Butirilcolinesterase/genética , Hidrolases de Éster Carboxílico/genética , Técnicas de Cultura de Células/métodos , Hepatócitos/citologia , Células-Tronco Embrionárias Humanas/citologia , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Perfilação da Expressão Gênica , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Hepatócitos/metabolismo , Células-Tronco Embrionárias Humanas/metabolismo , Humanos , Oseltamivir/farmacologia , Esferoides Celulares/citologia , Esferoides Celulares/metabolismo , Regulação para Cima/efeitos dos fármacos
6.
J Korean Med Sci ; 33(53): e343, 2018 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-30595684

RESUMO

BACKGROUND: Linkage of public healthcare data is useful in stroke research because patients may visit different sectors of the health system before, during, and after stroke. Therefore, we aimed to establish high-quality big data on stroke in Korea by linking acute stroke registry and national health claim databases. METHODS: Acute stroke patients (n = 65,311) with claim data suitable for linkage were included in the Clinical Research Center for Stroke (CRCS) registry during 2006-2014. We linked the CRCS registry with national health claim databases in the Health Insurance Review and Assessment Service (HIRA). Linkage was performed using 6 common variables: birth date, gender, provider identification, receiving year and number, and statement serial number in the benefit claim statement. For matched records, linkage accuracy was evaluated using differences between hospital visiting date in the CRCS registry and the commencement date for health insurance care in HIRA. RESULTS: Of 65,311 CRCS cases, 64,634 were matched to HIRA cases (match rate, 99.0%). The proportion of true matches was 94.4% (n = 61,017) in the matched data. Among true matches (mean age 66.4 years; men 58.4%), the median National Institutes of Health Stroke Scale score was 3 (interquartile range 1-7). When comparing baseline characteristics between true matches and false matches, no substantial difference was observed for any variable. CONCLUSION: We could establish big data on stroke by linking CRCS registry and HIRA records, using claims data without personal identifiers. We plan to conduct national stroke research and improve stroke care using the linked big database.


Assuntos
Bases de Dados Factuais , Armazenamento e Recuperação da Informação , Acidente Vascular Cerebral/patologia , Doença Aguda , Idoso , Big Data , Feminino , Humanos , Revisão da Utilização de Seguros , Masculino , Pessoa de Meia-Idade , Sistema de Registros
7.
Asian-Australas J Anim Sci ; 29(4): 564-70, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26949958

RESUMO

The Sal-like 1 gene (Sall1) is essential for kidney development, and mutations in this gene result in abnormalities in the kidneys. Mice lacking Sall1 show agenesis or severe dysgenesis of the kidneys. In a recent study, blastocyst complementation was used to develop mice and pigs with exogenic organs. In the present study, transcription activator-like effector nuclease (TALEN)-mediated homologous recombination was used to produce Sall1-knockout porcine fibroblasts for developing knockout pigs. The vector targeting the Sall1 locus included a 5.5-kb 5' arm, 1.8-kb 3' arm, and a neomycin resistance gene as a positive selection marker. The knockout vector and TALEN were introduced into porcine fibroblasts by electroporation. Antibiotic selection was performed over 11 days by using 300 µg/mL G418. DNA of cells from G418-resistant colonies was amplified using polymerase chain reaction (PCR) to confirm the presence of fragments corresponding to the 3' and 5' arms of Sall1. Further, mono- and bi-allelic knockout cells were isolated and analyzed using PCR-restriction fragment length polymorphism. The results of our study indicated that TALEN-mediated homologous recombination induced bi-allelic knockout of the endogenous gene.

8.
Microsc Microanal ; 20(6): 1798-804, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25347999

RESUMO

The dehydrogenated microstructure of the lithium borohydride-yttrium hydride (LiBH4-YH3) composite obtained at 350°C under 0.3 MPa of hydrogen and static vacuum was investigated by transmission electron microscopy combined with a focused ion beam technique. The dehydrogenation reaction between LiBH4 and YH3 into LiH and YB4 takes place under 0.3 MPa of hydrogen, which produces YB4 nano-crystallites that are uniformly distributed in the LiH matrix. This microstructural feature seems to be beneficial for rehydrogenation of the dehydrogenation products. On the other hand, the dehydrogenation process is incomplete under static vacuum, leading to the unreacted microstructure, where YH3 and YH2 crystallites are embedded in LiBH4 matrix. High resolution imaging confirmed the presence of crystalline B resulting from the self-decomposition of LiBH4. However, Li2B12H12, which is assumed to be present in the LiBH4 matrix, was not clearly observed.

9.
Asian-Australas J Anim Sci ; 27(11): 1644-51, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25358326

RESUMO

Transgenic animals have become important tools for the production of therapeutic proteins in the domestic animal. Production efficiencies of transgenic animals by conventional methods as microinjection and retrovirus vector methods are low, and the foreign gene expression levels are also low because of their random integration in the host genome. In this study, we investigated the homologous recombination on the porcine ß-casein gene locus using a knock-in vector for the ß-casein gene locus. We developed the knock-in vector on the porcine ß-casein gene locus and isolated knock-in fibroblast for nuclear transfer. The knock-in vector consisted of the neomycin resistance gene (neo) as a positive selectable marker gene, diphtheria toxin-A gene as negative selection marker, and 5' arm and 3' arm from the porcine ß-casein gene. The secretion of enhanced green fluorescent protein (EGFP) was more easily detected in the cell culture media than it was by western blot analysis of cell extract of the HC11 mouse mammary epithelial cells transfected with EGFP knock-in vector. These results indicated that a knock-in system using ß-casein gene induced high expression of transgene by the gene regulatory sequence of endogenous ß-casein gene. These fibroblasts may be used to produce transgenic pigs for the production of therapeutic proteins via the mammary glands.

10.
J Imaging ; 10(4)2024 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-38667991

RESUMO

The continuous monitoring of civil infrastructures is crucial for ensuring public safety and extending the lifespan of structures. In recent years, image-processing-based technologies have emerged as powerful tools for the structural health monitoring (SHM) of civil infrastructures. This review provides a comprehensive overview of the advancements, applications, and challenges associated with image processing in the field of SHM. The discussion encompasses various imaging techniques such as satellite imagery, Light Detection and Ranging (LiDAR), optical cameras, and other non-destructive testing methods. Key topics include the use of image processing for damage detection, crack identification, deformation monitoring, and overall structural assessment. This review explores the integration of artificial intelligence and machine learning techniques with image processing for enhanced automation and accuracy in SHM. By consolidating the current state of image-processing-based technology for SHM, this review aims to show the full potential of image-based approaches for researchers, engineers, and professionals involved in civil engineering, SHM, image processing, and related fields.

11.
Appl Radiat Isot ; 207: 111250, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38457856

RESUMO

The purpose of this study is to evaluate the derived concentration guideline levels for unrestricted site reuse the Korea research reactor unit 1 and 2. Distribution coefficients for Co-60 and Sr-90 were derived, and site-specific values of the KRR soil were applied for the DCGLs for the seven target nuclide. The distribution coefficients of Co-60 and Sr-90 were 6,128 and 86.0 mL/g. The DCGLs derived from the dose by age group were 0.053 Bq/g for Co-60 and 45.0 Bq/g for H-3.

12.
Int J Biol Macromol ; 272(Pt 1): 132734, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38815950

RESUMO

Kraft lignin (KL) holds significant potential as a renewable resource for the development of innovative materials that are currently not fully utilized. In this study, a novel iminated lignin (IL) was synthesized by grafting primary amine lignin (N-KL) onto salicylaldehyde. The effects of the dosage and reaction temperature on the nitrogen content of N-KL were evaluated. The maximum nitrogen content in N-KL reached to 3.32 %. Characterization by spectroscopy techniques (FT-IR, XPS, and NMR), elemental analysis, and gel permeation chromatography confirmed the imination of lignin. Additionally, the antioxidant activity of the lignin samples was investigated using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability. Moreover, the DPPH radical scavenging capacity of IL-6 (IC50 = 38.6 ± 3.9 µg/mL) was close to that of commercial antioxidant butylated hydroxytoluene (BHT) (IC50 = 37.7 ± 4.5 µg/mL). Furthermore, the adsorption equilibrium results indicated that IL-6 had a maximum uptake of 115.6 mg/g Pb2+, which was 3.2-fold higher than that of KL. Kinetic adsorption experiments suggested that IL-6 adsorption follows a pseudo-second-order model. Therefore, the synthesized iminated lignin is a promising candidate for the development of environmentally friendly materials with applications as an antioxidant and lead-ion adsorbent.


Assuntos
Sequestradores de Radicais Livres , Chumbo , Lignina , Lignina/química , Chumbo/química , Sequestradores de Radicais Livres/química , Sequestradores de Radicais Livres/síntese química , Compostos de Bifenilo/química , Picratos/química , Cinética , Adsorção , Antioxidantes/química , Antioxidantes/síntese química
13.
Patient Educ Couns ; 125: 108289, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38631197

RESUMO

OBJECTIVES: This study explores patient perspectives (ideas, concerns, and expectations) in surgeon-patient consultations. METHODS: We examined 54 video-recorded consultations using applied conversation analysis. Consultations took place from 2012 to 2017 in an Australian metropolitan hospital clinic centre and involved seven surgeons across six specialties. RESULTS: Patient perspectives emerged in less than one third of consultations. We describe the initiation of and response to potential perspectives sequences, demonstrating how patients and surgeons co-construct these sequences when they do occur. CONCLUSIONS: Findings suggest a need for greater attention to supporting patient agency through explicit pursuit of patient perspectives. The implications extend to the Calgary-Cambridge Guide, suggesting that it may benefit from a focus on active pursuit and appropriate responsiveness to patient perspectives. PRACTICE IMPLICATIONS: This study highlights the need for surgeons to actively engage with the patient perspective offered in consultations, emphasising the importance of respect for the patient's knowledge and expectations to improve patient satisfaction and healthcare outcomes.


Assuntos
Comunicação , Satisfação do Paciente , Relações Médico-Paciente , Encaminhamento e Consulta , Cirurgiões , Humanos , Masculino , Feminino , Austrália , Pessoa de Meia-Idade , Cirurgiões/psicologia , Adulto , Gravação em Vídeo , Pesquisa Qualitativa , Participação do Paciente , Idoso
14.
Genes Genomics ; 46(2): 253-261, 2024 02.
Artigo em Inglês | MEDLINE | ID: mdl-38236352

RESUMO

BACKGROUND: Interactions of plants with biotic stress factors including bacteria, fungi, and viruses have been extensively investigated to date. Plasmodiophora brassicae, a protist pathogen, causes clubroot disease in Cruciferae plants. Infection of Chinese cabbage (Brassica rapa) plants with P. brassica results in the formation of root galls, which inhibits the roots from absorbing soil nutrients and water. Sugar, the major source of carbon for all living organisms including pathogens and host plants, plays an important role in plant growth and development. OBJECTIVE: To explore the roles of BrSWEET2, BrSWEET13, and BrSWEET14 in P. brassicae resistance, Arabidopsis thaliana T-DNA knockout mutants sweet2, sweet13, and sweet14 were employed. METHODS: To isolate total RNA from the collected root nodules, the root tissues washed several times with running water and frozen tissues with liquid nitrogen. Total RNA was extracted using the Spectrum™ Plant Total RNA Kit (SIGMA) and cDNA was synthesized in a 20 µl reaction volume using the ReverTra Ace-α-® kit (TOYOBO). Real-time PCR was performed in a 10 µl reaction volume containing 1 µl of template DNA, 1 µl of forward primer, 1 µl of reverse primer, 5 µl of 2× iQTM SYBR® Green Supermix (BioRad), and 2 µl of sterile distilled water. The SWEET genes were genotyped using BioFACT™ 2× TaqBasic PCR Master Mix 2. RESULTS: Both sweet2 and sweet14 showed strong resistance to P. brassicae compared with wild-type Arabidopsis and Chinese cabbage plants and sweet13 mutant plants. Pathogenicity assays indicated that the SWEET2 gene plays an important role in clubroot disease resistance in higher plants.


Assuntos
Brassica rapa , Brassica , Plasmodioforídeos , Brassica rapa/genética , Plasmodioforídeos/genética , Brassica/genética , Água , RNA
15.
Appl Radiat Isot ; 200: 110985, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37598561

RESUMO

For radiation protection optimization, ICRP proposed dose constraint as quantitative value for planned exposure situation based on representative person concept. The objective of this study is to derive dose constraints for the general public around nuclear power plants in Korea by applying representative person concept. The dose constraints for the general public around NPPs were derived through a total of six steps. The steps consisted of setting source terms, setting exposure pathways and scenarios, setting candidate groups for a critical group decision, setting habit data, calculating radiation doses, and proposing dose constraints. Through these steps, the radiation dose distribution of the general public around the NPPs was obtained, and dose constraints were proposed using the dose distribution. Radiation doses to the general public around all the Korea NPP sites ranged 1.63 × 10-2 to 1.32 × 10-1 mSv/y. Using the dose distribution, 0.15 mSv/y, 0.10 mSv/y, and 0.08 mSv/y were proposed as dose constraints. The dose constraint values derived in this study are proposals. Therefore, it is judged that the dose constraints should need furthermore discussion with regulators, licensees, and radiation protection experts considering societal and economic factors for radiation protection. The proposal for dose constraints developed in this study can be used to optimize radiation protection for the general public around the NPPs.

16.
Chem Commun (Camb) ; 59(46): 6987-7003, 2023 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-37165690

RESUMO

Polymer membranes represent an attractive platform for energy-efficient gas separation, but they are known to suffer from plasticization during continuous gas-separation processes. This phenomenon is caused by the spontaneous relaxation of individual polymer chains arising from the swelling effect induced by high-pressure highly soluble gases such as CO2, and it weakens the stability of the membrane, leading to a significant loss of selectivity during the separation of mixed gases. Thus, minimizing the disadvantages of polymer membranes is essential to ensure reliable gas-separation performance for practical applications. This feature article summarizes the theory underlying the plasticization of polymer membranes and introduces covalent and non-covalent approaches to suppress plasticization behaviour on a molecular level.

17.
Stem Cell Res ; 69: 103081, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-37001365

RESUMO

PDGFRB encodes platelet-derived growth factor receptor beta (PDGFR-ß), a cell surface tyrosine kinase receptor for members of the platelet-derived growth factor family. It is required for the normal development of the vascular and nervous systems and rearrangement of the actin cytoskeleton. PDGFR-ß plays an essential role in early liver diseases, including liver fibrosis. Here, we generated a human induced pluripotent stem cell (iPSC) line, KITi001-A-1, using CRISPR/Cas9. This reporter iPSC line and its derivatives are useful for tracing PDGFR-ß-expressing cells and for screening for liver fibrosis-inducing compounds.


Assuntos
Células-Tronco Pluripotentes Induzidas , Receptor beta de Fator de Crescimento Derivado de Plaquetas , Humanos , Receptor beta de Fator de Crescimento Derivado de Plaquetas/genética , Receptor beta de Fator de Crescimento Derivado de Plaquetas/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Linhagem Celular , Sistemas CRISPR-Cas/genética , Fator de Crescimento Derivado de Plaquetas/metabolismo , Diferenciação Celular
18.
Artigo em Inglês | MEDLINE | ID: mdl-36527808

RESUMO

In this study, an automated online micro-solid-phase extraction (µSPE)-liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the detection of metabolites of cannabidiol (CBD), Δ8-tetrahydrocannabinol (Δ8-THC), and Δ9-tetrahydrocannabinol (Δ9-THC), particularly 7-carboxy- cannabidiol (7-COOH-CBD), 11-nor-9-carboxy-Δ8-tetrahydrocannabinol (Δ8-THCCOOH), 11-nor-9-carboxy-Δ9-tetrahydrocannabinol (Δ9-THCCOOH), and 11-nor-9-carboxy-Δ9- tetrahydrocannabinol-glucuronide (Δ9-THCCOOH-glu) in urine. An instrument top sample preparation (ITSP) cartridge was introduced to increase the sensitivity toward analytes and decrease the matrix effect of the urine. LC-MS/MS analysis was performed in the multiple-reaction monitoring mode, and the analytes were separated using an Acquity UPLC HSS T3 (2.1 × 100 mm, 1.8 µm) column and gradient elution with water containing 0.05 % acetic acid and methanol as the mobile phase. The calibration range was 0.5-200 ng/mL for all the analytes, with a correlation coefficient (r) of ≥0.996 and a weighting factor of 1/x2. The limits of detection for 7-COOH-CBD, Δ8-THCCOOH, Δ9-THCCOOH, and Δ9-THCCOOH-glu were 0.06, 0.02, 0.03, and 0.1 ng/mL, respectively. The intra- and inter-day accuracy ranged from -8.0 to 6.2 % and -7.3 to 7.8 % with a precision of ≤7.2 % and ≤6.2 %, respectively. The method was also validated for selectivity, recovery, matrix effect, stability, and dilution integrity. The developed method was successfully applied to the analysis of 78 urine samples, and 7-COOH-CBD, Δ8-THCCOOH, Δ9-THCCOOH, and Δ9-THCCOOH-glu were detected in 54 urine samples at normalized concentrations of 1.1, 0.6-939.1, 0.9-2595.0, and 1.3-527.6 ng/mg creatinine, respectively.


Assuntos
Canabidiol , Canabinoides , Canabidiol/análise , Dronabinol/análise , Cromatografia Líquida/métodos , Canabinoides/análise , Espectrometria de Massas em Tandem/métodos , Limite de Detecção , Extração em Fase Sólida
19.
Int J Biol Macromol ; 225: 1384-1393, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36435473

RESUMO

The development of eco-friendly, sustainable, biodegradable, and biocompatible green biopolymer composites is becoming increasingly important. In this study, acetoacetylated lignin (ATL) was obtained via an eco-friendly, facile one-step synthesis reaction, and chitosan (CS)-containing ATL films (CSL) were prepared. The chemical structural analysis of ATL confirmed that the acetoacetyl groups were successfully grafted onto kraft lignin (KL). ATL with adequate acetoacetyl groups exhibited enhanced molecular weight and antioxidant and ultraviolet (UV)-shielding properties. In particular, ATL, with a half maximal inhibitory concentration (IC50) of 23.8 µg·mL-1, exhibited superior antioxidant activity than butylated hydroxytoluene (38.3 µg·mL-1) and KL (50.0 µg·mL-1). When ATL was incorporated into the CS solution to prepare biofilms, the antioxidant activity, UV-shielding property, water resistance, and thermal stability of the CSL greatly improved. Notably, the UV-A and UV-B shielding properties of the 2 % CSL were 130 % and 78 % higher than those of the pure CS film, respectively. Therefore, ATL designed with lignin-derived multifunctional properties has potential applications as an antioxidant and UV-shielding bio-additive and shows significant prospects in food packaging and biomedical applications.


Assuntos
Quitosana , Quitosana/química , Lignina/química , Antioxidantes/farmacologia , Antioxidantes/química , Biopolímeros , Água
20.
Stem Cell Res ; 66: 102986, 2023 02.
Artigo em Inglês | MEDLINE | ID: mdl-36493708

RESUMO

CRISPR/Cas9-based transcriptional regulation systems can induce the site-specific activation or repression of endogenous genes. p300 is a transcriptional co-activator that functions as a histone acetyltransferase that regulates gene transcription via chromatin remodeling. Here, we generated a human embryonic stem cell line stably expressing catalytically dead Cas9 (dCas9) fused to the catalytic core domain of human p300 via lentiviral transduction. This cell line can be used for locus-specific histone acetylation in combination with guide RNAs, and is a valuable tool for gene regulation in stem cell research.


Assuntos
Proteína 9 Associada à CRISPR , Células-Tronco Embrionárias Humanas , Humanos , Proteína 9 Associada à CRISPR/genética , Sistemas CRISPR-Cas/genética , Regulação da Expressão Gênica , Fatores de Transcrição/genética , Linhagem Celular , Ativação Transcricional
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA