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1.
Dement Geriatr Cogn Disord ; 53(3): 162-167, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38593753

RESUMO

INTRODUCTION: The relationship between cognitive function and subsequent sarcopenia remains unclear. Therefore, this study aimed to examine the associations of performance on multiple cognitive domains with sarcopenia in the middle-aged and older adults. METHODS: This longitudinal analysis (wave 2011-2013) included 2,934 participants from the CHARLS study. Sarcopenia was defined by the Asian Sarcopenia Working Group 2019 criteria. Cognitive function was measured by the Chinese version of the Mini-Mental State Examination (MMSE). Three interpretable techniques, namely SHapley Additive exPlanations (SHAP) and two built-in methods (coefficients of logistic regression and Gini importance of random forest), were used to assess the relationship between MMSE, its components (orientation, attention, episodic memory, and visuospatial ability) and sarcopenia. In addition, the association of MMSE score and its components with sarcopenia was further validated using stepwise regression. RESULTS: All interpretable methods showed that MMSE score was important predictors of sarcopenia, especially the SHAP (MMSE score ranked top one). For its components, episodic memory, visuospatial ability, and attention showed high predictive value compared with orientation. Stepwise regression analyses showed that MMSE score and its components of episodic memory and visuospatial ability were correlated with sarcopenia, with their odds ratios of 0.93 (95% CI: 0.91-0.96, p < 0.001), 0.87 (95% CI: 0.82-0.93, p < 0.001), and 1.32 (95% CI: 1.05-1.65, p = 0.016), respectively. CONCLUSIONS: Better cognitive function especially episodic memory and visuospatial ability was negatively associated with incident sarcopenia among community middle-aged and older adults.


Assuntos
Cognição , Sarcopenia , Humanos , Sarcopenia/psicologia , Masculino , Feminino , Idoso , Pessoa de Meia-Idade , Estudos Longitudinais , Cognição/fisiologia , Memória Episódica , Testes de Estado Mental e Demência , Disfunção Cognitiva/psicologia , China/epidemiologia , Testes Neuropsicológicos , Idoso de 80 Anos ou mais , Atenção/fisiologia
2.
Nano Lett ; 23(13): 5877-5885, 2023 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-37040490

RESUMO

Nanoneedles are a useful tool for delivering exogenous biomolecules to cells. Although therapeutic applications have been explored, the mechanism regarding how cells interact with nanoneedles remains poorly studied. Here, we present a new approach for the generation of nanoneedles, validated their usefulness in cargo delivery, and studied the underlying genetic modulators during delivery. We fabricated arrays of nanoneedles based on electrodeposition and quantified its efficacy of delivery using fluorescently labeled proteins and siRNAs. Notably, we revealed that our nanoneedles caused the disruption of cell membranes, enhanced the expression of cell-cell junction proteins, and downregulated the expression of transcriptional factors of NFκB pathways. This perturbation trapped most of the cells in G2 phase, in which the cells have the highest endocytosis activities. Taken together, this system provides a new model for the study of interactions between cells and high-aspect-ratio materials.


Assuntos
Endocitose , Proteínas , Membrana Celular
3.
Psychogeriatrics ; 24(3): 645-654, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38514389

RESUMO

BACKGROUND: Older adults with hypertension have a high risk of disability, while an accurate risk prediction model is still lacking. This study aimed to construct interpretable disability prediction models for older Chinese with hypertension based on multiple time intervals. METHODS: Data were collected from the Chinese Longitudinal Healthy Longevity and Happy Family Study for 2008-2018. A total of 1602, 1108, and 537 older adults were included for the periods of 2008-2012, 2008-2014, and 2008-2018, respectively. Disability was measured by basic activities of daily living. Least absolute shrinkage and selection operator (LASSO) was applied for feature selection. Five machine learning algorithms combined with LASSO set and full-variable set were used to predict 4-, 6-, and 10-year disability risk, respectively. Area under the receiver operating characteristic curve was used as the main metric for selection of the optimal model. SHapley Additive exPlanations (SHAP) was used to explore important predictors of the optimal model. RESULTS: Random forest in full-variable set and XGBoost in LASSO set were the optimal models for 4-year prediction. Support vector machine was the optimal model for 6-year prediction on both sets. For 10-year prediction, deep neural network in full variable set and logistic regression in LASSO set were optimal models. Age ranked the most important predictor. Marital status, body mass index, score of Mini-Mental State Examination, and psychological well-being score were also important predictors. CONCLUSIONS: Machine learning shows promise in screening out older adults at high risk of disability. Disability prevention strategies should specifically focus on older patients with unfortunate marriage, high BMI, and poor cognitive and psychological conditions.


Assuntos
Atividades Cotidianas , Pessoas com Deficiência , Hipertensão , Humanos , Feminino , Masculino , Idoso , Estudos Longitudinais , Hipertensão/epidemiologia , China/epidemiologia , Atividades Cotidianas/psicologia , Pessoas com Deficiência/estatística & dados numéricos , Pessoas com Deficiência/psicologia , Aprendizado de Máquina , Idoso de 80 Anos ou mais , Longevidade , Avaliação da Deficiência , Medição de Risco , Avaliação Geriátrica/métodos , Avaliação Geriátrica/estatística & dados numéricos , Pessoa de Meia-Idade , População do Leste Asiático
4.
J Am Chem Soc ; 145(30): 16924-16937, 2023 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-37466996

RESUMO

The genomes of myxobacteria harbor a variety of biosynthetic gene clusters encoding numerous secondary metabolites, including ribosomally synthesized and post-translationally modified peptides (RiPPs) with diverse chemical structures and biological activities. However, the biosynthetic potential of RiPPs from myxobacteria remains barely explored. Herein, we report a novel myxobacteria lanthipeptide myxococin identified from Myxococcus fulvus. Myxococins represent the first example of lanthipeptides, of which the characteristic multiple thioether rings are installed by employing a Class II lanthipeptide synthetase MfuM and a Class I lanthipeptide cyclase MfuC in a cascaded way. Unprecedentedly, we biochemically characterized the first M61 family aminopeptidase MfuP involved in RiPP biosynthesis, demonstrating that MfuP showed the activity of an endopeptidase activity. MfuP is leader-independent but strictly selective for the multibridge structure of myxococin A and responsible for unwrapping two rings via amide bond hydrolysis, yielding myxococin B. Furthermore, the X-ray crystal structure of MfuP and structural analysis, including active-site mutations, are reported. Finally, myxococins are evaluated to exhibit anti-inflammatory activity in lipopolysaccharide-induced macrophages without detectable cytotoxicity.


Assuntos
Myxococcales , Peptídeos/química , Processamento de Proteína Pós-Traducional
5.
Anal Chem ; 95(48): 17438-17443, 2023 12 05.
Artigo em Inglês | MEDLINE | ID: mdl-37991715

RESUMO

Real-time biomolecular monitoring requires biosensors based on affinity reagents, such as aptamers, with moderate to low affinities for the best binding dynamics and signal gain. We recently reported Pro-SELEX, an approach that utilizes parallelized SELEX and high-content bioinformatics for the selection of aptamers with predefined binding affinities. The Pro-SELEX pipeline relies on an algorithm, termed AptaZ, that can predict the binding affinities of selected aptamers. The original AptaZ algorithm is computationally complex and slows the overall throughput of Pro-SELEX. Here, we present Apta FastZ, a rapid equivalent of AptaZ. The Apta FastZ algorithm considers the spare nature of the sequences from SELEX and is coded to avoid unnecessary comparison between sequences. As a result, Apta FastZ achieved a 10 to 40-fold faster computing speed compared to the original AptaZ algorithm while maintaining identical outcomes, allowing the bioinformatics to be completed within 1-10 h for large-scale data sets. We further validated the affinity of myeloperoxidase aptamers predicted by Apta FastZ by experiments and observed a high level of linear correlation between predicted scores and measured affinities. Taken together, the implementation of Apta FastZ could greatly accelerate the current Pro-SELEX workflow, allowing customized aptamers to be discovered within 3 days using preselected DNA libraries.


Assuntos
Aptâmeros de Nucleotídeos , Aptâmeros de Nucleotídeos/química , Técnica de Seleção de Aptâmeros , Biblioteca Gênica , Biologia Computacional
6.
Metab Eng ; 75: 131-142, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36528227

RESUMO

FK228 (romidepsin) is the only natural histone deacetylases (HDACs) inhibitor approved by FDA to treat cutaneous and peripheral T-cell lymphoma. However, the limited supply and severe cardiotoxicity of FK228 underscore the importance to develop an effective synthetic biology platform for the manufacturing and fine-tuning of this drug lead. In this work, we constructed a Burkholderia chassis for the high-yield production of FK228-family (unnatural) natural products. By virtue of the optimized Burkholderia-specific recombineering system, the biosynthetic gene cluster (BGC) encoding the FK228-like skeleton thailandepsins (tdp) in Burkholderia thailandensis E264 was replaced with an attB integration site to afford the basal chassis KOGC1. The tdp BGC directly captured from E264 was hybridized with the FK228-encoding BGC (dep) using the versatile Red/ET technology. The hybrid BGC (tdp-dep) was integrated into the attB site of KOGC1, resulting in the heterologous expression of FK228. Remarkably, the titer reached 581 mg/L, which is 30-fold higher than that of native producer Chromobacterium violaceum No. 968. This success encouraged us to further engineer the NRPS modules 4 or 6 of hybrid tdp-dep BGC by domain units swapping strategy, and eight new FK228 derivatives (1-8) varying in the composition of amino acids were generated. Especially, the titers of 2 and 3 in KOGC1 were up to 985 mg/L and 453 mg/L, respectively. 2 and 3 displayed stronger cytotoxic activity than FK228. All in all, this work established a robust platform to produce FK228 and its new derivatives in sufficient quantities for anticancer drug development.


Assuntos
Burkholderia , Depsipeptídeos , Depsipeptídeos/genética , Depsipeptídeos/química , Depsipeptídeos/farmacologia , Burkholderia/genética , Burkholderia/química , Proteínas de Ligação a DNA
7.
Nano Lett ; 22(12): 4774-4783, 2022 06 22.
Artigo em Inglês | MEDLINE | ID: mdl-35639489

RESUMO

Magnetic cell sorting is an enabling tool for the isolation of specific cellular subpopulations for downstream applications and requires the cells to be labeled by a sufficient number of magnetic nanoparticles to leverage magnetophoresis for efficient separation. This requirement makes it challenging to target weakly expressed biomarkers. Here, we developed a new approach that selectively and efficiently amplifies the magnetic labeling on cells through sequentially connected antibodies and nanoparticles delivered to the surface or interior of the cell. Using this approach, we achieved amplification up to 100-fold for surface and intracellular markers. We also demonstrated the utility of this assay for enabling high-performance magnetic cell sorting when it is applied to the analysis of rare tumor cells for cancer diagnosis and the purification of transfected CAR T cells for immunotherapy. The data presented demonstrate a useful tool for the stratification of rare cell subpopulations.


Assuntos
Magnetismo , Nanopartículas , Separação Celular , Fenômenos Magnéticos , Fenômenos Físicos
8.
Int J Mol Sci ; 24(7)2023 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-37047328

RESUMO

In the past decade, immense progress has been made in advancing personalized medicine to effectively address patient-specific disease complexities in order to develop individualized treatment strategies. In particular, the emergence of 3D bioprinting for in vitro models of tissue and organ engineering presents novel opportunities to improve personalized medicine. However, the existing bioprinted constructs are not yet able to fulfill the ultimate goal: an anatomically realistic organ with mature biological functions. Current bioprinting approaches have technical challenges in terms of precise cell deposition, effective differentiation, proper vascularization, and innervation. This review introduces the principles and realizations of bioprinting with a strong focus on the predominant techniques, including extrusion printing and digital light processing (DLP). We further discussed the applications of bioprinted constructs, including the engraftment of stem cells as personalized implants for regenerative medicine and in vitro high-throughput drug development models for drug discovery. While no one-size-fits-all approach to bioprinting has emerged, the rapid progress and promising results of preliminary studies have demonstrated that bioprinting could serve as an empowering technology to resolve critical challenges in personalized medicine.


Assuntos
Bioimpressão , Engenharia Tecidual , Humanos , Engenharia Tecidual/métodos , Medicina de Precisão , Bioimpressão/métodos , Impressão Tridimensional , Medicina Regenerativa , Alicerces Teciduais
9.
Int J Mol Sci ; 24(4)2023 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-36835532

RESUMO

MSX1 is an important member of the muscle segment homeobox gene (Msh) family and acts as a transcription factor to regulate tissue plasticity, yet its role in goat endometrium remodeling remains elusive. In this study, an immunohistochemical analysis showed that MSX1 was mainly expressed in the luminal and glandular epithelium of goat uterus, and the MSX1 expression was upregulated in pregnancy at days 15 and 18 compared with pregnancy at day 5. In order to explore its function, goat endometrial epithelial cells (gEECs) were treated with 17 ß-estrogen (E2), progesterone (P4), and/or interferon-tau (IFNτ), which were used to mimic the physiological environment of early pregnancy. The results showed that MSX1 was significantly upregulated with E2- and P4-alone treatment, or their combined treatment, and IFNτ further enhanced its expression. The spheroid attachment and PGE2/PGF2α ratio were downregulated by the suppression of MSX1. The combination of E2, P4, and IFNτ treatment induced the plasma membrane transformation (PMT) of gEECs, which mainly showed the upregulation of N-cadherin (CDH2) and concomitant downregulation of the polarity-related genes (ZO-1, α-PKC, Par3, Lgl2, and SCRIB). The knockdown of MSX1 partly hindered the PMT induced by E2, P4, and IFNτ treatment, while the upregulation of CDH2 and the downregulation of the partly polarity-related genes were significantly enhanced when MSX1 was overexpressed. Moreover, MSX1 regulated the CDH2 expression by activating the endoplasmic reticulum (ER) stress-mediated unfolded protein response (UPR) pathway. Collectively, these results suggest that MSX1 was involved in the PMT of the gEECs through the ER stress-mediated UPR pathway, which affects endometrial adhesion and secretion function.


Assuntos
Endométrio , Cabras , Gravidez , Feminino , Animais , Cabras/metabolismo , Endométrio/metabolismo , Progesterona/metabolismo , Membrana Celular , Células Epiteliais/metabolismo , Epitélio
10.
Small ; 18(17): e2106097, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35344274

RESUMO

Circulating tumor cell (CTC) clusters are associated with increased metastatic potential and worse patient prognosis, but are rare, difficult to count, and poorly characterized biophysically. The PillarX device described here is a bimodular microfluidic device (Pillar-device and an X-magnetic device) to profile single CTCs and clusters from whole blood based on their size, deformability, and epithelial marker expression. Larger, less deformable clusters and large single cells are captured in the Pillar-device and sorted according to pillar gap sizes. Smaller, deformable clusters and single cells are subsequently captured in the X-device and separated based on epithelial marker expression using functionalized magnetic nanoparticles. Clusters of established and primary breast cancer cells with variable degrees of cohesion driven by different cell-cell adhesion protein expression are profiled in the device. Cohesive clusters exhibit a lower deformability as they travel through the pillar array, relative to less cohesive clusters, and have greater collective invasive behavior. The ability of the PillarX device to capture clusters is validated in mouse models and patients of metastatic breast cancer. Thus, this device effectively enumerates and profiles CTC clusters based on their unique geometrical, physical, and biochemical properties, and could form the basis of a novel prognostic clinical tool.


Assuntos
Neoplasias da Mama , Células Neoplásicas Circulantes , Animais , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Separação Celular , Feminino , Humanos , Dispositivos Lab-On-A-Chip , Camundongos , Células Neoplásicas Circulantes/patologia , Prognóstico
11.
Artigo em Inglês | MEDLINE | ID: mdl-35316174

RESUMO

Two novel, designated strains 29W222T and 2943T, were isolated from the marine sediment from Aoshan Bay, Jimo, PR China. Growth was observed at pH 6.0-8.5 (optimum, pH 7.5) for strain 29W222T, and pH 5.5-8.5 (pH 7.0) for strain 2943T. Both strains displayed growth in 0.5-6 % NaCl with an optimum at 1 % for 29W222T; 0.5 % for 2943T. Both strains grew optimally at 33 °C. The results of phylogenetic analyses based on 16S rRNA gene sequences indicated that 29W222T and 2943T represented members of the genus Fulvivirga and strain 29W222T was most closely related to Fulvivirga kasyanovii KMM 6220T (97.9 % sequence similarity) and Fulvivirga imtechensis AK7T (95.0 %), and 2943T to Fulvivirga imtechensis AK7T (95.7 %) and Fulvivirga kasyanovii KMM 6220T (94.8 %). The genomic DNA G+C contents of 29W222T and 2943T were 39.9 and 37.7 mol%, respectively. The results of chemotaxonomic analysis indicated that the sole respiratory quinone was menaquinone 7 (MK-7), and the major fatty acid was iso-C15 : 0 for both strains. Average nucleotide identity and average amino acid identity values between strain 29W222T and Fulvivirga kasyanovii KMM 6220T were 78.9 and 83.6 %, respectively; the corresponding values between 2943T and Fulvivirga imtechensis AK7T were 69.8 and 63.6 %, respectively. Therefore, strains 29W222T and 2943T represent to two novel species of the genus Fulvivirga, for which the names Fulvivirga marina sp. nov. (29W222T=KCTC 62848T=MCCC 1K05194T) and Fulvivirga sediminis sp. nov. (2943T=KCTC 62847T= MCCC 1K05144T) are proposed, respectively.


Assuntos
Ácidos Graxos , Água do Mar , Técnicas de Tipagem Bacteriana , Bacteroidetes , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Sedimentos Geológicos , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
12.
Anal Chem ; 93(4): 2327-2335, 2021 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-33432815

RESUMO

Rare CD19+ leukemic B cells present in purified T cell populations can cause disease relapse and even the failure of CD19-targeting CAR-T therapy as these rare cells have the ability to self-mask their surface CD19 and escape from the recognition of T cells. It is therefore critical to efficiently detect and robustly deplete rare leukemic B cells in samples of therapeutic T cells. Here, we present a novel microfluidic approach to address the challenges specific to quality control of therapeutic T cells - CAR-QC. CAR-QC utilizes immunomagnetic labeling with a highly selective microfluidic device to rank and isolate rare leukemic B cells in T cell populations. CAR-QC offers ultrasensitive detection of leukemic B cells at single-cell resolution and robust depletion efficiency up to 99.985%. We demonstrate that CAR-QC outperforms flow cytometry and magnetic-activated cell sorting for detecting or purifying spiked samples. In addition, we prove that the improved performance of CAR-QC helps to avoid the occurrence and possibly relapse of rare leukemic B cells in vitro.


Assuntos
Linfócitos B/fisiologia , Linfócitos T/fisiologia , Linhagem Celular , Humanos , Separação Imunomagnética , Leucemia de Células B , Técnicas Analíticas Microfluídicas
13.
Acc Chem Res ; 53(8): 1445-1457, 2020 08 18.
Artigo em Inglês | MEDLINE | ID: mdl-32662263

RESUMO

Cellular heterogeneity in biological systems presents major challenges in the diagnosis and treatment of disease and also complicates the deconvolution of complex cellular phenomena. Single-cell analysis methods provide information that is not masked by the intrinsic heterogeneity of the bulk population and can therefore be applied to gain insights into heterogeneity among different cell subpopulations with fine resolution. Over the last 5 years, an explosion in the number of single-cell measurement methods has occurred. However, most of these methods are applicable to pure populations of cultured cells and are not able to handle high levels of phenotypic heterogeneity or a large background of nontarget cells. Microfluidics is an attractive tool for single cell manipulation as it enables individual encasing of single cells, allowing for high-throughput analysis with precise control of the local environment. Our laboratory has developed a new microfluidics-based analytical strategy to meet this unmet need referred to as magnetic ranking cytometry (MagRC). Cells expressing a biomarker of interest are labeled with receptor-coated magnetic nanoparticles and isolated from nontarget cells using a microfluidic device. The device ranks the cells according to the level of bound magnetic nanoparticles, which corresponds to the expression level of a target biomarker. Over the last several years, two generations of MagRC devices have been developed for different applications. The first-generation MagRC devices are powerful tools for the quantitation and analysis of rare cells present in heterogeneous samples, such as circulating tumor cells, stem cells, and pathogenic bacteria. The second-generation MagRC devices are compatible with the efficient recovery of cells sorted on the basis of protein expression and can be used to analyze large populations of cells and perform phenotypic CRISPR screens. To improve analytical precision, newer iterations of the first-generation and second-generation MagRC devices have been integrated with electrochemical sensors and Hall effect sensors, respectively. Both generations of MagRC devices permit the isolation of viable cells, which sets the stage for a wide range of applications, such as generating cell lines from rare cells and in vitro screening for effective therapeutic interventions in cancer patients to realize the promise of personalized medicine. This Account summarizes the development and application of the MagRC and describes a suite of advances that have enabled single-cell tumor cell analysis and monitoring tumor response to therapy, stem cell analysis, and detection of pathogens.


Assuntos
Biomarcadores/metabolismo , Nanopartículas de Magnetita/química , Análise de Célula Única/métodos , Anticorpos/química , Anticorpos/imunologia , Aptâmeros de Nucleotídeos/química , Aptâmeros de Nucleotídeos/metabolismo , Bactérias/metabolismo , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/metabolismo , Farmacorresistência Bacteriana/genética , Molécula de Adesão da Célula Epitelial/metabolismo , Humanos , Dispositivos Lab-On-A-Chip , Células Neoplásicas Circulantes/metabolismo , Proteínas de Ligação às Penicilinas/imunologia , Proteínas de Ligação às Penicilinas/metabolismo , RNA Mensageiro/metabolismo , Receptor ErbB-2/imunologia , Receptor ErbB-2/metabolismo , Análise de Célula Única/instrumentação , Células-Tronco/citologia , Células-Tronco/metabolismo
14.
J Am Chem Soc ; 142(35): 14805-14809, 2020 09 02.
Artigo em Inglês | MEDLINE | ID: mdl-32786736

RESUMO

Profiling the heterogeneous phenotypes of live cancer cells is a key capability that requires single-cell analysis. However, acquiring information at the single-cell level for live cancer cells is challenging when small collections of cells are being targeted. Here, we report single-cell analysis for low abundance cells enabled by fluorescent droplet cytometry (FDC), an approach that uses a biomarker-specific enzymatic fluorescent assay carried out using a droplet microfluidic platform. FDC utilizes DNA-functionalized antibodies in droplets to achieve specific on-cell target detection and enables characterization and profiling of live cancer cells with single-cell resolution based on their surface phenotype. Using this approach, we achieve live-cell phenotypic profiling of multiple surface markers acquired with small (<40 cells) collections of cells.


Assuntos
Citometria de Fluxo , Corantes Fluorescentes/química , Técnicas Analíticas Microfluídicas , Neoplasias/patologia , Análise de Célula Única , Linhagem Celular Tumoral , Humanos , Masculino , Imagem Óptica , Tamanho da Partícula , Fenótipo , Propriedades de Superfície
15.
J Cell Physiol ; 234(10): 18666-18678, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30927262

RESUMO

In ruminant, adequate endometrial function is a major factor affecting implantation and economic efficiency. However, the precise mechanisms regulating goat endometrial function during the peri-implantation period of pregnancy are still unclear. Here, we investigated the functional role and signal transduction of the fifth component of the constitutive photomorphogenic-9 signalosome (COPS5) in the regulation of endometrial function in endometrial epithelial cells (EECs). Our results showed that hormones decreased COPS5 expression, and COPS5-mediated regulation of endometrial function. We also found that knockdown of COPS5 hindered EECs proliferation by the G1-phase cell cycle arrest. Hormones affected the activity of COPS5 through hormones receptors, while feedback from the expression of COPS5 regulated the transcription of the receptor. Moreover, knockdown of endoplasmic reticulum (ER) to nucleus signaling 1 (ERN1) via si-ERN1 partly inhibited endometrial function in shCOPS5 EECs. In addition, blocking the mTOR pathway by rapamycin promoted endometrial function in si-ERN1-transfected shCOPS5 EECs. Overall, these results suggest that COPS5 negatively regulates goat endometrial function via the ERN1 and mTOR-autophagy pathways and provide new insights into the mechanistic pathways of COPS5 during female reproductive development.


Assuntos
Autofagia , Complexo do Signalossomo COP9/metabolismo , Endométrio/metabolismo , Cabras/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Animais , Pontos de Checagem do Ciclo Celular , Células Epiteliais/metabolismo , Feminino , Fase G1 , Regulação da Expressão Gênica , Gravidez , Transdução de Sinais
16.
Nano Lett ; 18(11): 7188-7193, 2018 11 14.
Artigo em Inglês | MEDLINE | ID: mdl-30335391

RESUMO

Cell morphology and geometry affect cellular processes such as stem cell differentiation, suggesting that these parameters serve as fundamental regulators of biological processes within the cell. Hierarchical architectures featuring micro- and nanotopographical features therefore offer programmable systems for stem cell differentiation. However, a limited number of studies have explored the effects of hierarchical architectures due to the complexity of fabricating systems with rationally tunable micro- and nanostructuring. Here, we report three-dimensional (3D) nanostructured microarchitectures that efficiently regulate the fate of human mesenchymal stem cells (hMSCs). These nanostructured architectures strongly promote cell alignment and efficient neurogenic differentiation where over 85% of hMSCs express microtubule-associated protein 2 (MAP2), a mature neural marker, after 7 days of culture on the nanostructured surface. Remarkably, we found that the surface morphology of nanostructured surface is a key factor that promotes neurogenesis and that highly spiky structures promote more efficient neuronal differentiation. Immunostaining and gene expression profiling revealed significant upregulation of neuronal markers compared to unpatterned surfaces. These findings suggest that the 3D nanostructured microarchitectures can play a critical role in defining stem cell behavior.


Assuntos
Diferenciação Celular , Células-Tronco Mesenquimais , Nanoestruturas/química , Neurônios , Técnicas de Cultura de Células/métodos , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Neurônios/citologia , Neurônios/metabolismo , Fatores de Tempo
17.
Nano Lett ; 18(10): 6222-6228, 2018 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-30188727

RESUMO

Cytotoxic chemotherapeutics are important tools for the clinical treatment of a variety of solid tumors. However, their use is often complicated by multidrug resistance that can develop in patients, limiting the potencies of these agents. New strategies are needed to provide versatile systems that can respond to and disable resistance mechanisms. We demonstrate the use of a new family of materials, programmable metal/semiconductor nanostructures, for drug delivery and mRNA sensing in drug-resistant cells. These materials are composed of a central core gold nanoparticle surrounded by a layer of DNA-capped quantum dots. The modularity of these "core-satellite" assemblies allows for the construction of superstructures with controlled size and the incorporation of multiple functionalities for drug delivery. The DNA sequence within the nanoparticle specifically binds to an mRNA encoding an important drug resistance factor, MRP1, inside cancer cells, releasing a potent anticancer drug doxorubicin. This event triggers a turn-on fluorescence emission along with a downregulation of the MRP1 drug efflux pump, a main resistance factor for doxorubicin, yielding a remarkable improvement in therapeutic efficacy against drug-resistant cancer cells. This work paves the way for the development of programmable materials with multiple synergistic functionalities for biomedical applications.


Assuntos
Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Neoplasias/tratamento farmacológico , Pontos Quânticos/uso terapêutico , Sistemas de Liberação de Medicamentos , Técnicas de Transferência de Genes , Ouro/química , Humanos , Nanopartículas Metálicas/química , Nanopartículas Metálicas/uso terapêutico , Proteínas Associadas à Resistência a Múltiplos Medicamentos/química , Proteínas Associadas à Resistência a Múltiplos Medicamentos/genética , Nanoestruturas/química , Nanoestruturas/uso terapêutico , Neoplasias/genética , Neoplasias/patologia , Pontos Quânticos/química , RNA Mensageiro/química , RNA Mensageiro/genética , RNA Mensageiro/uso terapêutico , Semicondutores
18.
Angew Chem Int Ed Engl ; 58(41): 14519-14523, 2019 10 07.
Artigo em Inglês | MEDLINE | ID: mdl-31389126

RESUMO

In living systems, interfacial molecular interactions control many biological processes. New stimuli-responsive strategies are desired to provide versatile model systems that can regulate cell behavior in vitro. Described here are potential-responsive surfaces that control cell adhesion and release as well as stem cell differentiation. Cell adhesion can be modulated dynamically by applying negative and positive potentials to surfaces functionalized with tailored monolayers. This process alters cell morphology and ultimately controls behavior and the fate of the cells. Cells can be detached from the electrode surface as intact clusters with different geometries using electrochemical potentials. Importantly, morphological changes during adhesion guide stem cell differentiation. The higher accessibility of the peptide under a positive applied potential causes phenotypic changes in the cells that are hallmarks of osteogenesis, whereas lower accessibility of the peptide promoted by negative potentials leads to adipogenesis.


Assuntos
Fibroblastos/fisiologia , Animais , Biomarcadores/metabolismo , Adesão Celular , Diferenciação Celular , Linhagem Celular , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Fenômenos Eletrofisiológicos , Regulação da Expressão Gênica , Camundongos , Osteogênese/fisiologia , Osteonectina/genética , Osteonectina/metabolismo , Osteopontina/genética , Osteopontina/metabolismo , Propriedades de Superfície
20.
Int J Syst Evol Microbiol ; 67(5): 1275-1280, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28109202

RESUMO

Strain NH1T, a pink-pigmented, facultatively anaerobic, heterotrophic, catalase-positive and oxidase-negative, Gram-stain-negative marine bacterium, was isolated from marine sediment on the coast of Weihai, China. Cells of strain NH1T were rod-shaped, 0.8-2.0 µm in length and 0.5-1.0 µm in width. The strain was able to grow at 13-37 °C, pH 5.5-8.5, in the presence of 0.0-8.0 % (w/v) NaCl. Optimal growth was observed at 28 °C, with 3.0 % (w/v) NaCl and pH 6.5-7.0. Nitrate was reduced. The G+C content of the DNA was 41.9 mol%. The major isoprenoid quinone was MK-7 and the main cellular fatty acids (>10 %) were summed feature 3 (33.6 %) comprising iso-C15 : 0 2-OH and/or C16 : 1ω7c, and iso-C15:0 (19.2%). The major polar lipids in strain NH1T were phosphatidylethanolamine, unidentified lipids, phospholipid and aminolipids. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain NH1T was highly related to the type strains of Algoriphagus antarcticus (97.87 % 16SrRNA gene sequence similarity) and Algoriphagus ratkowskyi (97.56 %). On basis of the phenotypic and phylogenetic data, strain NH1T should be classified as representing a novel species of the genus Algoriphagus, for which the name Algoriphagus resistens sp. nov. is proposed. The type strain is NH1T (=MCCC 1H00140T=KCTC 52228T).


Assuntos
Bacteroidetes/classificação , Sedimentos Geológicos/microbiologia , Filogenia , Água do Mar/microbiologia , Técnicas de Tipagem Bacteriana , Bacteroidetes/genética , Bacteroidetes/isolamento & purificação , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química
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