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1.
Nano Lett ; 24(6): 2063-2070, 2024 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-38299886

RESUMO

On-chip integrated meta-optics promise to achieve high-performance and compact integrated photonic devices. To arbitrarily engineer the optical trajectory along the propagation path in an on-chip integrated scheme is of significance in fundamental physics and various emerging applications. Here, we experimentally demonstrate an on-chip metasurface integrated on a waveguide to enable predefined arbitrary optical trajectories in the visible regime. By transformation of the transverse phase to generate longitudinal mapping, the guided waves are extracted and molded into any different optical trajectories (parabola, hyperbola, and cosine). More intriguingly, predefined polarization states with longitudinal variation are also successfully imparted along the trajectory. Owing to the on-chip propagation scheme, the trajectories are uniquely free from zero-order diffraction interference, naturally having a higher signal-to-noise ratio beyond conventional free-space forms. Overall, such on-chip optical trajectory engineering allows for miniaturized integration and can find paths in potential applications of complex optical manipulation, advanced laser fabrication, and microscopic imaging.

2.
Nano Lett ; 22(5): 2059-2064, 2022 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-35201771

RESUMO

Despite various advances in achieving arbitrary optics steering, one of the longstanding challenges is to achieve optical merging for combining multidirectional beams through single-time reflection/transmission in free space. Typically, dual-directional beam merging is conducted by combining half-transmission and half-reflection using beam splitters; however, it leads to a bulky system with stray light and low merging efficiency. The difficulty of free-space beam merging lies in imparting respective distinct wavevectors to different directional beams. Herein, we originally proposed and successfully demonstrated the free-space optical merging (FOM) functionality based on the inverse-designed meta-grating architecture in the visible regime. By utilizing the inverse problem solver, two proposed meta-grating schemes experimentally enable merging of dual-directional beams into the same outgoing angle for the first time merely through single-time reflection. We envision that the creation of free-space merging performance can be widely applicable to the future optical system and facilitate the miniature optical devices and integration.

3.
Nano Lett ; 22(24): 9990-9996, 2022 12 28.
Artigo em Inglês | MEDLINE | ID: mdl-36490382

RESUMO

Devices used for meta-optics display are currently undergoing a revolutionary transition from static to dynamic. Despite various tuning strategy demonstrations such as mechanical, electrical, optical, and thermal tunings, a longstanding challenge for their practical application has been the achievement of a conveniently accessible real-life tuning scheme for realizing versatile functionality dynamics outside the laboratory. In this study, we demonstrate a practical tuning strategy to realize a dynamic color printing with a switchable meta-holography exhibition based on hydrogel-based nanocavities. On the basis of the inflation sensitivity of a hydrogel to humidity alteration, its transmissive color was notably tuned from 450 to 750 nm. More intriguingly, by controlling the sample dry/immersed states in real time, we successfully enabled dual-channel switchable meta-holography. With the advantages of facile architecture, daily stimulus with large-area modulation, and high chromaticity, our proposed hydrogel-based nanocavities provide a promising path toward tunable display/encryption, optical sensors, and next-generation display technology.


Assuntos
Holografia , Hidrogéis , Eletricidade , Umidade , Impressão Tridimensional
4.
Opt Express ; 28(16): 23652-23659, 2020 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-32752358

RESUMO

A new type of diffractive lens based on hybridized Fabry-Perot (FP) cavities with high-NA and achromatic features for arbitrary dual-wavelengths is theoretically proposed and demonstrated. We utilize the subwavelength-scale metal-insulator-metal nanocavity to form a Fresnel zone plate (MIM-FZP) that benefits from both spectral selectivity and high numerical aperture (NA > 0.9) to enable lensing functionality. By taking advantage of the different transmission orders from MIM, any arbitrary dual-wavelength achromatic focusing design is achieved. Using this approach, we merge two independent MIM-FZP designs and realize achromatic focusing performance at the selected dual-wavelength of 400/600 nm. Furthermore, the achromatic lens also exhibits a crucial potential for dynamically tuning of the operation wavelengths and focusing lengths as actively scaling the core layer thickness of MIM. The unique MIM-FZP design can be practically fabricated using a grayscale lithography technique. We believe such high-NA and achromatic optical devices enjoy great simplicity for structural design and can easily find applications including high-resolution imaging, new-generation integrated optoelectronic devices, confocal collimation, and achromatic lens, etc.

5.
J Surg Res ; 205(2): 464-473, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27664897

RESUMO

BACKGROUND: Mesenchymal stem cell (MSC) transplantation is an effective treatment therapy for ischemic ulcers. However, in high-glucose microenvironment, the original inflammation-inhibiting function of MSCs leads to turns into secreting large amounts of inflammatory mediators, such as tumor necrosis factor alpha, for example, which decreases their capacity and becomes poor quality stem cells over inflammation cells for diabetic foot ulcers repair in the healing of diabetic foot ulcers. Erythropoietin (EPO) is an anti-inflammatory, proangiogenic cytokine. It is unclear whether EPO-activated MSCs with biomaterials can promote the effective healing of diabetic foot ulcers. METHODS: Cultivated MSCs in MSC-L, MSC-H, EPO-G, Akt-G, and mTOR-G, then separated the supernatant-conditioned medium of these groups to stimulate human umbilical vein endothelial cells on proliferation and migration experiments; a new type of biomaterial planted with the EPO-activated MSCs was applied to the diabetic foot ulcers of the C57 mice. RESULTS: In vitro experiments showed that EPO could stimulate MSCs to secrete vascular endothelial growth factor in high-glucose microenvironment. More importantly, EPO could reduce the damage to MSCs by high-glucose microenvironment, promote their proliferation and migration functions, and inhibit the high-glucose-induced MSCs from secreting the inflammatory mediator tumor necrosis factor alpha. In vivo experiments showed greater angiogenesis in EPO-MSC group than in control group, ulcer healing in EPO-MSC group was significantly better than that in control group, and MSCs partially differentiated into endothelial cells. EPO-activated MSCs could inhibit the monocyte invasion of localized diabetic foot ulcers. CONCLUSIONS: Our results indicate that EPO-activated MSCs can promote the effective healing of diabetic foot ulcers. The mechanism is that EPO can change stem cells from excessive inflammation into general inflammation and improved diabetic foot ulcers inflammatory microenvironment.


Assuntos
Microambiente Celular/fisiologia , Pé Diabético/terapia , Eritropoetina/metabolismo , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/metabolismo , Cicatrização/fisiologia , Animais , Biomarcadores/metabolismo , Movimento Celular , Proliferação de Células , Citocinas/metabolismo , Pé Diabético/metabolismo , Glucose/metabolismo , Células Endoteliais da Veia Umbilical Humana/fisiologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neovascularização Fisiológica/fisiologia , Resultado do Tratamento
6.
ACS Nano ; 18(28): 18693-18700, 2024 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-38958405

RESUMO

Facing the challenge of information security in the current era of information technology, optical encryption based on metasurfaces presents a promising solution to this issue. However, most metasurface-based encryption techniques rely on limited decoding keys and struggle to achieve multidimensional complex encryption. It hinders the progress of optical storage capacity and puts encryption security at a disclosing risk. Here, we propose and experimentally demonstrate a multidimensional encryption system based on chip-integrated metasurfaces that successfully incorporates the simultaneous manipulation of three-dimensional optical parameters, including wavelength, direction, and polarization. Hence, up to eight-channel augmented reality (AR) holograms are concealed by near- and far-field fused encryption, which can only be extracted by correctly providing the three-dimensional decoding keys and then vividly exhibit to the authorizer with low crosstalk, high definition, and no zero-order speckle noise. We envision that the miniature chip-integrated metasurface strategy for multidimensional encryption functionalities promises a feasible route toward the encryption capacity and information security enhancement of the anticounterfeiting performance and optically cryptographic storage.

7.
Mol Ther ; 20(8): 1501-7, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22692496

RESUMO

Duchenne muscular dystrophy (DMD) is a fatal, X-linked muscle disease caused by mutations in the dystrophin gene. Adeno-associated viral (AAV) vector-mediated gene replacement strategies hold promise as a treatment. Studies in animal models and human trials suggested that immune responses to AAV capsid proteins and transgene products prevented efficient gene therapy. In this study, we used widespread intramuscular (i.m.) injection to deliver AAV6-canine micro-dystrophin (c-µdys) throughout a group of skeletal muscles in dystrophic dogs given a brief course of commonly used immunosuppressants. Robust c-µdys expression was obtained for at least two years and was associated with molecular reconstitution of the dystrophin-glycoprotein complex (DGC) at the muscle membrane. Importantly, c-µdys expression was maintained for at least 18 months after discontinuing immunosuppression. The results obtained in a relevant preclinical model of DMD demonstrate feasibility of widespread AAV-mediated muscle transduction and transgene expression in the presence of transient immunosuppression to achieve molecular reconstitution that can be directly translated to human trials.


Assuntos
Distrofina/metabolismo , Distrofia Muscular de Duchenne/metabolismo , Distrofia Muscular de Duchenne/terapia , Animais , Western Blotting , Linhagem Celular , Cães , Distrofina/genética , Ensaio de Imunoadsorção Enzimática , Humanos , Microscopia Eletrônica , Microscopia de Fluorescência , Distrofia Muscular de Duchenne/genética
8.
Huan Jing Ke Xue ; 44(1): 395-404, 2023 Jan 08.
Artigo em Zh | MEDLINE | ID: mdl-36635827

RESUMO

Available selenium (Se) in soil was the predominant factor affecting the content of Se in crops. In order to reasonably delineate the Se-rich soil range and propose theoretical guidance for the cultivation of natural Se-rich crops in a region where the surface soils had a high level of available-Se and a low level of total-Se, 8814 samples in surface soil and 195 root-crop matching samples were collected in Shizuishan in northern Ningxia. On the basis of the main line of analysis of available-Se, the following research was conducted: by synthetically studying the total-Se and available-Se in surface soil and root soil, the morphology of Se in surface soil, as well as Se in crops, deep and coordinated analyses of content among total-Se, available-Se, and Se in root-crop matching samples were carried out, and the suitable threshold for Shizuishan was confirmed. A multiple regression model of available-Se was established to determine the main physical and chemical indexes affecting available-Se, which were expected to improve the Se enrichment rate of crops through the enhancement of available-Se. The results demonstrated that ω(Se) and ω(Seavailable)in the surface soil in Shizuishan were 0.26 mg·kg-1 and 12.85 µg·kg-1, respectively, and the characteristics of Se and available-Se in root-crop matching samples could represent those in surface soil. Thus, it was recommended to use 0.24 mg·kg-1 as the suitable threshold of Se-rich soil. The multiple regression model of available-Se showed that the increase in total-Se and soil elements affecting soil fertility could promote the enrichment of available-Se.


Assuntos
Selênio , Poluentes do Solo , Solo/química , Produtos Agrícolas/química , Poluentes do Solo/análise
9.
Huan Jing Ke Xue ; 44(8): 4429-4439, 2023 Aug 08.
Artigo em Zh | MEDLINE | ID: mdl-37694637

RESUMO

In order to explore the environmental geochemistry characteristics of heavy metals (HMs) in soil-crop systems in an old industrial city, the concentration and fraction of HMs in the paddy, wheat, and maize root soil and their seeds were detected and analyzed. Subsequently, statistical methods, risk assessment coding (RAC), the bio-enrichment coefficient factor (BCF), influence index of comprehensive quality (IICQ), and ArcGIS spatial interpolation were used to conduct the translocation, accumulation, and comprehensive risk assessment of HMs in soil-crop systems. The results showed that the average concentrations of As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn in root soil were ranked respectively as follows:12.56, 0.19, 63.48, 23.52, 0.038, 28.86, 21.68, and 69.47 mg·kg-1. HMs in root soil were accumulated to some extent in comparison with the soil background value in Ningxia, especially Cd and Hg, but did not exceed the soil environmental pollution screening value (GB 14618-2018). The average concentrations of the eight aforementioned elements in supporting crop seeds were 0.0149, 0.0112, 0.075, 6.7, 0.0015, 0.67, 0.0427, and 20.48 mg·kg-1 in turn. The over-limit ratio of As, Pb, and Cr in crop seeds was 4%, 3%, and 1%, respectively, relative to the national food safety standards (GB 2762-2017), whereas the other five elements were within the allowable range. In comparison to those in paddy and wheat, HMs hardly tended to translocate to maize seeds from root soil. According to the results of IICQ in soil-crop systems, the cultivated soil was in the state of slight sub-contamination regionally, and only 10% of sampling points showed slight (sub-)contamination-submoderate contamination, where we could replant maize to reduce HMs contamination risk.


Assuntos
Mercúrio , Metais Pesados , Cádmio , Chumbo , China , Medição de Risco , Solo , Triticum
10.
Huan Jing Ke Xue ; 43(8): 4179-4189, 2022 Aug 08.
Artigo em Zh | MEDLINE | ID: mdl-35971715

RESUMO

Shizuishan is a typical exhausted resources-based city in the northern area of the Ningxia Hui autonomous region in China. In order to develop the planting industry of selenium (Se)-rich agricultural products and promote green and sustainable urban development and transformation, investigations on the quality of Se-rich land were carried out in Shizuishan City, where 7399 surface soil (0-20 cm) samples of farmlands, 30 atmospheric precipitation samples, and nine parent rocks were collected. By means of semi variogram model construction by GS+, Kriging interpolation in ArcGIS and statistics via SPSS, such as correlation analysis and mean-value analysis, the content, spatial distribution, and enrichment factors of Se-soil were analyzed. Further, the enrichment characteristics of soil Se in alkaline conditions were summarized. The results indicated that ω(Se) in surface soil was (0.26±0.12) mg·kg-1, and its spatial distribution was highly auto-correlated. The variation in Se content was related to natural factors. Along Helan Mountain, the content of Se in the surface soil was comparatively higher than that where coal mines were located. The parent rock was the principal factor that controlled the enrichment of soil Se. The physical and chemical properties of soil such as organic material, pH, and iron and manganese oxides had crucial effects on the enrichment of soil Se in a surficial environment. Compared to a strong alkaline environment, alkaline conditions were beneficial for the enrichment of Se in the surface soil.


Assuntos
Selênio , Poluentes do Solo , China , Cidades , Fazendas , Solo/química , Poluentes do Solo/análise
11.
Cell Prolif ; 55(11): e13316, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35869570

RESUMO

OBJECTIVES: To determine the therapeutic effect of tetrahedral framework nucleic acids (tFNAs) on diabetic wound healing and the underlying mechanism. MATERIALS AND METHODS: The tFNAs were characterized by polyacrylamide gel electrophoresis (PAGE), atomic force microscopy (AFM), transmission electron microscopy (TEM), dynamic light scattering (DLS) and zeta potential assays. Cell Counting Kit-8 (CCK-8) and migration assays were performed to evaluate the effects of tFNAs on cellular proliferation and migration. Quantitative polymerase chain reaction (Q-PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect the effect of tFNAs on growth factors. The function and role of tFNAs in diabetic wound healing were investigated using diabetic wound models, histological analyses and western blotting. RESULTS: Cellular proliferation and migration were enhanced after treatment with tFNAs in a high-glucose environment. The expression of growth factors was also facilitated by tFNAs in vitro. During in vivo experiments, tFNAs accelerated the healing process in diabetic wounds and promoted the regeneration of the epidermis, capillaries and collagen. Moreover, tFNAs increased the secretion of growth factors and activated the Wnt pathway in diabetic wounds. CONCLUSIONS: This study indicates that tFNAs can accelerate diabetic wound healing and have potential for the treatment of diabetic wounds.


Assuntos
Diabetes Mellitus , Ácidos Nucleicos , Humanos , Ácidos Nucleicos/farmacologia , Via de Sinalização Wnt , Cicatrização , Proliferação de Células
12.
Mol Ther ; 18(6): 1165-72, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20372105

RESUMO

We evaluated the potential of lung-directed gene therapy for alpha1-antitrypsin (AAT) deficiency using an adeno-associated virus type 6 (AAV6) vector containing a human AAT (hAAT) complementary DNA (cDNA) delivered to the lungs of mice and dogs. The results in normal and immune-deficient mice showed that hAAT concentrations were much higher in lung fluid than in plasma, and therapeutic levels were obtained even in normal mice. However, in normal mice an immune response against the vector and/or transgene limited long-term gene expression. An AAV6 vector expressing a marker protein verified that AAV6 vectors efficiently transduced lung cells in dogs. Delivery of AAV6-hAAT resulted in low levels of hAAT in dog serum but therapeutic levels in the lung that persisted for at least 58 days to 4 months in three immunosuppressed dogs. Expression in the serum was not detectable after 45 days in one nonimmune suppressed dog. A lymphoproliferative response to AAV capsid but not to hAAT was detected even after immunosuppression. These results in mice and dogs show the feasibility of expression of therapeutic levels of AAT in the lungs after AAV vector delivery, and advocate for approaches to prevent cellular immune responses to AAV capsid proteins for persistence of gene expression in humans.


Assuntos
Dependovirus/genética , Técnicas de Transferência de Genes , Vetores Genéticos , Pulmão/metabolismo , alfa 1-Antitripsina/genética , Animais , Líquido da Lavagem Broncoalveolar , DNA Complementar , Cães , Ensaio de Imunoadsorção Enzimática , Terapia Genética , Humanos , Camundongos , Deficiência de alfa 1-Antitripsina/terapia
13.
Mol Ther ; 18(3): 617-24, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20040912

RESUMO

We previously demonstrated that direct intramuscular injection of rAAV2 or rAAV6 in wild-type dogs resulted in robust T-cell responses to viral capsid proteins, and others have shown that cellular immunity to adeno-associated virus (AAV) capsid proteins coincided with liver toxicity and elimination of transgene expression in a human trial of hemophilia B. Here, we show that the heparin-binding ability of a given AAV serotype does not determine the induction of T-cell responses following intramuscular injection in dogs, and identify multiple epitopes in the AAV capsid protein that are recognized by T cells elicited by AAV injection. We also demonstrate that noninvasive magnetic resonance imaging (MRI) can accurately detect local inflammatory responses following intramuscular rAAV injection in dogs. These studies suggest that pseudotyping rAAV vectors to remove heparin-binding activity will not be sufficient to abrogate immunogenicity, and validate the utility of enzyme-linked immunosorbent spot (ELISpot) assay and MRI for monitoring immune and inflammatory responses following intramuscular injection of rAAV vectors in preclinical studies in dogs. These assays should be incorporated into future human clinical trials of AAV gene therapy to monitor immune responses.


Assuntos
Dependovirus/metabolismo , Terapia Genética/métodos , Músculos/metabolismo , Animais , Cães , Ensaio de Imunoadsorção Enzimática/métodos , Epitopos/química , Proteoglicanas de Heparan Sulfato/química , Heparina/metabolismo , Sistema Imunitário/metabolismo , Inflamação , Imageamento por Ressonância Magnética/métodos , Microscopia de Fluorescência/métodos , Músculos/patologia , Peptídeos/química , Linfócitos T/imunologia
14.
ACS Appl Mater Interfaces ; 13(32): 38623-38628, 2021 Aug 18.
Artigo em Inglês | MEDLINE | ID: mdl-34369745

RESUMO

Two-dimensional (2D) metasurfaces hold great promise to enable multiplexing and multifunctional optical devices due to their artificial freedom in design, device miniaturization, etc. Various multiplexing and multifunctional metasurfaces have been extensively studied, including polarization multiplexing, wavelength multiplexing, and orbit angular momentum (OAM) multiplexing. However, due to the lack of angular encoding freedom, angular multiplexing switchable nanoprinting has rarely been studied or demonstrated yet to the best of our knowledge. Here, we realize angular multiplexing switchable nanoprinting functionality with independent amplitude encryption based on visible-frequency metasurfaces. By screening a large number of structural designs and breaking the angular correlation, we eventually obtain optimal metasurface designs to realize dual-channel arbitrary image encryption. Furthermore, we illustrate that the proposed scheme would serve as an optical information concealment/retrieval strategy by combining the structural color and amplitude modulation. Overall, we believe that angular multiplexing metasurfaces would easily find promising applications, including optical information encryption/concealment, multifunctional switchable devices, and advanced eyeglass-free three-dimensional (3D) stereoscopic displays.

15.
Br J Haematol ; 149(1): 101-10, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20067562

RESUMO

We retrospectively analyzed transfusion requirements within the first 100 d among allogeneic haematopoietic cell transplantation (HCT) recipients with haematological malignancies given either myeloablative (n = 1353) or nonmyeloablative conditioning (n = 503). We confirmed that myeloablative recipients required more platelet and red blood cell (RBC) transfusions than nonmyeloablative recipients (P < 0.0001 for both). Myeloablative patients given peripheral blood stem cells required less platelet transfusions (P < 0.0001) than those given marrow while RBC transfusion requirements did not differ significantly. Subsequent analyses were restricted to nonmyeloablative recipients. Platelet and RBC transfusions were less frequent among related compared to unrelated recipients (P < 0.0001 for both), with comparable median numbers of transfused units. Major/bidirectionally ABO-mismatched recipients required more RBC transfusions than ABO-matched recipients (P = 0.006). Rates of graft rejection/failure, grades II-IV acute and chronic graft-versus-host-disease (GVHD), 2-year relapse, 3-year survivals and non-relapse mortality were comparable among ABO-matched, minor-mismatched, and major/bidirectionally mismatched recipients (P = 0.93, 0.72, 0.57, 0.36, 0.17 and 0.79, respectively). Times to disappearance of anti-donor IgG and IgM isohemagglutinins among major/bidirectionally ABO-mismatched recipients were affected by magnitude of pre-HCT titres (P < 0.001 for both) but not GVHD (P = 0.71 and 0.78, respectively). In conclusion, nonmyeloablative recipients required fewer platelet and RBC transfusions and among them, both unrelated and major/bidirectionally ABO-mismatched recipients required more RBC transfusions. ABO incompatibility did not affect nonmyeloablative HCT outcomes.


Assuntos
Sistema ABO de Grupos Sanguíneos/imunologia , Transfusão de Componentes Sanguíneos/métodos , Incompatibilidade de Grupos Sanguíneos , Transplante de Células-Tronco Hematopoéticas , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Transfusão de Eritrócitos , Neoplasias Hematológicas/terapia , Humanos , Lactente , Pessoa de Meia-Idade , Transfusão de Plaquetas , Estudos Retrospectivos , Análise de Sobrevida , Condicionamento Pré-Transplante/métodos
16.
Hum Gene Ther ; 18(1): 18-26, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-17176210

RESUMO

Recombinant adeno-associated virus (rAAV)-mediated gene transfer has shown promise for treating diseases in various animal models including the mdx mouse model of Duchenne muscular dystrophy (DMD). In many cases, however, preclinical studies in inbred mice have not successfully predicted human clinical responses. To assess the potential clinical utility of treating human DMD patients by AAV-mediated gene delivery, we performed a series of direct intramuscular injections in random-bred wild-type dogs. AAV serotypes 2 and 6 carrying different promoter-transgene cassettes were produced as previously described for murine studies and administered intramuscularly. The injection sites were biopsied at various time points and analyzed for transgene expression and immunohistochemical analysis. In contrast to the generally nonimmunogenic nature of these vectors in murine studies, both AAV2 and AAV6 vectors elicited robust cellular immune responses regardless of the transgene expressed, the cellular specificity of the promoter, and the muscle type injected. Viral purification by various methods did not diminish T cell-mediated infiltration. Our data indicate that AAV2 and AAV6 capsid proteins can elicit primary cellular immune responses when injected into the skeletal muscle of random-bred dogs, and suggest the possibility of cellular immunity to AAV vectors in humans.


Assuntos
Dependovirus/imunologia , Terapia Genética , Vetores Genéticos/imunologia , Imunidade Celular , Distrofia Muscular Animal/terapia , Distrofia Muscular de Duchenne/imunologia , Distrofia Muscular de Duchenne/terapia , Animais , Modelos Animais de Doenças , Cães , Terapia Genética/efeitos adversos , Vetores Genéticos/administração & dosagem , Humanos , Camundongos , Camundongos Endogâmicos mdx , Distrofia Muscular Animal/imunologia , Distrofia Muscular Animal/patologia , Distrofia Muscular de Duchenne/patologia , Linfócitos T/imunologia , Linfócitos T/patologia , Transgenes/imunologia
17.
Hum Gene Ther Clin Dev ; 26(3): 194-201, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26252064

RESUMO

Cardiomyocytes derived from human induced pluripotent stem cells (iPSCs) show great promise as autologous donor cells to treat heart disease. A major technical obstacle to this approach is that available induction methods often produce heterogeneous cell population with low percentage of cardiomyocytes. Here we describe a cardiac enrichment approach using nonintegrating adeno-associated virus (AAV). We first examined several AAV serotypes for their ability to selectively transduce iPSC-derived cardiomyocytes. Results showed that AAV1 demonstrated the highest in vitro transduction efficiency among seven widely used serotypes. Next, differentiated iPSC derivatives were transduced with drug-selectable AAV1 expressing neomycin resistance gene. Selection with G418 enriched the cardiac cell fraction from 27% to 57% in 2 weeks. Compared with other enrichment strategies such as integrative genetic selection, mitochondria labeling, or surface marker cell sorting, this simple AAV method described herein bypasses antibody or dye labeling. These findings provide proof of concept for large-scale cardiomyocyte enrichment by exploiting AAV's intrinsic tissue tropism.


Assuntos
Diferenciação Celular , Dependovirus/genética , Vetores Genéticos/genética , Miócitos Cardíacos/citologia , Miócitos Cardíacos/metabolismo , Células-Tronco/citologia , Células-Tronco/metabolismo , Diferenciação Celular/genética , Linhagem Celular , Dependovirus/classificação , Expressão Gênica , Técnicas de Transferência de Genes , Genes Reporter , Humanos , Imuno-Histoquímica , Imunofenotipagem , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Sorogrupo , Transdução Genética , Tropismo Viral
18.
Am J Phys Med Rehabil ; 93(11 Suppl 3): S97-107, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25313664

RESUMO

The development of clinical vectors to correct genetic mutations that cause inherited myopathies and related disorders of skeletal muscle is advancing at an impressive rate. Adeno-associated virus vectors are attractive for clinical use because (1) adeno-associated viruses do not cause human disease and (2) these vectors are able to persist for years. New vectors are now becoming available as gene therapy delivery tools, and recent preclinical experiments have demonstrated the feasibility, safety, and efficacy of gene therapy with adeno-associated virus for long-term correction of muscle pathology and weakness in myotubularin-deficient canine and murine disease models. In this review, recent advances in the application of gene therapies to treat inherited muscle disorders are presented, including Duchenne muscular dystrophy and x-linked myotubular myopathy. Potential areas for therapeutic synergies between rehabilitation medicine and genetics are also discussed.


Assuntos
Terapia Genética/métodos , Distrofia Muscular de Duchenne/terapia , Miopatias Congênitas Estruturais/terapia , Medicina Física e Reabilitação/tendências , Medicina Regenerativa/tendências , Animais , Estudos de Coortes , Terapia Combinada , Modelos Animais de Doenças , Cães , Previsões , Vetores Genéticos , Humanos , Doenças Musculares/diagnóstico , Doenças Musculares/terapia , Distrofia Muscular de Duchenne/diagnóstico , Distrofia Muscular de Duchenne/mortalidade , Miopatias Congênitas Estruturais/diagnóstico , Miopatias Congênitas Estruturais/mortalidade , Medicina Física e Reabilitação/métodos , Prognóstico , Medicina Regenerativa/métodos , Análise de Sobrevida , Resultado do Tratamento
19.
Stem Cell Res ; 12(2): 467-80, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24434629

RESUMO

The ability to extract somatic cells from a patient and reprogram them to pluripotency opens up new possibilities for personalized medicine. Induced pluripotent stem cells (iPSCs) have been employed to generate beating cardiomyocytes from a patient's skin or blood cells. Here, iPSC methods were used to generate cardiomyocytes starting from the urine of a patient with Duchenne muscular dystrophy (DMD). Urine was chosen as a starting material because it contains adult stem cells called urine-derived stem cells (USCs). USCs express the canonical reprogramming factors c-myc and klf4, and possess high telomerase activity. Pluripotency of urine-derived iPSC clones was confirmed by immunocytochemistry, RT-PCR and teratoma formation. Urine-derived iPSC clones generated from healthy volunteers and a DMD patient were differentiated into beating cardiomyocytes using a series of small molecules in monolayer culture. Results indicate that cardiomyocytes retain the DMD patient's dystrophin mutation. Physiological assays suggest that dystrophin-deficient cardiomyocytes possess phenotypic differences from normal cardiomyocytes. These results demonstrate the feasibility of generating cardiomyocytes from a urine sample and that urine-derived cardiomyocytes retain characteristic features that might be further exploited for mechanistic studies and drug discovery.


Assuntos
Distrofina/deficiência , Células-Tronco Pluripotentes Induzidas/patologia , Distrofia Muscular de Duchenne/patologia , Distrofia Muscular de Duchenne/urina , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Adulto , Animais , Estudos de Casos e Controles , Diferenciação Celular/fisiologia , Células Cultivadas , Descoberta de Drogas , Feminino , Humanos , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/biossíntese , Fatores de Transcrição Kruppel-Like/urina , Masculino , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Miócitos Cardíacos/citologia , Proteínas Proto-Oncogênicas c-myc/biossíntese , Proteínas Proto-Oncogênicas c-myc/urina , Telomerase/urina , Adulto Jovem
20.
Int J Cardiovasc Imaging ; 29(4): 819-26, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23070737

RESUMO

Myocardial perfusion studies using dynamic contrast-enhanced cardiac magnetic resonance imaging (CMRI) could provide valuable, quantitative information regarding heart physiology in diseases such as Duchenne muscular dystrophy (DMD), that lead to diffuse myocardial damage. The goal of this effort was to develop an intuitive but physiologically meaningful method for quantifying myocardial perfusion by CMRI and to test its ability to detect global myocardial differences in a dog model of DMD. A discrete-time model was developed that parameterizes contrast agent kinetics in terms of an uptake coefficient that describes the forward flux of contrast agent into the tissue, and a retention coefficient that describes the rate of decay in tissue concentration due to contrast agent efflux. This model was tested in 5 dogs with DMD and 6 healthy controls which were imaged using a perfusion sequence on a 3T clinical scanner. CINE and delayed-enhancement CMRI acquisitions were also used to assess cardiac function and the presence of myocardial scar. Among functional parameters measured by CMRI, no significant differences were observed. No myocardial scar was observed. Increased perfusion in DMD was observed with an uptake coefficient of 6.76 ± 2.41 % compared to 2.98 ± 1.46 % in controls (p = 0.03). Additionally, the retention coefficient appeared lower at 82.2 ± 5.8 % in dogs with DMD compared to 90.5 ± 6.6 % in controls (p = 0.12). A discrete-time kinetic model of uptake and retention of contrast agent in perfusion CMRI shows potential for the study of DMD.


Assuntos
Meios de Contraste/farmacocinética , Gadolínio DTPA/farmacocinética , Imagem Cinética por Ressonância Magnética , Distrofia Muscular de Duchenne/diagnóstico , Distrofia Muscular de Duchenne/metabolismo , Imagem de Perfusão do Miocárdio/métodos , Miocárdio/metabolismo , Animais , Circulação Coronária , Modelos Animais de Doenças , Cães , Modelos Biológicos , Distrofia Muscular de Duchenne/patologia , Distrofia Muscular de Duchenne/fisiopatologia , Miocárdio/patologia , Valor Preditivo dos Testes , Distribuição Tecidual
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