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Investigation of the dynamic structural changes in the actin cytoskeleton during cell migration provides crucial information about the physiological conditions of a stem cell during in-vitro culture. Here we proposed a quantitative analytical model associated with texture extraction with cell tracking techniques for in situ monitoring of the cytoskeletal density change of stem cells in phase-contrast microscopy without fluorescence staining. The reliability of the model in quantifying the texture density with different orientation was first validated using a series of simulated textural images. The capability of the method to reflect the spatiotemporal regulation of the cytoskeletal structure of a living stem cell was further proved by applying it to a set of 72 h phase-contrast microscopic video of the growth dynamics of mesenchymal stem cells in vitro culture.
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The quality of stem cells obtained through serial subcultivation is the pivotal factor determining the therapeutic effectiveness of regenerative medicine. However, an effective quality monitoring system for cell culture is yet to be established. Detailed parameter studies of the migratory behavior of stem cells at different passages may provide insight into the deterioration of stemness. Thus, this study aimed to evaluate the feasibility of quantitative bioimage analysis for monitoring stem cell quality during in vitro culture and to explore the passaging effects on stem cell migration. An image-based analytical tool using cell tracking, cytometric analyses, and gating with time-lapse microscopy was developed to characterize the migratory behavior of human mesenchymal stem cells (hMSCs) isolated from human adipose tissue (hADAS) and placenta (hPDMC) cultured on chitosan membranes. Quantitative analysis was performed for the single cells and assembled spheroids selected from 15 videos of Passages 3, 5, and 11 for hADAS and those from 12 videos of Passages 7, 11, and 16 for hPDMC. These passages were selected to represent the young, matured, and degenerated stem cells, respectively. Migratory behavior varied with cell passages. The mobility of single hMSCs decreased at degenerated passages. In addition, enhancement of mobility, due to transformation from single cells to spheroids, occurred at each passage. The young hMSCs seemed more likely to move as single cells rather than as aggregates. Once matured, they tended to aggregate with strong 3D spheroid formability and increased mobility. However, the spheroid formability and mobility decreased at late passage. The increase in aggregation rate with passaging may be a compensatory mechanism to enhance the declining mobility of hMSCs through cell coordination. Our findings regarding the passaging effects on stem-cell migratory behavior agree with biochemical reports, suggesting that the developed imaging method is capable of monitoring the cell-culture quality effectively. © 2020 International Society for Advancement of Cytometry.
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Células Madre Mesenquimatosas , Tejido Adiposo , Técnicas de Cultivo de Célula , Diferenciación Celular , Movimiento Celular , Células Cultivadas , Femenino , Humanos , Embarazo , Células MadreRESUMEN
In this paper, we introduce a newly developed multi-scale wavelet model for the interpretation of surface electromyography (SEMG) signals and validate the model's capability to characterize changes in neuromuscular activation in cases with myofascial pain syndrome (MPS) via machine learning methods. The SEMG data collected from normal (Nâ¯=â¯30; 27 women, 3 men) and MPS subjects (Nâ¯=â¯26; 22 women, 4 men) were adopted for this retrospective analysis. SMEGs were measured from the taut-band loci on both sides of the trapezius muscle on the upper back while he/she conducted a cyclic bilateral backward shoulder extension movement within 1â¯min. Classification accuracy of the SEMG model to differentiate MPS patients from normal subjects was 77% using template matching and 60% using K-means clustering. Classification consistency between the two machine learning methods was 87% in the normal group and 93% in the MPS group. The 2D feature graphs derived from the proposed multi-scale model revealed distinct patterns between normal subjects and MPS patients. The classification consistency using template matching and K-means clustering suggests the potential of using the proposed model to characterize interference pattern changes induced by MPS.
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Electromiografía/métodos , Aprendizaje Automático , Modelos Neurológicos , Músculo Esquelético/fisiopatología , Síndromes del Dolor Miofascial/fisiopatología , Adulto , Femenino , Humanos , Masculino , Músculo Esquelético/fisiología , Hombro/fisiología , Hombro/fisiopatologíaRESUMEN
Dynamic behavior of stem cells during in vitro development is diverse. Previous cell tracking studies have focused more on cell proliferation than on cell aggregation. However, the enhancement of cell proliferation in association with cell aggregation has been reported. In a previous study, we also demonstrated that the aggregation of adult human mesenchymal stem cells to form three-dimensional (3D) cellular spheroids helped maintain the expression of stemness marker genes in the cells. However, the dynamic behavioral changes triggered by spheroid formation remain to be investigated. A scheme of image processing techniques is proposed to meet this need. A hybrid-thresholding technique was first developed for efficient segmentation of cell clusters, after which a cell tracking method based on pair-matching with topological constraints was designed. Two morphological indices were derived to track the timing of 3D spheroid formation during the cellular aggregation process. Five cell motility indices measured from single cells and 3D spheroids were then compared. After confirmation of more than 90% correspondence between the results obtained by manual tracking and the proposed methods, an analysis of cellular behavior reveals a significant increase in motility in association with spheroid formation, consistent with a previous report that used a gene expression approach. This study proposed a systematic image analysis method to quantify the dynamic behavior of stem cells for stemness evaluation during cell culturing in vitro. Results demonstrated the validity of the developed platform in investigation of the dynamic behavior of cell aggregation in stem cell cultures in vitro.
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Células Madre Adultas/fisiología , Movimiento Celular/fisiología , Procesamiento de Imagen Asistido por Computador/métodos , Células Madre Mesenquimatosas/fisiología , Esferoides Celulares/citología , Tejido Adiposo/citología , Adulto , Células Madre Adultas/citología , Agregación Celular , Técnicas de Cultivo de Célula , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Quitosano/farmacología , Biología Computacional/métodos , Humanos , Células Madre Mesenquimatosas/citología , Microscopía , Biología de Sistemas/métodosRESUMEN
PURPOSE: The aims of this study were to investigate the hypometabolic regions of FDG PET compared with the nigrostriatal dopamine pathway abnormalities in TRODAT-1 scan in patients with dementia with Lewy bodies (DLBs) at mild and dementia stages as well as to validate the correlation among networks being constructed with clinical data. MATERIALS AND METHODS: A total of 25 DLB patients were classified into 2 functional groups stratified by the Clinical Dementia Rating (CDR) Scale (CDR 0.5: n = 14, mild stage; CDR 1 or 2: n = 11, dementia stage) compared with 9 age-matched controls. Neuroimaging survey was applied using information derived from FDG PET by performing voxel-based analysis and a semiquantitative Tc-TRODAT-1 scan to correlate these results with the cognitive and Unified Parkinson's Disease Rating Scale. RESULTS: Compared with normal database, the patients with mild stage showed hypometabolism in the temporal regions, anterior cingulate cortex, inferior orbital region, thalamus, and caudate nucleus. Although at the dementia stage, more extensive cortical hypometabolism involving occipital region were found. The dopamine transporter levels derived from TRODAT-1 scan had excellent discrimination in diagnosing DLB compared with age-matched normal controls (1.58 [0.2] and 1.84 [0.1], P < 0.01) but without significant differences between mild and dementia stages. The sophisticated cortical-brainstem networks by FDG PET and TRODAT-1 yielded good clinical correlation. CONCLUSIONS: The networks yielded from FDG PET and TRODAT-1 revealed good correlation with clinical data and that nigrostriatal pathway abnormalities are preceded by typical occipital hypometabolism in mild stage of DLB. Dopamine transporter levels may serve as early diagnostic tool and FDG PET as staging indicator for DLB pathology.
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Fluorodesoxiglucosa F18 , Enfermedad por Cuerpos de Lewy/diagnóstico por imagen , Compuestos de Organotecnecio , Tomografía de Emisión de Positrones , Radiofármacos , Tropanos , Anciano , Anciano de 80 o más Años , Corteza Cerebral/diagnóstico por imagen , Corteza Cerebral/metabolismo , Corteza Cerebral/patología , Cuerpo Estriado/diagnóstico por imagen , Cuerpo Estriado/metabolismo , Cuerpo Estriado/patología , Femenino , Humanos , Masculino , Sustancia Negra/diagnóstico por imagen , Sustancia Negra/metabolismo , Sustancia Negra/patologíaRESUMEN
Novel peripheral nerve conduits containing the negatively charged Tremella fuciformis polysaccharide (TF) were prepared, and their efficacy in bridging a critical nerve gap was evaluated. The conduits were made of poly(D,L-lactide) (PLA) with asymmetric microporous structure. TF was immobilized on the lumen surface of the nerve conduits after open air plasma activation. The TF-modified surface was characterized by the attenuated total reflection Fourier-transformed infrared spectroscopy and the scanning electron microscopy. TF modification was found to enhance the neurotrophic gene expression of C6 glioma cells in vitro. TF-modified PLA nerve conduits were tested for their ability to bridge a 15 mm gap of rat sciatic nerve. Nerve regeneration was monitored by the magnetic resonance imaging. Results showed that TF immobilization promoted the nerve connection in 6 weeks. The functional recovery in animals receiving TF-immobilized conduits was greater than in those receiving the bare conduits during an 8-month period. The degree of functional recovery reached ~90% after 8 months in the group of TF-immobilized conduits.
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Quantification of regional (99m)Tc-TRODAT-1 binding ratio in the striatum regions in SPECT images is essential for differential diagnosis between Alzheimer's and Parkinson's diseases. Defining the region of the striatum in the SPECT image is the first step toward success in the quantification of the TRODAT-1 binding ratio. However, because SPECT images reveal insufficient information regarding the anatomical structure of the brain, correct delineation of the striatum directly from the SPECT image is almost impossible. We present a method integrating the active contour model and the hybrid registration technique to extract regions from MR T1-weighted images and map them into the corresponding SPECT images. Results from three normal subjects suggest that the segmentation accuracy using the proposed method was compatible with the expert decision but has a higher efficiency and reproducibility than manual delineation. The binding ratio derived by this method correlated well (R (2) = 0.76) with those values calculated by commercial software, suggesting the feasibility of the proposed method.
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Cuerpo Estriado/diagnóstico por imagen , Cuerpo Estriado/patología , Imagen por Resonancia Magnética/estadística & datos numéricos , Tomografía Computarizada de Emisión de Fotón Único/estadística & datos numéricos , Algoritmos , Enfermedad de Alzheimer/diagnóstico , Enfermedad de Alzheimer/diagnóstico por imagen , Enfermedad de Alzheimer/patología , Encéfalo/diagnóstico por imagen , Encéfalo/patología , Biología Computacional , Diagnóstico por Computador/estadística & datos numéricos , Sistemas Especialistas , Humanos , Compuestos de Organotecnecio , Enfermedad de Parkinson/diagnóstico , Enfermedad de Parkinson/diagnóstico por imagen , Enfermedad de Parkinson/patología , Interpretación de Imagen Radiográfica Asistida por Computador , Radiofármacos , Tomografía Computarizada por Rayos X/estadística & datos numéricos , TropanosRESUMEN
To solve the limitations in using the conventional parametric measures to define myofascial pain, a 3-D multi-scale wavelet energy variation graph is proposed as a way to inspect the pattern of surface electromyography (SEMG) variation between the dominant and nondominant sides at different frequency scales during a muscle contraction cycle and the associated changes with the upper-back myofascial pain. The model was developed based on the property of the wavelet energy of the SEMG signal revealing the degree of correspondence between the shape of the motor unit action potential and the wavelet waveform at a certain scale in terms of the frequency band. The characteristic pattern of the graph for each group (30 normal and 26 patient subjects) was first derived and revealed the dominant-hand effect and the changes with myofascial pain. Through comparison of individual graphs across subjects, we found that the graph pattern reveals a sensitivity of 53.85% at a specificity of 83.33% in the identification of myofascial pain. The changes in these patterns provide insight into the transformation between different fiber recruitment, which cannot be explored using conventional SEMG features. Therefore, this multi-scale analysis model could provide a reliable SEMG features to identify myofascial pain.
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Electromiografía/métodos , Fibromialgia/fisiopatología , Modelos Neurológicos , Contracción Muscular , Músculo Esquelético/fisiopatología , Unión Neuromuscular/fisiopatología , Transmisión Sináptica , Adulto , Algoritmos , Simulación por Computador , Femenino , Humanos , Masculino , Persona de Mediana Edad , Modelos Estadísticos , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Optoelectronics techniques to induce protoporphyrin IX fluorescence with topically applied 5-aminolevulinic acid on the oral mucosa have been developed to noninvasively detect oral cancer. Fluorescence imaging enables wide-area screening for oral premalignancy, but the lack of an adequate fluorescence enhancement method restricts the clinical imaging application of these techniques. This study aimed to develop a reliable fluorescence enhancement method to improve PpIX fluorescence imaging systems for oral cancer detection. Three contrast features, red-green-blue reflectance difference, R∕B ratio, and R∕G ratio, were developed first based on the optical properties of the fluorescence images. A comparative study was then carried out with one negative control and four biopsy confirmed clinical cases to validate the optimal image processing method for the detection of the distribution of malignancy. The results showed the superiority of the R∕G ratio in terms of yielding a better contrast between normal and neoplastic tissue, and this method was less prone to errors in detection. Quantitative comparison with the clinical diagnoses in the four neoplastic cases showed that the regions of premalignancy obtained using the proposed method accorded with the expert's determination, suggesting the potential clinical application of this method for the detection of oral cancer.
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Diagnóstico por Imagen/métodos , Aumento de la Imagen/métodos , Neoplasias de la Boca/diagnóstico , Neoplasias de la Boca/metabolismo , Protoporfirinas/metabolismo , Ácido Aminolevulínico , Diagnóstico por Imagen/instrumentación , Fluorescencia , Humanos , Aumento de la Imagen/instrumentación , Fenómenos Ópticos , Fármacos FotosensibilizantesRESUMEN
The performance of an asymmetric conduit made of microporous polylactic acid (PLA) in promoting the long-term peripheral nerve regeneration across a 20-mm-long sciatic nerve gap was evaluated by a rabbit sciatic nerve transection model. Magnetic resonance imaging (MRI) was employed to monitor the nerve regeneration process. The extents of nerve regeneration and conduit degradation were quantified by image analysis. Functional and histological analyses were followed to assess nerve reinnervation. MR images showed that the transected nerve was connected at about 4 months. The diameter of the regenerated nerve continued to increase while the conduit was gradually degraded. The conduit was completely degraded in 18 months. The degradation kinetics in vivo was estimated based on MR images. The functional recovery after 18 months was â¼82% based on electrophysiology. The extension range of the operated limb was slowly recuperated to â¼81% at 18 months. Histology showed that nerve bundles were self-assembled after 16-18 months, but the morphologies were still different from those of normal sciatic nerve. This was the first work on the long-term evaluation of peripheral nerve regeneration in a rabbit model, and the first to report the use of MRI to obtain the real-time images of regenerated nerve in a biomaterial conduit as well as to define the degradation rate of the conduit in vivo. The platform established in this study serves to evaluate the regeneration of larger-diameter (>3-mm) nerve across a long-gap bridged by a conduit.
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Materiales Biocompatibles/química , Ácido Láctico/química , Regeneración Nerviosa , Polímeros/química , Nervio Ciático/fisiología , Animales , Imagen por Resonancia Magnética , Masculino , Poliésteres , Porosidad , ConejosRESUMEN
This study proposes a VR-based tool, named "Virtual City," to evaluate wayfinding abilities that are reported to decline with some cognition dysfunctions, such as dementia. This platform was designed according to cognition theories and took advantage of convenience of real-time recording by using VR techniques to make the evaluations less subjective and more accurate. Our previous study has shown the feasibility of using VR techniques to evaluate the sense of direction; we further use this newly developed tool to assess the effect of "screen type" and "reference landmark" on wayfinding performances. Experimental results show that subjects perform better with true-size 3D visual displays and reference landmarks, suggesting the factors that need to be considered for clinical application by using VR tools to detect cognition impairment in wayfinding.
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Cognición , Interfaz Usuario-Computador , Adulto , Ciudades , Gráficos por Computador , Simulación por Computador , Computadores , Femenino , Humanos , Masculino , Orientación , Desempeño Psicomotor/fisiología , Programas Informáticos , Percepción Espacial , Taiwán , Percepción VisualRESUMEN
This paper presents the development of a computer-aided image management tool, named CAIMT, used for patient setup error measurement in radiation treatment planning. The CAIMT incorporates user interaction to facilitate contour detection from both portal film and simulation film. The detected contours were then used as features for image registration through application of the generalized Hough transform (GHT). Positioning error was measured when the optimal registration was achieved. The CAIMT has been applied to register the image pairs of a rando(TM) phantom and five real subjects, in order to evaluate its reliability and stability. The promising results suggest that the CAIMT can assist radiotherapists to align the simulation film and the portal film precisely and steadily and therefore adjust patient positioning to the optimum in a more efficient way.
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Neoplasias de Cabeza y Cuello/radioterapia , Interpretación de Imagen Radiográfica Asistida por Computador/métodos , Planificación de la Radioterapia Asistida por Computador/métodos , Algoritmos , Humanos , Variaciones Dependientes del Observador , Fantasmas de Imagen , Reproducibilidad de los Resultados , Interfaz Usuario-ComputadorRESUMEN
This study intends to explore the wavelet denoising for optimal MUAP detection through the wavelet analysis of surface electromyographic (SEMG) signals. We first derive an estimator for signal to noise ratio and show that this estimator correlates to the quality of the reconstructed simulated signal. When applying this estimator to evaluate the SEMG signal, we find that the reconstructed signal is insensitive to the selection of denoising methods. This finding is further confirmed by the identical plots of those reconstructed SEMG data. In addition, the close correspondence of MUAP occurrences in the reconstructed signal and those in the original signal suggests that the denoising procedure can preserve the features of MUAP in the original SEMG signals.
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Simulación por Computador , Interpretación Estadística de Datos , Electromiografía/instrumentación , Electrofisiología/instrumentación , Procesamiento de Señales Asistido por Computador/instrumentación , Algoritmos , Artefactos , Computadores , Electromiografía/métodos , Electrofisiología/métodos , Diseño de Equipo , Humanos , Modelos Estadísticos , Proyectos de InvestigaciónRESUMEN
This paper presents a computer-aided platform, named Virtual Hospital (VH), to evaluate the wayfinding ability that is found impaired in senile people with early dementia. The development of the VH takes the advantage of virtual reality technology to make the evaluation of the sense of direction more convenient and accurate then the conventional way. A pilot study was carried out to test its feasibility in differentiating the sense of direction between different genders. The results with significant differences in the response time (p<<0.05) and the pointing error (p<<0.01) between genders suggest the potential of the VH for clinical uses. Further improvement on the human-machine interface is necessary to make it easy for geriatric people to use.