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1.
J Prosthet Dent ; 129(3): 478-485, 2023 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-36621357

RESUMEN

STATEMENT OF PROBLEM: Support structures are essential for the quality of resin-based prostheses made by the digital light processing (DLP), but few studies have evaluated the effect of support structure on the accuracy of zirconia-based anatomic contour prostheses. PURPOSE: The purpose of this in vitro study was to evaluate the effect of maximum support attachment angle (MSA) on the intaglio surface trueness of anatomic contour prostheses made by DLP and compare the trueness of 2-unit anatomic contour prostheses with that of those produced by milling. MATERIAL AND METHODS: Anatomic contour single-unit prostheses were manufactured using DLP and a suspension with 3-mol% yttria-stabilized zirconia. Four different conditions of MSA values to the vertical axis of the object (50, 55, 60, and 65 degrees) were applied (n=10). After printing, postprocessing, and sintering, all successfully produced prostheses were evaluated for intaglio surface trueness by considering the root mean square (RMS). Using the MSA showing the highest trueness, the 2-unit prostheses made by DLP (DLP group) were compared with milled (MIL group) prostheses in terms of intaglio accuracy (n=10). One-way analysis of variance and a post hoc pairwise comparison or independent t test were used for trueness analysis (α=.05). RESULTS: Three MSA groups (50, 55, and 60 degrees) were successfully produced with significant differences between the trueness of the single-unit prostheses for the groups with different MSA values (P<.05). The highest trueness was in the 50-degree MSA group. The 2-unit prostheses of the DLP group with 50-degree MSA showed significantly lower trueness than those of the MIL group (P<.05); however, the RMS values of both groups were lower than 50 µm. CONCLUSIONS: The intaglio surface trueness of anatomic contour DLP-generated prostheses can be improved by changing the MSA. The 50-degree MSA was beneficial for the accuracy of both single-unit and 2-unit DLP-generated prostheses, produced within clinically acceptable limits.


Asunto(s)
Diseño Asistido por Computadora , Implantes Dentales , Circonio , Dentadura Completa
2.
Obstet Gynecol Surv ; 74(2): 111-125, 2019 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-30756125

RESUMEN

IMPORTANCE: Pregnancy is getting more and more complex due to increasing number of complications that may affect fetal outcomes. The introduction of newer "proteomics and metabolomics" technologies in the field of obstetrics and gynecology may allow physicians to identify possible associated etiologies that affect the mother during pregnancy and lead to associated complications affecting the offspring. OBJECTIVE: The principal objective of this review article is to provide a comprehensive evaluation of the use of proteomics and metabolomics in complicated pregnancies. Future studies that incorporate data from multiple technologies may allow the development of an integrated biological system approach to maternal genomes, proteomes, and metabolomes in pregnancy. EVIDENCE ACQUISITION AND RESULTS: We conducted a substantial MEDLINE, EBSCOhost, and Cochrane database search for all the relevant articles containing use of "omics" technologies in pregnancy. We identified 197 relevant articles, following standardized systematic review process along with grading systems; 69 eligible articles were identified. CONCLUSION/RELEVANCE: We sought to provide a comprehensive review in this emerging field of "omics" in pregnancy and associated complications. This article focuses mainly on use of proteomics and metabolomics identification techniques and possible interventions for early pregnancy complications to improve neonatal outcomes.


Asunto(s)
Metabolómica/métodos , Complicaciones del Embarazo/metabolismo , Proteómica/métodos , Femenino , Humanos , Embarazo
3.
Biomed Microdevices ; 20(4): 87, 2018 10 05.
Artículo en Inglés | MEDLINE | ID: mdl-30291460

RESUMEN

Selective cell enrichment technologies can play an important role in both diagnostic and therapeutic areas. However, currently used cell sorting techniques have difficulties in rapidly isolating only the desired target cells from a large volume of body fluids. In this work, we developed a filtering system that can quickly separate and highly concentrate cells from a large volume of solution, depending on their size, using a silicon membrane filter. To overcome the problems caused by material limitations of the brittle silicon, we designed a novel membrane filter with various pore designs. From these designs, the most optimal design with high pore density, while preventing crack formation was derived by applying fluid dynamics simulation and near-field stress analysis. The membrane filter system using the selected design was fabricated, and cell filtration performance was evaluated. The LNCaP cell in horse blood was recovered up to 86% and enriched to 187-fold compared to initial cell populations after filtration at a flow rate of 5 mL/min. The results demonstrate that the filter presented in this study can rapidly and selectively isolate target cells from a large volume of body fluid sample.


Asunto(s)
Separación Celular/instrumentación , Filtración/instrumentación , Hidrodinámica , Membranas Artificiales , Silicio/química , Estrés Mecánico , Diseño de Equipo , Humanos , Células Jurkat
4.
Biomaterials ; 35(26): 7501-10, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24917030

RESUMEN

Circulating tumor cells (CTCs) are rare cells and the presence of these cells may indicate a poor prognosis and a high potential for metastasis. Despite highly promising clinical applications, CTCs have not been investigated thoroughly, due to many technical limitations faced in their isolation and identification. Current CTC detection techniques mostly take the epithelial marker epithelial cell adhesion molecule (EpCAM), however, accumulating evidence suggests that CTCs show heterogeneous EpCAM expression due to the epithelial-to-mesenchymal transition (EMT). In this study, we report that a microchip filter device incorporating slit arrays and 3-dimensional flow that can separate heterogeneous population of cells with marker for CTCs. To select target we cultured breast cancer cells under prolonged mammosphere culture conditions which induced EMT phenotype. Under these conditions, cells show upregulation of caveolin1 (CAV1) but down-regulation of EpCAM expression. The proposed device which contains CAV1-EpCAM conjugated bead has several tens of times increased throughput. More importantly, this platform enables the enhanced capture yield from metastatic breast cancer patients and obtained cells that expressed various EMT markers. Further understanding of these EMT-related phenotypes will lead to improved detection techniques and may provide an opportunity to develop therapeutic strategies for effective treatment and prevention of cancer metastasis.


Asunto(s)
Antígenos de Neoplasias/metabolismo , Neoplasias de la Mama/sangre , Caveolina 1/metabolismo , Moléculas de Adhesión Celular/metabolismo , Separación Celular/instrumentación , Proteínas Inmovilizadas/metabolismo , Técnicas Analíticas Microfluídicas/instrumentación , Células Neoplásicas Circulantes/metabolismo , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular Tumoral , Molécula de Adhesión Celular Epitelial , Transición Epitelial-Mesenquimal , Diseño de Equipo , Femenino , Filtración/instrumentación , Humanos , Células Neoplásicas Circulantes/patología
5.
Anal Chem ; 86(8): 3735-42, 2014 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-24641782

RESUMEN

Full automation with high purity for circulating tumor cell (CTC) isolation has been regarded as a key goal to make CTC analysis a "bench-to-bedside" technology. Here, we have developed a novel centrifugal microfluidic platform that can isolate the rare cells from a large volume of whole blood. To isolate CTCs from whole blood, we introduce a disc device having the biggest sample capacity as well as manipulating blood cells for the first time. The fully automated disc platform could handle 5 mL of blood by designing the blood chamber having a triangular obstacle structure (TOS) with lateral direction. To guarantee high purity that enables molecular analysis with the rare cells, CTCs were bound to the microbeads covered with anti-EpCAM to discriminate density between CTCs and blood cells and the CTCs being heavier than blood cells were only settled under a density gradient medium (DGM) layer. To understand the movement of CTCs under centrifugal force, we performed computational fluid dynamics simulation and found that their major trajectories were the boundary walls of the DGM chamber, thereby optimizing the chamber design. After whole blood was inserted into the blood chamber of the disc platform, size- and density-amplified cancer cells were isolated within 78 min, with minimal contamination as much as approximately 12 leukocytes per milliliter. As a model of molecular analysis toward personalized cancer treatment, we performed epidermal growth factor receptor (EGFR) mutation analysis with HCC827 lung cancer cells and the isolated cells were then successfully detected for the mutation by PCR clamping and direct sequencing.


Asunto(s)
Separación Celular/instrumentación , Dispositivos Laboratorio en un Chip , Células Neoplásicas Circulantes/patología , Automatización , Células Sanguíneas , Línea Celular Tumoral , Centrifugación por Gradiente de Densidad , Análisis Mutacional de ADN , Receptores ErbB/genética , Humanos , Microfluídica , Reacción en Cadena de la Polimerasa , Medicina de Precisión
6.
J Craniofac Surg ; 24(5): 1565-8, 2013 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-24036727

RESUMEN

The zygomatic nerve (ZN), which originates from the maxillary nerve at the pterygopalatine fossa, enters the orbit through the inferior orbital fissure. Within the lateral region of the orbit, the ZN divides into the zygomaticofacial (ZF) and zygomaticotemporal (ZT) nerves. The ZF and ZT nerves then pass on to the face and temporal region through the zygomaticoorbital foramen and enter their own bony canals within the zygomatic bone. However, multiple zygomaticofacial and zygomaticotemporal canals (ZFCs and ZTCs, respectively) can be observed, and their detailed intrabony courses are unknown. The aim of this study was clarify the three-dimensional intrabony courses and running patterns of the ZFCs and ZTCs, both to obtain a detailed anatomical description and for clinical purposes. Fourteen sides of the zygomatic bones were scanned as two-dimensional images using a micro-computed tomography (CT), with 32-µm slice thickness. Intrabony structures of each canals were three-dimensionally reconstructed and analyzed using Mimics computer software (Version 10.01; Materialise, Leuven, Belgium). We found that some ZTC was originated from ZFC. In 71.4% of the specimens, the ZTC(s) divided from the intrabony canal along the course of the ZFC(s). In other cases, 28.6% of ZTCs were opened through each corresponding ZT foramen. Zygomaticofacial canal originates from zygomaticoorbital foramen, divided into some of ZTCs, and is finally opened as ZF foramen. This new anatomical description of the intrabony structures of the ZFC(s) and ZTC(s) within the zygomatic bone by micro-CT technology provided helpful information to surgeons performing clinical procedures such as Le Fort osteotomy and reconstructive surgeries in the midface region.


Asunto(s)
Pueblo Asiatico , Imagenología Tridimensional , Nervio Maxilar/patología , Órbita/inervación , Hueso Temporal/inervación , Hueso Temporal/cirugía , Microtomografía por Rayos X , Cigoma/inervación , Cigoma/cirugía , Anciano de 80 o más Años , Bélgica , Femenino , Humanos , Interpretación de Imagen Asistida por Computador , Masculino , Reconstrucción Mandibular , Órbita/cirugía , Osteotomía Le Fort , Procedimientos de Cirugía Plástica , Programas Informáticos , Hueso Temporal/patología , Tomografía Computarizada por Rayos X , Cigoma/patología
8.
Biosens Bioelectron ; 47: 508-14, 2013 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-23628845

RESUMEN

Using hybrid nanoparticles (HNPs), we demonstrate simultaneous capture, in situ protein expression analysis, and cellular phenotype identification of circulating tumor cells (CTCs). Each HNP consists of three parts: (i) antibodies that bind specifically to a known biomarker for CTCs, (ii) a quantum dot that emits fluorescence signals, and (iii) biotinylated DNA that allows capture and release of CTC-HNP complex to an in-house developed capture & recovery chip (CRC). To evaluate our approach, cells representative of different breast cancer subtypes (MCF-7: luminal; SK-BR-3: HER2; and MDA-MB-231: basal-like) were captured onto CRC and expressions of EpCAM, HER2, and EGFR were detected concurrently. The average capture efficiency of CTCs was 87.5% with identification accuracy of 92.4%. Subsequently, by cleaving the DNA portion with restriction enzymes, captured cells were released at efficiencies of 86.1%. Further studies showed that these recovered cells are viable and can proliferate in vitro. Using HNPs, it is possible to count, analyze in situ protein expression, and culture CTCs, all from the same set of cells, enabling a wide range of molecular- and cellular-based studies using CTCs.


Asunto(s)
Biomarcadores de Tumor/sangre , Neoplasias de la Mama/sangre , Nanopartículas/química , Células Neoplásicas Circulantes/inmunología , Anticuerpos/química , Anticuerpos/inmunología , Antígenos de Neoplasias/sangre , Antígenos de Neoplasias/aislamiento & purificación , Biotina/química , Neoplasias de la Mama/diagnóstico , Moléculas de Adhesión Celular/sangre , Moléculas de Adhesión Celular/aislamiento & purificación , ADN/química , Molécula de Adhesión Celular Epitelial , Receptores ErbB/sangre , Receptores ErbB/aislamiento & purificación , Femenino , Fluorescencia , Regulación Neoplásica de la Expresión Génica , Humanos , Células MCF-7 , Puntos Cuánticos/química , Receptor ErbB-2/sangre , Receptor ErbB-2/aislamiento & purificación
9.
Oncotarget ; 4(3): 474-93, 2013 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-23562889

RESUMEN

This study reports on probing the utility of in situ chromatin texture features such as nuclear DNA methylation and chromatin condensation patterns - visualized by fluorescent staining and evaluated by dedicated three-dimensional (3D) quantitative and high-throughput cell-by-cell image analysis - in assessing the proliferative capacity, i.e. growth behavior of cells: to provide a more dynamic picture of a cell population with potential implications in basic science, cancer diagnostics/prognostics and therapeutic drug development. Two types of primary cells and four different cancer cell lines were propagated and subjected to cell-counting, flow cytometry, confocal imaging, and 3D image analysis at various points in culture. Additionally a subset of primary and cancer cells was accelerated into senescence by oxidative stress. DNA methylation and chromatin condensation levels decreased with declining doubling times when primary cells aged in culture with the lowest levels reached at the stage of proliferative senescence. In comparison, immortal cancer cells with constant but higher doubling times mostly displayed lower and constant levels of the two in situ-derived features. However, stress-induced senescent primary and cancer cells showed similar levels of these features compared with primary cells that had reached natural growth arrest. With regards to global DNA methylation and chromatin condensation levels, aggressively growing cancer cells seem to take an intermediate level between normally proliferating and senescent cells. Thus, normal cells apparently reach cancer-cell equivalent stages of the two parameters at some point in aging, which might challenge phenotypic distinction between these two types of cells. Companion high-resolution molecular profiling could provide information on possible underlying differences that would explain benign versus malign cell growth behaviors.


Asunto(s)
Proliferación Celular , Senescencia Celular/fisiología , Cromatina/metabolismo , Metilación de ADN , Ciclo Celular/efectos de los fármacos , Ciclo Celular/genética , Ciclo Celular/fisiología , División Celular/efectos de los fármacos , División Celular/genética , División Celular/fisiología , Línea Celular Tumoral , Núcleo Celular/genética , Núcleo Celular/metabolismo , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/genética , Supervivencia Celular/fisiología , Células Cultivadas , Senescencia Celular/efectos de los fármacos , Senescencia Celular/genética , Cromatina/genética , Humanos , Peróxido de Hidrógeno/farmacología , Inmunohistoquímica , Microscopía Confocal , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/patología , Oxidantes/farmacología , Estrés Oxidativo/fisiología , Factores de Tiempo
10.
Analyst ; 138(9): 2558-66, 2013 May 07.
Artículo en Inglés | MEDLINE | ID: mdl-23478433

RESUMEN

A fully automated point-of-care testing (POCT) system with a surface acoustic wave (SAW) immunosensor was developed for rapid and sensitive detection of cardiac troponin I (cTnI) in body fluid (plasma and whole blood). The assay, based on gold nanoparticle sandwich immunoassay and subsequent gold staining, was performed on the SAW immunosensor packaged inside a disposable microfluidic cartridge. The entire fluidic process, including plasma separation, reagent transport, metering, and mixing, was carried out by controlling the centrifugal force acting on the rotating cartridge and laser-irradiated ferrowax microvalves. On investigation of sensor response to various cTnI concentrations, the system exhibited a high performance with a detection limit of 6.7 pg mL(-1), and the coefficient of variation was less than 10% over the entire test range (10 pg mL(-1) to 25 ng mL(-1)). On comparing this POCT system with a clinically utilized system in a physical laboratory (Centaur® XP; Siemens), a correlation coefficient of 0.998 was found, validating the diagnostic capability of the SAW immunosensor.


Asunto(s)
Inmunoensayo/instrumentación , Técnicas Analíticas Microfluídicas/instrumentación , Sistemas de Atención de Punto , Troponina I/sangre , Diseño de Equipo , Oro/química , Humanos , Límite de Detección , Nanopartículas/química , Sonido
11.
Small ; 9(18): 3103-10, 2013 Sep 23.
Artículo en Inglés | MEDLINE | ID: mdl-23401221

RESUMEN

Circulating tumor cells (CTCs), though exceedingly rare in the blood, are nonetheless becoming increasingly important in cancer diagnostics. Despite this keen interest and the growing number of potential clinical applications, there has been limited success in developing a CTC isolation platform that simultaneously optimizes recovery rates, purity, and cell compatibility. Herein, a novel tracheal carina-inspired bifurcated (TRAB) microfilter system is reported, which uses an optimal filter gap size satisfying both 100% theoretical recovery rate and purity, as determined by biomechanical analysis and fluid-structure interaction (FSI) simulations. Biomechanical properties are also used to clearly discriminate between cancer cells and leukocytes, whereby cancer cells are selectively bound to melamine microbeads, which increase the size and stiffness of these cells. Nanoindentation experiments are conducted to measure the stiffness of leukocytes as compared to the microbead-conjugated cancer cells, with these parameters then being used in FSI analyses to optimize the filter gap size. The simulation results show that given a flow rate of 100 µL min(-1), an 8 µm filter gap optimizes the recovery rate and purity. MCF-7 breast cancer cells with solid microbeads are spiked into 3 mL of whole blood and, by using this flow rate along with the optimized microfilter dimensions, the cell mixture passes through the TRAB filter, which achieves a recovery rate of 93% and purity of 59%. Regarding cell compatibility, it is verified that the isolation procedure does not adversely affect cell viability, thus also confirming that the re-collected cancer cells can be cultured for up to 8 days. This work demonstrates a CTC isolation technology platform that optimizes high recovery rates and cell purity while also providing a framework for functional cell studies, potentially enabling even more sensitive and specific cancer diagnostics.


Asunto(s)
Técnicas Analíticas Microfluídicas/métodos , Microscopía de Fuerza Atómica/métodos , Células Neoplásicas Circulantes/metabolismo , Tráquea , Humanos , Microfluídica
12.
BMC Pharmacol Toxicol ; 14: 11, 2013 Feb 11.
Artículo en Inglés | MEDLINE | ID: mdl-23394161

RESUMEN

BACKGROUND: The spatial organization of the genome is being evaluated as a novel indicator of toxicity in conjunction with drug-induced global DNA hypomethylation and concurrent chromatin reorganization. 3D quantitative DNA methylation imaging (3D-qDMI) was applied as a cell-by-cell high-throughput approach to investigate this matter by assessing genome topology through represented immunofluorescent nuclear distribution patterns of 5-methylcytosine (MeC) and global DNA (4,6-diamidino-2-phenylindole = DAPI) in labeled nuclei. METHODS: Differential progression of global DNA hypomethylation was studied by comparatively dosing zebularine (ZEB) and 5-azacytidine (AZA). Treated and untreated (control) human prostate and liver cancer cells were subjected to confocal scanning microscopy and dedicated 3D image analysis for the following features: differential nuclear MeC/DAPI load and codistribution patterns, cell similarity based on these patterns, and corresponding differences in the topology of low-intensity MeC (LIM) and low in intensity DAPI (LID) sites. RESULTS: Both agents generated a high fraction of similar MeC phenotypes across applied concentrations. ZEB exerted similar effects at 10-100-fold higher drug concentrations than its AZA analogue: concentration-dependent progression of global cytosine demethylation, validated by measuring differential MeC levels in repeat sequences using MethyLight, and the concurrent increase in nuclear LIM densities correlated with cellular growth reduction and cytotoxicity. CONCLUSIONS: 3D-qDMI demonstrated the capability of quantitating dose-dependent drug-induced spatial progression of DNA demethylation in cell nuclei, independent from interphase cell-cycle stages and in conjunction with cytotoxicity. The results support the notion of DNA methylation topology being considered as a potential indicator of causal impacts on chromatin distribution with a conceivable application in epigenetic drug toxicology.


Asunto(s)
Antineoplásicos/farmacología , Azacitidina/farmacología , Citidina/análogos & derivados , Metilación de ADN , 5-Metilcitosina/inmunología , Anticuerpos Monoclonales/farmacología , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Citidina/farmacología , Humanos , Neoplasias Hepáticas , Masculino , Fenotipo , Neoplasias de la Próstata
13.
Anal Chem ; 84(17): 7400-7, 2012 Sep 04.
Artículo en Inglés | MEDLINE | ID: mdl-22881997

RESUMEN

Isolation of circulating tumor cells (CTCs) by size exclusion can yield poor purity and low recovery rates, due to large variations in size of CTCs, which may overlap with leukocytes and render size-based filtration methods unreliable. This report presents a very sensitive, selective, fast, and novel method for isolation and detection of CTCs. Our assay platform consists of three steps: (i) capturing CTCs with anti-EpCAM conjugated microbeads, (ii) removal of unwanted hematologic cells (e.g., leukocytes, erythrocytes, etc.) by selective sedimentation of CTCs within a density gradient medium, and (iii) simple microfiltration to collect these cells. To demonstrate the efficacy of this assay, MCF-7 breast cancer cells (average diameter, 24 µm) and DMS-79 small cell lung cancer cells (average diameter, 10 µm) were used to model CTCs. We investigated the relative sedimentation rates for various cells and/or particles, such as CTCs conjugated with different types of microbeads, leukocytes, and erythrocytes, in order to maximize differences in the physical properties. We observed that greater than 99% of leukocytes in whole blood were effectively removed at an optimal centrifugal force, due to differences in their sedimentation rates, yielding a much purer sample compared to other filter-based methods. We also investigated not only the effect of filtration conditions on recovery rates and sample purity but also the sensitivity of our assay platform. Our results showed a near perfect recovery rate (~99%) for MCF-7 cells and very high recovery rate (~89%) for DMS-79 cells, with minimal amounts of leukocytes present.


Asunto(s)
Separación Inmunomagnética , Células Neoplásicas Circulantes , Anticuerpos Inmovilizados/inmunología , Antígenos de Neoplasias/inmunología , Sedimentación Sanguínea , Moléculas de Adhesión Celular/inmunología , Línea Celular Tumoral , Molécula de Adhesión Celular Epitelial , Eritrocitos/citología , Humanos , Leucocitos/citología , Células MCF-7 , Microesferas
14.
Yonsei Med J ; 45(3): 501-9, 2004 Jun 30.
Artículo en Inglés | MEDLINE | ID: mdl-15227738

RESUMEN

In this paper, an emergency telemedicine system was designed for the transmission of real-time multimedia for remote consultation, including radiological images, patient records, video-conferencing, full-quality video, ECG, BP, respiration, temperature, SpO(2), systolic and diastolic pressures and heart rate. The standardized, modular, software-based design architecture, without resorting to external hardware compression boards, enables the low-cost implementation of the telemedicine system, using the unified, systematic and compact integration of multimedia on general personal computers. Experimental tests on local networks analyze the technical aspects of designed systems, and inter-hospital experiments demonstrate its clinical usefulness.


Asunto(s)
Sistemas de Computación , Servicios Médicos de Urgencia/organización & administración , Servicio de Urgencia en Hospital/organización & administración , Consulta Remota/organización & administración , Telemedicina/organización & administración , Computadores , Diseño de Equipo , Humanos , Proyectos Piloto , Programas Informáticos
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