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J Prosthet Dent ; 2021 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-33468316


STATEMENT OF PROBLEM: Anterior tooth selection is an important step in complete denture treatment as it plays a pivotal role not only in esthetics but also in mastication and pronunciation. However, conventional methods for tooth selection are not well established and rely on facial measurements and proportions, which vary among different ethnicities. PURPOSE: The purpose of this clinical study was to investigate the relationship between interalar width and intercanine distance and to compare different clinical methods for determining the position of the canine tooth. MATERIAL AND METHODS: Two hundred Thai participants (100 men and 100 women) aged 18 to 25 years with 6 full maxillary anterior teeth were enrolled in this study. The interalar width and intercanine distance were measured with digital vernier calipers and compared by using the paired-samples t test. To determine the canine position, 2 reference lines-the alar line (A line) and the inner canthus of the eye to alar line (IA line)-were drawn through the canine on both sides. The horizontal distances from each reference line to the canine cusp tip and distal contact point were evaluated and then analyzed using the 1-sample t test. RESULTS: All measurements were significantly different between men and women (P<.01). Interalar width was greater than intercanine distance in both sexes. In men, the A line coincided with the canine distal contact point (P>.05). In contrast, the IA line was distal to the canine distal contact point by 3.5 ±3.6 mm on the left side and by 3.9 ±3.4 mm on the right side. In women, the A line was situated between the canine cusp tip and distal contact point. It was mesial to the distal contact point by 2.0 ±2.0 mm on the left side and by 1.8 ±2.0 mm on the right side. The IA line was distal to the canine distal contact point by 1.2 ±2.6 mm on the left side and by 1.6 ±2.7 mm on the right side. CONCLUSIONS: The interalar width is greater than the intercanine distance in both sexes. The A line is more clinically relevant than the IA line for predicting canine position. The A line can directly determine the distal contact point of the canine in edentulous male patients. However, in women, a distance of approximately 2 mm should be added distal to the A line to locate the distal contact point of the canine on both sides.

Toxicol Pathol ; 43(6): 883-9, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26157038


Tissue localization of immune cells is critical to the study of disease processes in mouse models of human diseases. However, immunohistochemistry (IHC) for immune cell phenotyping in mouse tissue sections presents specific technical challenges. For example, CD4 and CD8 have been difficult to detect using IHC on formalin-fixed and paraffin-embedded mouse tissue, prompting alternative methods. We investigated the use of formalin-free zinc-salt fixation (ZN) and optimized IHC protocols for detecting a panel of immune cell-related markers (CD3, CD4, CD8, Foxp3, B220, F4/80, CD68, and major histocompatibility complex [MHC] class-I, MHC class-II, and Gr-1). The IHC results for these markers were compared on mouse spleen tissue treated with neutral buffered formalin (NBF) or ZN with or ZN without antigen retrieval (AR). Whereas CD4 and CD8 were not detected in NBF-treated tissue, all markers were detected in ZN-treated tissue without AR. Thus, the use of ZN treatment for IHC staining can be a good tool for studying immunoreactive lesions in tissues.

Biomarcadores/análise , Imunidade Celular/efeitos dos fármacos , Imuno-Histoquímica/métodos , Fixação de Tecidos/métodos , Zinco/química , Animais , Antígenos/análise , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Fixadores , Formaldeído , Camundongos , Baço/citologia , Baço/imunologia
PLoS One ; 9(10): e108978, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25279759


Electron tomography (ET) of biological samples is used to study the organization and the structure of the whole cell and subcellular complexes in great detail. However, projections cannot be acquired over full tilt angle range with biological samples in electron microscopy. ET image reconstruction can be considered an ill-posed problem because of this missing information. This results in artifacts, seen as the loss of three-dimensional (3D) resolution in the reconstructed images. The goal of this study was to achieve isotropic resolution with a statistical reconstruction method, sequential maximum a posteriori expectation maximization (sMAP-EM), using no prior morphological knowledge about the specimen. The missing wedge effects on sMAP-EM were examined with a synthetic cell phantom to assess the effects of noise. An experimental dataset of a multivesicular body was evaluated with a number of gold particles. An ellipsoid fitting based method was developed to realize the quantitative measures elongation and contrast in an automated, objective, and reliable way. The method statistically evaluates the sub-volumes containing gold particles randomly located in various parts of the whole volume, thus giving information about the robustness of the volume reconstruction. The quantitative results were also compared with reconstructions made with widely-used weighted backprojection and simultaneous iterative reconstruction technique methods. The results showed that the proposed sMAP-EM method significantly suppresses the effects of the missing information producing isotropic resolution. Furthermore, this method improves the contrast ratio, enhancing the applicability of further automatic and semi-automatic analysis. These improvements in ET reconstruction by sMAP-EM enable analysis of subcellular structures with higher three-dimensional resolution and contrast than conventional methods.

Tomografia com Microscopia Eletrônica/métodos , Processamento de Imagem Assistida por Computador/métodos , Algoritmos , Artefatos
PLoS One ; 9(10): e108948, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25299706


Cellular uptake of clustered α2ß1-integrin induces the formation of membrane compartments that subsequently mature into a multivesicular body (MVB). Enhanced internalization mediated by clustered integrins was observed upon infection by the picornavirus echovirus 1 (EVI). We elucidated the structural features of virus-induced MVBs (vMVBs) in comparison to antibody-induced control MVBs (mock infection) by means of high-pressure cryo fixation of cells followed by immuno electron tomography during early entry of the virus. Three-dimensional tomograms revealed a marked increase in the size and complexity of these vMVBs and the intraluminal vesicles (ILVs) at 2 and 3.5 hours post infection (p.i.), in contrast to the control MVBs without virus. Breakages in the membranes of vMVBs were detected from tomograms after 2 and especially after 3.5 h suggesting that these breakages could facilitate the genome release to the cytoplasm. The in situ neutral-red labeling of viral genome showed that virus uncoating starts as early as 30 min p.i., while an increase of permeability was detected in the vMVBs between 1 and 3 hours p.i., based on a confocal microscopy assay. Altogether, the data show marked morphological changes in size and permeability of the endosomes in the infectious entry pathway of this non-enveloped enterovirus and suggest that the formed breakages facilitate the transfer of the genome to the cytoplasm for replication.

Integrina alfa2beta1/metabolismo , Corpos Multivesiculares/metabolismo , Corpos Multivesiculares/virologia , Infecções por Picornaviridae/metabolismo , Infecções por Picornaviridae/fisiopatologia , Picornaviridae/metabolismo , Picornaviridae/fisiologia , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Membrana Celular/fisiologia , Membrana Celular/virologia , Citoplasma/metabolismo , Citoplasma/fisiologia , Tomografia com Microscopia Eletrônica/métodos , Endossomos/metabolismo , Endossomos/fisiologia , Endossomos/virologia , Humanos , Microscopia Confocal , Corpos Multivesiculares/fisiologia , Permeabilidade , Infecções por Picornaviridae/virologia
J Virol ; 84(21): 11145-51, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20739526


Membrane glycoproteins of alphavirus play a critical role in the assembly and budding of progeny virions. However, knowledge regarding transport of viral glycoproteins to the plasma membrane is obscure. In this study, we investigated the role of cytopathic vacuole type II (CPV-II) through in situ electron tomography of alphavirus-infected cells. The results revealed that CPV-II contains viral glycoproteins arranged in helical tubular arrays resembling the basic organization of glycoprotein trimers on the envelope of the mature virions. The location of CPV-II adjacent to the site of viral budding suggests a model for the transport of structural components to the site of budding. Thus, the structural characteristics of CPV-II can be used in evaluating the design of a packaging cell line for replicon production.

Alphavirus/fisiologia , Membrana Celular/virologia , Glicoproteínas/metabolismo , Montagem de Vírus , Liberação de Vírus , Animais , Transporte Biológico , Linhagem Celular , Cricetinae , Tomografia com Microscopia Eletrônica , Vacúolos , Proteínas Virais/metabolismo
Mol Ther ; 17(5): 828-36, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19240693


The development of techniques to efficiently deliver genes using nonviral approaches can broaden the application of gene delivery in medical applications without the safety concerns associated with viral vectors. Here, we designed a clustered integrin-binding platform to enhance the efficiency and targetability of nonviral gene transfer to HeLa cells with low and high densities of alpha(v)beta(3) integrin receptors. Arg-Gly-Asp (RGD) nanoclusters were formed using gold nanoparticles functionalized with RGD peptides and used to modify the surface of DNA/poly(ethylene imine) (PEI) polyplexes. DNA/PEI polyplexes with attached RGD nanoclusters resulted in either 5.4- or 35-fold increase in gene transfer efficiency over unmodified polyplexes for HeLa cells with low- or high-integrin surface density, respectively. The transfection efficiency obtained with the commercially available vector jetPEI-RGD was used for comparison as a vector without clustered binding. JetPEI-RGD exhibited a 1.2-fold enhancement compared to unmodified jetPEI in cells with high densities of alpha(v)beta(3) integrin receptors. The data presented here emphasize the importance of the RGD conformational arrangement on the surface of the polyplex to achieve efficient targeting and gene transfer, and provide an approach to introduce clustering to a wide variety of nanoparticles for gene delivery.

Técnicas de Transferência de Genes , Vetores Genéticos/química , Integrina alfaVbeta3/química , Integrina alfaVbeta3/metabolismo , Citometria de Fluxo , Células HeLa , Humanos , Iminas/química , Modelos Biológicos , Nanopartículas/química , Oligopeptídeos/química , Polietilenos/química