Constitutive plasma membrane targeting and microdomain localization of Dok5 studied by single-molecule microscopy

In the present study, single-molecule fluorescence microscopy was used to examine the characteristics of plasma membrane targeting and microdomain localization of enhanced yellow fluorescent protein (eYFP)-tagged wild-type Dok5 and its variants in living Chinese hamster ovary (CHO) cells. We found that Dok5 can target constitutively to the plasma membrane, and the PH domain is essential for this process. Furthermore, single-molecule trajectories analysis revealed that Dok5 can constitutively partition into microdomain on the plasma membrane. Finally, the potential mechanism of microdomain localization of Dok5 was discussed. This study provided insights into the characteristics of plasma membrane targeting and microdomain localization of Dok5 in living CHO cells. (C) 2008 Elsevier B.V. All rights reserved.

Heterodimerization of integrin Mac-1 subunits studied by single-molecule imaging

Heterodimerization of integrin Mac-1 (alpha(M) beta(2)) Subunits plays important role on regulating leukocytes adhesion to extracellular matrix or endothelial cells. Here, using total internal reflection microscopy, we investigated the heterodimerization of integrin Mac-1 subunits at the single-molecule level in live cells. Individual alpha(M) subunit fused to the enhanced yellow fluorescent protein (eYFP) was imaged at the basal plasma membrane of live Chinese hamster ovary (CHO) cells. Through analysis of mean square displacement (MSD), diffusion coefficient, the size of restricted domain and fraction of molecules undergoing restricted diffusion, we found that as compared with the diffusion in the absence of beta(2) subunit, the diffusion of single-molecule of alpha(M)-YFP was suppressed significantly in the presence of beta(2) subunit. Thus, based on the oligomerization-induced trapping model, we suggested that in the presence of beta(2) subunit, the am subunit may form heterodimer with it. (C) 2008 Elsevier Inc. All rights reserved.

Detection of constitutive heterodimerization of the integrin Mac-1 subunits by fluorescence resonance energy transfer in living cells

Macrophage differentiation antigen associated with complement three receptor function (Mac-1) belongs to beta(2) subfamily of integrins that mediate important cell-cell and cell-extracellular matrix interactions. Biochemical studies have indicated that Mac-1 is a constitutive heterodimer in vitro. Here, we detected the heterodimerization of Mac-1 subunits in living cells by means of two fluorescence resonance energy transfer (FRET) techniques (fluorescence microscopy and fluorescence spectroscopy) and our results demonstrated that there is constitutive heterodimerization of the Mac-1 subunits and this constitutive heterodimerization of the Mac-1 subunits is cell-type independent. Through FRET imaging, we found that heterodimers of Mac-1 mainly localized in plasma membrane, perinuclear, and Golgi area in living cells. Furthermore, through analysis of the estimated physical distances between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) fused to Mac-1 subunits, we suggested that the conformation of Mac-1 subunits is not affected by the fusion of CFP or YFP and inferred that Mac-1 subunits take different conformation when expressed in Chinese hamster ovary (CHO) and human embryonic kidney (HEK) 293T cells, respectively. (c) 2006 Elsevier Inc. All rights reserved.

Detection of constitutive homomeric associations of the integrins Mac-1 subunits by fluorescence resonance energy transfer in living cells

Integrins alpha(M)beta(2) plays important role on leukocytes, such as adhesion, migration, phagocytosis, and apoptosis. It was hypothesized that homomeric associations of integrin subunits provide a driving force for integrins activation, and simultaneously inducing the formation of integrins clusters. However, experimental reports on homomeric associations between integrin subunits are still controversial. Here, we proved the homomeric associations of the isolated Mac-1 subunits in living cells using three-channel fluorescence resonance energy transfer (FRET) microscopy and FRET spectra methods. We found that the extent of homomeric associations between beta(2) subunits is higher than alpha(M) subunits. Furthermore, FRET imaging indicated that the extent of homomeric associations of the Mac-1 subunits is higher along the plasma membrane than in the cytoplasm. Finally, we suggested that homomeric associations of the transmernbrane domains or/and cytoplasmic domains may provide the driving force for the formation of constitutive homomeric associations between alpha(M) or beta(2) subunits. (c) 2006 Elsevier Inc. All rights reserved.

The autocorrelation of speckles in deep Fresnel diffraction region and characterizations of random self-affine fractal surfaces

This paper studies the correlation properties of the speckles in the deep Fresnel diffraction region produced by the scattering of rough self-affine fractal surfaces. The autocorrelation function of the speckle intensities is formulated by the combination of the light scattering theory of Kirchhoff approximation and the principles of speckle statistics. We propose a method for extracting the three surface parameters, i.e. the roughness w, the lateral correlation length xi and the roughness exponent alpha, from the autocorrelation functions of speckles. This method is verified by simulating the speckle intensities and calculating the speckle autocorrelation function. We also find the phenomenon that for rough surfaces with alpha = 1, the structure of the speckles resembles that of the surface heights, which results from the effect of the peak and the valley parts of the surface, acting as micro-lenses converging and diverging the light waves.

R Gizarte-zientzietarako. Datuen eta eskalen analisia Rcommander-ekin

Datuen analisi kuantitatiboa egin behar duen edonork, gaur egun, aukera zabala du. Programa ugari daude, komertzialak zein doakoak, edozein analisi mota, sinpleena zein konplexuena, egiteko. Programa komertzialen artean, SPSS, SAS, STATISTICA, Systat, Stata edo GenStat aipa daitezke. Programa horiek merkatu osoaz jabetu dira, eta Europako eta Amerikako unibertsitateetan analisi-tresna moduan erabiltzen dira. Programa horiek joan dira, lehenengo bertsioetatik gaurkoetaraino, interfaze grafikoak hobetuz (GUI Graphical User Interfaces) eta analisi-eredu gero eta konplexuagoak erantsiz. Leihoen kudeaketan oinarrituriko testuinguru-menuen eta goitik beherako menuen garapenari esker, programak erabilerrazagoak dira, eta analisi zein eredu formal konplexuenekin lan egiteko aukera izan du erabiltzaileen komunitateak.

Broadband infrared luminescence from bismuth-doped GeS2-Ga2S3 chalcogenide glasses

Near-infrared luminescence is observed from bismuth-doped GeS2-Ga2S3 chalcogenide glasses excited by an 808 nm laser diode. The emission peak with a maximum at about 1260 nm is observed in 80GeS(2)-20Ga(2)S(3):0.5Bi glass and it shifts toward the long wavelength with the addition of Bi gradually. The full width of half maximum (FWHM) is about 200 nm. The broadband infrared luminescence of Bi-doped GeS2-Ga2S3 chalcogenide glasses may be predominantly originated from the low valence state of Bi, such as Bi+. Raman scattering is also conducted to clarify the structure of glasses. These Bi-doped GeS2-Ga2S3 chalcogenide glasses can be applied potentially in novel broadband optical fibre amplifiers and broadly tunable laser in optical communication system.

Effects of process on microstructure and properties of translucent Dy-alpha-SiAlON sintered at lower temperatures

Hot pressing (HP) at higher sintering temperature has been a traditional and prevalent technique for the fabrication of alpha-SiAlON. In order to prepare translucent SiAlON more easily, LiF was used as a non-oxide sintering additive to lower the sintering temperature to <= 1650 degrees C. As a result, all of the samples possessed a good hardness and fracture toughness. At the same time, the lower temperature sintered samples showed a higher optical transmittance in the range of 2.5-5.5 mu m wavelength (0.5 mm in thickness). The maximum infrared transmission reached 68% at a wavelength of 3.3 mu m. The present work shows that the sintering process has a strong effect on microstructure and property of alpha-SiAlON. To be exact, a lower sintering temperature and longer holding time can produce some fully-developed microstrcture, which is beneficial for the optical transmittance. (C) 2008 The Ceramic Society of Japan. All rights reserved.

Hot-pressed translucent aluminum oxynitride (AlON) ceramics

AlON with a composition of Al23O27N5 was prepared by hot pressing at temperatures lower than 1900°C. The microstructures and final properties, including both mechanical properties and optical properties, of the sintered specimens were studied. The results showed that sintering temperature had a great influence on the densification of specimens and could lead to very different properties, especially the optical transmittance and the maximum infrared transmission.

Desenvolvimento de interfaces gráficas para estatística Bayesiana aplicada à comparação mista de tratamentos

A partir dos avanços obtidos pela industria farmacêutica surgiram diversos medicamentos para o combate de enfermidades. Esses medicamentos possuem efeito tópico similar porém com suaves modificações em sua estrutura bioquímica, com isso a concorrência entre as industrias farmacêuticas se torna cada vez mais acirrada. Como forma de comparar a efetividade desses medicamentos, surgem diversas metodologias, com o objetivo de encontrar qual seria o melhor medicamento para uma dada situação. Uma das metodologias estudadas é a comparação mista de tratamentos, cujo objetivo é encontrar a efetividade de determinadas drogas em estudos e/ou ensaios clínicos que abordem, mesmo que de maneira indireta, os medicamentos estudados. A utilização dessa metodologia é demasiadamente complexa pois requer conhecimento de linguagens de programação em ambientes estatísticos além do domínio sobre as metodologias aplicadas a essa técnica. O objetivo principal desse estudo é a criação de uma interface gráfica que facilite a utilização do MTC para usuários que não possuam conhecimento em linguagens de programação, que seja de código aberto e multiplataforma. A expectativa é que, com essa interface, a utilização de técnicas mais abrangentes e avançadas seja facilitada, além disso, venha tornar o ensinamento sobre o tema mais facilitado para pessoas que ainda não conhecem o método