4 resultados para Division of cells

em Repositório Científico do Instituto Politécnico de Lisboa - Portugal


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Seismic recordings of IRIS/IDA/GSN station CMLA and of several temporary stations in the Azores archipelago are processed with P and S receiver function (PRF and SRF) techniques. Contrary to regional seismic tomography these methods provide estimates of the absolute velocities and of the Vp/Vs ratio up to a depth of similar to 300 km. Joint inversion of PRFs and SRFs for a few data sets consistently reveals a division of the subsurface medium into four zones with a distinctly different Vp/Vs ratio: the crust similar to 20 km thick with a ratio of similar to 1.9 in the lower crust, the high-Vs mantle lid with a strongly reduced VpNs velocity ratio relative to the standard 1.8, the low-velocity zone (LVZ) with a velocity ratio of similar to 2.0, and the underlying upper-mantle layer with a standard velocity ratio. Our estimates of crustal thickness greatly exceed previous estimates (similar to 10 km). The base of the high-Vs lid (the Gutenberg discontinuity) is at a depth of-SO km. The LVZ with a reduction of S velocity of similar to 15% relative to the standard (IASP91) model is terminated at a depth of similar to 200 km. The average thickness of the mantle transition zone (TZ) is evaluated from the time difference between the S410p and SKS660p, seismic phases that are robustly detected in the S and SKS receiver functions. This thickness is practically similar to the standard IASP91 value of 250 km. and is characteristic of a large region of the North Atlantic outside the Azores plateau. Our data are indicative of a reduction of the S-wave velocity of several percent relative to the standard velocity in a depth interval from 460 to 500 km. This reduction is found in the nearest vicinities of the Azores, in the region sampled by the PRFs, but, as evidenced by SRFs, it is missing at a distance of a few hundred kilometers from the islands. We speculate that this anomaly may correspond to the source of a plume which generated the Azores hotspot. Previously, a low S velocity in this depth range was found with SRF techniques beneath a few other hotspots.

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The conventional methods used to evaluate chitin content in fungi, such as biochemical assessment of glucosamine release after acid hydrolysis or epifluorescence microscopy, are low throughput, laborious, time-consuming, and cannot evaluate a large number of cells. We developed a flow cytometric assay, efficient, and fast, based on Calcofluor White staining to measure chitin content in yeast cells. A staining index was defined, its value was directly related to chitin amount and taking into consideration the different levels of autofluorecence. Twenty-two Candida spp. and four Cryptococcus neoformans clinical isolates with distinct susceptibility profiles to caspofungin were evaluated. Candida albicans clinical isolate SC5314, and isogenic strains with deletions in chitin synthase 3 (chs3Δ/chs3Δ) and genes encoding predicted Glycosyl Phosphatidyl Inositol (GPI)-anchored proteins (pga31Δ/Δ and pga62Δ/Δ), were used as controls. As expected, the wild-type strain displayed a significant higher chitin content (P < 0.001) than chs3Δ/chs3Δ and pga31Δ/Δ especially in the presence of caspofungin. Ca. parapsilosis, Ca. tropicalis, and Ca. albicans showed higher cell wall chitin content. Although no relationship between chitin content and antifungal drug susceptibility phenotype was found, an association was established between the paradoxical growth effect in the presence of high caspofungin concentrations and the chitin content. This novel flow cytometry protocol revealed to be a simple and reliable assay to estimate cell wall chitin content of fungi.

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Microbial adhesion is a field of recognized relevance and, as such, an impressive array of tools has been developed to understand its molecular mechanisms and ultimately for its quantification. Some of the major limitations found within these methodologies concern the incubation time, the small number of cells analyzed, and the operator's subjectivity. To overcome these aspects, we have developed a quantitative method to measure yeast cells' adhesion through flow cytometry. In this methodology, a suspension of yeast cells is mixed with green fluorescent polystyrene microspheres (uncoated or coated with host proteins). Within 2 h, an adhesion profile is obtained based on two parameters: percentage and cells-microsphere population's distribution pattern. This flow cytometry protocol represents a useful tool to quantify yeast adhesion to different substrata in a large scale, providing manifold data in a speedy and informative manner.

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Introduction: The samples obtained from fine needle aspiration in liquid base cytology (FNAC) are often limited by scarce cellularity compared to the amount of colloid and presence of blood. Accordingly, it was important to test alternative technical procedures so as to maximize the cellularity of each sample. Objective: To compare the morphological features and cellularity of the three procedures in the FNAC cytodiagnosis of the thyroid. Methods: A total of 31 cases were each subjected to a cell block and ThinPrep preparation as well as a routine smear. The observation and analysis was performed using an optical microscope. Cytological diagnosis of each cell block case was objectively analysed for cellularity, presence of background and cellular preservation. Each smear and ThinPrep case was analysed for the presence or absence of cells. The data was analysed with Microsoft Excel (Office 2010) and SPSS (Statistical Package of Social Science) version 15.0 for Windows. Results: Of 31 cases, only 20 had thyroid cells in the cell block and ThinPrep preparations, however, all smear cases contained thyroid cells. Some background was found in 30 Cell block cases with only 5 of these containing well preserved cells for cytodiagnosis. Conclusions: As indicated by the results, smear is the most appropriate procedure for FNAC of the thyroid.