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1.
Cristian A. Acevedo Elizabeth Y. Sanchez Juan G. Reyes Manuel E. Young 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2010,878(3-4):449-455
It is known that skin releases volatile organic compounds to the environment, and also that its emission pattern changes with aging of the skin. It could be considered, that these compounds are intermediaries in cell metabolism, since many intermediaries of metabolic pathways have a volatile potential. In this work, a simple and non-destructive method consisting of SPME sampling and GC/MS analysis was developed to identify volatile organic emanations from cell cultures. This technique, applied to skin cells culture, indicates that the cells or cell metabolism produce several skin emissions. Chemometric analysis was performed in order to explore the relationship between a volatile profile and the senescence of cell cultures. Volatile profiles were different for cell cultures in different degrees of senescence, indicating that volatile compound patterns could be used to provide information about the age of skin cells. 相似文献
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The existence of duodeno-gastric reflux was evaluated in a group of 15 healthy subjects, in fasting and for 24 hours. The assessment of duodeno-gastric reflux was made by quantitation of the bile acids (BA) present in the gastric juice. The individual free and conjugated BA were separated and quantified by means of thin-layer chromatography and in situ spectrofluorometry. In 7 of the subjects studied no BA were detected, and in the other 8 subjects the BA levels were below 6 mumol reflux/hour. There were no free BA detected in any of the subjects. The levels of BA in gastric juice increased progressively with age, but there were no differences between sex. The chromatographic technique used is highly sensitive for the analysis of BA in biological samples. The study of BA in the gastric juice provides a quantitative and reliable assessment of the degree of duodeno-gastric reflux. 相似文献
4.
Tanner Miest Dyana Saenz Anne Meehan Manuel Llano Eric M. Poeschla 《Methods (San Diego, Calif.)》2009,47(4):298-303
RNAi is a powerful technology for analyzing gene function in human cells. However, its utility can be compromised by inadequate knockdown of the target mRNA or by interpretation of effects without rigorous controls. We review lentiviral vector-based methods that enable transient or stable knockdowns to trace mRNA levels in human CD4+ T cell lines and other targets. Critical controls are reviewed, including rescue of the pre-knockdown phenotype by re-expression of the targeted gene. The time from thinking about a potential knockdown target to analysis of phenotypes can be as short as a few weeks. 相似文献
5.
Carmen de Sena-Tomás Mónica Navarro-González Ursula Kües José Pérez-Martín 《Genetics》2013,195(1):47-57
The fungal fruiting body or mushroom is a multicellular structure essential for sexual reproduction. It is composed of dikaryotic cells that contain one haploid nucleus from each mating partner sharing the same cytoplasm without undergoing nuclear fusion. In the mushroom, the pileus bears the hymenium, a layer of cells that includes the specialized basidia in which nuclear fusion, meiosis, and sporulation occur. Coprinopsis cinerea is a well-known model fungus used to study developmental processes associated with the formation of the fruiting body. Here we describe that knocking down the expression of Atr1 and Chk1, two kinases shown to be involved in the response to DNA damage in a number of eukaryotic organisms, dramatically impairs the ability to develop fruiting bodies in C. cinerea, as well as other developmental decisions such as sclerotia formation. These developmental defects correlated with the impairment in silenced strains to sustain an appropriated dikaryotic cell cycle. Dikaryotic cells in which chk1 or atr1 genes were silenced displayed a higher level of asynchronous mitosis and as a consequence aberrant cells carrying an unbalanced dose of nuclei. Since fruiting body initiation is dependent on the balanced mating-type regulator doses present in the dikaryon, we believe that the observed developmental defects were a consequence of the impaired cell cycle in the dikaryon. Our results suggest a connection between the DNA damage response cascade, cell cycle regulation, and developmental processes in this fungus. 相似文献
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W. Kelley Thomas J. T. Vida Linda M. Frisse Manuel Mundo James G. Baldwin 《Journal of nematology》1997,29(3):250-254
To effectively integrate DNA sequence analysis and classical nematode taxonomy, we must be able to obtain DNA sequences from formalin-fixed specimens. Microdissected sections of nematodes were removed from specimens fixed in formalin, using standard protocols and without destroying morphological features. The fixed sections provided sufficient template for multiple polymerase chain reaction-based DNA sequence analyses. 相似文献
8.
Maria João Feio Trefor B. Reynoldson Verónica Ferreira Manuel Augusto S. Graça 《Hydrobiologia》2007,579(1):55-68
We sampled macroinvertebrates at 75 locations in the Mondego river catchment, Central Portugal, and developed a predictive
model for water quality assessment of this basin, based on the Reference Condition Approach. Sampling was done from June to
September 2001. Fifty-five sites were identified as “Reference sites” and 20 sites were used as “Test sites” to test the model.
At each site we also measured 40 habitat variables to characterize water physics and chemistry, habitat type, land use, stream
hydrology and geographic location. Macroinvertebrates were generally identified to species or genus level; a total of 207
taxa were found. By Unweighted Pair Group Method with Arithmetic mean (UPGMA) clustering and analysis of species contribution
to similarities percentage (SIMPER), two groups of reference sites were established. Using Discriminant Analysis (stepwise
forward), four variables correctly predicted 78% of the reference sites to the appropriate group: stream order, pool quality,
substrate quality and current velocity. Test sites’ environmental quality was established from their relative distance to
reference sites, in MDS ordination space, using a series of bands (BEAST methodology). The model performed well at upstream
sites, but at downstream sites it was compromised by the lack of reference sites. As with the English RIVPACS predictive model,
the Mondego model should be continually improved with the addition of new reference sites. The adaptation of the Mondego model
methodology to the Water Framework Directive is possible and would consist mainly of the integration of the WFD typology and
increasing the number of ellipses that define quality bands.
Handling editor: K. Martens 相似文献
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Luís M. S. Loura Fábio Fernandes Manuel Prieto 《European biophysics journal : EBJ》2010,39(4):589-607
Lateral membrane heterogeneity, in the form of lipid rafts and microdomains, is currently implicated in cell processes including signal transduction, endocytosis, and cholesterol trafficking. Various biophysical techniques have been used to detect and characterize lateral membrane domains. Among these, Förster resonance energy transfer (FRET) has the crucial advantage of being sensitive to domain sizes smaller than 50-100 nm, below the resolution of optical microscopy but, apparently, similar to those of rafts in cell membranes. In the last decade, several formalisms for the analysis of FRET in heterogeneous membrane systems have been derived and applied to the study of microdomains. They are critically described and illustrated here. 相似文献