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  • 11
    ISSN: 1573-4994
    Keywords: Fluorogenic probes ; microspectrofluorometry ; HL60 cell line ; oxidative stress ; thiol status
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract Development of microspectrofluorometric methods using specific fluorogenic probes has provided precious help in studying in situ oxidative stress and cellular protective systems. The aim of this study was to determine ROS production concomitantly with a modification of the intracellular thiol pool after applying an oxidative stress to a nonadherent cell model represented by the HL60 cell line. The dichlorodihydrofluorescein diacetate (H2DCFDA) probe assessed the kinetic production of ROS by cells submitted to the chemical oxidant t-butylhydroperoxide with a high signal/noise ratio. The probe sensitivity permitted us to detect endogenous ROS production in HL60 cells and the protective effect of N-acetyl cysteine against ROS. The chloromethylfluorescein diacetate probe (CMFDA) permitted us to evaluate the thiol depleting effect of N-ethyl maleimide. Complete thiol depletion was associated with a moderate increase in ROS production. The cell viability was determined with calcein-AM, which gave results similar to those with the tetrazolium dye. This probe was not affected by intracellular pH and did not required an extraction step, unlike tetrazolium dye. In conclusion, cell-permeant fluorogenic probes are useful and sensitive tools to determine in situ ROS production concomitantly with consecutive change in the thiol system in a living and non-adherent cell model.
    Type of Medium: Electronic Resource
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  • 12
    ISSN: 1572-9788
    Keywords: mapping ; morphology ; QTL ; RFLP ; spike density ; Triticeae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Quantitative trait loci (QTLs) for three traits related to ear morphology (spike length, number of spikelets, and compactness as the ratio between number of spikelets and spike length) in wheat (Triticum aestivum L.) were mapped in a doubled-haploid (DH) population derived from the cross between the cultivars Courtot and Chinese Spring. A molecular marker linkage map of this cross that had previously been constructed based on 187 DH lines and 380 markers was used for QTL mapping. The genome was well covered (85%) except chromosomes 1D and 4D and a set of anchor loci regularly spaced (one marker each 15.5 cM) were chosen for marker regression analysis. The presence of a QTL was declared at a significance threshold α = 0.001. The population was grown in one location under field conditions during three years (1994, 1995 and 1998). For each trait, 4 to 6 QTLs were identified with individual effects ranging between 6.9% and 21.8% of total phenotypic variation. Several QTLs were detected that affected more than one trait. Of the QTLs 50% were detected in more than one year and two of them (number of spikelets on chromosome 2B, and compactness on chromosome 2D) emerged from the data from the three years. Only one QTL co-segregated with the gene Q known to be involved in ear morphology, namely the speltoid phenotype. However, this chromosome region explained only a minor part of the variation (7.5–11%). Other regions had a stronger effect, especially two previously unidentified regions located on chromosomes 1A and 2B. The region on the long arm of chromosome 1A was close to the locus XksuG34-1A and explained 12% of variation in spike length and 10% for compactness. On chromosome 2B, the QTL was detected for the three traits near the locus Xfbb121-2B. This QTL explained 9% to 22% of variation for the traits and was located in the same region as the gene involved in photoperiod response (Ppd2). Other regions were located at homoeologous positions on chromosomes 2A and 2D.
    Type of Medium: Electronic Resource
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