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排序方式: 共有454条查询结果,搜索用时 15 毫秒
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Efficient flow cytometric assay for platelet-leukocyte aggregates in whole blood using fluorescence signal triggering. 总被引:10,自引:0,他引:10
BACKGROUND: Platelet-leukocyte aggregates (PLAs) may be important in thrombotic and inflammatory disease states, but accurate assessment of PLA formation in vivo is hampered by the propensity for in vitro artefacts caused by sample manipulation. A whole blood flow cytometric assay for circulating PLAs, based on minimal sample manipulation, was thus developed. METHODS: Citrated whole blood was labeled with a RPE-CD45 MAb (leukocyte marker) and an FITC-CD42a (GPIX) MAb (platelet marker). The latter was used to avoid possible influences of platelet glycoprotein proteolysis by neutrophil-derived proteases. The samples were mildly fixed with 0.5% formaldehyde saline. The cytometer was triggered by RPE-CD45 fluorescence. Leukocyte subpopulations were separated according to their typical light scattering and CD45 expression. RESULTS: Minimal sample manipulation and mild sample fixation resulted in minor in vitro artefacts and good sample stability. Fluorescence triggering increased the efficiency of the flow cytometric analysis approximately 5-fold compared with triggering with light scatter, and allowed discrimination of leukocyte subpopulations. The majority of PLAs involved monocytes and neutrophils, rather than lymphocytes, both without and with in vitro stimulation by ADP or thrombin. A cocktail of blocking MAbs to CD62P, CD15, GPIIb/IIIa and the CD11b/CD18 complex had no effect on unstimulated samples, whilst totally inhibiting aggregation induced by 10(-5) M ADP, suggesting that the PLAs in unstimulated blood were preformed in vivo. CONCLUSIONS: This whole blood flow cytometric assay for PLAs is simple and efficient, and appears to reflect closely platelet-leukocyte aggregates in circulating blood in vivo. 相似文献
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The histology of the gonads of free and captive summer whiting, Sillago ciliata , were compared to assess the potential of this fish for aquaculture. There were no qualitative differences between the free and captive males: both completed spermatogenesis through to spermiation. The captive females were non-reproductive, reaching only the vitellogenic stage before the eggs were resorbed by atresia.
A reproductive score was devised to assess reproductive potential in the comparison of the two populations. The females in the captive population had lower reproductive potentials, as shown by the score, and started development a month later than the normal free population. The captive males had higher reproductive scores than the free fish but they were also not spawning and, therefore, the higher score indicated that this potential was not realized. The score provides a standard against which experimental results can be compared and subsequently assessed. 相似文献
A reproductive score was devised to assess reproductive potential in the comparison of the two populations. The females in the captive population had lower reproductive potentials, as shown by the score, and started development a month later than the normal free population. The captive males had higher reproductive scores than the free fish but they were also not spawning and, therefore, the higher score indicated that this potential was not realized. The score provides a standard against which experimental results can be compared and subsequently assessed. 相似文献
5.
Thomas Bell Michael B. Bonsall Angus Buckling Andrew S. Whiteley Timothy Goodall Robert I. Griffiths 《Biology letters》2010,6(5):639-642
Productivity and predation are thought to be crucial drivers of bacterial diversity. We tested how the productivity–diversity of a natural bacterial community is modified by the presence of protist predators with different feeding preferences. In the absence of predators, there was a unimodal relationship between bacterial diversity and productivity. We found that three protist species (Bodo, Spumella and Cyclidium) had widely divergent effects on bacterial diversity across the productivity gradient. Bodo and Cyclidium had little effect on the shape of the productivity–diversity gradient, while Spumella flattened the relationship. We explain these results in terms of the feeding preferences of these predators. 相似文献
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Kari O. Raivio Cheri S. Lazar Michael A. Becker 《Biochimica et Biophysica Acta (BBA)/General Subjects》1981,678(1):58-64
During the growth cycle of normal fibroblasts and of fibroblasts deficient in glucose-6-phosphate dehydrogenase activity, the concentration of 5-phosphoribosyl-1-pyrophosphate and of Pi, as well as the activity of 5-phosphoribosyl-1-pyrophosphate synthetase, decreased to stable values in confluent cultures. A high degree of correlation (0.89 and 0.91 for two normal and 0.69 for one glucose-6-phosphate dehydrogenase-deficient cell strain, respectively) was shown between intracellular Pi and 5-phosphoribosyl-1-pyrophosphate concentrations under varying culture and incubation conditions. 5-phosphoribosyl-1-pyrophosphate concentrations were elevated in normal fibroblasts incubated with methylene blue only if intracellular Pi levels were high. Neither methylene blue nor 6-aminonicotinamide, singly, affected intracellular Pi concentrations. However, when normal cells were pretreated with 6-aminonicotinamide and then with methylene blue, intracellular Pi decreased, 5-phosphoribosyl-1-pyrophosphate was depleted, and its rate of generation decreased. Under similar conditions, glucose-6-phosphate dehydrogenase-deficient fibroblasts maintained unaltered Pi levels, and 5-phosphoribosyl-1-pyrophosphate concentration and generation were slightly increased. The decrease in intracellular Pi in normal cells after the combined treatment was commensurate with an accumulation of 6-phosphogluconate, which did not take place in mutant cells. The changes in 5-phosphoribosyl-1-pyrophosphate synthesis, whether due to the stage of growth or various experimental manipulations, were always concordant with changes in intracellular Pi level. The regulatory role of Pi is consistent with the known enzymic properties of 5-phosphoribosyl-1-pyrophosphate synthetase. 相似文献
9.
I T Norton D M Goodall S A Frangou E R Morris D A Rees 《Journal of molecular biology》1984,175(3):371-394
The thermally induced order-disorder transition of xanthan (extracellular bacterial polysaccharide from Xanthomonas campestris) has been investigated by optical rotation, differential scanning calorimetry, stopped-flow reaction kinetics and low-angle laser light scattering, and the results have been analysed in terms of Zimm -Bragg helix-coil transition theory. The reciprocal of the transition midpoint temperature (Tm) varies linearly with the logarithm of cation (K+) the salt dependence of Tm, is in agreement with Manning polyelectrolyte theory the ordered structure. The associated increase in cation binding, calculated from the salt dependence of tm, is in agreement with the Manning polyelectrolyte theory for one of the candidate structures from X-ray diffraction, a 5(1) single helix stabilized by packing of side-chains along the polymer backbone, but not for the alternative double-helix structure that has also been proposed. At each salt concentration, the two fundamental parameters of the Zimm -Bragg theory, s and sigma, were calculated. The equilibrium constant for growth of the ordered structure (s) is derived directly from calorimetric measurement of transition enthalpy (delta Hcal ), and sigma, which quantifies the relative instability of the helix nucleus, is derived from the ratio of delta Hcal to the apparent transition enthalpy (delta Happ ) obtained by van't Hoff analysis of the optical rotation data. The temperature course of conformational ordering calculated theoretically is in good quantitative agreement with experimental results from both optical rotation and scanning calorimetry. The calculated average length of stable, ordered chain-sequences increases with decreasing temperature, but equals or exceeds the total chain length from light scattering only at temperatures more than approximately equal to 70 K below Tm, suggesting that ordered and disordered regions may co-exist within the same xanthan molecule. Consistent with this interpretation, the observed rate of conformational ordering increases sharply under conditions where the starting solution for dynamic measurements is partially ordered, suggesting that ordered sequences within each chain may act as helix nuclei for adjacent disordered regions, so that helix growth, rather than the slower nucleation process, becomes rate limiting. 相似文献
10.
Archd. L. Goodall 《BMJ (Clinical research ed.)》1941,2(4225):926-927