Optimization of a method for simultaneous analysis of cell surface phenotype and cell cycle in human bone marrow and peripheral blood leukocytes by flow cytometry: the value of both internal and external standards

Three different methods for the simultaneous analysis of surface phenotype and DNA quantification were compared. One method, involving the fixation of cells in 70% ethanol, was convincingly superior, both with regard to the CV of the G0G1 peak and the intensity of the DNA labelling. Furthermore, the correlation between the surface antigen densities before and after fixation were high. Experiments evaluating the intraday and the interday variation of the DNA ratio (the mean channel of the G0G1 peak of the sample divided by the mean channel of the G0G1 peak of chicken erythrocytes), documented the former to be small, with S.D. values varying from 0.0 to 0.016, while the latter were considerably higher with S.D. values varying from 0.077 to 0.123. Since the intraday variation of the DNA ratio was consistently low and the interday variation strongly correlated to the position of the red fluorescence test beads, it was possible to minimize the interday variation of the DNA ratio, by calculating the DNA index as the ratio between the DNA ratio of the sample and that of an external control (buffy coat leukocytes). Analyzing normal bone marrow and calculating the DNA index (DI) on the basis of these ratios, the confidence limits of the DI were decreased by more than half the values obtained when DI calculation was based solely on an internal standard, thereby making subsequent ploidy determinations of patient samples more precise. We conclude that this setup of internal and external standards allows accurate determinations of DNA aneuploidy even in an assay where whole cells labelled for surface antigen and DNA content are analyzed.     

Implant exudate leucocyte response to anti-inflammatory drug treatment

Implantation of artificial materials elicits a local inflammatory response. In this study a chamber model technique, allowing sampling of the inflammatory exudate for further analysis in vitro, was used. Male Sprague-Dawley rats were injected daily with two different anti-inflammatory drugs, betamethasone and indomethacin, and the local cellular response was compared with a control group. The retrieved exudate was evaluated with respect to the number of leucocytes, cell viability, differential counts and serum-opsonized zymosan stimulated chemiluminescence (CL). In all groups the majority of cells were polymorphonuclear granulocytes (PMNGs). Betamethasone and high-dose indomethacin (1.92 mg kg^-1 body weight day^-1) treatment caused a marked reduction in the number of accumulated leucocytes 6 days after implantation. A substantial inhibition of the CL response was observed 6 days after treatment with betamethasone (4.23 mg kg^-1 body weight day^-1). An increased CL responsiveness was observed after 24 h with low-dose indomethacin (0.03 mg kg^-1 body weight day^-1) and after 6 days with high-dose indomethacin (1.92 mg kg^-1 body weight day^-1) treatment. In summary, depending on the anti-inflammatory drug treatment, dose and time after implant surgery, either an inhibition or stimulation of leucocyte accumulation and activation was observed. This study shows the possibilities of sampling the inflammatory exudate adjacent to a biomaterial implanted in vivo. This chamber model may be useful for the analysis of the inflammatory reaction around an implanted biomaterial during pharmacological treatment.     

Prolonged release Matrix Pellets Prepared by Melt Pelletization I. Process Variables

A melt pelletization process was investigated in an 8 litre laboratory scale high shear mixer using a formulation with paracetamol, glyceryl monostearate 40-50, and microcrystalline wax. The effects of jacket temperature, product temperature during massing, product load, massing time and impeller speed were investigated by means of factorially designed experiments. The maximum yield of pellets in the range of 500-1400μm was found to approx. 90%. For process conditions preventing deposition of moist mass, the process was found to be reproducible. Impeller speed and massing time were found to be important process variables. Remarkably low in vitro drug release rates were observed in USP-dissolution tests.     

Absence of tolerance to the aversive stimulus properties of ethanol following oral ethanol self-administration

Aluminum has been reported to inhibit long-term potentiation (LTP) following in vivo administration and decrease glutamate release following in vitro exposure. Because glutamate release is critical for synaptic transmission and the development and maintenance of LTP in the hippocampus, we examined the effects of aluminum chloride (AlCl3) on depolarization-induced glutamate release and LTP in rat hippocampal slices. The effects of AlCl3 on [14C]glutamate release were examined by incubation of slices in depolarizing (56 mM)K+ buffer solution in the absence or presence of 2 mM CaCl2. After 15 min depolarization, AlCl3 (100-1000 microM) did not significantly affect Ca(2+)-dependent [14C]glutamate release from slices, whereas a known Ca2+ channel blocker (100 microM CdCl2) decreased Ca(2+)-dependent [14C]glutamate release by approximately 50%. In contrast to a previous report, acute exposure to AlCl3 was without effect on depolarization-dependent glutamate release. LTP of the population spike (PS) in CA1 of hippocampus was induced by the delivery of stimulus trains to the stratum radiatum. LTP of the PS was observed in both control slices and slices bathed in solution containing 100 microM AlCl3. Neither the magnitude nor longevity (measured up to 1 h posttrain) of LTP distinguished control from aluminum-exposed slices. The lack of sensitivity in rat to the encephalopathic changes induced by aluminum, or methodological differences in exposure conditions may account for the lack of effect of aluminum on in vitro LTP in rat hippocampus.     

方兴未艾的智能网技术

智能网(IN)的概念于1984年一被提出,便在世界范围内迅速掀起了IN研究的热潮,虽然至今已有十个年头,但仍可以说,IN技术的发展才刚刚起步。本文就是基于当前国际上对IN的研究,来介绍智能网的基本概念和结构,进而阐述其设计思想及主要特征,并对IN的未来发展趋势进行了探讨,最后为我国发展IN技术提出了可行建议。     

Characterization of 1.55-μm pulses from a self-seededgain-switched Fabry-Perot laser diode using frequency-resolved opticalgating

The intensity and frequency chirp of picosecond pulses from a self-seeded gain-switched Fabry-Perot laser diode have been directly measured using the technique of frequency-resolved optical gating. Measurements over an output sidemode suppression ratio (SMSR) range of 15-35 dB show that higher SMSR's are associated with an increasingly linear frequency chirp across the output pulses. This complete pulse characterization allows the conditions for optimum pulse compression to be determined accurately, and indicates that transform-limited, pedestal free pulses can be obtained at an SMSR of 35 dB     

低荧光阳离子防塌剂WFT─666的合成与性能

ＷＦＴ－６６６是由磺甲基腐殖酸钾、有机硅聚合物、低分子阳离子有机化合物、苯酚、甲醛缩聚而成的一种新型阳离子低荧光防塌剂。本文介绍了ＷＦＴ－６６６的合成及室内性能评定结果。ＷＦＴ－６６６可抗盐至饱和盐水，抗钙和镁，耐２００℃高温，荧光显示低。用该剂处理的钻井液性能稳定，流变参数合理，页岩回收率高，页岩相对膨胀率低，页岩稳定指数高，与其它处理剂的配伍性好。