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91.
Inhibition of sickle beta-chain (betaS)-dependent polymerization by nonhuman alpha-chains. A superinhibitory mouse-horse chimeric alpha-chain 总被引:1,自引:0,他引:1
P Nacharaju RP Roy SP White RL Nagel AS Acharya 《Canadian Metallurgical Quarterly》1997,272(44):27869-27876
Horse alpha-chain inhibits sickle beta-chain-dependent polymerization; however, its inhibitory potential is not as high as that of mouse alpha-chain. Horse alpha-(1-30) and alpha-(31-141) segments make, respectively, minor and major contributions to the inhibitory potential of horse alpha-chain. The sum of the inhibitory potential of the two segments does not account for the inhibitory potential of the full-length horse alpha-chain. Although the polymerization inhibitory potential of horse alpha-chain is lower than mouse alpha-chain, the inhibitory potential of horse alpha-(31-141) is comparable to that of mouse alpha-(31-141). When mouse alpha-(1-30) is stitched to horse alpha-(31-141), the product is a chimeric alpha-chain with an inhibitory potential greater than mouse alpha-chain. In contrast, the stitching of horse alpha-(1-30) with mouse alpha-(31-141) had no additional inhibitory potential. Molecular modeling studies of HbS containing the mouse-horse chimeric alpha-chain indicate altered side-chain interactions at the alpha1beta1 interface when compared with HbS. In addition, the AB/GH corner perturbations facilitate a different stereochemistry for the interaction of the epsilon-amino group of Lys-16(alpha) with the beta-carboxyl group of Asp-116(alpha), resulting in a decrease in the accessibility of the side chain of Lys-16(alpha) to the solvent. Based on molecular modeling, we speculate that these perturbations by themselves, or in synergy with the altered conformational aspects of the alpha1beta1 interactions, represent the molecular basis of the superinhibitory potential of the mouse-horse chimeric alpha-chains. 相似文献
92.
CW Taylor LM Wang AF List D Fernandes GD Paine-Murrieta CS Johnson RL Capizzi 《Canadian Metallurgical Quarterly》1997,33(10):1693-1698
The objectives of this study were to evaluate the protective effects of amifostine against paclitaxel-induced toxicity to normal and malignant human tissues. Haematopoietic progenitor colony assays were used to establish the number of CFU-GEMM and BFU-E colonies after incubation with WR-1065 alone, Amifostine alone, paclitaxel (2.5 or 5 microM) +/- WR-1065 or amifostine. MTT and alkaline elution assays evaluated the in vitro growth inhibitory and DNA damaging effects, respectively, of paclitaxel with or without amifostine against normal human fibroblasts and human non-small cell lung cancer (NSCLC) cells. This combination was also evaluated in vivo using severe combined immune deficient (scid) mouse models of early (non-palpable tumours) and advanced (palpable tumours) human ovarian cancer. Human 2780 ovarian cancer cells were inoculated subcutaneously while paclitaxel and amifostine were administered intraperitoneally. A brief exposure (15 min) to amifostine not only protected human haematopoietic progenitor colonies from paclitaxel toxicity, but stimulated the growth of CFU-GEMM and BFU-E beyond control values. Amifostine protected normal human lung fibroblasts from paclitaxel-induced cytotoxicity and DNA single-strand breaks. However, paclitaxel cytotoxicity and DNA single-strand breaks were actually enhanced by pretreatment with amifostine in the NSCLC model. Importantly, amifostine did not interfere with paclitaxel antitumour activity even with prolonged exposure (24.5 h) of the lung cancer cells to high concentrations (1.2 mM) in vitro or following five repetitive high doses (200 mg/kg) given to scid mice with human ovarian cancer xenografts. Indeed, under certain circumstances, amifostine resulted in sensitisation of tumour cells to paclitaxel. Our results confirm previous reports of the ability of amifostine to protect normal tissues from the toxic effects of chemotherapy drugs and now extend these observations to paclitaxel. 相似文献
93.
The expression of the NMDA subtype of glutamate receptors was investigated by Western blot analysis and RT-PCR in cultured chick Bergmann and Müller glial cells. Using subunit-specific antibodies directed to the carboxy terminus of the rat NMDAR2A/B we detected the expression of the NMDAR2 subunit in both kinds of culture. The functional subunit of the NMDA receptor, NMDAR1, was detected by means of RT-PCR. These results, together with our previous functional characterization of NMDA receptors in radial glia, provide conclusive evidence for the expression of functional NMDA receptor/channels in Bergmann and Muller glia cells. Our findings strengthen the notion of a modulatory role of glial cells in synaptic transmission. 相似文献
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RR Bosch AM Patel SE Van Emst-de Vries RL Smeets JJ De Pont PH Willems 《Canadian Metallurgical Quarterly》1998,346(2-3):345-351
We investigated the effects of nitric oxide (NO) donors, S-nitroso-N-acetylpenicillamine and sodium nitroprusside on basal and K+-evoked release of [3H]noradrenaline from superfused synaptosomes from the rat cerebral cortex. Both substances produced concentration-dependent increases in the release of the labeled transmitter under basal and depolarized conditions. The effects of the donors on basal release were Ca2+-independent but were not inhibited by the carrier-uptake blocker, desipramine; the effects were abolished by hemoglobin (an NO scavenger). Thirty-five minutes after stimulation with sodium nitroprusside, the synaptosomes were still responsive to KCl stimulation, indicating that the donor's effects were not caused by damage to the synaptosome membrane. The cGMP analogue, 8-bromo-cGMP, had no effect on basal release, and the enhanced release produced by sodium nitroprusside was not inhibited by the specific inhibitor of soluble guanylate cyclase, 1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one, indicating that NO's effects on basal release of the neurotransmitter are guanylate cyclase-independent. Both of the NO donors had more marked effects on release of [3H]noradrenaline during K+-stimulated depolarization. The NO-mediated increase in this case was partially antagonized by 10 microM LH-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one, and 8-Br-cGMP was also capable of producing concentration-dependent increases in the K+-stimulated release of the transmitter. These findings indicate that the effects of the NO donors on [3H]noradrenaline release during depolarization are partially mediated by the activation of guanylate cyclase. 相似文献
96.
The classical action of the hormone 1,25-dihydroxyvitamin D3 (VD) is the regulation of calcium metabolism. In contrast, the peptide hormone atrial natriuretic factor (ANF) is one of the few known nonclassical VD responding genes. We screened the promoter of the rat ANF gene and identified a typical VD receptor (VDR) binding site formed by a direct repeat of two hexameric core binding motifs spaced by three nucleotides, between positions -907 and -891. Like most of the DR3-type VD response elements this sequence is bound with high affinity (Kd = 0.53 nM) by a heterodimer formed by VDR and retinoid X receptor. In a heterologous promoter context one copy of this sequence mediated an about fourfold gene activation by VD and a half-maximal activation (EC50) value of 0.48 nM VD. This characterizes the identified sequence as one of the most potent VD response elements. 相似文献
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98.
99.
RL Vimal 《Canadian Metallurgical Quarterly》1998,15(7):1756-1766
Threshold-elevation (TE-) versus-mask-spatial-frequency (SF) curves and TE-versus-mask-contrast curves, produced by the oblique-masking technique, were reported for uncrossed stimuli (color-test-on-color-mask and luminance-test-on-luminance-mask) [Invest. Ophthalmol. Visual Sci. Suppl. 34, 751 (1993) and Vision. Res. 23, 873 (1983)]. The technique minimizes the artifacts that are due to spatial phase effects, spatial beats, spatial probability summation, and local cues. My goal was to measure these curves for crossed stimuli (color-test-on-luminance-mask and luminance-test-on-color-mask) by this oblique-masking technique and to compare the curves with those reported in previous studies. For this purpose threshold contrasts were measured by a yes-no procedure with randomized double staircases. Test targets were vertical spatially localized (D6) patterns, and masks were oblique sinusoidal patterns; both the test and the mask were presented simultaneously, for 2 s (Gaussian window), on a color monitor interfaced with an ATVista system and a Powell achromatizing lens. The test SF's were 0.125, 0.5, 2, 4, and 8 cycles per degree (cpd); mask SF's were 0.031-16 cpd; and mask contrasts were 6.25%-50%. Furthermore, the Red-Green channel was defined by the minimum flicker and the hue cancellation techniques. Results show mostly masking effect (TE > 1) at contrasts above threshold; sometimes, separability (TE = 1) and above-threshold facilitation (TE < 1) effects were also observed, depending on the test SF, the mask SF, the mask contrast, and the subject. In general, the magnitudes of TE's are smaller and the TE-versus-mask-SF curves are slightly narrower for the oblique-cross-masking conditions than those for the respective oblique uncross masking. In addition, the TE-versus-mask-contrast curves for the crossed conditions are mostly shallower than those for the respective uncrossed conditions. Furthermore, mostly the color-luminance asymmetry (color masks luminance more than luminance masks color) is found, in mild form, for SF's > or = 0.5 cpd. For the lower SF of 0.125 cpd, there is either a lack of asymmetry or a very mild asymmetry of the opposite kind (luminance masks color slightly more than color masks luminance) seems to prevail. In general, the oblique-masking data shows mild asymmetry and reduced facilitation; both are consistent with reduced local cues, similar to those shown by randomized phase data, thus making the data suitable for SF analysis; moreover, at high contrast, the masking data are consistent with those reported in previous studies. 相似文献
100.