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2.
Softening of the flesh and the rise in ethylene evolution and respiration associated with ripening in pear (Pyrus communis L.) fruit was delayed when mannose was vacuum infiltrated into intact fruit. The extent of delay could be modified by altering the concentration or the volume of mannose applied to the fruit. Inhibition of ripening was associated with phosphorylation of mannose to mannose 6-phosphate (M6P), and accumulation of M6P was associated with lowered levels of inorganic phosphate (Pi), glucose 6-phosphate (G6P), and ATP in the fruit tissue. Subsequently, however, as the M6P was metabolized, the levels of Pi, G6P, and ATP increased and ripening processes were concomitantly released from inhibition. Hence, the degree of inhibition by mannose or the release from inhibition was related to the level of M6P in the fruit and its rate of metabolism. The data provide correlative evidence to support a view that one inhibitory effect of mannose is depletion of Pi in the cell as a result of phosphorylation of mannose to M6P. Inhibition of ripening by mannose was not alleviated by co-application of glucose as a competitive substrate for the hexokinase(s), or by Pi, presumably the depleted metabolite. Also, incubation of tissue disks with M6P resulted in inhibition of ethylene production and respiration. The structural analogs of mannose, glucosamine, and 2-deoxyglucose, which have been shown to mimic mannose action in several plant tissues, did not cause inhibition of ripening of pear fruit comparable with that associated with mannose. Both analogs stimulated respiration, and glucosamine caused only a small inhibition of softening and ethylene evolution. Another mannose analog, α-methylmannoside, did inhibit fruit ripening though to a lesser extent than mannose. Its influence was also associated with accumulation of M6P and a decrease of Pi levels. We conclude that the mannose effect may, in part, be due to M6P toxicity, as well as by depletion of Pi. 相似文献
3.
A procedure for large-scale isolation of RNA-free plasmid and phage DNA without the use of RNase 总被引:1,自引:0,他引:1
Zeev Lev 《Analytical biochemistry》1987,160(2):332-336
A preparative procedure for the large-scale isolation of plasmid DNA without the use of RNAse is described. Crude plasmid DNA is prepared using a standard boiling method. High-molecular-weight RNA is removed by precipitation with LiCl, and low-molecular-weight RNA is removed by sedimentation through high-salt solution. The procedure is inexpensive, rapid, simple, and particularly suitable for processing several large-scale preparations simultaneously. A similar procedure has been developed for preparation of lambda-phage DNA. 相似文献
4.
Effect of membrane cholesterol on dimyristoylphosphatidylcholine-induced vesiculation of human red blood cells 总被引:1,自引:0,他引:1
E J Frenkel F A Kuypers J A Op den Kamp B Roelofsen P Ott 《Biochimica et biophysica acta》1986,855(2):293-301
During incubation of intact human erythrocytes with sonicated dimyristoylphosphatidylcholine (DMPC) vesicles, the cells change their discoid morphology to form echinocytes and finally give rise to the release of membrane vesicles. In this process, the red cell membrane accumulates DMPC and loses up to 15% of its cholesterol. On the other hand, replacement of 25% of the endogenous phosphatidylcholine species by DMPC without affecting the cholesterol level of the erythrocytes can be achieved by incubation with DMPC/cholesterol (1:1, mol/mol) sonicated vesicles in the presence of the phosphatidylcholine-specific phospholipid-transfer protein from bovine liver. This replacement also gives rise to an echinocytic cell morphology, but no membrane vesiculation can be observed. However, the vesiculation process can as yet be initiated upon a subsequent decrease of the cholesterol level, by incubation of those modified cells in the presence of sonicated vesicles of pure egg phosphatidylcholine. Incubation of native erythrocytes with pure egg phosphatidylcholine vesicles, on the other hand, results in cholesterol depletion, but does neither induce the formation of echinocytes nor the release of membrane vesicles. Cellular ATP levels are not affected during these incubations. From these results, it can be concluded that a decrease in cholesterol content of the erythrocyte membrane is essential for the DMPC-induced vesiculation of those cells. 相似文献
5.
A previous publication [Frenkel, Cain, and Chao, Biochem. Biophys. Res. Commun. 127, 849-856 (1985)] described the observation that double-stranded DNA which was briefly exposed to methyl mercury (MeHg) and purified to remove free methyl mercury was transcribed at a higher rate by RNA polymerase II from wheat germ. The specificity of this phenomenon has now been investigated by examining the activity of this MeHg-exposed DNA as a template-primer for DNA polymerases. DNA synthesis by the bacteriophage T4-induced DNA polymerase was higher with the MeHg-exposed DNA as a template-primer than with control DNA. In contrast, the rate of DNA synthesis by E. coli DNA polymerase I was lower with the MeHg-exposed DNA as template-primer. With both enzymes (as well as with RNA polymerase II), after denaturation of the MeHg-exposed and control DNAs the differences in template activity were either eliminated or markedly reduced. The enzymes are thus able to detect a MeHg-induced alteration in DNA. In contrast, circular dichroism, a physical method that is sensitive to conformational changes in DNA, did not detect any difference between the MeHg-exposed and control DNAs. 相似文献
6.
Zeev Ronai Judith Shaham Joseph Stein Dov Sulitzeanu 《Cancer immunology, immunotherapy : CII》1986,22(1):72-75
Summary A polyethylene glycol precipitation technique was used to determine the levels of circulating immune complexes (CIC) in breast cancer and melanoma patients. All patients in the study had undergone surgery and were free of distant metastatic disease. CIC were measured at two to four time intervals, of 3 to 6 months each, over an average follow-up period of 13.5 months (range 7–20 months). In both groups of patients, metastatic disease developed with a higher frequency in patients who had undetectable CIC levels throughout the follow-up period or had become negative at the time metastases were discovered. 相似文献
7.
Eviatar Nevo Zeev Gerechter-Amitai Avigdor Beiles Edward M. Golenberg 《Plant Systematics and Evolution》1986,153(1-2):13-30
From 114 accessions of wild emmer wheat from 11 sites in Israel, known for their allozymic variation (Nevo & al. 1982), individual genotypes were tested for resistance to one isolate of stripe rust both in the seedling stage in a growth chamber and in the adult plant stage in the field. The results indicate that resistance to stripe rust in seedlings and adults are significantly correlated (rs = 0.40, p < 0.001). Genetic polymorphisms of resistance to stripe rust vary geographically and are predictable by climatic, as well as allozymic markers. Three variable combinations of rainfall, evaporation, and temperature explain significantly 0.40–0.53 of the spatial variance in disease resistance to stripe rust, suggesting the operation of natural selection. Several allozyme genotypes are significantly associated with disease resistance. We conclude that natural populations of wild emmer wheat in Israel contain large amounts of disease resistance genes. These populations could be effectively screened and then utilized by the phytopathologist for identifying resistant genotypes and producing new resistant cultivars.Patterns of Resistance of Wild Wheat to Pathogens in Israel II. 相似文献
8.
Sea urchins have elaborated multiple defenses to assure monospermic fertilization. In this work, we have concentrated on a study of the mechanism(s) by which hydrogen peroxide (H2O2) prevents polyspermy in Arbacia punctulata. We found that it is not H2O2 but probably hypochlorous acid/hypochlorite (HOCl/OCl?) derived from H2O2 that is toxic to the supernumerary sperm. The spermicidal activity of H2O2 is potentiated by at least one order of magnitude by cupric ions (Cu2+). This increased toxicity is not due to the formation of hydroxyl radicals (·OH) because ·OH scavengers did not counteract the activity of Cu2+. More-over, substitution of Cu2+ by ferrous ions (Fe2+), which are known to cause formation of ·OH from H2O2, had no effect on fertilization even at 102?103 times higher concentrations. In contrast, 3-amino-1,2,4-triazole (AT), an HOCl/OCl? scavenger, totally reversed the toxic effects of Cu2+. Furthermore, we found that HOCl/OCl? is generated in solutions of H2O2 and Cu2+ in the presence of 0.5 M NaCl and that its accumulation is abolished by AT. Thus it is possible that the antifertility properties of copper are due to its ability to mediate formation of HOCl/OCl?. HOCl/OCl? generated by Cu2+ from H2O2 and Cl?, a low concentration of exogenously added HOCl/OCl?, or increased concentrations of H2O2 has similar inhibitory effects on the fertilization process in sea urchins. Therefore, we suggest that polyspermy is prevented by the action of a myeloperoxidase that affects the formation of HOCl/OCl? from the Cl? present in sea water through reaction with H2O2 generated by the newly fertilized egg. 相似文献
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