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Preparation of pancreatic mRNA: cell-free translation of an insulin-immunoreactive polypeptide. 总被引:23,自引:0,他引:23 下载免费PDF全文
Total nucleic acid extraction and selection of poly A-containing molecules yield preparative quantities of undegraded mRNA from adult and fetal pancreas. Using a stringent immunoassay, this mRNA is found to direct the synthesis of an immunoreactive insulin polypeptide in the wheat germ translation system. On sodium dodecyl sulfate polyacrylamide gels, this polypeptide (12,000-13,000 daltons) is larger than proinsulin (9,000 daltons). 相似文献
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Plasma albumin levels were measured in partially hepatectomized, sham operated and control rats. The levels fell in both the partially hepatectomized and sham operated groups; while the latter group returned to normal within a few days, the low plasma albumin in the partially hepatectomized animals was sustained. Albumin synthesis rates in the isolated perfused rat liver were measured in the three groups of animals at varying intervals after partial hepatectomy. There was a significant depression of albumin synthesis rate in terms of both liver and whole animal weights when compared to the sham operated and control animals. This depression was almost completely reversed by the addition of arginine, asparagine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, threonine, tryptophan and valine added together to 10 times their normal plasma concentrations. The addition of hydrocortisone had no effect on the albumin synthesis rate after partial hepatectomy. Studies in vivo in the three groups of animals (partially hepatectomized, sham operated and control animals) revealed a fall in the albumin catabolic rate after partial hepatectomy coinciding with the fall in the albumin synthesis rate. An hypothesis whereby the amino acids may have their stimulatory effect is proposed. 相似文献
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Summary The rapid increase in acidic gibberellin-like (GA-like) activity occurring in etiolated wheat (Triticum aestivum L.); leaf tissue in response to red light is phytochrome controlled. The production of some of the GA-like activity appears to possess a similar escapetime from far-red reversibility to leaf unrolling and is sensitive to Amo 1618. Application of these GA-like substances to leaf sections maintained in the dark causes unrolling. Increased GA-like activity can also be detected in non-irradiated portions of partly irradiated leaves.Abbreviations GA
gibberellin
- Amo 1618
2-isopropyl-4-(trimethylammonium chloride)-5-methylphenyl piperidine carboxylate
- CCC
-chloroethyltrimethyl ammonium chloride 相似文献
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Selecting the appropriate rodent diet for endocrine disruptor research and testing studies 总被引:6,自引:0,他引:6
Thigpen JE Setchell KD Saunders HE Haseman JK Grant MG Forsythe DB 《ILAR journal / National Research Council, Institute of Laboratory Animal Resources》2004,45(4):401-416
Selecting the optimum diet for endocrine disruptor (ED) research and testing studies in rodents is critical because the diet may determine the sensitivity to detect or properly evaluate an ED compound. Dietary estrogens can profoundly influence many molecular and cellular event actions on estrogen receptors and estrogen-sensitive genes. The source, concentration, relative potency, and significance of dietary estrogens in rodent diets are reviewed, including dietary factors that focus specifically on total metabolizable energy and phytoestrogen content, which potentially affect ED studies in rodents. Research efforts to determine dietary factors in commercially available rodent diets that affect uterotrophic assays and the time of vaginal opening in immature CD-1 mice are summarized. A checklist is provided of important factors to consider when selecting diets for ED research and testing studies in rodents. Specific metabolizable energy levels are recommended for particular bioassays. Discussions include the between-batch variation in content of the phytoestrogens daidzein and genistein, the effects of total metabolizable energy and phytoestrogens on the timing (i.e., acceleration) of vaginal opening, and increased uterine weight in immature CD-1 mice. It is concluded that rodent diets differ significantly in estrogenic activity primarily due to the large variations in phytoestrogen content; therefore animal diets used in all ED studies should ideally be free of endocrine-modulating compounds. 相似文献
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Summary Two sugarbeet (Beta vulgaris L.) genotypes, REL-1 and REL-2, were used to measure the level of somatic embryo and shoot production from hormone-autonomous
callus plated under varied nutrient medium combinations of abscisic acid with the growth regulators 6-benzyladenine, 1-naphthaleneacetic
acid, or 2,4-dichlorophenoxyacetic acid, with eight sole nitrogen sources, or with different sucrose concentrations. Clone
REL-2 produced embryos up to 35-fold more frequently than clone REL-1. Inclusion of abscisic acid at some concentrations consistently
improved embryo production in all experiments and was observed to stimulate shoot production. At some concentrations, 1-naphthaleneacetic
acid as well as urea and glutamine stimulated greater embryo production over the control, but only for REL-1, for which there
was greater room for improvement. Three and five percent sucrose were superior to 1, 7, and 9%. Higher initial 6-benzyladenine
concentration [in the range 0, 0.1−1.0 mg/L (0.44−4.44 μM)] was associated with lower embryo production but greater shoot regeneration for both clones. REL-2 was significantly better
than REL-1 in shoot regeneration. The range of embryo production was more than 35-fold between genotypes, whereas the range
of physiological effects was no greater than 10-fold. REL-2 has been released to sugarbeet researchers because of its superior
embryogenic and shoot regeneration abilities for application in biotechnology. 相似文献
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Janet E. Saunders Craig C. Beeson Rick G. Schnellmann 《Journal of bioenergetics and biomembranes》2013,45(1-2):87-99
Mitochondrial stress results in changes in mitochondrial function, morphology and homeostasis (biogenesis, fission/fusion, mitophagy) and may lead to changes in mitochondrial subpopulations. While flow cytometric techniques have been developed to quantify features of individual mitochondria related to volume, Ca2+ concentration, mtDNA content, respiratory capacity and oxidative damage, less information is available concerning the identification and characterization of mitochondrial subpopulations, particularly in epithelial cells. Mitochondria from rabbit kidneys were stained with molecular probes for cardiolipin content (nonyl acridine orange, NAO) and membrane potential (tetramethylrhodamine, TMRM) and analyzed using flow cytometry. We validated that side scatter was a better indicator of volume and that as side scatter (SSC) decreased mitochondrial volume increased. Furthermore, those mitochondria with the highest NAO content had greater side scattering and were most highly charged. Mitochondria with average NAO content were of average side scattering and maintained an intermediate charge. Those mitochondria with low NAO content had minimal side scattering and exhibited minimal charge. Upon titration with the uncoupler carbonylcyanide-4-(trifluoromethoxy)-phenylhydrazone (FCCP), it was found that the high NAO content subpopulations were more resistant to uncoupling than lower NAO content populations. Ca2+-induced swelling of mitochondria was evaluated using probability binning (PB) analyses of SSC. Interestingly, only 30 % of the mitochondria showed changes in response to Ca2+, which was blocked by cyclosporine A. In addition, the small, high NAO content mitochondria swelled differentially in response to Ca2+ over time. Our results demonstrate that flow cytometry can be used to identify mitochondrial subpopulations based on high, mid and low NAO content and/or volume/complexity. These subpopulations showed differences in membrane potential, volume, and responses to uncoupling and Ca2+-induced swelling. 相似文献
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