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131.
Summary Chromosomes were isolated in a preparative scale by synchronisation of CHO cells with a double Thymidine block followed by an arrest in the metaphase by addition of Colcemid. Under proper cultivation conditions a mitotic index of 77% total cells could be routinely achieved. Bulk chromosome preparations free of nuclei and other subcellular particles have been obtained by low speed centrifugation followed by a 60 transfer countercurrent distribution using aqueous two phase systems composed of polyethylenglycol and dextran. The partition of CHO chromosomes previously purified in aqueous two phase systems were studied further to develop a protocol for the separation and isolation of individual chromosomes. Partition experiments with chromosomes changing the electrostatic phase potential by addition of charged PEG-derivatives suggest the existence of relatively highly charged chromosome groups. Most promising results with regard to separation were obtained using two PEG-derivatives, which interact specifically with the bases in DNA. For this affinity partitioning a GC- and AT-specific macroligand were employed. Comparing CCD's using each of these ligands information on the GC and AT content of exposed DNA in the chromosomes groups could be derived, demonstrating that specific sequences of DNA are accessible at the surface of metaphase chromosomes. 相似文献
132.
Gabriele Mezzetti Mariastella Moruzzi Giorgio Piccinini Maria G. Monti Bruno Barbiroli 《Molecular and cellular biochemistry》1986,70(2):141-149
Summary Chick duodenal mucosa contains an endogenous factor which is capable to inhibit selectively a homologous polyamine-sensitive protein kinase. The inhibitor was partially purified and characterized, and it was found to contain typical mucopolysaccharidic components.Glycosidases digestion studies, selective degradation analysis and spectrophotometric titrations with metachromatic dyes indicated that the inhibitor preparation contained two major moieties identified as heparin-like and heparan sulfate-like structures. In chick intestine the inhibitor was specific for polyamine-sensitive protein kinase since selectively interacted with it and was inert towards other cAMP-independent and cAMP-dependent protein kinases. The inhibitory effect of the endogenous factor was counteracted by naturally occurring polyamines such as spermine. The order of potency of various polyamines was: spermine > thermine spermidine diamines. The release of inhibition by addition of physiological concentrations of spermine was also apparent when using cytosolic proteins as endogenous phosphate acceptors. These results suggest that a possible role of polyamine in the regulation of polyamine-sensitive protein kinase in the intestine is to protect the enzyme from the inhibitory action of endogenous heparinoids. 相似文献
133.
Rat liver casein kinase TS (Ck-TS) having quarternary structure α2β2, autophosphorylates at its 25 kDa, β-subunits, incorporating up to 1.2 mol P/mol enzyme. According to their effects on the autophosphorylation pattern the effectors of Ck-TS activity can be grouped into 3 classes: (i) inhibitors, like heparin, which also prevent the autophosphorylation of the β-subunit; (ii) stimulators possessing several amino groups (like spermine) which increase the autophosphorylation at the β-subunit; (iii) stimulators possessing several guanido groups, like protamines and related peptides, which prevent the phosphorylation of the β-subunit, while promoting the autophosphorylation of the 38 kDa α-subunit. In the presence of such polyarginyl effectors the 130 kDa Ck-TS is converted into forms with higher sedimentation coefficient. 相似文献
134.
The effects of methadone (METH) on serum levels of prolactin (PRL), growth hormone (GH), corticosterone (CS) and TSH were determined in developing rats. METH increased PRL, GH and CS and decreased TSH at all ages tested, but the time course and magnitude of these effects changed during ontogeny. METH effects on day 10 were lower in magnitude than those observed in adults. In 20 day old pups, METH effects on GH and CS were comparable to those of adults, but TSH effects were still blunted. METH effects on hormone secretion in both 10 and 20 day old pups lasted longer than those observed in adults. Naloxone blocked all hormonal responses in adults, but did not completely block METH effects on CS secretion in 10 day old pups. 相似文献
135.
Epinephrine: A Potential Neurotransmitter in Retina 总被引:17,自引:13,他引:4
Abstract: Dopamine (DA), norepinephrine (NE), and epinephrine (EPI) are present in rat retina. DA is the major catecholamine, whereas NE and EPI represent ∼5% of the DA content. DA is contained in a subpopulation of amacrine cells and has been the subject of numerous studies. We investigated the origin and properties of NE and EPI in retina. Following superior cervical ganglionectomy, there was a decrease in NE content, but no decrease in EPI or phenylethanolamine- N -methyltransferase (PNMT) activity. PNMT in retina has many of the substrate-specificity and inhibitor-sensitivity characteristics of other tissues. Enzyme activity is enhanced in newborn rats by treatment with dexamethasone. Exposure to a lighted environment increases retinal EPI in normal and superior cervical ganglionectomized rats. EPI content increased for more than 2 h in a lighted environment. We conclude that most of the NE is contained within the sympathetic neurons that innervate the eye from the superior cervical ganglion, whereas EPI is contained in retinal elements that are responsive to photic stimulation. 相似文献
136.
Anna Maria Tagliasacchi Laura Maria Costantina Forino Manuela Frediani Silvana Avanzi 《Protoplasma》1983,115(2-3):95-103
Summary The pattern of DNA and RNA puffs in pair VII of polytene chromosomes has been investigated in the suspensor ofPhaseolus coccineus during early embryo development. The pattern of3H-TdR and3H-U incorporation has been also detected. Collected data indicate that: 1. both heterochromatic regions, p11 and q(111+112), of chromosome pair VII, organize large DNA puffs; 2. DNA puffs of both regions are specific of different embryo differentiation steps; 3. a seasonal influence on the DNA puffing seems also to be present, as demonstrated by the comparison of the results collected in two different crops; 4. the incorporation experiment by3H-TdR evidences that not all DNA puffs show clustered labeling; 5. the RNA puffing of the two regions seems also to be specific of determined embryo stages. 相似文献
137.
138.
Summary The development of a synthetic medium that supports growth and differentiation of insect embryonic tissues afforded the possibility
of studying the interactions between nerve and other cell types in long term cultures. The mechanical dissociation of embryonic
nerve tissues results in survival of nerve cells but not of glial cells. The dissociated glial-free neurons produce a dense
fibrillar network in the presence, but not in the absence, of foregut explants or other tissues from same donors. Nerve fiber
bundles outgrowing from dissociated neurons enter foregut segments and establish synaptic connections with muscle cells. Foregut
explants undergo differentiation and become contractile in long term cultures when innervated by dissociated nerve cells.
The progressive deterioration of similar foregut tissues cultured alone contrasts with the excellent condition of innervated
explants and suggests that this is due to trophic factors released by nerve fibers. The same in vitro systems provided the
opportunity of studying the interaction between nerve fibers produced by the autonomic ingluvial ganglion, which adheres to
the surface of the alimentary tract, and muscle cells. Multiple esophagus explants from cockroach embryos become interconnected
by fibers emerging from ingluvial ganglia, when the explants are combined in vitro at short distance from each other. Muscle
cells migrating from the esophagi line up on axons branching out in the medium, or form contractile ribbons which, in turn,
establish connections with nerve fibers. The thigmotropism of muscle cells and strong affinity for nerve fibers reveal a new
aspect of muscle cells-to-fibers interaction, amenable to further analysis in vitro.
This work was supported in part by United States Public Health Service grant NS-03777 and grant GB-16330 X from the National
Science Foundation 相似文献
139.
Summary The adventitious roots of onion (Allium cepa L.) or onion bulb scales were treated either with growth retardant Phosfon-D (2,4-dichlorobenzyltributylphosfonium chloride), or with low temperature (5° C). Low temperature, likewise Phosfon-D treatment, resulted in the appearance of membranous structures within the cytoplasm of the root cells fixed in chromaceto-formalin (CrAF) and in the occurrence of minute vacuoles in the living onion bulb scale cells. The cytochemical analysis revealed the membrane-like elements to contain basic and acid proteins as well as nucleic acids.It seems that the observed effect of Phosfon-D and that of low temperature were due to the local accumulation of protein resulting in changes of the physical properties of the cytoplasm. The fact that membranous structures do contain nucleic acid may suggest their important, metabolic role especially in the process of cold acclimation of plant tissue.This work was partly supported by Grant No. FG-PO-225 from U.S. Department of Agriculture, Agricultural Research Program, Public Law 480. 相似文献
140.
The fate of newly made DNA in Escherichia coli mutants thermosensitive in DNA synthesis 总被引:5,自引:0,他引:5
Summary
E. coli mutants exist in which DNA synthesis is thermosensitive. In one class of these mutants DNA synthesis stops immediately if a critical temperature (42°C) is reached. When DNA replication in such mutants is followed by 3H thymidine incorporation at 33°C, it is found that 1. only the newly made DNA is degraded at 42°C, 2. the discontinuously replicated DNA is lost predominantly at 42°C, 3. 1–3% of the chromosomal DNA is rendered acid soluble at 42°C without concomitant loss of viability of the cells at 33°C.Replication of phage DNA is inhibited in the same mutant at 42°C. However, when DNA synthesis is followed in infected cells at 33°C it is found that 1. no degradation of specific DNA seems to occur at 42°C in the early phase of infection, 2. replicating DNA molecules in the late phase of infection are completed at 42°C before DNA synthesis comes to a halt. 相似文献