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41.
Charles M. Lent Joyce Ono Kent T. Keyser Harvey J. Karten 《Journal of neurochemistry》1979,32(5):1559-1563
Abstract— We have measured serotonin (5-HT) within large and small neurosomata which are vitally stained by Neutral Red dye. A micro-radioenzymatic technique which is sensitive to 50fmol of 5-HT was employed on intact ganglia, 75 μm Retzius Cells (RZ) and a 10 μm ventro-lateral cell (VL) taken from the leech Macrobdella decora. The stain does not affect the levels of 5-HT in either ganglia or RZ. The VL cell body contains 5-HT at concentrations of at least 100 m m . Microspectrofluorometry of all the ganglionic neurosomata which fluoresce following the Falck-Hillarp formaldehyde condensation reaction detected rapidly-fading emission peaks of 509–523 nanometers. We conclude that all seven fluorescent neurons in the leech ganglion very probably contain serotonin. 相似文献
42.
Enzyme-linked immunosorbent analysis for aflatoxin B1. 总被引:1,自引:0,他引:1
An enzyme-linked immunosorbent analysis (ELISA) permitted the detection of less than 10 pg of aflatoxin B1 per ml. The antitoxin was most specific for aflatoxins B1 and B2alpha, and least specific for aflatoxin G1. 相似文献
43.
44.
Joyce M. Fisher Marco Rabinovitz 《Biochemical and biophysical research communications》1982,104(2):851-853
A specific copper chelator, 2,9-dimethyl-4,7-diphenyl-1,10-phenanthrolinedisulfonic acid substituted for mercaptoethanol to support growth of a L1210 lymphoma in primary culture. Added Cu++, but not Zn++ or Fe++ interfered with growth promotion by the chelator. It also can protect an established L1210 culture, which does not require mercaptoethanol, from cytotoxicity of two bis-thiosemicarbazones. Since these are known to require copper for cytotoxicity, the results indicate that 2,9-dimethyl-4,7-diphenyl-1,10-phenanthrolinedisulfonic acid acts by removing a source of endogenous copper in the tissue culture medium which prevents growth of the primary culture. 相似文献
45.
Lambdacyhalothrin cattle ear tags controlled horn fly, Haematobia irritans (L.), for 14 wks or longer during 1986-1988 in Georgia, USA. In 1989 and 1990, control of < 50 horn flies per side of cow was achieved for < or = 4 wk because of high levels of pyrethroid resistance in horn flies selected with lambdacyhalothrin. The highest resistance ratios (RRs) were seen in 1989. These were 498 for lambdacyhalothrin; 92,000 for fenvalerate; and 54 for permethrin. RRs for cypermethrin as high as 8,800 were estimated in 1990 when the RR for fenvalerate was only 1,060. No cross-resistance to diazinon was detected. These high levels of pyrethroid resistance seem to have a large component of metabolic resistance. Synergistic coefficients as high as 3,600 were determined by addition of nonlethal amounts of piperonyl butoxide. Resistance development in a no-pyrethroid-use area indicates movements of > or = 3km by sufficient numbers of horn flies can significantly change the RR. 相似文献
46.
Mark A. Yorek Joyce A. Dunlap Mark R. Stefani Eric P. Davidson 《Journal of neurochemistry》1992,58(5):1626-1636
It has been proposed that abnormal myo-inositol metabolism may be a factor in the development of diabetic complications. Studies with animal models of diabetes and cultured cells have suggested that hyperglycemia by an unknown mechanism may alter myo-inositol metabolism and content. Recently, we have shown that L-fucose, a 6-deoxy sugar whose content has been reported to be increased in diabetes, is a potent inhibitor of myo-inositol transport. To examine the effect of L-fucose on myo-inositol metabolism, neuroblastoma cells were cultured in medium supplemented with L-fucose. L-Fucose is a competitive inhibitor of Na(+)-dependent, high-affinity myo-inositol transport. The Ki for inhibition of myo-inositol transport by L-fucose is about 3 mM. L-Fucose is taken up and accumulates in neuroblastoma cells. The uptake of L-fucose is inhibited by Na+ depletion, D-glucose, glucose analogues, phloridzin, and cytochalasin B. In contrast, neither myo-inositol nor L-glucose inhibits L-fucose uptake. Chronic exposure of neuroblastoma cells to 1-30 mM L-fucose causes a decrease in myo-inositol accumulation and incorporation into inositol phospholipids, intracellular free myo-inositol content, and phosphatidylinositol levels. Na+,K(+)-ATPase transport activity is decreased by about 15% by acute or chronic exposure of neuroblastoma cells to L-fucose. Similar defects occur when neuroblastoma cells are exposed chronically to 30 mM glucose. Cell myo-inositol metabolism and Na+/K(+)-pump activity are maintained when 250 microM myo-inositol is added to the L-fucose-supplemented medium. Unlike the effect of chronic exposure of neuroblastoma cells to medium containing 30 mM glucose, the resting membrane potential of neuroblastoma cells is not altered by chronic exposure of the cells to 30 mM L-fucose. The effect of L-fucose on cultured neuroblastoma cell properties occurs at concentrations of L-fucose which may exist in the diabetic milieu. These data suggest that increased concentrations of L-fucose may have a role in myo-inositol-related defects in mammalian cells. 相似文献
47.
Two-dimensional (2-D) gel analysis of replication intermediates in the Chinese hamster dihydrofolate reductase domain has suggested that nascent chains can initiate at any of a large number of sites scattered throughout a ~50 kb “initiation locus” (although the level of initiation detected at any given site within this region was relatively low). This result contrasts markedly with data from anin vitro strand switching assay suggesting that >80% of initiations occur within a single 500 bp fragment lying within the initiation locus. In an effort to reconcile these two disparate views of the initiation reaction, we have questioned the validity of our 2-D gel data in several ways. We show here that: 1) the number of replication bubbles detected in the DHFR locus in the early S period is markedly increased when the cells are released from a synchronizing agent that inhibits initiationper se, rather than from aphidicolin, which is a chain elongation inhibitor; 2) initiation in the DHFR domain occurs only during the first 90 min of the S period, as would be expected of an early-firing origin; 3) a pulse of3H-thymidine moves through the structures observed on 2-D gels with the kinetics expected ofbonafide replication intermediates; and 4) preparations of replication intermediates that are subsequently analyzed on 2-D gels appear, by electron microscopy, to represent the typical theta structures and single-forked molecules expected of bidirectional origins of replication; no unusual structures (e.g., microbubbles) were seen. 相似文献
48.
Directed excision of a transgene from the plant genome 总被引:40,自引:0,他引:40
Sandra H. Russell Joyce L. Hoopes Joan T. Odell 《Molecular & general genetics : MGG》1992,234(1):49-59
Summary The effectiveness of loxP-Cre directed excision of a transgene was examined using phenotypic and molecular analyses. Two methods of combining the elements of this system, re-transformation and cross pollination, were found to produce different degrees of excision in the resulting plants. Two linked traits, -glucuronidase (GUS) and a gene encoding sulfonylurea-resistant acetolactate synthase (ALSr), were integrated into the genome of tobacco and Arabidopsis. The ALSr gene, bounded by loxP sites, was used as the selectable marker for transformation. The directed loss of the ALST gene through Cre-mediated excision was demonstrated by the loss of resistance to sulfonylurea herbicides and by Southern blot analysis. The -glucuronidase gene remained active. The excision efficiency varied in F1 progeny of different lox and Cre parents and was correlated with the Cre parent. Many of the lox × Cre F1 progeny were chimeric and some F2 progeny retained resistance to sulfonylureas. Re-transformation of lox/ALS/lox/GUS tobacco plants with cre led to much higher efficiency of excision. Lines of tobacco transformants carrying the GUS gene but producing only sulfonylurea-sensitive progeny were obtained using both approaches for introducing cre. Similarly, Arabidopsis lines with GUS activity but no sulfonylurea resistance were generated using cross pollinations. 相似文献
49.
Mechanism of DNA replication fidelity for three mutants of DNA polymerase I: Klenow fragment KF(exo+), KF(polA5), and KF(exo-) 总被引:2,自引:0,他引:2
B T Eger R D Kuchta S S Carroll P A Benkovic M E Dahlberg C M Joyce S J Benkovic 《Biochemistry》1991,30(5):1441-1448
Inhibition of the pre-steady-state burst of nucleotide incorporation by a single incorrect nucleotide (nucleotide discrimination) was measured with the Klenow fragment of DNA polymerase I [KF(exo+)]. For the eight mispairs studied on three DNA sequences, only low levels of discrimination ranging from none to 23-fold were found. The kinetics of dNTP incorporation into the 9/20-mer at low nucleotide concentrations was also determined. A limit of greater than or equal to 250 s-1 was placed on the nucleotide off-rate from the KF(exo+)-9/20-dTTP complex in accord with nucleotide binding being at equilibrium in the overall kinetic sequence. The influence of the relatively short length of the 9/20-mer on the mechanism of DNA replication fidelity was determined by remeasuring important kinetic parameters on a 30/M13-mer with high homology to the 9/20-mer. Pre-steady-state data on the nucleotide turnover rates, the dATP(alpha S) elemental effect, and the burst of dAMP misincorporation into the 30/M13-mer demonstrated that the kinetics were not affected by the length of the DNA primer/template. The effects on fidelity of two site-specific mutations, KF(polA5) and KF(exo-), were also examined. KF(polA5) showed an increased rate of DNA dissociation and a decreased rate of polymerization resulting in less processive DNA synthesis. Nevertheless, with at least one misincorporation event, that of dAMP into the 9/20-mer, KF(polA5) displays an increased replication fidelity.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
50.
Martha C. Willcox Sandra M. Reed Joyce A. Burns J. C. Wynne 《American journal of botany》1990,77(10):1257-1259
The stage of pollen development at the time of anther culture is an important factor in the production of haploids. The objectives of the current study were to develop a staining procedure for peanut (Arachis hypogaea L., ssp. hypogaea) microspores, to describe and document the stages of microsporogenesis in peanut, and to confirm a previous report concerning correlations of peanut floral bud shape with stage of microspore development. A staining procedure using propionic carmine provided adequate staining of pollen mother cells, microspores, and pollen. Pollen mother cells and microspores could easily be differentiated by their size and cell wall structure. Plants grown in a controlled environment were found to have highly synchronized microspore development, both within an anther and among anthers contained in the same bud. In addition, floral bud shape was confirmed as a reliable indicator of anther stage in peanuts. 相似文献