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At least two types of glucose transporter exist in cultured renal epithelial cells, a Na(+)-glucose cotransporter (SGLT), capable of interacting with D-glucose but not 2-deoxy-D-glucose (2dglc) and a facilitated transporter (GLUT) capable of interacting with both D-glucose and 2dglc. In order to examine the polarity of transport in cultured renal epithelia, 2dglc and D-glucose uptakes were measured in confluent cultures of LLC-PK1 cells grown on collagen-coated filters that permitted access of medium to both sides of the monolayer. The rates of basolateral uptake of both 1 mM glucose (Km 3.6 mM) and 1 mM 2dglc (Km 1.5 mM) were greater than apical uptake rates and the (apical-to-basolateral)/(basolateral-to-apical) flux ratio was high for glucose (9.4) and low for 2dglc (0.8), thus, confirming the lack of interaction of 2dglc with the apical SGLT. Specific glucose transport inhibitor studies using phlorizin, phloretin and cytochalasin B confirmed the polarised distribution of SGLT and GLUT in LLC-PK1 cells. Basolateral sugar uptake could be altered by addition of insulin (1 mU/ml) which increased 2dglc uptake by 72% and glucose uptake by 50% and by addition of 20 mM glucose to the medium during cell culture which decreased 2dglc uptake capacity at confluence by 30%. During growth to confluence, 2dglc uptake increased to a maximum, then decreased at the time of confluence, coincident with a rise in uptake capacity for alpha-methyl-D-glucoside, a hexose that interacts only with the apical SGLT. It was concluded that the non-metabolisable sugar 2dglc was a useful, specific probe for GLUT in LLC-PK1 cells and that GLUT was localised at the basolateral membrane after confluence. 相似文献
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N E Miller 《Biofeedback and self-regulation》1985,10(1):3-24
This paper will deal with a variety of topics such as dangers and opportunities from the developing crisis in health care costs, a cooperative study of the cost-effectiveness of treatments for headaches, the need for a federation of related societies, exploiting remarkable electronic advances, the wide range of adaptive functions of visceral learning (including its role in homeostasis), why maladjustments occur and their implications for biofeedback, and the need for analytic experiments involving adequate amounts of training. 相似文献
15.
Werner E.C. Muller Jürgen Conrad Rudolf K. Zahn Renate Steffen Gerhard Uhlenbruck Isabel Miller 《Differentiation; research in biological diversity》1984,26(1-3):30-35
Abstract. The Hexactinellida sponge Aphrocallistes vastus contains a soluble aggregation factor (AF) whose purification has been described in this communication. It is characterized by a S°20.w value of 37 and a buoyant density of 1.45 g/cm3 . The AF is a glycoporteinaceous particle composed of three major protein species; no core structure could be visualized. In the presence of Ca2+ , the AF causes secondary aggregation of single cells. The aggregation process is temperature, pH, and ionic strength independent within a broad range. Evidence is presented indicating that two (or more) AF molecules are required for the establishment of a stable cell: cell interaction. In contrast to the AFs from demosponges, the hexactinellid AF functions species-unspecifically. 相似文献
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Overproduction from a cellulase gene with a high guanosine-plus-cytosine content in Escherichia coli 总被引:1,自引:0,他引:1
G P O'Neill D G Kilburn R A Warren R C Miller 《Applied and environmental microbiology》1986,52(4):737-743
A recombinant exoglucanase was expressed in Escherichia coli to a level that exceeded 20% of total cellular protein. To obtain this level of overproduction, the exoglucanase gene coding sequence was fused to a synthetic ribosome-binding site, an initiating ATG, and placed under the control of the leftward promoter of bacteriophage lambda contained on the runaway replication plasmid vector pCP3 (E. Remaut, H. Tsao, and W. Fiers, Gene 22:103-113, 1983). With the exception of an inserted asparagine adjacent to the initiating ATG, the highly expressed exoglucanase is identical to the native exoglucanase. The overproduced exoglucanase can be isolated easily in an enriched form as insoluble aggregates, and exoglucanase activity can be recovered by solubilization of the aggregates in 6 M urea or 5 M guanidine hydrochloride. Since the codon usage of the exoglucanase gene is so markedly different from that of E. coli genes, the overproduction of the exoglucanase in E. coli indicates that codon usage may not be a major barrier to heterospecific gene expression in this organism. 相似文献
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Although the central and stable position of female rhesus monkeys in semi-free-ranging bands has been observed and described, the position of the developing male in the social group is not as well delineated. The present study is an attempt to describe interactions of 18 males with members of their families. Quantitative observations were made on one band (E) of rhesus monkeys on Cayo Santiago. Incidences of behavior recorded were joining, grooming, and threatening. The resulting data were analyzed by comparing the behavior of the males studied toward members of their genealogy as well as unrelated members of the social group. Behavior of family and non-relatives toward the males was also revealed. Statistical analysis revealed that the male subjects engaged in more positive social interactions within the matrifocal family than with other members of the genealogy, and in turn, more such behavior was observed within the genealogy than with unrelated monkeys. Threatening behavior within the genealogies was infrequent. Age effects were noted in that family members directed more interactions toward younger males and as males matured, they groomed more but joined family members less frequently. These data suggest mechanisms of socialization within the genealogy that may be different from those employed within the unrelated social group. 相似文献
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