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1.
The precision of extrachromosomal homologous recombination and gene targeting in plant cells was investigated. Recombination was directed to introns of selectable marker genes where potential changes could persist without affecting the function and therefore the selectability of the genes. Approximately 9 kb of crossover regions was rescued and sequenced. Changes were detected at a frequency below one point mutation per 1000 bp, indicating that extrachromosomal recombination and gene targeting both appear to occur with high fidelity. 相似文献
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Mei-Ling Siu-Caldera Jeffrey W. Clark Anabela Santos-Moore Sara Peleg Yan Yun Liu Milan R. Uskokovi Surendra Sharma G. Satyanarayana Reddy 《The Journal of steroid biochemistry and molecular biology》1996,59(5-6)
1α,25(OH)2-16-ene-D3, a synthetic analog of the steroid hormone, 1α,25(OH)2D3, has great potential to become a drug in the treatment of leukemia and other proliferative disorders, because of its minimal in vivo calcemic activity associated with a potent inhibitory effect on cell growth. However, at present, the mechanisms through which 1α,25(OH)2-16-ene-D3 expresses its biological activities are still not completely understood. Our previous in vitro study in a perfused rat kidney indicated for the first time that 1α,25(OH)2-16-ene-D3 and 1α,25(OH)2D3 are metabolized differently. 1α,25(OH)2-24-oxo-16-ene-D3, an intermediary metabolite of 1α,25(OH)2-16-ene-D3 formed through the C-24 oxidation pathway, accumulated significantly in the perfusate when compared to 1α,25(OH)2-24-oxo-D3, the corresponding intermediary metabolite of 1α,25(OH)2D3. In a subsequent in vivo study, we also reported that 1α,25(OH)2-24-oxo-16-ene-D3 exerted immunosuppressive activity equal to its parent, without causing significant hypercalcemia. In order to establish further the critical role of 1α,25(OH)2-24-oxo-16-ene-D3, in generating some of the key biological activities ascribed to its parent, we performed the present in vitro study using a human myeloid leukemic cell line (RWLeu-4) as a model. Comparative target tissue metabolism studies indicated that 1α,25(OH)2-16-ene-D3 and 1α,25(OH)2D3 are metabolized differently in RWLeu-4 cells, and the differences were similar to the ones we previously observed in the rat kidney. The significant finding was the accumulation of 1α,25(OH)2-24-oxo-16-ene-D3 in RWLeu-4 cells because of its resistance to further metabolism. Biological activity studies indicated that both 1α,25(OH)2-16-ene-D3 and its 24-oxo metabolite produced growth inhibition and promoted differentiation of RWLeu-4 cells to the same extent, and these activities were several fold higher than those exerted by 1α,25(OH)2D3. In addition, the genomic action of each vitamin D compound was assessed in a rat osteosarcoma cell line (ROS 17/2.8) by measuring its ability to transactivate a gene construct containing the vitamin D response element of the osteocalcin gene linked to the growth hormone reporter gene. In these studies, both 1α,25(OH)2-16-ene-D3 and its 24-oxo metabolite exerted similar but potent transactivation activity which was several fold greater than that exerted by 1α,25(OH)2D3 itself. In summary, our results indicate that the production and slow clearance of the bioactive intermediary metabolite, 1α,25(OH)2-24-oxo-16-ene-D3, in RWLeu-4 cells contributes significantly to the final expression of the enhanced biological activities ascribed to its parent analog, 1α,25(OH)2-16-ene-D3. 相似文献
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Neonatal transplantation tolerance to the products of theH-2
b
complex was induced in B10.A (H-2
a
) mice. On the basis of the survival of skin allografts it was found that antigens determined by theD region of theH-2
b
complex (of the B10.A(2R) strain) were most easily overcome and that tolerance to the products of theD end of theH-2 complex (of the B10.A(4R) strain) was also easy to induce. The antigens produced by theK end ofH-2 (of the B10.A(5R) and B10.A(3R) strains) represented a stronger incompatibility barrier and a difference in the entireH-2
b
complex caused strongest resistance to tolerance induction. When tolerance to the products of the entireH-2
b
complex was induced in newborn B10.A mice, and the neonatally treated animals were grafted simultaneously with five different grafts, those disparate at theK end ofH-2 and in the entireH-2 region were rejected in some animals, while the grafts disparate at theD end of H-2 remained intact in the same mice. No dependence on theI-J subregion was observed in this system. Furthermore, tolerance was more easily inducible in male than in female B10.A mice. 相似文献
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Summary A membrane potential (inside negative) across the plasma membrane of the obligatory aerobic yeastRhodotorula gracilis is indicated by the intracellular accumulation of the lipid-soluble cations tetraphenylphosphonium and triphenylmethylphosphonium. The uptake of these ions is inhibited by anaerobic conditions, by uncouplers, by addition of diffusible ions, or by increase of the leakiness of the membrane caused by the polyene antibiotic nystatin. The membrane potential is strongly pH-dependent, its value increasing with decreasing extracellular proton concentration. Addition of transportable monosaccharides causes a depolarization of the electrical potential difference, indicating that the H+-sugar cotransport is electrogenic. The effect on the membrane potential is enhanced by increasing the sugar concentration. The half-saturation constants of depolarization ford-xylose andd-galactose were comparable to those of the corresponding transport system for the two sugars. All agents that depressed the membrane potential inhibited monosaccharide transport; hence the membrane potential provides energy for active sugar transport in this strain of yeast. 相似文献
7.
Milan Korcok 《CMAJ》1983,128(3):254-256
The quadriceps tendon and patellar ligament rupture rarely, even when under great stress or trauma, but can rupture spontaneously in patients with chronic diseases such as gout, rheumatoid arthritis and renal disease. Several factors probably combine to weaken the tendon, including an impoverished local vascular supply, repeated microtrauma and secondary hypoparathyroidism with osteodystrophy. In the three cases reported here, one of which was bilateral, the patients were being treated for chronic renal disease; surgical repair of the tendons led to sound healing and a return to normal function of the joints. 相似文献
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