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Summary Methods are described which demonstrate the use of unidirectional influx of14C-tetraphenylphosphonium (14C-TPP+) into isolated intestinal epithelial cells as a quantitative sensor of the magnitude of membrane potentials created by experimentally imposed ion gradients. Using this technique the quantitative relationship between membrane potential () and Na+-dependent sugar influx was determined for these cells at various Na+ and -methylglucoside (-MG) concentrations. The results show a high degree of dependence for the transport Michaelis constant but a maximum velocity for transport which is independent of . No transinhibition by intracellular sugar (40mm) can be detected. Sugar influx in the absence of Na+ is insensitive to 1.3mm phlorizin and independent of . The mechanistic implications of these results were evaluated using the quality of fit between calculated and experimentally observed kinetic constants for rate equations derived from several transport models. The analysis shows that for models in which translocation is the potential-dependent step the free carrier cannot be neutral. If it is anionic, the transporter must be functionally asymmetric. A model in which Na+ binding is the potential-dependent step (Na+ well concept) also provides an appropriate kinetic fit to the experimental data, and must be considered as a possible mechanistic basis for function of the system. 相似文献
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Annarosa Selli Franca Crociani Diego Matteuzzi Giorgio Crisetig 《Current microbiology》1986,13(1):33-38
Feedback inhibition of crude and purified extracts of homoserine dehydrogenase and threonine deaminase activities in the genusBifidobacterium was studied. Homoserine dehydrogenase was partially or completely inhibited byl-threonine, and a marked inhibitory effect byl-isoleucine on threonine deaminase was observed. In the speciesBifidobacterium cuniculi high levels ofl-valine reversed the inhibitory effect ofl-isoleucine. The -aminobutyric acid-resistant mutant Ru 326/106 of the speciesB. ruminale, overproducer ofl-isoleucine, had a derepressed homoserine dehydrogenase and a lesser feedback inhibition byl-threonine. Homoserine dehydrogenase appeared to be in bifids specifically NAD dependent. The regulatory mechanisms of aspartate family amino acid biosynthesis in bifidobacteria was discussed. 相似文献
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Characterization of a pollen-specific gene family from Brassica napus which is activated during early microspore development 总被引:13,自引:0,他引:13
Diego Albani Laurian S. Robert Pauline A. Donaldson Illimar Altosaar Paul G. Arnison Steven F. Fabijanski 《Plant molecular biology》1990,15(4):605-622
In this paper we describe the isolation and characterization of a genomic clone (Bp4) from Brassica napus which contains three members of a pollen-specific multigene family. This family is composed of 10 to 15 closely related genes which are expressed in early stages of microspore development. The complete nucleotide sequence of the clone Bp4 and of three homologous cDNA clones is reported. One of the genes (Bp4B) contained in the genomic clone is believed to be non-functional because of sequence rearrangements in its 5 region and intron splicing sites. The remaining genes (Bp4A and Bp4C), as well as the cDNA clones, appear to code for small proteins of unique structure. Three different types of proteins can be predicted as a result of the deletion of carboxy or amino terminal portions of a conserved core protein. These proteins all share a common alternation of hydrophobic and hydrophilic domains. A fragment of the genomic clone containing the gene Bp4A, as well as the non-functional gene Bp4B, was introduced into tobacco plants via Agrobacterium-mediated transformation. The functional gene Bp4A is expressed in transgenic tobacco plants and shows spatial and temporal regulation consistent with the expression patterns seen in Brassica napus. 相似文献
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A preliminary report on the use of transfer factor for treating stage D3 hormone-unresponsive metastatic prostate cancer 总被引:4,自引:0,他引:4
Dr. Giancarlo Pizza Caterina De Vinci Diego Cuzzocrea Domenico Menniti Ernesto Aiello Paolo Maver Giuseppe Corrado Piero Romagnoli Ennio Dragoni Giuseppe LoConte Umberto Riolo Aldopaolo Palareti Paolo Zucchelli Vittorio Fornarola Dimitri Viza 《Biotherapy》1996,9(1-3):123-132
As conventional treatments are unsuccessful, the survival rate of stage D3 prostate cancer patients is poor. Reports have
suggested the existence of humoral and cell-mediated immunity (CMI) against prostate cancer tumour-associated antigens (TAA).
These observations prompted us to treat stage D3 prostate cancer patients with an in vitro produced transfer factor (TF) able
to transfer, in vitro and in vivo, CMI against bladder and prostate TAA. Fifty patients entered this study and received one
intramuscular injection of 2–5 units of specific TF monthly. Follow-up, ranging from 1 to 9 years, showed that complete remission
was achieved in 2 patients, partial remission in 6, and no progression of metastatic disease in 14. The median survival was
126 weeks, higher than the survival rates reported in the literature for patients of the same stage. 相似文献
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Maurizio Varnier Patrizio Sarto Diego Martines Liliana Lora Francesco Carmignoto Graham Peter Leese Remo Naccarato 《European journal of applied physiology and occupational physiology》1994,69(1):26-31
The aim of this study was to investigate whether, when muscle glycogen is reduced, a pre-exercise infusion of branched-chain amino acids (BCAA) modifies exercise performance or the metabolic and respiratory responses to incremental exercise. Six moderately trained volunteers took part in the following protocol on two occasions. On day 1, at 9 a.m. in the postabsorptive state, they performed a graded incremental exercise (increases of 35 W every 4 min) to exhaustion (Ex-1). A meal of 1,000 kcal (4,200 kJ; 60% protein, 40% fat) was consumed at 12 p.m. No food was then allowed until the end of the experiment (20–21 h later). A 90-min period of exercise at alternating high and moderate intensities, designed to deplete muscle glycogen, was performed between 6 p.m. and 7.30 p.m. The morning after (day 2), the subjects randomly received either a mixed solution of BCAA (260 mg × kg–1 × h–1 for 70 min), or saline. They then repeated the graded incremental exercise to exhaustion (Ex-2). Metabolic and respiratory measurements suggested a muscle glycogen-depleted state had been achieved. No significant differences were observed in total work performed, maximal oxygen uptake or plasma ammonia, alanine, and blood pyruvate concentrations in the two treatments. After BCAA infusion, higher blood lactate concentrations were observed at maximal power output in comparison with those during saline [BCAA 4.97 (SEM 0.41) mmol × l–1, Saline 3.88 (SEM 0.47) mmol × l–1,P < 0.05]. In summary, in conditions of reduced muscle glycogen content, after a short period of fasting, BCAA infusion had no significant effect on the total work that could be performed during a graded incremental exercise. 相似文献
20.
DNA supercoiling and thermal regulation of unsaturated fatty acid synthesis in Bacillus subtilis 总被引:4,自引:1,他引:3
Roberto Grau Daniela Gardiol Gerardo Claudio Glikin Diego de Mendoza 《Molecular microbiology》1994,11(5):933-941
Bacillus subtilis growing at 37° C synthesizes, almost exclusively, saturated fatty acids. However, when a culture growing at 37°C is transferred to 20°C, the synthesis of unsaturated fatty acids is induced. The addition of the DNA gyrase inhibitor novobiocin specifically prevented the induction of unsaturated fatty acid synthesis at 20° C. Furthermore, it was determined that plasmid DNA isolated from cells growing at 20°C was significantly more negatively supercoiled than the equivalent DNA isolated from cells growing at 37°C. The overall results agree with the hypothesis that an increase in DNA supercoiling associated with a temperature downshift could regulate the unsaturated fatty acids synthesis in B. subtilis. 相似文献