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11.
Peter Fürst Lena Zimmerman Raymond Oulès Veronique Yahiel Christina Johnson Jonas Bergström 《Analytical biochemistry》1982,122(2):394-403
A system for automatic analyses of medium-sized compounds (middle molecules-MM) is described, applying gel filtration combined with gradient ion-exchange chromatography and using an automatic sample injector, motoric selector valves, and a minicomputer, which controls the system. Quantitation is performed by integrating the peak areas obtained. Using the automatic analyzer 7 MM solutes could be separated, detected, and quantified. The method permits a sampling rate of 28 samples per week. The precision of the method was 9.5%. The error in a single determination based on duplicate samples was 10% and in duplicate determinations was 7%. The automatic, analytical system is a prerequisite for performing sereening studies within a reasonable period of time. The present automatic, analytical system has been tested during a period of 3–5 years, and used in more than 4000 determinations. 相似文献
12.
Masaru Kamishohara Susan Kenney Renee Domergue David T. Vistica Edward A. Sausville 《Experimental cell research》2000,256(2):468
KRN5500 is a semisynthetic spicamycin analogue consisting of a seven-carbon amino sugar linked to a C14 unsaturated fatty acid through glycine and to the amino group of adenine. The drug inhibits cell growth potently and has antitumor activity in in vivo models. The mechanism of the antiproliferative effect of KRN5500 remains to be elucidated. We have found that acute exposure of drug-sensitive HT-29 colon adenocarcinoma cells to the drug results initially in swelling of the Golgi apparatus. Continuous exposure to the drug resulted in the emergence of a resistant population of cells characterized by numerous intracellular vacuoles. These KRN5500-resistant tumor cells exhibited increased staining with the Golgi stain NBD C6–ceramide and the ER–Golgi fluorescent dye BODIPY–brefeldin A, which, unlike the parental drug-sensitive cells, was dispersed throughout the cytoplasm. Marker enzymes associated with the ER (glucose 6-phosphatase) and cis-Golgi (GalNAc transferase) were elevated >2-fold and nearly 4-fold, respectively, in drug-resistant cell lines while the trans-Golgi marker enzyme, galactosyltransferase, was not. The additional findings that the KRN5500-resistant cells have a >2-fold elevation in ERGIC-53, a cis-Golgi marker protein of the ER–Golgi intermediate compartment (ERGIC), as well as increased 58K, a 58-kDa microtubule-binding protein with formiminotransferase cyclodeaminase activity, and tubulin indicate that the cellular secretory pathway is a primary determinant of sensitivity to KRN5500, as resistance to this agent corresponds with accumulation of several components relatable to ER and cis-Golgi function. Further support for this conclusion is provided by studies which demonstrate that KRN5500 alters the distribution of newly synthesized carcinoembryonic antigen within the secretory pathway, including arrest of this N-glycosylated protein in the Golgi of LS-174T colon carcinoma cells. 相似文献
13.
《Bioorganic & medicinal chemistry》2014,22(2):827-833
RCAI-147 is one of the hydroxylated analogues of KRN7000 which is known as a ligand for the activation of CD1d mediated invariant natural killer T cells (iNKT cells) and releases both T helper 1 (Th1) cytokines such as IFN-γ and T helper 2 (Th2) cytokines such as IL-4. KRN7000 has been anticipated as an antitumor drug or an adjuvant for viral infection such as influenza, because of its strong secretion of IFN-γ. In an interesting twist, it has been obvious in our previous paper that RCAI-147 induces much more Th2 cytokines (IL-4) than Th1 cytokines (IFN-γ) from iNKT cells compared to KRN7000, and shows fairly good result in the experimental autoimmune encephalomyelitis (EAE) test. Therefore, synthesis of RCAI-172 (C6-OH epimer of RCAI-147) was attempted to examine the biological activity. As a result, RCAI-172 was synthesized and its biological activity biased to Th2 response largely compared to that of KRN7000. However, this level decreased to approximately 61% compared to that of RCAI-147. And the clinical score of RCAI-172 for EAE suppression was disappointing. There exist seven chiral centers in the aglycon part of RCAI-172, and even though the change of configuration is just one position (C6-OH), the effect on both Th1/Th2 response and EAE test is fairly large. 相似文献
14.
Eiji Yoshioka Michiyo Morino Hideji Tanaka Hiroshi Shinkai 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1997,691(2):5178
A sensitive reliable method for the extraction, separation and quantitation of KRN5500 (I), a spicamycin derivative, from rat plasma was developed. It involves solid-phase extraction of the drug using a Bond Elut C18 cartridge and reversed-phase HPLC on a YMC-Pack ODS column with an ultraviolet detector. The intra- and inter-assay coefficients of variation by manual (n=10) and automatic (n=5) extraction were less than 9 and 13% and 6 and 8%, respectively. The limit of quantitation of each extraction procedure was 2 ng potency/ml. This extraction method may thus be considered useful for monitoring I in animals following its administration. 相似文献
15.
G. Harzer H. Rokos M.K. Otto A. Bacher S. Gishla S. Ghisla 《Biochimica et Biophysica Acta (BBA)/General Subjects》1978,540(1):48-54
Phosphotransferase from carrot is shown to catalyze the phosphorylation of 6,7-dimethyl-8-ribityllumazine specifically at position 5′ of the ribityl side chain. The lumazine 5′-phosphate is neither a substrate nor an inhibitor of riboflavin synthase from Bacillus subtilis and Escherichia coli. It follows that the obligatory product of riboflavin synthase is riboflavin and not FMN. 相似文献