We describe non‐radiative energy‐transfer experiments to measure the rates of polymer interdiffusion in P(MMA‐co‐BA) latex films formed in the presence of poly(vinyl alcohol) (PVOH). PVOH had relatively little effect on the initial efficiency of energy transfer, even when the amount of PVOH was large enough to form the continuous phase. Since ΦET(0) is a measure of the interfacial area between D‐ and A‐labeled cells in the film, we conclude that under these circumstances the dispersed P(MMA‐co‐BA) copolymer is in the form of clusters with many contacts between particles containing D and A labels. These large amounts of PVOH also reduce the amount of polymer diffusion that takes place when the films are annealed. When smaller amounts of PVOH are present, the effects are measurable but much smaller. In the presence of 2 to 17 wt.‐% PVOH, the polymer diffusion rate is retarded. The magnitude of the effect increases with the amount of PVOH present, and the effect is larger at 45 °C than at 63 °C. We show that the PVOH has its largest influence at the very early stages of polymer diffusion.
Schematic representation of an energy transfer experiment, which monitors polymer interdiffusion in a latex film. 相似文献
We studied pacemaker current (if), the inward current activated by hyperpolarization in rabbit sinoatrial (SA) node myocytes, with the permeabilized-patch-clamp
technique. The tyrosine kinase inhibitors genistein (50 μM) or herbimycin A (35 μM) reduced the amplitude of if in response to step hyperpolarizations in the diastolic range of potentials. A two-step voltage-clamp protocol revealed that
the reduction in if is due to a decrease in maximal if conductance. The observed effects are due to tyrosine kinase inhibition since an inactive analog of genistein did not reduce
if. To further examine the mechanism of action, we added 2 mM chlorophenylthio cAMP (CPTcAMP, a membrane-permeant cAMP analog)
to the bathing Tyrode, which increased if. Genistein still reduced if in the presence of CPTcAMP. This suggests that the pathway mediating the actions of tyrosine kinase inhibition on if is independent of cAMP- or protein-kinase-A-mediated phosphorylation.
Received: 28 January 1997 / Received after revision: 21 April 1997 / Accepted: 22 April 1997 相似文献
Sequences at the 3'-ends of both positive and negative strands of Hepatitis C virus (HCV) RNA harbor cis-acting elements required for RNA replication. However, little is known about the properties of the negative RNA strand as a template for the synthesis of positive RNA strand. In this study, a purified recombinant HCV RNA-dependent RNA polymerase (RdRp) was used to investigate the synthesis of positive RNA strand using the 3'-terminal region of negative RNA strand ((-)3'T RNA) as template. A mutagenesis analysis was performed to evaluate the role of the 3'-proximal stem-loop and the first 3'-cytidylate (3'C) of the negative RNA strand in the synthesis of the positive RNA strand. A negative RNA strand of wild type (wt) HCV as template was able to direct the synthesis of a full-length positive RNA strand. Deletion of the 3'-proximal stem-loop resulted in an approximately 90% decrease in RNA synthesis. Disruption of the 3'-proximal stem-loop structure by nucleotide substitutions led to a 70-80% decrease in RNA synthesis. However, the restoration of the stem-loop by compensatory mutations in the stem region restored also the RNA synthesis. Likewise, the deletion or substitution of the first 3'C by guanylate (G) led to a 90% decrease in the RNA synthesis; while the substitution by adenylate (A) or uridylate (U) resulted in a 60-80% decrease in the RNA synthesis only. These findings demonstrate that the 3'-proximal stem-loop and the first 3'C of the negative RNA strand of HCV are two cis-acting elements involved in the synthesis of the positive RNA strand. 相似文献
One cause of congenital lactic acidosis is a mutation in the E1 alpha-subunit of the pyruvate dehydrogenase multienzyme complex. Little is known about the consequences of these mutations at the enzymatic level. Here we study the A199T mutation by expressing the protein in Escherichia coli. The specific activity is 25% of normal and the K(m) for pyruvate is elevated by 10-fold. Inhibitors of lactate dehydrogenase might be a useful therapy for patients with such mutations. 相似文献
A fully automated immunoassay for total plasma homocysteine assay was evaluated at four centers. To measure total homocysteine, oxidized forms of homocysteine in serum and plasma were reduced by dithiothreitol and assayed by a competitive fluorescence polarization technique. The assay had within-run precision from 0.9 to 3.0% and total precision from 2.8 to 4.1% for control materials with homocysteine concentrations of approximately 7, 12.5, and 25 micromol/L, a sensitivity of 0.35 micromol/L, good parallelism upon dilution, and analytical recovery ranging from 97.4 to 103.8%. The immunoassay correlated with four different HPLC assays for homocysteine, yielding a slope of 0.98, an intercept of -0.19 micromol/L, and a correlation coefficient of 0.966 for 440 paired samples. The reference range, determined with plasma samples from 609 males and 600 females, yielded a mean of 9.17+/-2.86 micromol/L, with a central 95% range of 4.78-15.43 micromol/L. The immunoassay is a suitable alternative to HPLC and may be useful in screening persons with high risk of coronary artery disease. 相似文献