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71.
72.
The effects of a thiazolidinone derivative, 3-[(3-trifluoromethyl)phenyl]-5-[(4-carboxyphenyl)methylene]-2-thioxo-4-thiazolidinone (or CFTRinh-172), on cystic fibrosis transmembrane conductance regulator (CFTR) gating were studied in excised inside-out membrane patches from Chinese hamster ovary cells transiently expressing wild-type and mutant CFTR. We found that the application of CFTRinh-172 results in an increase of the mean closed time and a decrease of the mean open time of the channel. A hyperbolic relationship between the closing rate and [CFTRinh-172] suggests that CFTRinh-172 does not act as a simple pore blocker. Interestingly, the potency of inhibition increases as the open time of the channel is increased with an IC50 in the low nanomolar range for CFTR channels locked in an open state for tens of seconds. Our studies also provide evidence that CFTRinh-172 can bind to both the open state and the closed state. However, at least one additional step, presumably reflecting inhibitor-induced conformational changes, is required to shut down the conductance after the binding of the inhibitor to the channel. Using the hydrolysis-deficient mutant E1371S as a tool as the closing rate of this mutant is dramatically decreased, we found that CFTRinh-172-dependent inhibition of CFTR channel gating, in two aspects, mimics the inactivation of voltage-dependent cation channels. First, similar to the recovery from inactivation in voltage-gated channels, once CFTR is inhibited by CFTRinh-172, reopening of the channel can be seen upon removal of the inhibitor in the absence of adenosine triphosphate (ATP). Second, ATP induced a biphasic current response on inhibitor-bound closed channels as if the ATP-opened channels "inactivate" despite a continuous presence of ATP. A simplified six-state kinetic scheme can well describe our data, at least qualitatively. Several possible structural mechanisms for the effects of CFTRinh-172 will be discussed. 相似文献
73.
Yu. V. Kovtun A. I. Skibenko E. I. Skibenko Yu. V. Larin A. N. Shapoval E. D. Volkov V. B. Yuferov 《Plasma Physics Reports》2010,36(12):1065-1071
The parameters of a dense (1013–1014 cm−3) plasma produced by ionization of a H2 + Ti mixture in a moderate-power (W ≤ 10 MW) pulsed reflective discharge are investigated. The dynamics of the plasma density, the elemental composition of the
generated plasma, the radial distribution of the electron density, the rotation velocity and frequency of the plasma layer
with n
p
≥ n
cr, the radial electric field, the coefficient of plasma particle separation, and the coefficient of plasma recombination in
the stage of plasma decay are determined. 相似文献
74.
Human artificial chromosomes (HACs) are autonomous molecules that can function and segregate as normal chromosomes in human cells. De novo HACs have successfully been used as gene expression vectors to complement genetic deficiencies in human cultured cells. HACs now offer the possibility of studying the regulation and expression of large genes in a variety of cell types from different tissues and correcting gene deficiencies caused by human inherited diseases. Complementary gene expression studies in mice, especially in mouse models of human genetic diseases, are also important in determining if large human transgenes can be expressed appropriately from artificial chromosomes. Toward this aim we are establishing artificial chromosomes in murine cells as novel gene expression vectors. Initially we transferred HAC vectors into murine cells, but were unable to generate de novo HACs at a reasonable frequency. We then transferred HACs previously established in human HT1080 cells to three different murine cell types by microcell fusion, followed by positive selection. We observed that the HACs in murine cells bound centromere protein C (CENP-C), a marker of active centromeres, and were detected under selection but rapidly lost when selection was removed. These results suggest that the HACs maintain at least a partially functional centromere complex in murine cells, but other factors are required for stability and segregation. Artificial chromosomes containing mouse centromeric sequences may be required for better stability and maintenance in murine cells. 相似文献
75.
Canevali C Orlandi M Zoia L Scotti R Tolppa EL Sipila J Agnoli F Morazzoni F 《Biomacromolecules》2005,6(3):1592-1601
The radicalization of unbleached lignocellulosic fibers obtained from thermomechanical (TMP) and chemothermomechanical (CTMP) pulps was performed in heterogeneous phase by reaction with dioxygen in the presence of N,N'-ethylenebis(salicylideneiminato)cobalt(II), [Co(salen)], as catalyst. Phenoxy cobalt radicals immobilized in fibers were observed by electron paramagnetic resonance (EPR) spectroscopy; their amount depends on the fiber swelling induced by reaction medium. The absolute concentration of such radicals in fibers, about 10(16) spin/g, reaches values 10 times higher than that of phenoxy radicals formed in similar oxidative reactions catalyzed by laccase. The generation of phenoxy cobalt radicals in fibers was related to structural changes of lignin units, detected by mono- and bidimensional nuclear magnetic resonance ((13)C NMR and 2D-HSQC) investigations, and to morphological modifications in fibers observed by scanning electron microscopy (SEM). 相似文献
76.
Sala G Beretta S Ceresa C Mattavelli L Zoia C Tremolizzo L Ferri A Carrì MT Ferrarese C 《Neurochemistry international》2005,46(3):227-234
Human neuroblastoma SH-SY5Y cells transfected with either familial amyotrophic lateral sclerosis-typical G93A mutant or wild-type copper/zinc superoxide dismutase were compared to untransfected cells in term of glutamate transport. Vmax of glutamate uptake was reduced in mutant cells, with no change in Km. No difference in EAAT1, EAAT2 and EAAT3 glutamate transporter mRNAs and immunoreactive proteins was found, suggesting that one or more transporters are functionally inactivated, possibly due to increased oxidative stress induced by the G93A mutation. Mutant cells showed a marked sensitivity to oxidants, resulting in a more pronounced reduction of glutamate uptake. Short-term antioxidant treatment did not reverse the impairment of glutamate uptake in G93A cells. Interestlingly, N-acetylcysteine was partially effective in preventing glutamate uptake reduction due to exogenous oxidative insults. Since the inhibition of the EAAT2 transporter subtype had no effect on glutamate re-uptake in this model, our study suggests an impaired function of the EAAT1/3 transporter subtypes, possibly due to oxidative inactivation, in the presence of mutant copper/zinc superoxide dismutase. Therefore, this model might prove to be a valuable tool to study the effects of mutant copper/zinc superoxide dismutase associated with amyotrophic lateral sclerosis on glutamate transport in neuronal cells, without the specific contribution of glial cells. These findings might lead to the identification of new therapeutic strategies aimed at preventing the damage associated with ALS. 相似文献
77.
The development of YAC cloning technology has directly enhanced the relationship among genetic, physical, and functional mapping
of genomes. Because of their large size, YACs have enabled the rapid construction of physical maps by ordered clone mapping
and contig building, and they complement other molecular approaches for mapping complex genomes. Large insert libraries are
constructted by size fractionating large DNA embedded in agarose and protecting DNA from degradation with polyamines. 相似文献
78.
Owing to the investigation of changeability variants of the left adrenal, anastomosis of the inferior diaphragmatic and ovarian (testicular) veins--tributaries of the human left renal vein, three variants in inflow of the left adrenal vein in comparison to the left ovarian (testicular) vein have been revealed. In the first variant, that occurs in 70% of cases, the left adrenal vein flows into the left renal vein by 15 cm more medially from the ostium of the ovarial (testicular) vein. The second variant occurs in 11%. The left adrenal and ovarian (testicular) ostia are situated at the same level. The distance from the hilus renalis to the place, where the ostia mentioned get into the renal vein, is about 39 mm. In the third variant (19%) the left adrenal vein gets into the left renal vein nearer to the hilus renalis than the ovarian (testicular) veins. An average distance from the hilus renalis to the ostium of the left adrenal vein is 33 mm. The second variant, when the left adrenal vein gets into the left renal vein, contributes to appearance of the primary varicocele at the left. When the operation for nephrectomy is made at the left, our data make it possible to recommend a purposeful ligation of the left renal vein more laterally to the ostium of the ovarian (testicular) vein. In 80% this ligation will not disturb the function of the left adrenal vein. 相似文献
79.
80.
Membrane interactions of myogenic cells of the 7-8 old chick embryos were investigated by electron microscopy. Tannic acid was used as specific fixative for biological membranes. Three types of specialized contacts can be revealed in fused myogenic cells: the first is like gap junctions, the second represents a pentalamellar structure, and the third is a so far non-described type of contact showing a membrane complex consisting of two parallel membranes arranged at a distance of approximately 15 nm and connected by electron-dense "bridges". The third type contact is revealed only by tannin-fixation. It is suggested that the "bridge" contact precedes the pentalamellar structure. A transition from the first type of contact to the second one is possible. The pentalamellar structure can be considered as an initial phase of fusion. 相似文献