Increasing the flexibility of the LANCE cAMP detection kit.

Plasma membranes of 6-h differentiated Dictyostelium discoideum cells contain a cAMP-binding protein with the properties ascribed to the chemotaxis receptor present on these cells. We have purified this cAMP-binding protein using DEAE-Sephadex chromatography, hydrophobic chromatography on decylagarose and preparative polyacrylamide gel electrophoresis in nonionic detergent. Photoaffinity labeling of the DEAE-purified material with 8-azido-[32P] cAMP shows that only an Mr = 70,000 species on sodium dodecyl sulfate gels contains a cAMP-binding site. Two-dimensional polyacrylamide gel electrophoresis of material eluted from decyl-agarose and photoaffinity labeled indicates that the cAMP-binding protein is the most acidic of many Mr = 70,000 proteins present. This method is readily scaled up to process up to 10(11) cells which yield from 25 to 100 micrograms of cAMP-binding protein. Nucleotide specificity studies established that the cAMP-binding site of the protein is similar to that of the cAMP receptor assayed on intact cells and membranes. The rates of association and dissociation of the cAMP-binding protein are extremely rapid as found for the receptor, and its affinity for cAMP is comparable. The cAMP-binding protein is a concanavalin A binding glycoprotein, and is resistant to proteolysis by trypsin, but not chymotrypsin. Like the cAMP receptor in membranes and crude detergent extracts, this cAMP-binding protein is inhibited by phenylmethylsulfonyl fluoride. The purified binding protein exists in solution largely as a monomeric species, with some dimer being detected on gel filtration. Based on these criteria, we conclude that this cAMP binding protein represents the binding subunit of the cAMP chemotaxis receptor.

Influence of freeze-thaw cycles on change of arsenic speciation and toxic effects to bacteria in paddy soil.

The effect of microstructural evolution on the permeability of sandstone under freeze-thaw cycles

Quantification of hepatitis B virus (HBV) DNA and hepatitis C virus (HCV) RNA often is performed in specimens that have been frozen and thawed more than once. To ensure optimal therapeutic and prognostic value, it is important to establish whether viral load measurements are affected by repeated freeze-thaw (FT) cycles. We therefore evaluated the effect of multiple FT cycles on HBV DNA and HCV RNA quantification by testing serum specimens subjected to one (baseline), two, four, and eight FT cycles with the appropriate Chiron Quantiplex assay. Linear regression analysis showed minor increases of 1.7% per FT cycle for both HBV DNA and HCV RNA. The rise in HCV RNA levels was more pronounced among low-concentration samples, since further analysis revealed an increase of 3.2% per FT cycle among samples with 0.2 to 3.86 Meq of HCV RNA per ml. Given that the coefficient of variation for the Quantiplex assays is generally 10 to 15%, the minor increases in HBV DNA and HCV RNA levels with progressive FT cycles for the specimens tested were recognized only because analysis of variance revealed a statistically significant trend (P < 0.05). Due to the minor statistical trend, the clinical impact for individual patient specimens is likely to be limited, but it may deserve further study. In conclusion, the concentration of HBV DNA and HCV RNA in serum specimens subjected to up to eight short-term FT cycles was stable.

Uniaxial compressive mechanical properties and stress–strain model for roller-compacted concrete with initial damage subjected to freeze–thaw cycles

Evolution of soil-water and shrinkage characteristics of an expansive clay during freeze-thaw and drying-wetting cycles

Cyclic AMP-binding protein and estrogen receptor: Antagonism during nuclear translocation in a hormone-dependent mammary tumor

Expansion and contraction of clogged open graded friction course exposed to freeze-thaw cycles and degradation of mechanical performance

Cracked rock masses in cold regions are subjected to freeze–thaw cycles over extended periods, resulting in freeze–thaw deformation. The combined effects of freeze–thaw cycling and the depth of cracks significantly influence the stability and durability of underground rock engineering in these regions. In some cold regions with minimal annual rainfall, rock masses are unable to absorb external water during freeze–thaw cycles. As freeze–thaw deformation progresses, the rock transitions naturally from a saturated state to an unsaturated state. To investigate the deformation damage mechanisms and evolution patterns of saturated red sandstone with initial non-penetrating cracks of varying depths (20 mm, 30 mm, 40 mm) under freeze–thaw cycling conditions without external water replenishment and with naturally varying saturation levels, relevant freeze–thaw cycle experiments and strain monitoring were conducted. The results indicate that cracked red sandstone experiences residual strain in each freeze–thaw cycle, which gradually accumulates, leading to irreversible freeze–thaw damage deformation. The cumulative residual strain of the rock specimen after 45 freeze–thaw cycles was 40.69 times greater than the residual strain from the first cycle. Additionally, the freeze–thaw strain characteristic values exhibited a clear correlation with crack depth. These findings provide experimental methods and data references for analyzing the deformation and failure mechanisms of cracked rock induced by freeze–thaw damage in cold regions.

The continuous increase in the emissions of iron ore tailings (IOTs) has caused serious pollution to the environment and raised costly IOT management costs. For protecting the environment and decreasing the economic cost, the fiber-reinforced tailing mortar was prepared with 50% IOTs, polyacrylonitrile fiber (PANF), and basalt fiber (BF). The mass loss, porosity, ultrasonic velocity, mechanical strength, and pore structure of tailing mortar after different treatments, including 100 freeze–thaw (FT) cycles and 90 sulfate attack (SA) cycles were analyzed to investigate the effect of PANF and BF on the durability. The test results indicated that BF and PANF have great bridging effects and cracking resistance to improve the durability properties. The B0.25P0.05 (BF: 0.25% and PANF: 0.05%) specimens possess the minimum mechanical strength loss, particularly the uniaxial compressive strength (UCS) only decreased by 5.58% after 100 FT cycles and 2.73% after 90 SA cycles. The B0.25P0.05 specimens also have the smallest dynamic elastic modulus loss, and the reduction was only 7.24% after 100 FT cycles and 0.76% after 90 SA cycles. For 120-day long-term tests, the B0.25P0.05 specimens have the maximum increment in the UCS (22.04%) and dynamic elastic modulus (5.60%). Besides, the mechanical strength of tailing mortar undergoing FT and SA cycles can be predicted accurately by dynamic elastic modulus (prediction error ≤5%). From a microscopic view, the nuclear magnetic resonance (NMR) test demonstrated that fibers significantly decrease the large pores and improve the pore microstructure, and thus raise the mechanical properties. Finally, the ranking of durability is B0.25P0.05 > B1.00P0 > B0P0.08 > B0P0. The hybrid fibers of rigid BF (6 mm) and flexible PANF (12 mm) in tailing mortar cross-climb to generate a denser network structure. Besides, BF and PANF in the mortars can effectively fill the pores and bear expansion stress caused by FT and SA cycles. Therefore, these effects remarkably limit the deformation with different sizes.

Effect of freeze-thaw cycles on engineering properties of biocemented sand under different treatment conditions

Induction of Tolerance to Freeze-Thaw (FT) Damage in Mammalian Cells by Pre-FT Hypothermia Treatment

In color gamut mapping of pictorial images, the lightness rendition of the mapped images plays a major role in the quality of the final image. For color gamut mapping tasks, where the goal is to produce a match to the original scene, it is important to maintain the perceived lightness contrast of the original image. Typical lightness remapping functions such as linear compression, soft compression, and hard clipping reduce the lightness contrast of the input image. Sigmoidal remapping functions were utilized to overcome the natural loss in perceived lightness contrast that results when an image from a full dynamic range device is scaled into the limited dynamic range of a destination device. These functions were tuned to the particular lightness characteristics of the images used and the selected dynamic ranges. The sigmoidal remapping functions were selected based on an empirical contrast enhancement model that was developed for the result of a psychophysical adjustment experiment. The results of this study showed that it was possible to maintain the perceived lightness contrast of the images by using sigmoidal contrast enhancement functions to selectively rescale images from a source device with a full dynamic range into a destination device with a limited dynamic range.© (1998) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Freeze-thaw cycles intensified the synergistic inhibitory effects of enrofloxacin and UV-328 on rye seedling growth.

Cytotoxicity assays in 96-well tissue culture plates allow rapid sample handling for multicondition experiments but have a limited dynamic range. Using DIMSCAN, a fluorescence digital image system for quantifying relative cell numbers in tissue culture plates, we have developed a 96-well cytotoxicity assay with a >4-log dynamic range. To overcome background fluorescence that limits detection of viable cells with fluorescein diacetate, we used 2'4'5'6'-tetrabromofluorescein (eosin Y) to quench background fluorescence in the medium and in nonviable cells to enhance the reduction of background fluorescence achieved with digital image thresholding. The sensitivity and linearity of the new assay were tested with serial dilutions of neuroblastoma and leukemia cell lines. DIMSCAN was compared with other in vitro cytotoxicity assays: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, colony formation, and trypan blue dye exclusion. Without background fluorescence reduction, scans produced a nearly flat curve across various cell concentrations from 100 to 10(6) cells per well. Either digital image thresholding or eosin Y dramatically reduced background fluorescence, and combining them achieved a linear correlation (r > 0.9) of relative fluorescence to viable cell number over >4 logs of dynamic range, even in the presence of 4 x 10(4) nonviable cells per well. Cytotoxicity of deferoxamine for neuroblastoma cell lines measured by the DIMSCAN assay achieved dose-response curves similar to data obtained by manual trypan blue counts or colony formation in soft agar but with a wider dynamic range. Long-term cultures documented the clonogenic ability of viable cells detected by DIMSCAN over the entire dynamic range. The cytotoxicity of two drug combinations (buthionine sulfoximine + melphalan or fenretinide + safingol) was tested using both DIMSCAN and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, and the wider dynamic range of DIMSCAN facilitated detection of synergistic interactions. DIMSCAN offers the ability to rapidly and efficiently conduct cytotoxicity assays in 96-well plates with a dynamic range of >4 logs. This assay enables rapid testing of anticancer drug combinations in microplates.