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Values Published (value + published)
Selected AbstractsUse of a blocking antibody method for the flow cytometric measurement of ZAP-70 in B-CLLCYTOMETRY, Issue 4 2006Mark Shenkin Abstract Background: In this study we developed a method to measure the amount of ZAP-70 [zeta accessory protein] in B-CLL cells without relying on the ZAP-70 expression of patient B or T cells to normalize fluorescence intensity. Methods: B-CLL cells were fixed with formaldehyde before surface staining with gating antibodies CD19PC5 and CD5FITC. The cells were permeabilized with saponin, and the ZAP-70 antigen was blocked in one tube with unlabeled antibody to ZAP-70 [clone 1E7.2]. Zap-70-PE was then added to this tube. ZAP-70-PE was added to a second tube without unlabeled antibody to ZAP-70. The mean fluorescence intensity of the ZAP-70 in the tube without unlabeled antibody divided by the mean fluorescence intensity of the ZAP-70 in the tube with unlabeled antibody equals the RATIO of total fluorescence to non-specific ZAP-70 fluorescence in the B-CLL cells. In a second method of analysis, a region is created in the histogram showing ZAP-70 fluorescence intensity in the tube with unlabeled antibody to ZAP-70. This region is set to 0.9% positive cells. This same region is then used to measure the % positive [%POS] ZAP-70 cells in the tube without unlabeled antibody to ZAP-70. The brighter the ZAP-70 fluorescence above the non-specific background, the higher the %POS. Results: Due to the varying amount of non-specific staining between patient B-CLL cells and other cells, the blocking antibody method yielded a more quantitative and reproducible measure of ZAP-70 in B-CLL cells than other methods, which use the ratio of B-CLL fluorescence to normal B or T-cell fluorescence. Using this improved method, ZAP-70 was determined to be negative if the RATIO was less than 2:1 and positive if the RATIO was greater than 2:1. ZAP-70 was determined to be negative if the %POS was less than 5% and positive if the %POS was greater than 5%, a cut-off value lower than previous values published, due to exclusion of non-specific staining. Both cut-offs were based upon patient specimen distribution profiling. Conclusions: Use of a blocking antibody resulted in a robust, reproducible clinical B-CLL assay that is not influenced by the need to measure the amount of ZAP-70 in other cells. ZAP-70 results segre gate patients into indolent and aggressive groups suggested by published clinical outcomes. © 2006 International Society for Analytical Cytology [source] Reinvestigating hyperpolarized 129Xe longitudinal relaxation time in the rat brain with noise considerationsNMR IN BIOMEDICINE, Issue 3 2008X. Zhou Abstract The longitudinal relaxation time of hyperpolarized (HP) 129Xe in the brain is a critical parameter for developing HP 129Xe brain imaging and spectroscopy and optimizing the pulse sequences, especially in the case of cerebral blood flow measurements. Various studies have produced widely varying estimates of HP 129Xe T1 in the rat brain. To make improved measurements of HP 129Xe T1 in the rat brain and investigate how low signal-to-noise ratio (SNR) contributes to these discrepancies, we developed a multi-pulse protocol during the washout of 129Xe from the brain. Afterwards, we applied an SNR threshold theory to both the multi-pulse protocol and an existing two-pulse protocol. The two protocols yielded mean,±,SD HP 129Xe T1 values in the rat brain of 15.3,±,1.2 and 16.2,±,0.9,s, suggesting that the low SNR might be a key reason for the wide range of T1 values published in the literature, a problem that might be easily alleviated by taking SNR levels into account. Copyright © 2007 John Wiley & Sons, Ltd. [source] Bond lengths in organic and metal-organic compounds revisited: X,H bond lengths from neutron diffraction dataACTA CRYSTALLOGRAPHICA SECTION B, Issue 3 2010Frank H. Allen The number of structures in the Cambridge Structural Database (CSD) has increased by an order of magnitude since the preparation of two major compilations of standard bond lengths in mid-1985. It is now of interest to examine whether this huge increase in data availability has implications for the mean bond-length values published in the late 1980s. Those compilations reported mean X,H bond lengths derived from rather sparse information and for rather few chemical environments. During the intervening years, the number of neutron studies has also increased, although only by a factor of around 2.25, permitting a new analysis of X,H bond-length distributions for (a) organic X = C, N, O, B, and (b) a variety of terminal and homometallic bridging transition metal hydrides. New mean values are reported here and are compared with earlier results. These new overall means are also complemented by an analysis of X,H distances at lower temperatures (T, 140,K), which indicates the general level of librational effects in X,H systems. The study also extends the range of chemical environments for which statistically acceptable mean X,H bond lengths can be obtained, although values from individual structures are also collated to further extend the chemical range of this compilation. Updated default `neutron-normalization' distances for use in hydrogen-bond and deformation-density studies are also proposed for C,H, N,H and O,H, and the low-temperature analysis provides specific values for certain chemical environments and hybridization states of X. [source] Dynamic binding capacity of plasmid DNA in histidine,agarose chromatographyBIOMEDICAL CHROMATOGRAPHY, Issue 9 2007F. Sousa Abstract The use of histidine,agarose chromatography in the purification of supercoiled (sc) plasmid DNA (pDNA) from Escherichia coli lysates has been reported recently. In the current work we describe a set of breakthrough experiments which were designed to study the effect of parameters such as flow-rate, temperature, concentration and conformation on the dynamic binding capacity of pDNA to the histidine support. One of the most striking results shows that the dynamic binding capacity for sc pDNA decreases linearly from 250.8 to 192.0 µg sc pDNA/mL when the temperature is varied from 5 to 24°C. This behaviour was attributed to temperature-induced, pre-denaturation conformational changes which promote the removal of negative superhelical turns in sc pDNA molecules and decrease the interaction of DNA bases with the histidine ligands. The capacity for sc pDNA was highly improved when using feeds with higher pDNA concentrations, a phenomenon which was attributed to the fact that pDNA molecules in more concentrated solutions are significantly compressed. A maximum capacity of 530.0 µg pDNA/mL gel was obtained when using a 125 µg/mL pDNA feed at 1 mL/min and 5°C, a figure which is comparable to the plasmid capacity values published for other chromatographic supports. Finally, a more than 2-fold increase in capacity was obtained when changing from open circular to sc pDNA solutions. Overall, the results obtained provide valuable information for the future development and implementation of histidine chromatography in the process scale purification of pDNA. Copyright © 2007 John Wiley & Sons, Ltd. [source] | ||||||||||