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31.
Xylose reductase (XR) from Neurospora crassa was purified to homogeneity and was found to be specific to NADPH (nicotinamide adenine dinucleotide phosphate). The purified enzyme showed M(r) of 60 and 29 kDa by gel filtration and SDS-PAGE indicating the presence of two subunits. The kinetic mechanism of xylose reductase is 'iso-ordered bi bi'. Inactivation of XR by N-bromosuccinimide (NBS) was found to be biphasic with second-order rate constants of 2.5 x 10(2) and 80 M-1S-1 for the fast (kf) and slow phase (ks), respectively. NADPH protected 90% of XR activity against inhibition by NBS. The fluorescence and circular dichroism (CD) studies revealed that inactivation was not due to gross conformational change in the enzyme. Analysis of the modified Stern-Volmer plot indicated that 49% of the tryptophanyl fluorescence was available for quenching which was completely abolished in the presence of NADPH confirming the involvement of tryptophan at the coenzyme binding site. Experimental evidence presented here serves to implicate the involvement of a tryptophan residue at the low-affinity NADPH binding site and the nature of this site has been assessed by using the hydrophobic probe ANS. 相似文献
32.
A new technique for understanding the organization of complex circuits in the vertebrate brain, scanning laser photostimulation, is described. This approach is based on the photolysis of a caged form of the excitatory neurotransmitter glutamate. Computer-controlled photostimulation and whole cell recording in brain slices allow the construction of detailed maps of the position, strength, sign and number of inputs converging on a single postsynaptic neuron. Scanning laser photostimulation offers many advantages over current techniques: spatial resolution is superb, fibers of passage are not activated, and thousands of presynaptic locations can be stimulated. This review describes the technique of photostimulation, outlines the instrumentation, necessary to implement it, and discusses the interpretation of photostimulation-derived data. Several examples of applications, ranging from mapping circuits in the mammalian visual cortex to determining receptor distributions on single neurons are considered. Although still in its early stages, scanning laser photostimulation offers neuroscientists a powerful tool for determining the organization and function of local brain circuits. 相似文献
33.
MB Spellerberg CJ Chapman CI Mockridge DA Isenberg FK Stevenson 《Canadian Metallurgical Quarterly》1995,6(2):52-56
The VH4-21 (V4-34) gene segment, a member of the VH4 family, is expressed early in B-cell maturation and is utilized by approximately 6% of normal adult B lymphocytes. This prevalence indicates an importance of VH4-21 in the B-cell repertoire. The gene also encodes certain autoantibodies being mandatory for pathological IgM anti-red cell antibodies directed against the I/i antigen, and also capable of encoding anti-DNA antibodies. Recognition of I/i antigen or DNA appears to be via two distinct sites on VH, with I/i binding mediated by sequences in the framework region, and DNA binding correlating with the presence of positively charged amino acids in complementarity-determining region 3. However, these positively charged residues appear to suppress the ability of the framework region to interact with I/i, rendering a single sequence monospecific for I/i or DNA. The IgM anti-DNA antibodies also recognize bacterial lipid A, whereas the anti-I/i antibodies do not, indicating that CDR3 may be involved in binding the negatively charged lipid A. Structural similarities between the DNA backbone and lipid A provide a possible explanation for this cross-reactivity. This dual recognition of bacterial antigen and autoantigen provides a potential link between infection and autoimmunity. 相似文献
34.
F Davodeau MA Peyrat A Necker R Dominici F Blanchard C Leget J Gaschet P Costa Y Jacques A Godard H Vie A Poggi F Romagné M Bonneville 《Canadian Metallurgical Quarterly》1997,158(12):5603-5611
Several studies have demonstrated the existence of a murine NK1.1+ alphabeta T cell subset expressing V alpha14+ TCR alpha-chains with highly conserved invariant junctional sequences and able to secrete Th2 cytokines when exposed to CD1+ stimulator cells. In humans, alphabeta T cells carrying invariant V alpha24+ TCR alpha-chains highly homologous to those expressed by murine NK1.1 cells have been recently described. Here we show that these cells (referred to as V alpha24inv T cells) and murine NK1.1+ alphabeta T cells resemble each other in several ways. First, like their murine counterparts, T cells expressing high levels of V alpha24inv TCRs can be either CD4- CD8- double negative (DN) or CD4+, but they never express heterodimeric CD8 molecules. Second, most V alpha24inv T cells are brightly stained by NKRP1-specific mAb but not by mAb directed against other type II transmembrane proteins of the NK complex. Third, DN and particularly CD4+ V alpha24inv T cells are greatly enriched for IL-4 producers. The concomitant expression of highly conserved TCRs of a particular set of NK markers and of Th2 cytokines in human and murine alphabeta T cells suggests a coordinate acquisition of these phenotypic and functional properties. Furthermore, the relatively high frequency of human V alpha24inv T cells, which are presently shown to represent on average 1/500 PBL, and the high interindividual variations of the size of this cell subset under physiologic conditions go for a major role played by alphabeta T cells carrying invariant TCR in a large array of immune responses. 相似文献
35.
Six measures of divergent thinking were administered to 825 men ranging in age from 17 to 101 over the period from 1959 to 1972; repeat administrations were given to a subset of 278 men after a 6-year interval. Cross-sectional analyses showed curvilinear trends, with an increase in scores for men under 40 and a decline thereafter. Repeated measures analyses on subjects initially aged 33 to 74 generally replicated this finding, whereas cross-sequential analyses suggested a decline for all cohorts tested at a later time. Additional analyses suggested that not all of the decline could be attributed to reduced speed of response production. These longitudinal findings confirm earlier cross-sectional reports of decline in divergent thinking abilities with age. (PsycINFO Database Record (c) 2010 APA, all rights reserved) 相似文献
36.
Pursuing optimal solutions for large scale transmission network planning problems is a formidable task due to their combinatorial nature and also due to the nonconvexities involved. Successful approaches using hierarchical Benders decomposition incur in a high computational cost mainly due to the need to solve a large integer program (the investment sub-problem) for every Benders iteration. In this work the authors propose to use heuristics within the decomposition framework, therefore avoiding to solve to optimality each integer sub-problem. The global computational effort is substantially reduced, and allows coping with large problems that would be intractable using classical combinatorial techniques. Case studies with the 6 bus Garver test system and a reduced Southeastern Brazilian power network are presented and discussed 相似文献
37.
H Komuro MB Valentine ST Rowe VJ Kidd S Makino GM Brodeur SL Cohn AT Look 《Canadian Metallurgical Quarterly》1998,33(11):1695-1698
Artificial illumination is an important factor in the management of layers. In this study, a new monochromatic light system was developed for egg layers. Prelaying pullets (Lohmann) were marked and housed in nine light and temperature control rooms (15 battery cages, 3 hens per cage; n = 45), divided into three light treatments: 0.1 and 0.01 W/m2 light intensity using light emitting diode (LED) lamps and 0.1 W/m2 using mini-fluorescent bulbs (PL) (control). In each of the LED rooms, three wavelengths were tested: 560 (n = 9), 660 (n = 9), 880 (n = 6), and 660 intermitted lighting (15 min light 45 min dark, 660IN) (n = 9). Birds were exposed to 12 h light and 12 h of darkness using PL lamps. At 21 wk of age, the light period was increased to 12.75 h by using 5.5 h of LED lamps and 7.25 of PL light source for Groups 1 and 2, the third group received 12.75 h of PL light. Until 28 wk of age, light hours increased by 0.5 h/w using LED light for Groups 1 and 2 and PL source for the third group, reaching 16 h of light at 28 wk of age. Egg production and feed consumption were recorded daily; egg components were recorded weekly for 10 mo. A significant reduction in egg production was observed in all 880nm groups; no differences in egg production and quality were found in the other groups. Feed consumption was significantly lower by 7% in all 0.01 W/m2 groups. We suggest that an important reduction in rearing costs of laying hens may be obtained by using this system. 相似文献
38.
Dominguez M. Pons-Nin J. Ricart J. Bermejo A. Costa E.F. Morata M. 《IEEE transactions on circuits and systems. I, Regular papers》2005,52(11):2286-2297
The aim of this paper is the analysis, simulation, and experimental verification of the /spl Sigma/-/spl Delta/ pulsed digital oscillator (PDO) topology. As it has been shown in previous works, the oscillation frequency and output spectrum in the PDO depend on the sampling frequency, the natural frequency of the microelectromechanical systems (MEMS) resonator and its damping factor. Here, extensive discrete-time simulations have been carried out which show that the normalized oscillation frequency as a function of the normalized natural frequency of the resonator is very similar to a distorted Devil's Staircase fractal. This nonlinear behavior is a direct consequence of the damping losses of the MEMS resonator. Analytical conditions for a perfect oscillation at the natural frequency of the resonator are also calculated. For this set of what we call "perfect" frequencies, it is also shown that the energy transfer from the electrical to the mechanical domain is maximum. Then a more generalized structure of the oscillator is considered and the drawn conclusions are tested against experimental results obtained from an oscillator prototype which uses a MEMS resonator with thermoelectric actuation and piezoresistive position sensing. 相似文献
39.
L. D. Monti P. E. Sandoli S. Costa V. C. Phan P. M. Piatti 《Journal of Automated Methods and Management in Chemistry》1993,15(5):177-181
Intermediate products of the metabolism of glucose, fat and amino-acid are important in the evaluation of such metabolic disorders as diabetes mellitus, liver disease and metabolic acidosis. In the present study, methods for the measurement of intermediate metabolites (lactate, pyruvate, alanine, β-hydroxybutyrate and glycerol) have been adapted to a fast centrifugal analyzer: the COBAS FARA. Correlation coeffcients rangedfrom 0.90 to 0.99, compared to established manual spectrophotometric methods. Within-run coeffcients of variation (CVs) ranged between 2.9 and 8.8% at low levels, between 1.5 and 5.7% at medium levels and between 1.2 and 5.6% at high levels. Between-run CVs were between 4.0 and 15.0% at low levels, between 1.7 and 7.0% at medium levels and between 1.3 and 2.7% at high levels. These fluorimetric assays for the determination of intermediate metabolites on COBAS FARA (Roche) have a good sensitivity and precision, are less costly than manual methods and can be used on a routine basis. 相似文献
40.