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31.
32.
Retroviral display of antibody fragments; interdomain spacing strongly influences vector infectivity 总被引:1,自引:0,他引:1
S Ager BH Nilson FJ Morling KW Peng FL Cosset SJ Russell 《Canadian Metallurgical Quarterly》1996,7(17):2157-2164
Five different single-chain antibody fragments (scFv) against human cell-surface antigens were displayed on murine ecotropic retroviral vectors by fusing them to the Moloney SU envelope glycoprotein. The spacing between the scFv and the SU glycoprotein was varied by fusing the scFv to residue +7 or to residue +1 of Moloney SU and by inserting linker sequences of different lengths between the domains. All of the chimeric envelopes were efficiently incorporated into vector particles and could bind to human cells through their displayed antibody fragments, but did not infect them. The spacing between the scFvs and the SU glycoproteins had no significant effect on the efficiency of envelope expression or viral incorporation and did not affect the binding properties of the chimeric envelopes, nor did it influence the efficiency of targeted gene delivery to human cells by scFv-displaying vectors. However, on murine fibroblasts the infectivity of vectors incorporating the chimeric envelopes was strongly influenced by the length of the interdomain spacer. The titers were very low when the single-chain antibodies were fused through a tripeptide linker to SU residue +7 and were greatly enhanced (up to 10(5)-fold) when they were fused to SU residue +1 through a heptapeptide linker. These results point to the importance of steric interactions between the domains of chimeric envelope glycoproteins and may have implications for retroviral vector design for human gene therapy. 相似文献
33.
For viruses made of nucleic acid and protein, the structure of the protein outer shell has, in the past, been found to be uniquely determined by the viral genome. However, here, non-denaturing agarose gel electrophoresis of bacteriophage T7 reveals two states of the mature T7 capsid; the conditions of growth are found to alter the population by T7 of these two electrophoretically defined states. Both states have been previously observed for a genetically altered T7 and they are observed here for wild-type T7. The average electrical surface charge density of a bacteriophage particle (delta) determines its state; the delta of particles in both states is negative. For a given condition of growth, the population of these two states is influenced by the extent to which the major T7 outer shell protein, p10A, is accompanied by its minor readthrough variant, p10B. Comparison of the two electrophoretic states reveals the following. (1) No difference in radius is present in the outer shell (+/-2%). (2) As the pH of electrophoresis is either increased or decreased from neutrality, the state becomes more highly populated for which delta is greater in magnitude (state 1). By changing the pH, some T7 particles are made to change state. (3) Particles in state 1 adsorb less quickly to host cells than do the particles in the alternative state (state 2). This latter observation suggests the hypothesis that state 1 evolved to reduce the probability of re-initiating an infection when conditions are not favorable for growth. This hypothesis is supported by the observation that, as conditions of growth become apparently more unfavorable, progeny increasingly populate state 1. 相似文献
34.
Glycol ethers such as 2-ethoxyethanol (EE) are widely used as solvents because they are miscible in aqueous and organic solutions. Toxic effects of EE in rodents include teratogenicity, fetotoxicity, hematotoxicity, and testicular atrophy. The purpose of this study was to determine the effect of dose on the absorption, metabolism, and excretion of 2-ethoxy [U-14C]ethanol by F344/N rats after inhalation exposure. Rats were exposed to either 5 ppm EE for 5 hr 40 min or 46 ppm EE for 6 hr. The uptake and metabolism of EE were linear in the concentration range studied. Significant percentages of the retained doses were exhaled during (22%) and after exposure (16%) as 14CO2. Forty-six percent of the retained dose was excreted in the urine. Approximately 10% of the retained dose was detected in the carcass 66 hr after exposure. The major urinary metabolite was ethoxyacetic acid (EAA), the toxic metabolite of EE. The amount of EAA excreted was linearly related to exposure concentration. Ethylene glycol and N-ethoxyacetyl glycinate were identified as minor metabolites excreted in the urine. The results of this study suggest that the toxicity of inhaled EE should be directly proportional to the exposure concentration up to 46 ppm if the toxicity of EE is due to EAA. 相似文献
35.
A series of experiments were carried out on three commercial steels to explore the possibility for characterizing tempered martensite embrittlement (TME) by Rockwell, macro-, and micro-Vickers hardness tests. The results indicate distinct hardness peaks in two steels and an inflection in the other around the TME temperature. A new analytical approach for examining the slope of hardness-tempering temperature plots appears to reveal the TME phenomenon more sensitively. Dilatometric examinations substantiate that the temperature of hardness peak/inflection occurs beyond the second stage of tempering. 相似文献
36.
JM Karski SJ Teasdale P Norman J Carroll K VanKessel P Wong MF Glynn 《Canadian Metallurgical Quarterly》1995,110(3):835-842
This prospective, double-blind, randomized trial assessed the effectiveness of high-dose tranexamic acid given in the preoperative period on blood loss in patients undergoing cardiopulmonary bypass. One hundred fifty patients scheduled to undergo cardiac operations with cardiopulmonary bypass were randomized into three groups of equal size. The first group received 10 gm of tranexamic acid intravenously over 20 minutes before sternotomy and a placebo infusion over 5 hours. The second group received 10 gm of tranexamic acid over 20 minutes and then another 10 gm infused intravenously over 5 hours. The control group received a placebo bolus and a placebo infusion over 5 hours (0.9% normal saline solution). The blood loss after the operation was measured at 6 hours and 24 hours. The homologous blood and blood products given during and up to 48 hours after operation were recorded. Eighteen percent of the control group patients shed more than 750 ml blood in 6 hours compared with only 2% in both tranexamic acid groups. Patients who shed more than 750 ml blood required 93% more red blood cell transfusions than patients without excessive bleeding. Tranexamic acid (10 gm) given intravenously in the period before cardiopulmonary bypass reduced blood loss over 6 hours by 50% and over 24 hours by 35%. Continued tranexamic acid infusion (10 gm over 5 hours) did not reduce bleeding further. There was no difference in the coagulation profile before operation between patients with and without excessive bleeding. However, coagulation tests done in the postoperative period indicated ongoing fibrinolysis and platelet dysfunction in patients with excessive bleeding. 相似文献
37.
The concept of control signals is fundamental to digital design. It evokes the notion of one or more signals that influence computation on other signals in a system, and regulate the behavior of the system. A formal definition of control signals is presented. It is argued that knowledge of the control signals in a complex design such as a microprocessor or a computer is crucial to the comprehension and specification of the design and that knowledge of the control signals of a design leads to a more efficient hardware implementation. The identification of control signals in two microprocessors are presented as examples 相似文献
38.
The superplastic properties of a rapidly solidified, high strength P/M Al alloy and the same alloy reinforced with SiC particulates
(SiC
p
) have been studied. To prepare superplastic test materials, a matrix alloy powder of composition 7.2Zn-2.4Mg-2Cu-0.2Zr-0.12Cr-0.2Co
(Kaiser PM-64) and the powder mixed with 10 to 20 vol pct SiC
p
(~5 μm diameter) were thermomechanically processed to very fine equiaxed grain structures of ~6 μm and ~8 μm, respectively.
Superplasticity in these materials was evaluated by characterizing (1) high temperature stability, (2) dynamic grain growth,
(3) strain rate sensitivity, (4) flow stress behavior, (5) cavitation and cavitation control, and (6) total superplastic strain.
It was observed that the PM-64 alloy could achieve a total elongation of over 800 pct, while the SiCp reinforced alloy could attain an elongation greater than 500 pct before failure. Also, it was shown that with the use of
hydrostatic pressure during superplastic flow, cavitation could be controlled. Observations were made of the effect SiC
p
reinforcement particles had on the superplastic flow stress behavior. Interpretations are proposed to explain the role of
particulates during superplastic straining. 相似文献
39.
40.