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41.
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
42.
Autologous or glutaraldehyde treated bovine pericardial valved patch was utilized for widening of the right ventricular outflow tract in 20 patients with tetralogy of Fallot (autologous pericardium group in 10 patients and bovine pericardium group in 10). Pericardial valve function of the both materials was evaluated by postoperative cardiac catheterization performed 1 year after the operation. There were no significant differences in pulmonary arterial and right ventricular pressures, and right ventricular ejection fraction and end-diastolic volume between the 2 groups. Pulmonary angiogram in the autologous pericardium group patients demonstrated the pulmonary regurgitation (PR) of grade 1 in 5 patients, grade 2 in 4 and grade 3 in 1. On the other hand, 1, 3 and 6 patients in the bovine pericardium group demonstrated no-PR, grade 1 PR and grade 2 PR, respectively. It was concluded that there were no significant differences between autologous and glutaraldehyde treated bovine pericardium as a material of valved patch for widening of the right ventricular outflow tract of tetralogy of Fallot. 相似文献
43.
The enzyme steroid 5 alpha-reductase (EC 1.3.99.5) is a component of an intercellular signaling pathway that determines cell fate in the primordium of the mammalian reproductive tract. During male phenotypic sexual differentiation, the dihydrotestosterone product of this enzyme binds to the androgen receptor and initiates development of the external genitalia and prostate. Genes encoding two isozymes of steroid 5 alpha-reductase with different biochemical properties and tissue distributions have recently been isolated. In the current study, we utilize in situ hybridization analysis to determine cell-type-specific expression patterns of the 5 alpha-reductase isozyme mRNAs in two androgen target tissues (regenerating ventral prostate and epididymis) and a peripheral tissue (liver). In regenerating ventral prostate, the type 1 mRNA is expressed in basal epithelial cells whereas expression of the type 2 mRNA is largely confined to stromal cells. These results were confirmed by immunohistochemical analysis and are consistent with distinct roles played by the isozymes in the prostate. In the epididymis, both 5 alpha-reductase isozyme mRNAs are expressed in epithelial cells. Only the type 1 mRNA is present in the liver. This mRNA is distributed in a striking spatial gradient extending from hepatocytes surrounding the portal triad (high expression) to those surrounding the central vein (low to absent expression). These findings demonstrate cell-type-specific expression of the steroid 5 alpha-reductase isozymes and underscore their distinct and overlapping functions in androgen physiology. 相似文献
44.
T Naranda WB Strong J Menaya BJ Fabbri JW Hershey 《Canadian Metallurgical Quarterly》1994,269(20):14465-14472
Translation initiation factor eIF-4B promotes the binding of mRNA to 40 S preinitiation complexes and together with eIF-4A possesses RNA helicase activity. To elucidate structural features involved in its function, a series of internal and C-terminal deletions, as well as point mutations, were constructed in the eIF-4B cDNA. The mutated cDNAs were expressed in transiently transfected COS-1 cells, and mutant forms of the factor were overproduced up to about 25-fold over endogenous eIF-4B levels. Inhibition of dihydrofolate reductase (DHFR) synthesis by high levels of eIF-4B variants was determined in vivo, and the binding of the eIF-4B forms to biotinylated RNA was measured in vitro. The results indicate that the N-terminal region containing the RNA binding motif with its RNP1 and RNP2 consensus elements is sufficient for inhibition of DHFR synthesis. Deletion of the RNP1 sequence abrogates RNA binding, but amino acid substitutions at conserved residues do not always inhibit RNA binding. Deletion of the DRYG domain near the middle of eIF-4B results in inhibition of RNA binding, but not of DHFR synthesis. Up to 164 residues of the C terminus are not required for RNA binding, but removal of 226 or more residues completely inhibits RNA binding, perhaps by the loss of two arginine-rich regions. The results suggest that both the RNA recognition motif and the arginine-rich region are required for stable RNA binding but that both are not necessary for in vivo inhibition of protein synthesis. 相似文献
45.
46.
AF Minnick MJ Roberts WB Young RM Kleinpell RJ Marcantonio 《Canadian Metallurgical Quarterly》1997,35(4):399-409
OBJECTIVES: Market forces make it essential to know what policies and actions influence patients' reports of hospital services. No studies have examined the role of patient characteristics, labor quality and staff characteristics, nonlabor resources, managerial practices, and employee attitudes within a single investigation. METHODS: The authors collected, simultaneously, data about labor, management and service processes, nonlabor resources, and employee attitudes on 117 nonintensive medical-surgical inpatient units in 17 hospitals selected from a pool of 69 institutions within a metropolitan area by a stratified random sample. Of the 2,595 patients who agreed to participate, 2,051 (79%) completed telephone interviews regarding their experiences with physical care, education, and pain management services within 26 days of hospital discharge. RESULTS: A significant amount of variation in patients' service reports was explained (adjusted R2 = 0.41 physical care, 0.35 pain management, 0.44 education). Although the predictors varied for each service report, patient characteristics, especially those related to personal resources, had a large explanatory role. A labor assignment pattern that could explain why earlier studies found labor quality and staff characteristics to have only a weak role in the prediction of patients' service reports was noted. CONCLUSIONS: The results related to patient characteristics may indicate opportunities to improve care by confronting service design strategies that erroneously rely on a homogeneous patient population. Measurement challenges identified by this study must be addressed to determine the role of labor quantity and staff characteristics. 相似文献
47.
A comprehensive model of the work–family interface was developed and tested. The proposed model extended prior research by explicitly distinguishing between work interfering with family and family interfering with work. This distinction allowed testing of hypotheses concerning the unique antecedents and outcomes of both forms of work–family conflict and a reciprocal relationship between them. The influence of gender, race, and job type on the generalizability of the model was also examined. Data were obtained through household interviews with a random sample of 631 individuals. The model was tested with structural equation modeling techniques. Results were strongly supportive. In addition, although the model was invariant across gender and race, there were differences across blue- and white-collar workers. Implications for future research on the work–family interface are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved) 相似文献
48.
49.
As part of a study of the possible application of polymerisable Langmuir-Blodgett (LB) films as ultra-fine-line e-beam resists, an investigation of the variation of film structure of 22-tricosenoic acid with differing deposition conditions has been made. Unexpected effects with significant implications for deposition speed and resist sensitivity have been observed, and the new techniques for film characterisation developed during the investigation have resulted in a revised model of deposition explaining the observed independence of the disorder causing optical scattering and the macroscopic features observed by polarised microscopy. 相似文献
50.