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111.
N Raskin A Jakubowski ID Sizing DL Olson SL Kalled CA Hession CD Benjamin DP Baker LC Burkly 《Canadian Metallurgical Quarterly》1998,161(7):3474-3483
The IL receptor common gamma (gamma c) chain is required for the formation of high affinity cytokine receptor complexes for IL-2, IL-4, IL-7, IL-9, and IL-15, and for signals regulating cell survival, growth, and differentiation. Our current understanding of how gamma c chain associates with multiple ligands and receptor subunits is drawn largely from its structural homology to the human growth hormone (hGH) receptor and known structure of the hGH/hGH receptor complex. These receptors share distinct features in their extracellular portions and are believed to function by a mechanism of ligand-induced association of receptor subunits. Here, we report the first directed mutational analysis of the human gamma c chain by alanine scanning conducted across seven regions likely to contain residues required for intermolecular contact. Functionally distinct, neutralizing anti-gamma c mAbs were employed to define critical residues. One particular mAb, CP.B8, unique in its ability to inhibit IL-2-, IL-4-, IL-7-, and IL-15-induced proliferation and high affinity cytokine binding of normal T cells as an intact mAb and as a Fab fragment, localized critical residues to four noncontinuous stretches, namely residues in loops AB and EF of domain 1, in the interdomain segment, and in loop FG of domain 2. Notably, these residues form a contiguous patch on the gamma c chain surface in a three-dimensional structural model. These results provide functional evidence for the location of contact points on gamma c chain required for its association with multiple ligands. 相似文献
112.
JM Cho AJ LaPorta JR Clark MJ Schofield SL Hammond PL Mallory 《Canadian Metallurgical Quarterly》1994,8(12):1380-3; discussion 1383-4
The clinical observation that a laparoscopic cholecystectomy is a minimally invasive operation has not been demonstrated on a biochemical basis. Interleukin-6, a known endogenous pyrogen and hepatocyte-stimulating protein, correlates with the significance of surgical trauma. Utilizing the IL-6 immunoassay, we studied this biochemical parameter of trauma to compare its response in laparoscopic vs open cholecystectomy. Sixteen patients who underwent only laparoscopic cholecystectomy showed peak IL-6 concentrations of 51 pg/ml (22-86) vs a peak IL-6 concentration of 124 pg/ml (56-225) for open cholecystectomy. Six additional patients who underwent an ERCP followed by laparoscopic cholecystectomy showed a dramatic rise in peak IL-6 concentration to 315 pg/ml (15-634). These results biochemically confirm the true minimal invasiveness of laparoscopic cholecystectomy. The findings in the ERCP-followed-by-laparoscopic-cholecystectomy group support the theory that two invasive procedures in close proximity may prime the cytokine system in its response to surgical trauma. 相似文献
113.
A probabilistic identification matrix for campylobacteria, comprising 76 phenotypic characters and 37 taxa, is described. The accuracy and integrity of the matrix was evaluated using established computer-assisted methods. Certain taxa (for example, Campylobacter concisus and Camp. gracilis) demonstrated significant phenotypic diversity; previous data corroborated these findings. Differentiation between a few pairs of taxa proved difficult, although discriminatory characteristics were noted in each of these cases. The results indicate that most campylobacteria can be identified accurately and objectively with phenotypic tests when probabilistic methods of data assessment are employed. 相似文献
114.
OBJECTIVE: To determine the prevalence of antibodies to filaggrin in a cross sectional sample of patients with rheumatoid arthritis (RA). METHODS: Filaggrin from human skin was either extracted with 0.05% Nonidet P-40 and then partially purified by precipitating in ethanol and resuspending in water (Nonidet preparation) or extracted with 9 M urea and then purified by sequential fractionation on a DEAE Sephadex column and on a strong cation exchange column (purified preparation). Antibodies to filaggrin were detected using immunoblotting techniques with sera diluted 1:50. Antikeratin antibodies (AKA) were detected using indirect immunofluorescence microscopy on sections of rat esophagus. RESULTS: Antibodies to filaggrin were detected in 5 of 30 sera of patients with RA using filaggrin from the Nonidet preparation and 6 of 49 sera using filaggrin from the purified preparation. AKA were detected in 13 of 40 sera. A positive correlation existed between the presence of AKA and the presence of antibodies to filaggrin using the purified preparation (p=0.017). CONCLUSION: These data indicate that the reactivity of RA sera with filaggrin is not identical to the presence of AKA and is variable depending upon the preparation of filaggrin used. The diagnostic value of antibodies to filaggrin remains to be proven. 相似文献
115.
A healthy, young adolescent girl developed primary pneumococcal peritonitis, an infection rarely reported in this age group in North America. Her course was further complicated by exudative pleural effusion and pneumonia despite receiving 10 days of clindamycin therapy. Laparascopy proved useful in making the initial diagnosis, but may have contributed to the pathogenesis of the pulmonary process. Case presentation, management, and etiology are discussed. 相似文献
116.
The Drosophila Medea gene is required downstream of dpp and encodes a functional homolog of human Smad4 总被引:1,自引:0,他引:1
JB Hudson SD Podos K Keith SL Simpson EL Ferguson 《Canadian Metallurgical Quarterly》1998,125(8):1407-1420
The Transforming Growth Factor-beta superfamily member decapentaplegic (dpp) acts as an extracellular morphogen to pattern the embryonic ectoderm of the Drosophila embryo. To identify components of the dpp signaling pathway, we screened for mutations that act as dominant maternal enhancers of a weak allele of the dpp target gene zerkn?llt. In this screen, we recovered new alleles of the Mothers against dpp (Mad) and Medea genes. Phenotypic analysis of the new Medea mutations indicates that Medea, like Mad, is required for both embryonic and imaginal disc patterning. Genetic analysis suggests that Medea may have two independently mutable functions in patterning the embryonic ectoderm. Complete elimination of maternal and zygotic Medea activity in the early embryo results in a ventralized phenotype identical to that of null dpp mutants, indicating that Medea is required for all dpp-dependent signaling in embryonic dorsal-ventral patterning. Injection of mRNAs encoding DPP or a constitutively activated form of the DPP receptor, Thick veins, into embryos lacking all Medea activity failed to induce formation of any dorsal cell fates, demonstrating that Medea acts downstream of the thick veins receptor. We cloned Medea and found that it encodes a protein with striking sequence similarity to human SMAD4. Moreover, injection of human SMAD4 mRNA into embryos lacking all Medea activity conferred phenotypic rescue of the dorsal-ventral pattern, demonstrating conservation of function between the two gene products. 相似文献
117.
De novo protein design: fully automated sequence selection 总被引:1,自引:0,他引:1
The first fully automated design and experimental validation of a novel sequence for an entire protein is described. A computational design algorithm based on physical chemical potential functions and stereochemical constraints was used to screen a combinatorial library of 1.9 x 10(27) possible amino acid sequences for compatibility with the design target, a betabetaalpha protein motif based on the polypeptide backbone structure of a zinc finger domain. A BLAST search shows that the designed sequence, full sequence design 1 (FSD-1), has very low identity to any known protein sequence. The solution structure of FSD-1 was solved by nuclear magnetic resonance spectroscopy and indicates that FSD-1 forms a compact well-ordered structure, which is in excellent agreement with the design target structure. This result demonstrates that computational methods can perform the immense combinatorial search required for protein design, and it suggests that an unbiased and quantitative algorithm can be used in various structural contexts. 相似文献
118.
SJ Dowell AL Bishop SL Dyos AJ Brown MS Whiteway 《Canadian Metallurgical Quarterly》1998,150(4):1407-1417
The mating pathway of Saccharomyces cerevisiae is widely used as a model system for G protein-coupled receptor-mediated signal transduction. Following receptor activation by the binding of mating pheromones, G protein betagamma subunits transmit the signal to a MAP kinase cascade, which involves interaction of Gbeta (Ste4p) with the MAP kinase scaffold protein Ste5p. Here, we identify residues in Ste4p required for the interaction with Ste5p. These residues define a new signaling interface close to the Ste20p binding site within the Gbetagamma coiled-coil. Ste4p mutants defective in the Ste5p interaction interact efficiently with Gpa1p (Galpha) and Ste18p (Ggamma) but cannot function in signal transduction because cells expressing these mutants are sterile. Ste4 L65S is temperature-sensitive for its interaction with Ste5p, and also for signaling. We have identified a Ste5p mutant (L196A) that displays a synthetic interaction defect with Ste4 L65S, providing strong evidence that Ste4p and Ste5p interact directly in vivo through an interface that involves hydrophobic residues. The correlation between disruption of the Ste4p-Ste5p interaction and sterility confirms the importance of this interaction in signal transduction. Identification of the Gbetagamma coiled-coil in Ste5p binding may set a precedent for Gbetagamma-effector interactions in more complex organisms. 相似文献
119.
GS Ogg X Jin S Bonhoeffer PR Dunbar MA Nowak S Monard JP Segal Y Cao SL Rowland-Jones V Cerundolo A Hurley M Markowitz DD Ho DF Nixon AJ McMichael 《Canadian Metallurgical Quarterly》1998,279(5359):2103-2106
Although cytotoxic T lymphocytes (CTLs) are thought to be involved in the control of human immunodeficiency virus-type 1 (HIV-1) infection, it has not been possible to demonstrate a direct relation between CTL activity and plasma RNA viral load. Human leukocyte antigen-peptide tetrameric complexes offer a specific means to directly quantitate circulating CTLs ex vivo. With the use of the tetrameric complexes, a significant inverse correlation was observed between HIV-specific CTL frequency and plasma RNA viral load. In contrast, no significant association was detected between the clearance rate of productively infected cells and frequency of HIV-specific CTLs. These data are consistent with a significant role for HIV-specific CTLs in the control of HIV infection and suggest a considerable cytopathic effect of the virus in vivo. 相似文献
120.