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91.
Iterative optimization of high-affinity proteases inhibitors using phage display. 1. Plasmin 总被引:2,自引:0,他引:2
We generated a series of libraries having variants of the first Kunitz domain of human lipoprotein-associated coagulation inhibitor (LACI-D1, also known as tissue-factor pathway inhibitor-I) displayed on bacteriophage M13 as pIII-fusions. We varied LACI-DI iteratively in two regions: the P1 region (positions 10-21) and the "second loop", (positions 31-39), which together form one end of the domain. Display-phage library Lib#1 allows 31 200 amino-acid sequences in P1 region (residues 13, 16-19). Preliminary, we screened Lib#1 against human plasmin (PLA, EC 3.4.21.7) immobolized on agarose to enrich for phage displaying variants with PLA affinity. We introduced a 1600-fold increase in second-loop diversity (residues 31, 32, 34, 39) into the population of selectants from Lib#1, yielding Lib#2. Lib#2 (allowing approximately 50 million amino-acid sequences) was screened against PLA-agarose to isolate highest affinity binders. Protein EPI-P211, derived from the best isolate of Lib#2, inhibits PLA with Ki = 2 nM (at least 500-fold better than LACI-D1) and with high specificity. We used amino-acid sequences of PLA-binding selectants to design a PLA-biased library (Lib#3) which we screened against PLA. The protein EPI-P302 (derived from the best binder obtained from Lib#3) has Ki for PLA inhibition of 87 pM, which is 25-fold better than the first-round best binder and > or = 12 500-fold better than LACI-D1. EPI-P302 also shows very high specificity for PLA vs other human proteases and is resistant to inactivation by oxidants and extremes of temperature or pH. Thus, one can use selectants from one library to design target-tailored combinatorial libraries and obtain quite stable, highly specific, very high-affinity binding molecules while maintaining an essentially human framework. 相似文献
92.
GS Alexopoulos BS Meyers RC Young S Mattis T Kakuma 《Canadian Metallurgical Quarterly》1993,150(11):1693-1699
OBJECTIVE: The goals of this longitudinal investigation were 1) to study the rate of development of irreversible dementia in elderly depressed patients with a dementia syndrome that subsided after improvement of depression and 2) to compare it with that of depressed, never-demented patients. METHOD: The subjects were 57 elderly patients consecutively hospitalized for major depression. At entry into the study, 23 subjects also met criteria for "reversible dementia," while 34 were without dementia. After a systematic clinical evaluation, the subjects were followed up at approximately yearly intervals for an average of 33.8 months. RESULTS: Irreversible dementia developed significantly more frequently in the depressed group with reversible dementia (43%) than in the group with depression alone (12%). Survival analysis showed that the group with reversible dementia had a 4.69-times higher chance of having developed dementia at follow-up than the patients with depression alone. No clinical characteristics at entry into the study were found to discriminate the subjects who developed irreversible dementia during the follow-up period from those who remained nondemented. CONCLUSIONS: These findings suggest that geriatric depression with reversible dementia is a clinical entity that includes a group of patients with early-stage dementing disorders. Therefore, identification of a reversible dementia syndrome is an indication for a thorough diagnostic workup and frequent follow-ups in order to identify treatable neurological disorders. 相似文献
93.
94.
Ellison P 《Proceedings of the Institution of Mechanical Engineers. Part H, Journal of engineering in medicine》2012,226(3):246-255
Clinical studies indicate that range of motion until prosthetic impingement is important in understanding unexplained failures of hip resurfacings, yet the underlying biomechanical principles have received little attention. This study investigates the mathematical relationships between component design, position, patient bone geometry and range of motion in hip resurfaced prostheses. Variations in range of motion and impingement-free safe-zones for cup position were calculated using an established method of vector analysis that facilitated parametric analysis in a time efficient manner. The alpha angle, defined as the angle between the centreline of the femoral neck and the waist of the femoral head/neck junction, was used to represent the natural femoral neck. Range of motion and impingement-free safe-zones were inversely proportional to the alpha angle and cup inclusion angle. The size of the safe-zone was most sensitive to the alpha angle with a 6 degrees reduction, decreasing the range of cup positions without impingement by 80-100%. Lowering the upper limit of cup inclination from 55 degrees to 45 degrees reduced the range of cup positions that allow impingement-free motion by 47-94%. No common safe-zone was observed for the range of component sizes and positions investigated. This offers an explanation to why clinic studies have failed to associate outcome with standardised positioning criteria. 相似文献
95.
This investigation examined whether cognition serves as a direct factor, mediates, or moderates the relationship between stressful life events and Children's Depression Inventory (CDI; Kovacs, 1992) factor scores in urban, ethnic minority youth. Ninety-eight middle school students completed measures of stressful life events, cognition (cognitive triad and locus of control), and depressive symptoms. Results provided support for both mediating and moderating models of depression. Specifically, the cognitive triad mediated the relationship between stressful life events and depressive symptom categories of negative mood, ineffectiveness, and anhedonia, while it interacted with stressful life events to predict negative self-esteem. In addition, locus of control was found to interact with stressful life events to predict anhedonia. Finally, the diathesis-stress model was not supported when predicting the CDI Interpersonal Problems factor score, which taps externalizing behaviors. Implications for school psychology research and practice include the importance of including measures of stressful life events and cognitive styles in assessment of children's social-emotional functioning. In addition, results highlight the need to target children's perceptions for intervention since they appear to play a significant role in psychological reactions to stressors. (PsycINFO Database Record (c) 2011 APA, all rights reserved) 相似文献
96.
The purpose of this study was to see whether the social environment of the church is associated with changes in feelings of gratitude toward God over time. The theoretical thrust of the authors’ analyses is captured in the following conceptual linkages: (a) People who attend church frequently are more likely to believe that their congregation is highly cohesive; (b) individuals who worship in more cohesive congregations will receive more emotional support from fellow church members; (c) people who receive more emotional support from coreligionists will be more likely to feel they have a close spiritual connection with others; and (d) people who feel they are closely connected with other individuals will feel especially grateful to God. Data from a nationwide longitudinal survey of older adults provide support for each of these hypothesized relationships. (PsycINFO Database Record (c) 2010 APA, all rights reserved) 相似文献
97.
Anthony Moschetti Lucas N. Vine Kyle Lethcoe Ruben K. Dagda Patricia Ellison Robert O. Ryan 《Lipids》2020,55(2):141-149
Coenzyme Q10 (CoQ10) is a strongly hydrophobic lipid that functions in the electron transport chain and as an antioxidant. CoQ10 was conferred with aqueous solubility by incorporation into nanoparticles containing phosphatidylcholine (PtdCho) and apolipoprotein (apo) A-I. These particles, termed CoQ10 nanodisks (ND), contain 1.0 mg CoQ10/5 mg PtdCho/2 mg apoA-I (97% CoQ10 solubilization efficiency). UV/Vis absorbance spectroscopy of CoQ10 ND revealed a characteristic absorbance peak centered at 275 nm. Incorporation of CoQ10 into ND resulted in quenching of apoA-I tryptophan fluorescence emission. Gel filtration chromatography of CoQ10 ND gave rise to a single major absorbance peak and HPLC of material extracted from this peak confirmed the presence of CoQ10. Incubation of cultured cells with CoQ10 ND, but not empty ND, resulted in a significant increase in the CoQ10 content of mitochondria as well as enhanced oxidative phosphorylation, as observed by a ~24% increase in maximal oxygen consumption rate. Collectively, a facile method to solubilize significant quantities of CoQ10 in lipid nanoparticles has been developed. The availability of CoQ10 ND provides a novel means to investigate biochemical aspects of CoQ10 uptake by cells and/or administer it to subjects deficient in this key lipid as a result of inborn errors of metabolism, statin therapy, or otherwise. 相似文献
98.
CE Thomas DF Ohlweiler AA Carr TR Nieduzak DA Hay G Adams R Vaz RC Bernotas 《Canadian Metallurgical Quarterly》1996,271(6):3097-3104
alpha-Phenyl-tert-butyl nitrone (PBN) is a nitrone spin trap, which has shown efficacy in animal models of oxidative stress, including stroke, aging, sepsis, and myocardial ischemia/reperfusion injury. We have prepared a series of novel cyclic variants of PBN and evaluated them for radical trapping activity in vitro. Specifically, their ability to inhibit iron-induced lipid peroxidation in liposomes was assessed, as well as superoxide anion (O2(-.)) and hydroxyl radical ((.)OH) trapping activity as determined biochemically and using electron spin resonance (ESR) spectroscopy. All cyclic nitrones tested were much more potent as inhibitors of lipid peroxidation than was PBN. The unsubstituted cyclic variant MDL 101,002 was approximately 8-fold more potent than PBN. An analysis of the analogs of MDL 101,002 revealed a direct correlation of activity with lipophilicity. However, lipophilicity does not solely account for the difference between MDL 101,002 and PBN, inasmuch as the calculated octanol/water partition coefficient for MDL 101,002 is 1.01 as compared to 1.23 for PBN. This indicated the cyclic nitrones are inherently more effective radical traps than PBN in a membrane system. The most active compound was a dichloro analog in the seven-membered ring series (MDL 104,342), which had an IC50 of 26 mum, which was 550-fold better than that of PBN. The cyclic nitrones were shown to trap (.)OH with MDL 101,002 being 20 25 times more active than PBN as assessed using 2-deoxyribose and p-nitrosodimethylaniline as substrates, respectively. Trapping of (.)OH by MDL 101,002 was also examined by using ESR spectroscopy. When Fenton's reagent was used, the (.)OH adduct of MDL 101,002 yielded a six-line spectrum with hyperfine coupling constants distinct from that of PBN. Importantly, the half-life of the adduct was nearly 5 min, while that of PBN is less than 1 min at physiologic pH. MDL 101,002 also trapped the O2(-.) radical to yield a six-line spectrum with coupling constants very distinct from that of the (.)OH adduct. In mice, the cyclic nitrones ameliorated the damaging effects of oxidative stress induced by ferrous iron injection into brain tissue. Similar protection was not afforded by the lipid peroxidation inhibitor U74006F, thus implicating radical trapping as a unique feature in the prevention of cell injury. Together, the in vivo activity, the stability of the nitroxide adducts, and the ability to distinguish between trapping of (.)OH and O2(-.) suggest the cyclic nitrones to be ideal reagents for the study of oxidative cell injury. 相似文献
99.
P Mooney M Sarmiento JM Bishop N Biswas NM Cason L Dauwe J Godfrey VP Kenney R Pemper E Rojek RC Ruchti WD Shephard G Ginther RM Edelstein CP Forsyth K Gamarnik AE Kreymer RJ Lipton JM McQuade DM Potter JS Russ L Spiegel DE Johnson D Buchholz LM Cremaldi SW Delchamps HS Mao JL Rosen W Sakumoto RA Schluter SB Sontz C Winter 《Canadian Metallurgical Quarterly》1989,39(9):2494-2498