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81.
GH Perdew H Wiegand JP Vanden Heuvel C Mitchell SS Singh 《Canadian Metallurgical Quarterly》1997,36(12):3600-3607
Several oncogenic protein kinases including c-raf-1 and pp60(v-src) are known to directly interact with the 90 kDa heat shock protein (hsp90)/p50 complexes. Using a monoclonal antibody to detect p50 during a purification scheme, p50 was purified to homogeneity. Internal amino acid sequence information was obtained and used to clone a partial cDNA. Comparison of the p50 sequence to other cloned proteins revealed 89% homology with a glycosaminoglycan-binding protein and 54% homology with Drosophila cell cycle control protein (cdc) 37. Monoclonal and polyclonal antibodies were produced against a cleaved fusion protein that recognizes p50 with a high level of specificity. These antibodies recognize the 50 kDa protein present in c-raf-1 and pp60(v-src) complexes. No other proteins were recognized with these antibodies suggesting that p50 is a unique protein. Immunocytochemical visualization of p50 in NIH 3T3 cells indicates a primarily cytoplasmic localization around the nuclear membrane. A survey of p50 expression in murine tissues on a protein blot revealed the following relative levels of expression; thymus > spleen > brain > heart > kidney > liver > lung > skeletal muscle. These results link studies demonstrating complexation of certain kinases with hsp90/p50 in mammalian cells and a number of reports in yeast and Drosophila, demonstrating the importance of cdc37 in cell cycle and kinase function. 相似文献
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83.
The inherited retinal dystrophies represent a large and heterogenous group of hereditary neurodegenerations, for many of which, the molecular defect has been defined. However, the mechanism of cell death has not been determined for any form of retinal degeneration. The retinal degeneration slow (rds-/-) mutation of mice is associated with nondevelopment of photoreceptor outer segments and gradual death of photoreceptor cell bodies, attributed to the absence of the outer segment protein rds/peripherin. Here, we examined the effects of a transgene encoding normal rds/peripherin that had integrated into the X-chromosome in male and female rds-/- mutant retinas. In 2-month-old transgenic males and homozygous-transgenic females on rds-/-, we observed virtually complete rescue of both the outer segment nondevelopment and photoreceptor degeneration. In contrast, hemizygous-transgenic rds-/- female littermates showed patchy distributions of the transgene mRNA, by in situ hybridization analysis, and of photoreceptor cells that contain outer segments. This pattern is consistent with random inactivation of the X-chromosome and mosaic expression of the transgene. Surprisingly, we observed significant photoreceptor cell loss in both transgene-expressing and nonexpressing patches in hemizygous female retinas. These observations were supported by nuclease protection analysis, which showed notably lower than predicted levels of transgene mRNA in retinas from hemizygous females compared with male and homozygous female littermates. This phenotype suggests an important component of non-cell-autonomous photoreceptor death in rds-/- mutant mice. These results have significance to both the etiology and potential treatment of human inherited retinal degenerations. 相似文献
84.
KL Gillotte S Lund-Katz M de la Llera-Moya JS Parks LL Rudel GH Rothblat MC Phillips 《Canadian Metallurgical Quarterly》1998,39(10):2065-2075
African green monkeys fed fat-specific diets served as a model to investigate the effect of phospholipid acyl chain modification on high density lipoprotein (HDL)-mediated cellular cholesterol efflux. Diets enriched in saturated, monounsaturated, n-6 polyunsaturated, or n-3 polyunsaturated fats were provided during both low cholesterol and cholesterol-enriched stages; sera and HDL3 samples were obtained at specific points during the treatment period. Analysis of the HDL phospholipid composition revealed significant acyl chain modification, consistent with the respective fat-specific diet. Cholesterol efflux from mouse L-cell fibroblasts to HDL3 isolated from the specific diet groups was measured and revealed no differences in the abilities of the particles to accept cellular cholesterol; determination of the bidirectional flux of cholesterol between the cells and HDL3 species further demonstrated no effect of phospholipid acyl chain modification on this process. The effects of dietary modification of phospholipid acyl chains on cellular cholesterol efflux were directly examined by isolating the HDL phospholipid and combining it with human apolipoprotein A-I to form well-defined reconstituted HDL particles. These complexes did not display any differences with respect to their ability to stimulate cellular cholesterol efflux. Incubations with 5% sera further confirmed that the fat-specific diets do not influence cholesterol efflux. These results suggest that the established influences of specific dietary fats on the progression of atherosclerosis are due to effects on cholesterol metabolism other than the efflux of cellular cholesterol in the first step of reverse cholesterol transport. 相似文献
85.
V Papadopoulos EP Widmaier H Amri A Zilz H Li M Culty R Castello GH Philip R Sridaran K Drieu 《Canadian Metallurgical Quarterly》1998,24(3-4):479-487
In various steroidogenic cell models, mitochondrial preparations and submitochondrial fractions, the expression of the mitochondrial 18 kDa peripheral-type benzodiazepine receptor (PBR) protein confers the ability to take up and release, upon ligand activation, cholesterol. Thus, cholesterol becomes available to P450scc on the inner mitochondrial membrane. These in vitro studies were validated by in vivo experiments. Treatment of rats with ginkgolide B (GKB), specifically reduced the ligand binding capacity, protein, and mRNA expression of the adrenocortical PBR and circulating glucocorticoid levels. Treatment with GKB also resulted in inhibition of PBR protein synthesis and corticosterone production by isolated adrenocortical cells in response to ACTH. The ontogeny of both PBR binding capacity and protein directly paralleled that of ACTH-inducible steroidogenesis in rat adrenal cells and in rats injected with ACTH. In addition, the previously described suppression of luteal progesterone synthesis in the pregnant rat by continuous in vivo administration of a gonadotropin-releasing hormone agonist may be due to decreased luteal PBR ligand binding and mRNA. These results suggest that (i) PBR is an absolute prerequisite for adrenocortical and luteal steroidogenesis, (ii) regulation of adrenal PBR expression may be used as a tool to control circulating glucocorticoid levels and (iii) the stress hypo-responsive period of neonatal rats may result from decreased adrenal cortical PBR expression. 相似文献
86.
N.B.O.L. Pettit M. Willatzen L. Ploug-Sørensen 《International Journal of Refrigeration》1998,21(5):404-414
A mathematical model of an evaporator based on one-dimensional partial differential equations representing mass conservation, and tube wall energy has been formulated. These equations are then restructured and linked to a program data base of all major refrigerants and refrigerant mixtures. The result is a simulation model of an evaporator that is general and flexible. The model is tested over a wide range of operating conditions and a simple controller is implemented to demonstrate the effectiveness of the model for controller and systems design. 相似文献
87.
Food consumption was measured in six female and seven male hand-raised marsh harrier (Circus aeruginosus) nestlings. Females consumed on average 4,321 g and males consumed 3,571 g of food during the nestling stage from 0 to 36 d. Total consumption until 56 d was 6,960 g and 5,822 g for females and males, respectively. On the basis of Fisher's sex ratio theory, this food intake ratio of 0.46 (intake male/[intake male + female]) would explain the observed male-biased fledging sex ratio of 55% males in marsh harrier broods. Growth, gross energy intake, and metabolizable energy intake were measured, along with metabolism of the nestlings, enabling us to determine energy allocation. The assimilation quotient (Q = 0.72) did not differ systematically between the sexes. Differences in metabolic rates between males and females at 15 and 30 d of age were fully attributable to the difference in body mass. Sexual size dimorphism in marsh harriers (female body mass around 60 d of age is 1.28 times greater than male mass) did not fully explain the difference in food intake between male and female nestlings: an analysis of energy requirements for growth and body mass in 16 avian species shows that energy intake was less than proportional to the average body mass at release. The data presented in this study are in agreement with Fisher's theory of inverse proportionality between the sex-specific ratios of energy requirements for growth and of offspring numbers in the marsh harrier population. 相似文献
88.
89.
Analytical performance of the Tandem-R free PSA immunoassay measuring free prostate-specific antigen
DL Woodrum CM French TM Hill SJ Roman HL Slatore JL Shaffer LG York KL Eure KG Loveland GH Gasior PC Southwick LB Shamel 《Canadian Metallurgical Quarterly》1997,43(7):1203-1208
The analytical performance of the Tandem-R free PSA assay available from Hybritech Inc. was evaluated. Comparison of recoveries of purified free (unbound) prostate-specific antigen (PSA) diluted in female serum in the Tandem-R free PSA assay and the Tandem-R (total) PSA assay demonstrated a link in calibration between the assays and an accurate determination of percent free PSA. The cross-reactivity of the assay to purified PSA-alpha 1-antichymotrypsin was determined to be < 1%. The minimum-detectable concentration was < 0.05 microgram/L. The within-run and between-day CVs were < or = 5% for samples with > 0.3 microgram/L free PSA. Dilution and recovery showed no significant deviations from linearity across the assay range. The assay was insensitive to interference from blood components. The Tandem-R free PSA kit was shown to be an accurate, precise, and reliable assay for the measurement of free PSA. 相似文献
90.