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31.
We describe the principles of a new generation of sequential or simultaneous time-resolved fluoroimmunoassays, namely, simple, rapid, liquid-phase non-separation procedures which may be applied to the measurement of urinary steroid and drug metabolites. As an example, a method for the measurement of estrone-3-glucuronide in undiluted urine is reported. This method has a similar sensitivity, specificity and accuracy to a conventional separation fluoroimmunoassay or radioimmunoassay but in terms of speed, convenience, precision, reliability and clinical utility the new method has many advantages. The labelled antigen is a novel fluorescent europium chelate covalently linked to estrone-3-glucuronide. The antibody-binding reaction involves the incubation of the labelled antigen (2ng) with a limited concentration of polyclonal or monoclonal antibodies to estrone-3-glucuronyl-6-BSA and an aliquot of standard or sample (undiluted urine; 10 μl) in microtitre wells. After a 10 min incubation, the fluorescence which emanates from the antibody-free label is measured in a time-resolved fluorometer and is proportional to the concentration of estrone-3-glucuronide in the standard or sample. The method may be applied for the monitoring of ovarian function in women. 相似文献
32.
Metabolism of larger high density lipoproteins accumulating in some families of baboons fed a high cholesterol and high saturated fat diet 总被引:1,自引:0,他引:1
R S Kushwaha D M Foster V N Murthy K D Carey H C McGill 《Journal of lipid research》1989,30(8):1147-1159
Progeny of certain baboon sires accumulate lipoproteins in high density lipoprotein-1 (HDL1) when challenged with a high cholesterol, high saturated fat diet. These studies were conducted to determine the apoprotein composition and metabolic fate of HDL1 in the plasma. HDL1 particles containing apoA-I with and without apoE were detected. The majority of particles, however, contained apoA-I without any detectable apoE. To determine the metabolic fate of HDL1 in plasma, HDL1 labeled with iodinated apoA-I from animals with high levels of HDL1 and iodinated apoA-I-labeled autologous HDL were coinjected into both high and low HDL1 animals. The data for the decay of radioactivity in HDL1 and HDL were analyzed by multicompartment modelling. The radioactivity from HDL1 was cleared from the plasma either via direct removal (9.1 +/- 4.7% in low and 21.7 +/- 8.3% in high HDL1 animals) or via its conversion to HDL. A large proportion of radioactivity from HDL1 was rapidly transferred to HDL directly or metabolized via an intermediate compartment. Most of the radioactivity from apoE-poor HDL1, however, was transferred to HDL. Both high and low HDL1 animals catabolized HDL1 and HDL similarly. Low HDL1 animals transferred HDL1 radioactivity to HDL much faster. No detectable radioactivity from HDL was transferred to HDL1. Thus, HDL1 that accumulates in high HDL1 animals is mainly a precursor for HDL. Our hypothesis is that this accumulation of HDL1 is due to the slower cholesteryl ester transfer from HDL to lower density lipoproteins, thus affecting reverse cholesterol transport in high HDL1 baboons. 相似文献
33.
An amino-terminal fragment of GAL4 binds DNA as a dimer 总被引:51,自引:0,他引:51
M Carey H Kakidani J Leatherwood F Mostashari M Ptashne 《Journal of molecular biology》1989,209(3):423-432
34.
Detailed comparisons of aortic valvular flow using saline, with that using a glycerin-based blood analog in a pulse duplicator are reported. The experiments were carried out to determine whether exposure to glycerin caused stiffening of bioprosthetic valve leaflets. For two pericardial bioprostheses and for a mechanical valve we observed a fluid-dependent systolic volume flow, a fluid-dependent regurgitation volume, and fluid-dependent systolic pressure differences. Volume flow changes, both forward and reverse, are independent of valve type. The observed pressure differences, while proportional to fluid density for the mechanical valve, are fluid dependent in a more complicated way for the pericardial valves. However, no trend of changing valvular performance was observed over as much as 80 days of glycerin exposure, indicating that it is unlikely that the fluid-dependent performance was caused by glycerin absorption by the valve leaflets. We conclude that valid performance comparisons between mechanical and bioprosthetic valves may be made using a glycerin-based fluid. Furthermore, it appears that any detailed analysis of the physical mechanisms of valvular flow dissipation will require a properly matched blood analog. 相似文献
35.
The influence of tonically active submucosal neurons on basal ion transport was studied using sheets of guinea pig ileum set up in flux chambers. Tetrodotoxin evoked an immediate and sustained decrease in short-circuit current that was sustained for 60 minutes compared with control tissues in which basal currents gradually decreased over time. Time-dependent changes in basal short-circuit currents in tissues treated with atropine were not significantly different from control tissues. The decrease in short-circuit current after tetrodotoxin resulted from a greater increase in net chloride absorption than sodium absorption. Changes in net sodium and chloride transport were due to an increase in the mucosal-to-serosal fluxes of these ions. The results suggest that tonic activity of submucosal neurons limits the absorptive capacity of the guinea pig ileum. 相似文献
36.
Molecular genetic study of the frequency of monosomy 22q11 in DiGeorge syndrome 总被引:10,自引:2,他引:8
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A. H. Carey D. Kelly S. Halford R. Wadey D. Wilson J. Goodship J. Burn T. Paul A. Sharkey J. Dumanski M. Nordenskjold R. Williamson P. J. Scambler 《American journal of human genetics》1992,51(5):964-970
It is well established that DiGeorge syndrome (DGS) may be associated with monosomy of 22q11-pter. More recently, DNA probes have been used to detect hemizygosity for this region in patients with no visible karyotypic abnormality. However, DGS has also been described in cases where the cytogenetic abnormality does not involve 22q11; for instance, four cases of 10p- have been reported. In this study we have prospectively analyzed patients, by using DNA markers from 22q11, to assess the frequency of 22q11 rearrangements in DGS. Twenty-one of 22 cases had demonstrable hemizygosity for 22q11. Cytogenetic analysis had identified interstitial deletion in 6 of 16 cases tested; in 6 other cases no karyotype was available. When these results are combined with those from our previous studies, 33 of 35 DGS patients had chromosome 22q11 deletions detectable by DNA probes. 相似文献
37.
R. A. Newton S. L. Phipps T. P. Flanigan N. R. Newberry J. E. Carey †C. Kumar ‡B. McDonald ‡C. Chen J. M. Elliott 《Journal of neurochemistry》1996,67(6):2521-2531
Abstract: Stable transfection of the human neuroblastoma cell line SH-SY5Y with the human 5-hydroxytryptamine2A (5-HT2A) or 5-HT2C receptor cDNA produced cell lines demonstrating ligand affinities that correlated closely with those for the corresponding endogenous receptors in human frontal cortex and choroid plexus, respectively. Stimulation of the recombinant receptors by 5-HT induced phosphoinositide hydrolysis with higher potency but lower efficacy at the 5-HT2C receptor (pEC50 = 7.80 ± 0.06) compared with the 5-HT2A receptor (pEC50 = 7.30 ± 0.08). Activation of the 5-HT2A receptor caused a transient fourfold increase in intracellular Ca2+ concentration. Whole-cell recordings of cells clamped at ?50 mV demonstrated a small inward current (2 pA) in response to 10 µM 5-HT for both receptors. There were no differences in potency or efficacy of phosphoinositide hydrolysis among four hallucinogenic [d-lysergic acid diethylamide (LSD), 1-(4-iodo-2,5-dimethoxyphenyl)-2-aminopropane (DOI), 5-methoxy-N,N-dimethyltryptamine, and mescaline] and three nonhallucinogenic drugs (m-chlorophenylpiperazine, quipazine, and ergotamine). Comparison of equipotent doses producing 20% of the maximal response induced by 5-HT revealed selective activation of the 5-HT2A receptor by LSD and to a lesser degree by DOI, mescaline, and ergotamine. Quipazine and 5-methoxy-N,N-dimethyltryptamine were relatively nonselective, whereas m-chlorophenylpiperazine selectively activated the 5-HT2C receptor. It is unlikely therefore that hallucinosis is mediated primarily by activity at the 5-HT2C receptor, whereas activity at the 5-HT2A receptor may represent an important but not unique mechanism associated with hallucinogenic drug action. 相似文献
38.
Exclusion of epidermal growth factor and high-resolution physical mapping across the Rieger syndrome locus. 总被引:5,自引:1,他引:4
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E. V. Semina N. A. Datson N. J. Leysens B. U. Zabel J. C. Carey G. I. Bell P. Bitoun C. Lindgren T. Stevenson R. R. Frants G. van Ommen J. C. Murray 《American journal of human genetics》1996,59(6):1288-1296
We have evaluated the 4q25-4q26 region where the autosomal dominant disorder Rieger syndrome has been previously mapped by linkage. We first excluded epidermal growth factor as a candidate gene by carrying out SSCP analysis of each of its 24 exons using a panel of seven unrelated individuals with Rieger syndrome. No evidence for etiologic mutations was detected in these individuals, although four polymorphic variants were identified, including three that resulted in amino acid changes. We next made use of two apparently balanced translocations, one familial and one sporadic, to identify a narrow physical localization likely to contain the gene or to be involved in regulation of gene function. Somatic cell hybrids were established from individuals with these balanced translocations, and these hybrids were used as a physical mapping resource for, first, preliminary mapping of the translocation breakpoints using known sequence tagged sites from chromosome 4 and then, after creating YAC and cosmids contigs encompassing the region, for fine mapping of those breakpoints. A cosmid contig spanning these breakpoints was identified and localized the gene to within approximately 150 kb of D4S193 on chromosome 4. The interval between the two independent translocations is approximately 50 kb in length and provides a powerful resource for gene identification. 相似文献
39.
James H. Asher Jr. Ronald W. Harrison Robert Morell Melisa L. Carey Thomas B. Friedman 《Genomics》1996,34(3):285
Waardenburg syndrome type 1 is caused by mutations inPAX3.Over 50 humanPAX3mutations that lead to hearing, craniofacial, limb, and pigmentation anomalies have been identified. APAX3mutant allele, segregating in a family, can show reduced penetrance and variable expressivity that cannot be explained by the nature of the mutation alone. TheMus musculus Pax3mutationSpd(Splotch-delayed, Pax3[formula]), coisogenic on the C57BL/6J (B6) genetic background, produces in heterozygotes a white belly spot with 100% penetrance and very few other anomalies. By contrast, manySpd/+ BC1progeny [F1♀Spd/+ (♀Spd/+ B6× ♂ +/+Mus spretus) × ♂ +/+ B6] exhibit highly variable craniofacial and pigmentary anomalies. Of the BC1Spd/+ progeny, 23.9% are estimated to be nonviable, and 32.1% are nonpenetrant for the white belly spot. The penetrance and expressivity of theSpd/+ genotype are controlled in part by the genetic background and the sex of the individual. A minimum of two genes interact withSpdto influence the craniofacial features of these mice. One of these genes may be either X-linked or sex-influenced, while the other is autosomal. TheA-locus (Agouti) or a gene closely linked toAalso plays a role in determining craniofacial features. At least one additional gene, possibly theA-locus or a gene linked toA,interacts withSpdand determines the presence and size of the white belly spot. The viability of BC1mice is influenced by at least three factors:Spd,A-locus alleles or a gene closely linked to theA-locus, and the sex of the mouse. These BC1mice provide an opportunity to identify genes that interact with and modify the expression ofPax3and serve as a model to identify the genes that modify the expression of humanPAX3mutations. 相似文献
40.
C. Carey O. Dunin-Borkowski F. Leon-Velarde D. Espinoza C. Monge 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1994,163(8):649-656
Oxygen consumption, air cell gases, hematology, blood gases and pH of Puna teal (Anas versicolor puna) embryos were measured at the altitude at which the eggs were laid (4150 m) in the Peruvian Andes. In contrast to the metabolic depression described by other studies on avian embryos incubated above 3700 m, O2 consumption of Puna teal embryos was higher than even that of some lowland avian embryos at equivalent body masses. Air cell O2 tensions dropped from about 80 toor in eggs with small embryos to about 45 toor in eggs containing a 14-g embryo; simultaneously air cell CO2 tension rose from virtually negligible amounts to around 26 torr. Arterial and venous O2 tensions (32–38 and 10–12 toor, respectively, in 12- to 14-g embryos) were lower than described previously in similarly-sized lowland wild avian embryos or chicken embryos incubated in shells with restricted gas exchange. The difference between air cell and arterial O2 tensions dropped significantly during incubation to a minimum of 11 torr, the lowest value recorded in any avian egg. Blood pH (mean 7.49) did not vary significantly during incubation. Hemoglobin concentration and hematocrits rose steadily throughout incubation to 11.5 g · 100 ml-1 and 39.9%, respectively, in 14-g embryos.Abbreviations PO2
partial pressure gradient of O2
- BM
body mass
-
D
diffusion coefficient
-
G
gas conductance (cm3·s-1·torr-1)
-
conductance to water vapor
- IP
internal pipping of embryos
-
P
ACO2
partial pressure of carbon dioxide in air cell
-
P
AO2
partial pressure of oxygen in air cell
-
P
aCO2
partial pressure of carbon dioxide in arterial blood
-
P
aCO2
partial pressure of oxygen in arteries
-
P
H
barometric pressure (torr)
-
PCO2
partial pressure of carbon dioxide
-
P
IO2
partial pressure in ambiant air
-
PO2
partial pressure of oxygen
-
P
VCO2
venous carbon dioxide partial pressure
-
P
VO2
mixed venous oxygen partial pressure
- SE
standard error
-
VO
2
oxygen consumption 相似文献