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1.
2.
Molecular Forms of Acetylcholinesterase and Butyrylcholinesterase in Human Plasma and Cerebrospinal Fluid 总被引:1,自引:0,他引:1
John R. Atack Elaine K. Perry James R. Bonham Robert H. Perry 《Journal of neurochemistry》1987,48(6):1845-1850
The measurement of cholinesterase activities in either plasma or cerebrospinal fluid (CSF) may ultimately prove to be relevant in the diagnosis of neurological and neuropsychiatric disorders. However, studies to date have examined only total enzyme activities. Therefore in the present study we have examined the distribution of the individual molecular forms of both acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in plasma and CSF using sucrose density gradient centrifugation. Although the total activities of AChE were of the same order of magnitude in plasma and CSF, there was a considerable difference (120-500-fold) between total BChE activity in the CSF and the BChE-rich plasma. The analysis of the individual molecular forms revealed that the predominant molecular species of AChE and BChE in the CSF--both lumbar and ventricular--was the G4 form. The G4 form also constituted the majority of the plasma BChE activity and, on average, over half (56%) of the plasma AChE activity. The significance of the AChE and BChE molecular form compositions of both plasma and CSF and their possible relationship to pathological states are discussed. 相似文献
3.
Pedro J. N. Silva Richard K. Koehn Walter J. Diehl III Robin P. Ertl Elaine B. Winshell Mauro Santos 《Biochemical genetics》1989,27(7-8):451-467
Four samples of the musselMytilus edulis were taken between 1984 and 1987 from Stony Brook, New York, and used to study the glucose-6-phosphate isomerase (GPI) polymorphism
in this species.In vitro specific activity andin vivo flux measured in the same animals were found to be significantly correlated. A significant effect of GPI genotype on flux
was observed in one of the samples; overall, significant evidence of effect of genotype on enzyme activity was also obtained.
GPI activities of common genotypes tend to deviate less from the population mean than those of rare (frequency less than 5%)
genotypes. This suggests the possibility that rare GPI genotypes are rare as a consequence of having biochemical properties
that deviate from an optimum level and, therefore, having a lower fitness. In support of this hypothesis, we found in one
of our samples that shell length is a concave function of GPI activity with an intermediate optimum activity level.
The financial support provided to P.J.N.S. by the Luso-American Educational Commission (Fulbright Program), the Instituto
Nacional de Investigacao Científica (Portugal), and the Faculdade de Ciências da Universidade de Lisboa during several stages
of this research is gratefully acknowledged. Financial support from the Ministerio de Educatión y Ciencia (Spain) in the form
of a postdoctoral Fulbright/MEC fellowship to M.S. is also gratefully acknowledged. Research was supported by National Science
Foundation Grant BSR-8415060 to R.K.K. This is contribution No. 736 from the Program in Ecology and Evolution, State University
of New York at Stony Brook.
On leave from Departamento de Biologia Vegetal, Faculdade de Ciências, Universidade de Lisboa, Campo Grande C2, Lisboa, Portugal. 相似文献
4.
Oxidation of γ-Hydroxybutyrate to Succinic Semialdehyde by a Mitochondrial Pyridine Nucleotide-Independent Enzyme 总被引:2,自引:0,他引:2
Elaine E. Kaufman Thomas Nelson David Miller Noam Stadlan 《Journal of neurochemistry》1988,51(4):1079-1084
An antibody that inhibits over 95% of the cytosolic NADP+-dependent gamma-hydroxybutyrate (GHB) dehydrogenase activity of either rat brain or kidney was found to inhibit only approximately 50% of the conversion of [1-14C]GHB to 14CO2 by rat kidney homogenate. A similar result was obtained with sodium valproate, a potent inhibitor of GHB dehydrogenase. The mitochondrial fraction from rat brain and kidney was found to catalyze the conversion of [1-14C]GHB to 14CO2. The dialyzed mitochondrial fraction also catalyzed the oxidation of GHB to succinic semialdehyde (SSA) in a reaction that did not require added NAD+ or NADP+ and which was not inhibited by sodium valproate. The enzyme from the mitochondrial fraction which converts GHB to SSA appears to be distinct from the NADP+-dependent cytosolic oxidoreductase which catalyzes this reaction. 相似文献
5.
The expression of proteins that play a role in neuronal differentiation was examined in central nervous system (CNS) micromass embryo cell cultures and compared to expression at comparable developmental stages in vivo. The protein product of the src proto-oncogene (pp60c-src) has been postulated to have a specific role in development because, although it is expressed in many tissues, marked increases in amount and activity of pp60c-src
occur in neurons at the time of differentiation. Another protein of interest, high molecular weight neurofilament (NF) protein, is found in differentiated neurons. In the present study, changes over time in the expression of these two proteins in vitro and in vivo were examined. In the micromass cell cultures, primary cells from day 12 rat embryo CNS are plated at high density and differentiate into neurons during five days in culture. Tissues from embryos grown in vivo were assessed at 12 and 17 days post-coitum. Proteins were quantified by PAGE separation of equal amounts of total protein followed by transfer to membranes, immunoblotting, and densitometric scanning of blots. Increases in the amount of both proteins with neuronal differentiation was shown. Protein kinase activity of immunoprecipitated pp60c-src
also increased in cell cultures and in embryos. Similarity in patterns of expression between in vitro and in vivo tissue samples provides further evidence that the cultures closely simulate in vivo differentiation and are a useful system for examining expression of developmental genes in vitro.Abbreviations BCIP
5-bromo-4-chloro-3-indolylphosphate p-toluidine salt
- CNS
central nervous system
- DPBS
Dulbecco's phosphate-buffered saline
- GAM-AP
goat anti-mouse IgG alkaline phosphatase conjugate
- LB
limb bud
- NF
high molecular weight neurofilament protein
- NBT
nitroblue tetrazolium chloride
- SDS-PAGE
polyacrylamide gel electrophoresis
- PVDF
polyvinylidene difluoride
- RIPA
radioimmunoprecipitation
- TBS
Tris-buffered saline
- TTBS
TBS with 0.05% Tween-20
Presented in part at the 1989 and 1990 Teratology Society Meetings and the 1990 Society of Toxicology Meetings. 相似文献
6.
Elaine F. Smith Andrew V. Roberts John Mottley Susan Denness 《Plant Cell, Tissue and Organ Culture》1991,27(3):309-313
Plantlets of Dendranthema grandiflora Pennine Reel were grown from nodal sections on Sorbarods saturated with liquid medium containing 0–3 mg 1–1 of various growth retardants. After 4 weeks they were transferred to compost and maintained at a relative humidity of 42% at 27.5°C. Wilting was assessed over a period of 3 h. Plantlets treated with paclobutrazol, flurprimidol, triapenthenol, chlorphonium chloride, uniconazol and ancymidol showed dose-related reductions in wilting up to a concentration of 3 mg 1–1. Responses to tetcyclacis and mepiquat chloride were weaker, and no responses to chlormequat chloride, BTS 44584 or diaminozide could be detected. These observations are compatible with an hypothesis that resistance to wilting derives from inhibited synthesis of gibberellins. 相似文献
7.
Jean F. Emly Wendy A. Ratcliffe Elaine Green Sarah J. Bowden David A. Heath Ann Blight Susan Hughes John G. Ratcliffe 《生物化学与生物物理学报:疾病的分子基础》1992,1180(1):58-64
The molecular forms of parathyroid hormone-related protein (PTHRP) in conditioned media from the BEN human lung cancer cell line, rat parathyroid cells (PT-r) and human keratinocytes were studied by gel-filtraton chromatography with assay of PTHRP by immunoassays and bioassay. Immunoreactivity (1–86 and 1–34) and bioactivity (1–34) in conditioned media eluted as a coincident major peak (approx. molecular mass 19–22 kDa) and there was evidence of amino-terminal species in the molecular mass range 10–16 kDa in BEN and keratinocyte media. Western blotting of PTHRP affinity purified by monoclonal antibodies directed at regions 1–34 or 37–67, identified a major species in all cell cytosols and media with an apparent molecular mass of 24–25 kDa, consistently slighty larger than recombinant PTHRP(1–141) (mobility of 21 kDa) which may represent an intact or native form of PTHRP. Additional amino-terminal species were identified in medium from keratinocytes (16 and 7 kDa), BEN cells (18 and 14 kDa) and PT-R cells (17 kDa), suggesting that processing occurs at the C-terminus and within the mid-region to form a range of amino-terminal fragments. 相似文献
8.
9.
C. Michael Bowman Elaine M. Berger Elaine N. Butler Karen M. Toth John E. Repine 《In vitro cellular & developmental biology. Plant》1985,21(3):140-142
Summary Zwitterion buffers are often used to modulate the pH of cell culture medium but their effect on cultured cells is controversial.
We found that addition of 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) caused superoxide dismutase (SOD) inhibitable
increases in nitroblue tetrazolium dye reduction and SOD and catalase inhibitable decreases in the growth of cultured bovine
pulmonary artery endothelial cells. The findings suggest that HEPES stimulates endothelial cells to make toxic oxygen metabolites
that contribute to decreased cell growth.
This work was supported in part by the National Institutes of Health, Colorado and American Lung Associations, Colorado and
American Heart Associations, the Council for Tobacco Research, and the Kroc, Hill, Swan and Kleberg Foundations. Dr. Bowman
is a Clinician Scientist Awardee of the American Heart Association. 相似文献
10.