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  • 1975-1979  (2)
  • 1975  (2)
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  • 1975-1979  (2)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 41 (1975), S. 227-232 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A method for the measurement of cytochrome P450 in unfixed cryostat sections is described. The sections are incubated for 10 minutes at room temperature in a buffered solution containing polyvinyl alcohol and sodium dithionite. Two incubations are performed on serial sections, one in nitrogen and the other in carbon monoxide. Readings are taken on a Vickers M85 microdensitometer fitted with a high sensitivity photomultiplier amplifier system, the measurements being made on corresponding fields in the serial sections. Subtraction of the nitrogen values from the carbon monoxide values, after allowing for an absorption shift, gives the absolute spectrum of cytochrome P450. The subtraction corrects for the tissue content of other haem-containing proteins. The cytochrome P450 spectrum shows a sharp maximum at 450 nm, and two other minor components absorbing at 444 nm and 458 nm. The content of cytochrome P450 in animals fed with phenobarbitone was 2.4 times greater than in control animals.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Histochemistry and cell biology 41 (1975), S. 323-334 
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Biochemical evidence, based on supra-physiological concentrations of thyrotrophin (TSH), has indicated that this hormone increases the NADP concentration in thyroid follicle cells. The hormone is believed to act both to enhance the reoxidation of the reduced coenzyme and to generate NADP from NAD. The latter effect, mediated by NAD+-kinase, could be a major control mechanism in thyroid metabolism. In the present study increased sensitivity has been obtained by the use of maintenance culture and of microdensitometry for measuring enzyme activity in the follicle cells. Using low physiological (0.1 μU/ml), and sub-physiological concentrations of the hormone as are used in the cytochemical bioassay systems, it has been shown that the rate of reoxidation of NADPH is enhanced by TSH. Moreover the NAD+-kinase activity is also greatly increased, and shows a direct relationship to the concentration of the hormone acting on the segments of the gland in vitro. It is possible that this phenomenon could be used to assay TSH. The results indicate that NAD+-kinase activity may play a significant part in the control of pentose-shunt oxidation in thyroid follicle cells.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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