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  • Articles  (2,892)
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  • 1
    ISSN: 0884-3996
    Keywords: Estrone-3-glucuronide ; pregnanediol-3α-glucuronide ; chemiluminescent immunoassay ; ovarian function ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The determination of the concentration of estrone-3-glucuronide and pregnanediol-3α-glucuronide has been performed by a chemiluminescent immunoassay in early morning urine samples of 14 normal menstruating women and 11 women affected by luteal phase defect. The early morning urine samples were daily collected for an entire menstrual cycle. We have employed a timed and measured volume collection procedure as correction factor. The integrated values of the hormonal data in definite time intervals were used to create a nomogram. By means of this method, it was possible to completely separate normal from luteal insufficiency subjects and to distinguish two different types of luteal phase defects. Moreover, the same approach was applied to the study of the role and the frequency of luteal phase defect in 15 patients affected by habitual abortion and in 17 premenopausal women who had undergone quadrantectomy for T1a No Mo breast cancer. A luteal phase defect was detected in nine of the aborting patients (60%) and in eight women affected by breast cancer (47%). Finally estrone-3-glucuronide was measured in early morning urine samples of 96 prepubertal and pubertal girls in different pubertal stages and in one patient affected by precocious puberty, before and during an agonist GnRH treatment. The urinary test of ovarian function seems to be suitable for diagnostic purposes and for clinical studies.
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  • 2
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 580-586 
    ISSN: 0884-3996
    Keywords: Pregnanetriol-3α-glucuronide ; immunoassay ; chemiluminescence ; ACTH test ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Pregnanetriol-3α-glucuronide (PTG) is the majority urinary metabolite of 17-hydroxyprogesterone (17OHP) and it typically increases in the commonest form of congenital adrenal hyperplasia (CAH), due to 21 hydroxylase deficiency.We developed a simple chemiluminescent immunoassay for the direct measurement of PTG in diluted urine in order to avoid the preliminary hydrolysis and extraction steps that are usually employed in gas-liquid chromatographic methods. The immunogenic complex PTG-bovine-serum-albumin was used to induce the formation of specific antibodies in New Zealand rabbits. In addition, PTG was conjugated to aminoethylethylisoluminol and the resulting tracer was characterized by mass spectrometry and used to monitor the immunological reaction. The characteristics of the antibody were determined with regard to specificity and sensitivity. The precision of the assay method was also established.PTG excretion was studied before and after the ACTH stimulation test (1 mg synthetic ACTH i.m.) in 11 normal women and in one subject affected by CAH due to 21-hydroxylase deficiency. PTG levels well correlated with 17OHP plasma concentrations both under basal and stimulated conditions, in normal women as well as in the patient affected by CAH.
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  • 3
    ISSN: 0884-3996
    Keywords: Bacterial bioluminescence ; LDH ; flow-analysis ; nylon ; immobilized enzymes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The catalytic activity of serum L-lactate dehydrogenase (LDH), was determined by monitoring the NADH produced by LDH with bacterial bioluminescent enzymes immobilized on a nylon coil.The LDH reaction of L-lactate with NAD took place in a flow-through mixing coil that preceded the bioluminescent detector coil. The response was linear from 1 to 5000 U/l at 37°C and from 3 to 2000 U/l at 25°C. The intra- and inter-assay reproducibility (CV%) were less than 10% and recovery range was 92% to 110%. The results agreed well with those obtained with a spectrophotometric method.
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  • 4
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 59-65 
    ISSN: 0884-3996
    Keywords: Luminescence quenching ; luminol ; lucigenin ; oxygen ; active species ; scavengers ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The chemiluminescence of luminol and lucigenin is often used to detect the production of reactive oxygen derivatives by phagocytic cells. Also, several quenchers and enzyme inhibitors are used to determine which oxygen derivatives are responsible for the observed effects. In the present work we have assessed the reliability of dimethylthiourea and cysteamine (OH. quenchers), desferrioxamine (iron chelator) and diethyldithiocarbamate (superoxide dismutase inhibitor). They all react with CIO- and are also strong inhibitors of the luminescence of luminol catalysed by horseradish peroxidase (HRP); cysteamine and diethyldithiocarbamate also react with H2O2. NaN3 is an inhibitor of myeloperoxidase and a quencher of singlet O2, but we found that under certain conditions it can amplify the the luminescence of luminol triggered by CIO- or Fenton's reagent. A complex of copper and penicillamine that had been proposed as an \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm O}_{\rm 2} ^{\bar .} $\end{document} quencher, quenches all luminescent reactions studied. On the other hand, we were able to confirm the relative specificity of other quenchers: taurine for CIO-, benzoate for OH. and mannitol for both OH. and ‘crypto-OH.’.
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  • 5
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 71-74 
    ISSN: 0884-3996
    Keywords: Monocyte ; activation ; chemiluminescence ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Human mononuclear cells were separated from freshly obtained peripheral venous blood by density centrifugation and the number of monocytes present estimated by volume spectroscopy. The mononuclear cells were then placed directly into the wells of a microtitre plate and incubated for one hour at 37°C to promote adherence of the monocytes to the plastic wells. Non-adherent cells were then removed by washing, thus avoiding the need to treat the monocytes with EDTA or other reagents during cell preparation. The time course and dynamics of the chemiluminescence response of adherent monocytes towards opsonized zymosan was similar to those seen using non-adherent cells.The ability of adherent monocyte preparations to produce chemiluminescence following incubation for varying periods with T-lymphocyte conditioned medium was investigated. The use of a microtitre plate chemiluminescence reader allows several plates to be assayed over the 24-hour period and since small numbers of cells are required, many cultures can be analysed in one experiment. This technique (Patent applied for) promises to be a powerful tool for dissecting the cellular events which occur during macrophage activation and examining the effect of various lymphokines on the ability of monocytes to produce a chemiluminescence response.
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  • 6
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 79-83 
    ISSN: 0884-3996
    Keywords: Bioluminescence ; circadian rhythm ; luciferin binding protein ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A luciferin binding protein LBP involved in the bioluminescence reaction of Gonyaulax polyedra was purified and used for antibody production. Luciferin bound to LBP is fluorescent and can be used as a marker in living cells, allowing the localization of LBP in cortical organelles to be visualized. In cell sections, the same peripheral localization was observed using anti-LBP and immunofluorescence microscopy. The amount of LBP is ten-fold greater from cells from in night phase compared to those from in day phase, as determined both by immunoblots of cell extracts, and in vivo fluorescence. These changes correlate with the circadian changes in bioluminescence of living cells.
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  • 7
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989) 
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 8
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 221-226 
    ISSN: 0884-3996
    Keywords: Lactoferrin ; elastase ; infection ; immunoluminometric ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: An immunoluminometric assay for plasma lactoferrin has been developed and used to study the levels in children and neonates with viral and bacterial infections. The reference range for plasma lactoferrin was 50-250 μg/l. Lactoferrin levels were significantly higher in patients with bacterial versus viral infections.
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  • 9
    Electronic Resource
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    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 1-2 
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 10
    Electronic Resource
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    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 200-207 
    ISSN: 0884-3996
    Keywords: Singlet oxygen ; peroxidase ; oxygenase ; peroxy radical ; superoxide anion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Singlet oxygen generation is reported from (1) enzymatic reaction and (2) electron transfer reactions of the superoxide anion measured directly with an ultrasensitive near-IR emission spectrophotometer by monitoring the O2(1Δg) → O2 (3Σg-) transition at 1268 nm. Near-IR emission spectra from the myeloperoxidase and lactoperoxidase enzymatic systems show only emission of singlet oxygen at 1268nm. The lipoxygenase/Na-linoleate enzymatic reaction exhibits two emissions, 1268 nm and 1288 nm. The latter emission is identified as originating from a peroxy radical. Spectral and kinetic data giving evidence of singlet oxygen generation is obtained from the reaction of potassium superoxide solubilized by 18-crown-6-ether in acetonitrile with a series of organometallic coordination compounds.
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  • 11
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 342-345 
    ISSN: 0884-3996
    Keywords: Luminous bacteria ; toxicity test ; outer membrane ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Mutants of the luminescent bacterial strain NRRL B-11177 were isolated with pleiotropic hypersensitivity towards hydrophobic antimicrobial agents. SDS-PAGE analyses of outer membrane proteins and lipopolysaccharides revealed that the outer membrane structure of the ahs-mutants was altered. QSAR analysis showed that the inhibitory effect of chloro-substituted phenols on bioluminescence of the ahs-mutants depended on their hydrophobicity. The effect of chlorinated phenols and detergents on bioluminescence was increased in the ahs-mutants. The potential use of these mutants in bioluminescent toxicity tests was discussed.
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  • 12
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 346-350 
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We have constructed a chimeric aequorin consisting of a fragment of the anti-NP immunoglobulin gene fused to the aequorin gene. Expression in a myeloma cell line has produced a Fab′-like molecule that has the ability to bind NIP specifically and generate bioluminescent activity. It takes approximately 8 h at 4 °C in the presence of 2-mercaptoethanol and coelenterazine to regenerate luminescent activity. While the flash kinetics of this recombinant molecule are similar to native aequorin, its quantum efficiency is ten times lower. Preliminary studies have been conducted to ascertain its usefulness for immunoassays. We have shown for this chimeric aequorin 7 × 10-19 moles can be detected in solution, also it can be used in a solid-phase assay and is stably stored at -70°C for at least 2 months.
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  • 13
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 357-366 
    ISSN: 0884-3996
    Keywords: Chlamydia ; solution phase hybridization ; microtitre dish ; Chemiluminescence ; enzyme triggerable dioxetanes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: With an estimated 3-4 million new cases per year, human infections from Chlamydia trachomatis are probably the most prevalent sexually transmitted disease (STD) in the United States. Diagnosis of Chlamydia is usually conducted by tissue culture methods. Direct immunofluorescence and ELISA tests have become available, but there remains a need for a test with better specificity and sensitivity. In response to this need, we have developed a rapid DNA hybridization assay using synthetic oligonucleotide probes to detect the presence of the Chlamydia trachomatis specific 7.4 kb plasmid. The assay involves solution phase hybridization of unlabelled probes, rapid capture of the probe-target duplex onto a microtitre dish surface, a new signal amplification technique that employs chemically cross-linked oligonucleotides, and an alkaline phosphatase labelled probe. Signal is obtained by reacting the labelled probe-target complex with an enzyme triggerable dioxetane substrate. Detection of the chemiluminescent output is performed either with a luminometer or by exposure to instant film. All 15 serovars of Chlamydia trachomatis react positively while organisms known to co-inhabit the human urogenital tract react negatively.
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  • 14
    Electronic Resource
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    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 367-372 
    ISSN: 0884-3996
    Keywords: Luciferin ; luciferase ; luciferin-O-phosphate ; bioluminescence ; firefly ; Photinus pyralis ; protein blotting ; nucleic acid hybridization ; reporter gene ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A relatively simple, very sensitive bioluminescence-enhanced detection system for protein blotting and nucleic acid hybridization is described. The method utilizes antibodies conjugated with alkaline phosphatase or nucleotide probes complexed with alkaline phosphatase. Then the alkaline phosphatase takes part in a reaction by releasing D-luciferin (Photinus pyralis) from D-luciferin-O-phosphate. Liberated D-luciferin reacts with luciferase, ATP and oxygen under light emission. Light is measured using the Argus-100 a photon counting camera system or photographic films. Bound alkaline phosphatase conjugated antibodies or hybridized nucleotide probes can be visualized. The limit of detection is at present 5 to 50 fg of protein (IgG), corresponding, for example to 30 to 300 × 10-21 mol. This means a much higher sensitivity of the detection system in comparison to systems used at present. Experiments concerning nucleic acid hybridization and visualization of the emitted light by a photon counting camera (Argus-100) are under investigation.
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  • 15
    Electronic Resource
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    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 381-389 
    ISSN: 0884-3996
    Keywords: Urinary tract infection ; diagnostic methods ; rapid microbiology ; ATP ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Rapid tests for bacteriuria have the highest value, if the test result is available while the patient is with the doctor. At the bacteriological laboratory rapid testing of samples obtained by mail may be cost-effective but is of little clinical value. In a previous study performed at a health care centre using conventional urine culture as a reference the ATP test came out as the most reliable one among several rapid bacteriuria tests. The present study was performed to see how the ATP test could be fitted into the routine of the health care centre. Female patients with UTI symptoms were asked to deliver a urine sample to the health care centre laboratory and to wait for the result before seeing the doctor. After having the symptoms confirmed the doctor based the diagnosis on the ATP value. A low ATP value ruled out UTI and a high ATP value confirmed UTI. In patients with an intermediary ATP value (10-50 nmol/I) a positive nitrite test was used to confirm UTI. Only those patients with intermediary ATP values and negative nitrite test had to wait for conventional urine culture. Thus in most patients the decision on antibiotic therapy or not was based on clinical symptoms and ATP results only. Antibiotics (trimethoprim) were given as single dose or as a conventional 7-day regime in a double-blind comparison. The correlation between the ATP method and conventional culture was good. Although results of the present study are promising the ATP test as performed is too complicated to become widely accepted at health care centres. However, the dipstick version of the ATP test at present being developed will make the method ideally suited for rapid bacteriuria testing at health care centres and similar doctor's surgery situations.
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  • 16
    Electronic Resource
    Electronic Resource
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 406-418 
    ISSN: 0884-3996
    Keywords: Firefly luciferase ; luminescence ; enzyme kinetics ; bioluminescence spectra ; allosteric activators ; firefly mRNA translation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Biochemical properties, spectral parameters of bioluminescence and reaction kinetics for Luciola mingrelica firefly luciferase are described and analysed. The kinetic scheme of the enzymatic process is proposed and discussed. Allosteric regulation of luciferase activity by ATP and its analogues is considered and binding Mg2+ to luciferase shown to increase its activity. Regulation mechanism of luciferase activity by phospholipids is analysed and choline-containing phospholipids shown to be specific luciferase activators. Some properties of firefly luciferae and the luciferase synthesized during firefly mRNA translation in frog oocytes are compared.
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  • 17
    Electronic Resource
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    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 436-445 
    ISSN: 0884-3996
    Keywords: Bioluminescence ; chemiluminescence ; lipid metabolism ; LDL oxidation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Using bioluminescence assays for glycerol, free fatty acids, β-hydroxybutyrate and lactate, we were able to perform complex studies of human energy and lipid metabolism both in serum samples in vivo and in isolated fat cells in vitro. These studies would have been impossible without reliable, specific and highly sensitive luminescence methods. Oxidatively modified low density lipoprotein (LDL) has been implicated in the pathogenesis of atherosclerosis. Adaptation of a chemiluminescence assay for lipid hydroperoxides to LDL isolated by specific precipitation from serum makes it possible to measure LDL oxidation in vivo. Cell dependent chemiluminescence was used to investigate whether receptor mediated endocytosis of LDL by macrophages leads to oxygen radical production in these cells. No activation of the membrane NAD(P)H oxidase was observed.
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  • 18
    Electronic Resource
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    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 484-499 
    ISSN: 0884-3996
    Keywords: Luminescence ; chromatography ; detection ; quantitative analysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: An overview is presented of the physicochemical basis of luminescence, and its application to the detection of chemicals (drugs, biomedically important compounds, environmentally active substances) in liquid chromatographic systems.
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  • 19
    Electronic Resource
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    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 543-550 
    ISSN: 0884-3996
    Keywords: Fibre-optic ; biosensor ; bioluminescence ; chemiluminescence ; immobilized enzymes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The potential of immobilized enzyme membranes in biosensors has been explored in our group for several years. Although part of our work has been mainly devoted to electrochemical transducers and oxidases for the design of enzyme electrodes, the demand for ultrasensitive and highly selective sensors led us to consider the use of luminescent enzyme systems associated to optical transduction. When considering the need for operational and reliable biosensors in biotechnology, immobilization and stability of the sensing element still remain, in most cases, an unavoidable problem. We recently proposed a very fast and reliable procedure for preparing enzymatic membranes from Pall (Biodyne Immunoaffinity membranes) supplied in a pre-activated form. Both the firefly and bacterial systems as well as peroxidase for the chemiluminescent determination of various analytes, could be bound to such a support.Based on this approach, a fibre-optic sensor with immobilized enzymes has been designed which permits bio- or chemiluminescent analysis of ATP, NADH or H2O2 respectively. With the NADH-based system, other analytes could be detected using coupled dehydrogenases. This device appears very promising and includes the convenience of both the luminescence sensitivity as well as the handling of the biosensor design.
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  • 20
    Electronic Resource
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    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 351-356 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; acridinium ester ; labelling ; oligonucleotide DNA probes ; hybridization ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Chemiluminescent acridinium ester derivatives have been synthesized and covalently attached to suitably modified synthetic DNA oligonucleotides. Attachment of acridinium ester label to primary aliphatic amine group(s) present in the synthetic DNA probe molecule is rapid and efficient. Methods have been developed for efficient separation of acridinium ester-labelled DNA from unincorporated labelling reagent and underivatized DNA.The basic hydrogen peroxide detection reaction and photon counting conditions for measurement of chemiluminescence emission from acridinium ester-labelled DNA probes have been optimized. Under optimal conditions, the observed detection limit for the labelled DNA (1:1 mole ratio) is the same as for the free acridinium ester label, which is 2 attomole sensitivity in the best case studied.
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  • 21
    Electronic Resource
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    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 375-380 
    ISSN: 0884-3996
    Keywords: Rapid microbiology ; adenosine triphosphate ; luminometer ; bioluminescence ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Use of firefly luciferase to assay adenosine triphosphate (ATP) extracted from microorganisms provides an easy means to enumerate microbes within minutes. The small amount of light produced is proportional to ATP and thus microbial number.The average bacterium contains around 10-15 g ATP per cell. Present reagents permit detection of 103 cells per tube. Luminometers currently on the market detect about 10-12 g ATP.Proper extraction of ATP from the microbes is an essential part of any protocol, as is the removal of non-microbial ATP from, for example, somatic cells also present in samples. The technique may be applied to a wide range of samples, for example food and beverages and clinical samples such as urine.The ATP assay gives a global measure of microbial numbers, i.e. it is not species specific unless a species separation step is included in the protocol.
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  • 22
    Electronic Resource
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 390-398 
    ISSN: 0884-3996
    Keywords: ATP assay ; ATP extraction ; biomass ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: ATP methodology needs to be further standardized and improved in order to avoid the pitfalls that have sometimes hampered its application to biomass assays. The following steps have been reconsidered as far as the bacteriological applications is concerned: (a)destruction of free and somatic ATP: replacement of apyrase by mammalian ATPase, more readily accessible to specific inhibition;(b)extraction of bacterial ATP: protection of luciferase by lipids against inhibitory effect of cationic detergents with production of a constant light response.New methods are proposed for the calibration of luminometers and for the matching of sample holders in multichannel instruments.The limit of sensitivity of ATP assays is discussed in the light of currently available reagents and instruments.
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  • 23
    Electronic Resource
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 399-405 
    ISSN: 0884-3996
    Keywords: ATP ; semen analysis ; sperm antibodies ; male fertility ; artificial insemination ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The measurement of the ATP content of fresh semen is as accurate as the estimation of sperm motility by conventional methods in discriminating between semen of fertile versus subfertile men. The ATP content of frozen thawed donor semen is correlated with the probability of conception per cycle of insemination. Exact quantification of cytotoxic sperm antibodies in serum is possible with the adenosine-triphosphate-release-cytotoxicity test, since measurement is free of the bias of microscopic examination. The procedure has been simplified by testing only one serum dilution and calculating the ‘sperm toxicity index’.
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  • 24
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 419-422 
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Allowing for the lipid nature of firefly luciferase we have developed a new method for obtaining high-activity and high-stability enzyme preparations for bioluminescent microassay. The method includes the step of differential centrifugation in presence of stabilizing additives which entails a partial purification of the enzyme and its essential stabilization likely due to the fact that luciferase retains its lipid environment which plays an important role in catalysis. The resultant luciferase preparation is stable in solution at 4 °C for 2-3 months and allows the detection of down to 10-11M ATP.A new method has been offered for luciferase immobilization on film carriers precoated with a phospholipid layer. By sorption of the enzyme on such carriers, the samples of immobilized luciferase have been obtained suitable for constructing chemiluminescent biosensors, in the form of luciferase-containing films. There are many-fold applications for detection of ATP micro-quantities.
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  • 25
    Electronic Resource
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    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 423-435 
    ISSN: 0884-3996
    Keywords: Immobilized enzymes ; nylon ; bioluminescence ; chemiluminescence ; flow systems ; epoxy methacrylate ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Immobilized enzymes are widely used in the clinical laboratory in the assay of several analytes and enzymes. The use of immobilized enzymes makes these reagents recoverable and re-usable, and in most cases increases their stability and catalytic activity. In conjunction with bioluminescent enzymes (firefly and bacterial luciferases) and chemiluminescent catalyst (peroxidase) we set up high-sensitive flow methods based on the use of nylon tube coil or epoxy methacrylate column as solid support.All the NAD(P)/NAD(P)H-dependent dehydrogenases (bacterial luciferase), ATP-dependent enzymes (firefly luciferase) and oxidases producing H2O2 (peroxidase) can be immobilized and a large variety of analytes have been sensitively measured. As an alternative format we also reported a dry chemistry method in which all the enzymes, substrates and cofactors are ready to use, supported on dry cellulose disks. Methodological problems such as flow conditions, stability, pH, ionic strength and analytical performances are also reported.
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  • 26
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 454-462 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; NADH ; ATP ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Bioluminescent methods are widely used for the assay of the co-factors, NADH and ATP. Although the bioluminescent method is highly sensitive, the enzymes used are unstable and expensive. Therefore a chemiluminescent method would be valuable in clinical routine assay.We have developed a chemiluminescent method for the assay of NADH using the 1-methoxy-5-methylphenazinium methyl sulphate (1-MPMS)/isoluminol(IL)/microperox-idase(m-POD) system.In order to increase the sensitivity of this method, enzymatic cycling system was coupled to the chemiluminescent assay of NADH.Alcohol dehydrogenase and malate dehydrogenase were used as the cycling enzyme. The standard curve was obtained in the range from 3 × 10-14 to 5 × 10-12mol/assay. The detection limit of NADH was 30fmol/assay which was comparable to that of the bioluminescent method using bacterial luciferase.Two chemiluminescent methods for the assay of ATP have been developed. Method 1 is the system using hexokinase/G6PDH and 1-PMS/IL/m-POD, and method 2 is the system based on the enzymatic cycling reaction of ATP using hexokinase/pyruvate kinase. Method 2 is 1000/fold more sensitive than the method 1. The detection limit of ATP was 10 fmol/assay.
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  • 27
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 463-474 
    ISSN: 0884-3996
    Keywords: Photoproteins ; calcium ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 28
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 575-579 
    ISSN: 0884-3996
    Keywords: 3α-Diol-G ; immunoassay ; chemiluminescence ; hirsutism ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We described a chemiluminescent immunoassay (CIA) for 5α-androstane-3α, 17β-diol-glucuronide (3α-diol-G) in human diluted urine. This method allowed the direct measurement in 1μl of urine avoiding the hydrolysis and extraction steps for sample pretreatment commonly used in routine methods. The hapten 3α-diol-G was synthesized by a Koenigs-Knorr reaction. The immunogenic complex, 3α-diol-G conjugated to bovine serum albumin (BSA), was employed to induce the formation of specific antibodies in New Zealand rabbits. In addition, the required chemiluminescent (CL) tracer was prepared. The characteristics of the antibody was determined as regard to specificity and sensitivity and the precision of the assay methods established. In 22 hirsute women affected by policystic ovarian syndrome we found 3α-diol-G values significantly (p 〈 0.01) higher (146.28 ± 73.77μg/g of creatinine; mean ± SD) than those observed in normal women (72.1 ± 32.58 μg/g of creatinine; mean ± SD).
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  • 29
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 587-593 
    ISSN: 0884-3996
    Keywords: Immunochemiluminescence ; streptavidin ; prolactin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We have developed a two-site immunoassay for human prolactin using two monoclonal antibodies: one of them immobilized on the solid phase and the other labelled with biotin. The serum is incubated simultaneously with the antibody-coated bead, the biotinylated antibody and the tracer (streptavidin-isoluminol or avidin-peroxidase complex). Our experimental work has been directed towards a common set of reagents to capture the prolactin and then, with different tracers, towards obtaining on the calibration curve the same results for unknown samples.On the basis of the positive results we obtained, we have developed a kit that can be used by the customer or as an enzyme-immunoassay or as a chemiluminescent immunoassay, depending on instrumentation available, spectrophotometer or luminometer.
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  • 30
    ISSN: 0884-3996
    Keywords: ATP ; bioluminescence ; firefly luciferase ; platelets ; erythrocytes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Direct bioluminescent ATP determination in platelets and erythrocytes involves the study of different parameters which are discussed here. Some parameters are linked to the bioluminescent reaction and to the analyte (ATP); others have regard to the biological matrix. The composition of bioluminescent reagents and the preparation and conservation of the ATP standard, also in the presence of excipients, are among the first given. Matrix problems involve cell characteristics related to age and form, lysis resistance and the possible formation of aggregates (platelets) that may inhibit the complete release of ATP. For these reasons we used the most efficient ATP release agent with the lowest inhibitory effect on luciferase. The data obtained correlate well with a bioluminescent method requiring extraction with ethanol/EDTA, and therefore more time, for ATP determination in platelets and erythrocytes.
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  • 31
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 635-640 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; acridinium ester ; immunoassay ; paramagnetic particles ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The features that Magic Lite products offer as an immunoassay delivery system are discussed. The use of paramagnetic particles and acridinium ester labels confers advantages of speed, sensitivity, and stability. The technology has been used to measure analytes of widely varying molecular weights and serum concentrations, indicating its potential to detect the full range of clinically relevant analytes. Initial development efforts have indicated that the advantages of the system can be effectively exploited in an automated instrument.
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  • 32
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 602-608 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; polymorphonuclear leukocytes ; phagocytosis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Peripheral blood phagocytic cells (PMNLs) are activated by contact with opsonized particles. Metabolic activation of PMNLs is associated with a remarkable increase in the respiratory burst and generates high energy oxygen compounds which are responsible for the bactericidal activity of PMNLs and for their ability to produce luminol-dependent chemiluminescence (CL). The CL phenomenon is measured by an automated and computerized photoluminometer (Berthold LB950) in whole blood stimulated with opsonized zymosan.This whole blood method of CL measurement has been applied to the study of the phagocytic process and to the investigation of cellular and humoral abnormalities in several pathologies, indicating this assay as a simple, rapid and reliable test.
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  • 33
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 47-51 
    ISSN: 0884-3996
    Keywords: Chemiluminescent immunoassay ; somatomedin C ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We present a method for the measurement of the total Somatomedin C (SmC) content in human early morning urine samples after dialysis, extraction, and concentration. We modified a chemiluminescence immunoassay, previously developed for SmC determination in serum, for analysis of SmC in urine. Appropriate sensitivity was obtained by the preparation of a new chemiluminescent tracer (AEEI-COOH-SmC) and the optimization of a competitive non-equilibrium immunoassay system which had a detection limit of 0.24 fmol SmC per tube.
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  • 34
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. ii 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 35
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 201-205 
    ISSN: 0884-3996
    Keywords: Lux genes ; T7 phage promoter ; luxF ; flavoproteins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The lux genes from Photobacterium phosphoreum (NCMB844) have been cloned into Escherichia coli in a plasmid containing the T7-bacteriophage promoter. By specific expression in vivo under the T7 promoter, five structural genes (luxA-E) coding for the fatty acid reductase and luciferase polypeptides were identified as well as a new gene, designated as luxF, which codes for a 26kDa polypeptide. This new gene is located between luxB and luxE and thus disrupts the structural gene order of luxCDABE found in the Vibrio genus. The luxF gene and the protein it codes for have recently been identified in other Photobacterium species and so appears to be widely distributed within this genus. Nucleotide sequencing of the luxF gene has shown it to code for a protein homologous to the luciferase subunits, coded by the luxA and luxB genes. Although this gene is not necessary for light emission in all luminescent bacteria, it must play an essential role in the biochemistry, physiology, or ecology of the luminescent system in species of the Photobacterium genus.
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  • 36
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 207-212 
    ISSN: 0884-3996
    Keywords: Regulatory mutants ; autoinducer ; luminescence ; Vibrio harveyi ; luciferase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Regulatory mutants of the luminescent bacterium, Vibrio harveyi, have been isolated whose light emission can be stimulated by extracts of the growth media. Chloroform extracts of conditioned media in which V. harveyi has been grown can increase light emission in one of the dark mutants, D34, over 103-fold. An increase in the level of the mRNA and the enzymes associated with the lux system can also be demonstrated.Analysis of the expression of the lux system in Escherichia coli transformed with DNA from the D34 regulatory mutant demonstrates that the mutation resides outside the luciferase structural genes. The results suggest that the decrease in light emission in the regulatory mutants may be due to a mutation in synthesis of an autoinducer analogous to that found for the Vibrio fischeri lux system.
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  • 37
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 3-3 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 38
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 23-23 
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 39
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 26-28 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 40
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 12-19 
    ISSN: 0884-3996
    Keywords: Tetrapyrrole luciferin ; luc cDNA clones ; luciferase of dinoflagellates ; scintillons ; organelles ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Bioluminescence in the dinoflagellate Gonyaulax polyedra occurs as brief bright flashes, originating from many (∼400) small (∼0.5 μm) cytoplasmic organelles which protrude into the acidic vacuole, and are thus surrounded by the tonoplast. Biochemically, the substrate is unusual; it is an open chain tetrapyrrole, highly unstable to air but protected in the cell at pH̃ 8 by virtue of a luciferin binding protein (LBP). This molecule is a dimer of 72 kDa subunits which, upon a decrease in pH, releases luciferin to react with oxygen in the luciferase (∼140 kDa) catalysed luminescent reaction. cDNAs for both luciferase and LBP have been isolated and cloned, and the identity of LBP was confirmed by hybrid selection and in vitro translation of the message. The tenfold circadian (day to night) change in the amount of LBP, which parallels the in vivo rhythm of luminescence, is due to de novo synthesis and subsequent degradation of the protein each day. The LBP mRNA levels, as determined by in vitro translations and by Northern hybridizations, do not vary over the daily cycle, indicating that circadian control of bioluminescence in this species is mediated at the level of translation.
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  • 41
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 31-39 
    ISSN: 0884-3996
    Keywords: Firefly luciferase ; click beetle luciferases ; bioluminescence spectra ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: In studying beetle bioluminescence in the early 1960s, Dr McElroy and his colleagues found that the Jamaican click beetle, Pyrophorus plagiophthalamus, was capable of emitting different colours of light. They further found that the luciferin substrate used by this beetle was the same as that in the firefly, demonstrating that the different colours of bioluminescence were due to differences in the structure of the luciferases. We have recently cloned cDNAs from this beetle species which code for at least four different luciferases. The luciferases are distinguishable by their different colours of bioluminescence when expressed in Escherichia coli. The sequence differences between these different luciferases are few, so the amino acids responsible for the different colours of emission must also be few. Through the construction of hybrid luciferases, by rearranging fragments of the original cDNA clones, we have identified some of these amino acid determinants of colour.
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  • 42
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 79-87 
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 43
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 88-98 
    ISSN: 0884-3996
    Keywords: Immunoassay ; acridinium ester ; chemiluminescence ; liposomes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A new series of stable acridinium ester conjugates have been developed for use as non-isotopic labels in immunoassay. They have proved to be a flexible alternative to radioimmunoassay. We present data showing the successful development of immunoassays in sandwich, competitive and receptor formats. In addition, hydrophilic acridinium ester analogues have been synthesized, encapsulated in liposomes, and utilized as labels in immunoassay. The potential of this technology is discussed.
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  • 44
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 99-111 
    ISSN: 0884-3996
    Keywords: Chemiluminescent substrates ; enzymes ; immunoassays ; sensitive detection ; enhancement ; 1,2-dioxetane ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We have synthesized and studied two 1,2-dioxetane-based chemiluminescent enzyme substrates: 3-(2′-spiroadamantane)-4-methoxy-4-(3″-phosphoryloxy)phenyl-1,2-dioxetane (AMPPD), and, 3-(2′-spiroadamantane)-4-methoxy-4- (3″-β-D′-galactopyrano-yloxy)phenyl-1,2-dioxetane (AMPGD), which can be activated to chemiluminescence at 470 nm by alkaline phosphatase and βD-galactosidase, respectively. In addition, we observed that certain macromolecules enhance the luminescence of AMPPD. For example, the addition of 0.1% bovine serum albumin amplifies the luminescent signal and improves the detection limit for alkaline phosphatase by approximately one order of magnitude under certain conditions. This effect is due to the presence of a hydrophobic microenvironment provided by the enhancer which ‘stabilizes’ the dephosphorylated AMPPD emitter.Alkaline phosphatase catalysed chemiluminescence from AMPPD is constant for a prolonged period of time. Using AMPPD we were able to detect 0.01 attomole quantities of alkaline phosphatase immobilized on membrane supports and imaged on photographic film and, in solution, measured in a luminometer.AMPPD and AMPGD offer alternatives to colorimetric and fluorescent subsrates for alkaline phosphatase and β-D-galactosidase labels used in enzyme immunoassays. The simplicity and sensitivity of this chemiluminescent readout allowed the development of rapid clinical assays (e.g. β-hCG, LH, TSH and others).
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  • 45
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 119-128 
    ISSN: 0884-3996
    Keywords: Luciferin ; luciferin derivatives ; luciferase ; bioluminescence ; enzyme immunoassay ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Ultrasensitive bioluminescence immunoassays for the determination of peptides and proteins (illustrated with human urinary kallikrein, bradykinin and the determination of human urinary kallikrein antibody titres) have been developed. The usable ranges of the standard curves are from 5 pg to 5000 pg per litre. The relative intra-assay coefficients of variation of the tests were between 2% and 6%, and the inter-assay coefficients of variation between 4% and 12%.
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  • 46
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 136-139 
    ISSN: 0884-3996
    Keywords: Luminescent labels ; acridinium-9-thiocarboxylates ; acridinium-9-(N-sulphonyl)carboxamides ; emission kinetics ; tracer stabilities ; immunoassays ; hCG ; TSH ; AFP ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 47
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 149-153 
    ISSN: 0884-3996
    Keywords: Thyroxine (T4) ; thyroid stimulating hormone (TSH) ; luminescent immunoassay ; xanthine oxidase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The use of xanthine oxidase in immunoanalysis has never been reported. We describe here a procedure in which the xanthine oxidase dependent luminescence of luminol is enhanced in the presence of Fe-EDTA complex, providing an highly sensitive assay (3 amol of enzyme) and a long-term signal.This specific amplification has been applied to T4 and ultrasensitive TSH solid phase immunoassays, with T4-XO and anti-TSH monoclonal antibody-XO conjugates as tracers. The performances of these assays are at least equivalent to those obtained with iodinated tracers, using the same solid phases and the same calibrators. The major advantages of these immunoassays are: (1) the long-term signal which can be repeatedly recorded over several days, (2) the high detection sensitivity, (3) the long-term stability of the luminescence reagent and (4) the stability of the conjugates.
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  • 48
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 159-163 
    ISSN: 0884-3996
    Keywords: ELISA ; FITC ; chemiluminescence ; anti-HBs ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A chemiluminescent enzyme linked immunosorbent assay (ELISA) for the detection of antibody to hepatitis B virus surface antigen (anti-HBs) in human serum has been developed. Polystyrene microtitre plates were coated with recombinant, yeast-derived hepatitis B surface antigen (rec-HBsAg). Patient serum samples and appropriate controls were added to the rec-HBsAg-coated wells and incubated to bind anti-HBs. The wells were then washed and a fluorescein isothiocyanate (FITC) conjugate of a human plasma-derived hepatitis B surface antigen (HBsAg) was added. Following incubation and further washing the bound FITC-labelled HBsAg was detected after addition of a horseradish peroxidase (HRP) conjugate of a monoclonal anti-FITC antibody and assaying for the enzyme. The activity of the HRP was measured using luminol and hydrogen peroxide as substrates and iodophenol as a chemiluminescence enhancer. The luminescence was recorded using a camera luminometer.Preliminary tests have shown the assay to be suitable for the detection of antibody in sera from both vaccinees and also from individuals with a past hepatitis B virus infection. The use of the FITC-anti-FITC system together with the measurement of a chemiluminescence signal makes possible the completion of this assay in a few hours. The assay has been shown to be both specific and sensitive and provides a permanent photographic record.
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  • 49
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 446-453 
    ISSN: 0884-3996
    Keywords: Bioluminescence analysis ; coupled reactions ; enzymatic cycling ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Luminescence analysis may be applied to many substances by arranging a prior reaction producing a species entering the light-emitting reaction. Under favourable conditions the two consecutive reactions are carried out simultaneously as a one-step procedure. In a bioluminescence assay, luciferase stability is frequently a problem, making it desirable to develop analytical schemes where the analytical response becomes largely independent of any impaired luciferase activity. The value of maximal emission or an approached steady-state level is a convenient and usually well-defined analytical parameter. When recording this level it is important to design the participating reactions in a way that compensates for changes in luciferase activity.
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  • 50
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 101-104 
    ISSN: 0884-3996
    Keywords: Bioluminescent assay ; bacterial ATP ; blood cultures ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A bioluminescent assay of bacterial ATP for rapid detection of bacterial growth in 512 clinical aerobic blood cultures was evaluated. At the detection limit of bacterial ATP (10-10 mol/l) in the blood cultures 94.2% of the true positive blood cultures were detected (sensitivity) and the specificity was 85.8%. If the cut-off limit was increased the sensitivity decreased and the specificity increased and at 2 × 10-9 mol/l ATP the maximum correctly classified blood cultures was reached. At this cut-off limit the sensitivity was 82.9% and the specificity was 99.6%. In 54.3% of the true positive blood cultures bacterial growth was detected more rapidly with the bioluminescent assay than with macroscopic examination and subculture.
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  • 51
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 105-113 
    ISSN: 0884-3996
    Keywords: Human neutrophils ; chemiluminescence ; reactive oxygen metabolites ; superoxide anion ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The bioluminescent oxygen metabolite indicator protein pholasin was characterized with respect to the type and location of reactive oxygen metabolites detected in suspensions of stimulated human neutrophils. Whereas pholasin detected reactive oxygen metabolites from neutrophil suspensions stimulated with soluble agents, particulate stimulants were apparently not effective triggering agents for pholasin-dependent neutrophil chemiluminescence. Neutrophils stimulated with fMet-Leu-Phe (1 to 100 nmol/l) showed maximum pholasin-dependent chemiluminescence 45 to 60s after stimulation. The time of maximum chemiluminescence was virtually independent of fMet-Leu-Phe concentration. In contrast, the time to reach maximum light emission increased from 60s with 100 nmol/l phorbol ester to 295s with 1 nmol/l phorbol ester. Significant inhibition of stimulated chemiluminescence was caused by both superoxide dismutase (20 μg/ml, 80% inhibition) and reduction of the oxygen concentration in the incubation medium to less than 0.5 μmol/l (95% inhibition). In contrast, the myeloperoxidase inhibitor sodium azide (0.1 nmol/l) afforded only 50% inhibition of the pholasin-dependent neutrophil chemiluminescence. Our results show that pholasin detects superoxide radicals released from cells stimulated by soluble stimulants but not intracellular oxidative activity elicited by particulate stimulants.
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  • 52
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 507-511 
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Advent of the multichannel plate and position sensitive detector has made possible true single photon counting imaging tubes. We have investigated the application of these detectors in studies of the ultraweak light emission of biological materials. Initially, we focussed our efforts on two objectives: (1) obtaining single photon counting images of living tissues using only the light (chemiluminescence) emitted by the specimen and (2) developing means of obtaining well-resolved spectra of weakly emitting sources. We have obtained a variety of images. One striking result of this work is the first observation of tissue specific localization of photon emission in situ. Using this detector we have also obtained the first well-resolved spectra of some important ultraweak emission processes. These results illustrate the potential use of single photon imaging in bioluminescence and chemiluminescence research.
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  • 53
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 512-522 
    ISSN: 0884-3996
    Keywords: CCD-imaging luminometer ; chemiluminescence ; quantitative imaging immunoassays ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We describe the application of a CCD Imaging Luminometer (CCDIL) for the detection and quantitation of rapid, simultaneous, peroxidase-linked chemiluminescent immunoassays of multiple samples of human serum alphafetoprotein (AFP). Results from some analogous immunoassays (total IgE, and thyroid stimulating hormone (TSH)) are included for comparison.Values for precision and antigen concentration obtained using the CCDIL and colorimetric versions of the immunoassays, on human serum samples, were in good agreement.The flexibility of the CCDIL is demonstrated; its ability to detect and quantitate antigen (particularly AFP) on a variety of solid phases is indicated.The work on the AFP immunoassays illustrates not only the flexibility of the CCDIL for sample presentation on a variety of solid phase systems, but also some relative merits of such systems.
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  • 54
    ISSN: 0884-3996
    Keywords: Luminol-dependent chemiluminescence ; oxyradical visualization ; serine protease inhibitor ; chlorpromazine ; single photon imaging ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Reaction difference of oxyradical generation and luminol-dependent photoemission of zymosan- and phorbol ester-treated neutrophils were investigated using a conventional photomultiplier and ultrasensitive photonic imaging technique. Zymosan-treated cells released a concentrated photonic burst corresponding to the cellular distribution. In contrast, phorbol ester-treated cells produced a negligible level of photoemission, and the additional application of Ca2+ ionophore enhanced the photonic burst, which was gradually spread out into extracellular space. Serine protease inhibitors did not attenuate PMA-induced chemiluminescence but did attenuate zymosan-induced chemiluminescence. This suggests the involvement of serine protease in the respiratory burst of phagocytizing neutrophils.
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  • 55
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 523-530 
    ISSN: 0884-3996
    Keywords: Chemiluminometry ; flow injection analysis ; automatic analyser ; immobilized enzyme column ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A compact automated analyser which could analyse constituents in biological fluids with a small sample volume and in a short time has been developed. The instrument was composed of a flow injection analysis system equipped with chemiluminometric detection and an immobilized enzyme column reactor used in combination. Chemiluminescence has high sensitivity, and its reaction proceeds very quickly. Furthermore, an immobilized enzyme column reactor can produce a sufficient amount of hydrogen peroxide from compounds in serum in a short time. When enzymes are used as reagents for the analysis of substances in blood or blood serum, the final signals emitted by different enzyme reactions are usually not only hydrogen peroxide but also ammonia, NAD(P)H and so on. However, the practical chemiluminescence method for ammonia and NAD(P)H has not been established. We have discovered a new practical method for ammonia and NAD(P)H using an enzyme column reactor consisting of both immobilized L-glutamate dehydrogenase and L-glutamate oxidase. The determinations of glucose and uric acid in serum by chemiluminometry after production of hydrogen peroxide by the respective oxidases are presented. A newly chemiluminometric determination of ammonia, NAD(P)H and its applications to other enzymatic analyses that give ammonia and NAD(P)H as a final signal are also described.
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  • 56
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 535-542 
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A novel optical biosensor for homogeneous immunoassay has been developed on the basis of the finding that electrochemical luminescence of pyrene-labelled antigen is extremely inhibited by immunochemical complexation. Electrochemical luminescence homogeneous immunoassay for human serum albumin (HSA), as a model analyte, was performed with a platinum plate electrode which was located in the vicinity of an optical fibre tip. HSA was determined in the concentration range of 3-25 × 10-6 mol/I.To improve electrochemical luminescence measurement an optical fibre electrode has been developed by fabricating a transparent platinum film on the top of an optical fibre. The minimum detectable limit of luminol was 10-11 mol/l with the optical fibre electrode. Luminol was applied as a label for homogeneous immunoassay.
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  • 57
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    Keywords: Bioluminescence ; ATP ; flow-analysis ; immobilized luciferase ; oxirane acrylic beads ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Firefly luciferase was immobilized on epoxy methacrylate beads and used for a continuous-flow assay of ATP extracted from platelets. The immobilized luciferase had a half-life of 3 days at 25°C; there was a 25% recovery of luciferase activity upon immobilization, and ca 50 reactors were made from 1 mg of commercial enzyme. The sensitivity of the assay was 0.3 pmol of ATP, and the response was linear between 1 and 500 pmol of ATP. The content of platelets obtained with the present method correlated well with those obtained using soluble luciferase.
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  • 58
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 31-40 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 59
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    Journal of Bioluminescence and Chemiluminescence 3 (1989) 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 60
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    Keywords: Granulocytes ; neuroblastoma ; ADCC ; cell killing ; chemiluminescence ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We investigated whether polymorphonuclear leukocytes (PMN) are able to kill human neuroblastoma cells either directly or if coated with antibody MAb 14.18 that recognizes ganglioside GD2 present on the cell surface of most neuroblastoma cells. Neuroblastoma cells could not be destroyed directly, whereas in the antibody-dependent reaction (ADCC-reaction) they were easily eliminated. In order to answer the question whether reactive oxygen intermediates are involved in this process, chemiluminescence measurements were performed. Compared to the signals that could be measured using opsonized zymosan as stimulus, only weak CL-signals could be registered during the ADCC reaction. Pretreatment of PMN with granulocyte-macrophage colony stimulating factor (GM-CSF) enhanced the CL-signals, catalase and SOD reduced it; however, cell killing was only slightly influenced in the presence of catalase and superoxide dismutase. These data suggested that reactive oxygen compounds do not play a prominent role in the killing process. Definitive evidence for this suggestion could be obtained using PMN from a patient with chronic granulomatous disease (CGD): MAb 14.18 coated neuroblastoma cells could be killed effectively, but no CL-signal could be registered, either in the ADCC-reaction or using opsonized zymosan as stimulus.
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  • 61
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 125-129 
    ISSN: 0884-3996
    Keywords: Lipid oxidation ; bioluminescence ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A simple, rapid bioluminescence test (BT) for the determination of lipid oxidation is described. The test utilizes an aldehyde-requiring dark mutant of Vibrio harveyi (M42) that emits light in the presence of long chain (C8-C16) aliphatic aldehydes. The procedure consists of treating the oil or fat with Co2+ ion in ethanolic medium at alkaline pH. This treatment facilitates the decomposition of the hydroperoxides into long-chain aldehydes, part of which is used by the bacteria to produce light. The test was evaluated with corn, soybean and safflower oils, and shows excellent correlation with the commonly used peroxide value assay.
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  • 62
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 115-124 
    ISSN: 0884-3996
    Keywords: Luminescence ; free fatty acid (FFA) analysis ; acyl-CoA synthetase ; peroxidase ; luminol ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A sensitive, specific, and partly automatic method for the analysis of free fatty acids is described. The assay involves activation of free fatty acids by acyl-CoA synthetase (EC 6.2.1.3) followed by oxidation of the thioesters by acyl-CoA oxidase. The H2O2 formed is determined in a reaction catalysed by horseradish peroxidase (EC 1.11.1.7) using luminol as electron donor. The assay has a linear range of 0.05 to 5 nmol of different free fatty acids (C10-C18) in the original sample. The efficiency of the method toward capric, lauric, myristic, palmitic, palmitoleic, stearic, oleic, and linoleic acid measured as recovery of light emission compared to that of H2O2 standards, was over 90%. AffiGel 501 was used to covalently bind the free thiol group in CoASH eliminating interference of this substance in the peroxidase-luminol reaction.
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  • 63
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 147-154 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; absolute quantum yields ; pentachlorophenyl oxalate ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Absolute chemiluminescence quantum yields (φCL) for reactions of bis-(pentachlorophenyl) oxalate (PCPO), hydrogen peroxide (H2O2) and 9:10 diphenyl anthracene (DPA) have been determined. A fully corrected chemiluminescence monitoring spectrometer was calibrated for spectral sensitivity using the chemiluminescence of the bis-(pentachlorophenyl) oxalate system as a liquid light source, the total photon output of which had previously been determined by chemical actinometry. At high (PCPO)/(H2O2) ratios φCL was found to be independent of PCPO and H2O2 concentrations.
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  • 64
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    Keywords: Luminometer ; evaluation ; rapid microbiology ; ATP assay ; firefly luciferase ; photometer ; radiometer ; comparison ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: An assessment has been carried out of the relative performance of ten instruments for quantification of adenosine triphosphate (ATP) by the firefly luciferase assay. The instruments evaluated were Amersham Amerlite Analyser, Dynatech Tube Luminometer, Dynatech Multiplate Luminometer, Dynatech Camera Luminometer, Hamilton Lumicon, LKB 1250 Luminometer, LKB 1251 Luminometer, Lumac Biocounter M2010A, Turner 20 TD Luminometer and a prototype version of the CLEAR Speed Tech 2000. An 800-fold difference in sensitivity was found between the most sensitive (Lumac, Turner) and the least sensitive (Dynatech Tube) of the conventional instruments. The Dynatech Camera Luminometer which worked on a completely different principle to the other instruments was about 5000 times less sensitive than the best of the photomultiplier tube instruments. The relative sensitivity of the instruments was maintained regardless of whether solutions of ATP in water or trichloroacetic acid extracts of bacteria were analysed. An analysis of 960 ATP bioluminescence assays showed that data obtained from such measurements are normally distributed.
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  • 65
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. ii 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 66
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 175-179 
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    Keywords: Luminol ; chemiluminescent label ; heparin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Acylated derivatives of luminol (5-amino-2,3-dihydro-1,4-phthalazinedione) bearing a carboxyl or amino group can be linked by amide bonds to a macromolecule requiring labelling. Though themselves of low quantum yield these compounds are alkali-labile and can be detected at a similar level of sensitivity to the parent compound luminol. These cheap, readily accessible compounds are less hydrophobic than other currently employed chemiluminescent labels. They also lack a positively charged nitrogen atom which could complicate their covalent linkage to polyanionic compounds. They thus appear well suited for labelling heparin and other macromolecules which interact with the luminal surface of blood vessels.
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  • 67
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 181-190 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; excitation energy transfer ; oxidative polymerization ; eumelanins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The catechol oxidase-catalysed and autoxidative transformation of 3,4-dihydroxyphenylalanine (DOPA) to eumelanin have been studied by oxygen consumption, energy transfer, absorption and fluorescence spectroscopy. Formation of transient dopachrome (λmax = 480 nm) and dopalutin (λex = 423 nm, λem = 491 nm) have been found in the enzymatic and autoxidative reaction. In the enzymatic reaction, neither a photon emission with quantum yield Φ 〉 10-13 nor energy transfer to triplet and singlet energy acceptors (sensitizers such as anthracene derivatives, xanthene dyes and chlorophyll-a) in water and micellar solutions have been found. The autoxidative reaction is chemiluminescent (Φ = 10-9), the emission occurring in the 400-600 nm range. The excitation energy is not transferred to sensitizers. The effect of various enzymes and traps of active oxygen species as well as the spectral distribution of chemiluminescence indicate that there is no emission from oxygen dimoles. Carbonates and active species of oxygen are shown to participate in the chemiexcitation reaction.
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  • 68
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 267-271 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; microtitre-plate luminometer ; opsonophagocytosis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A protocol for polymorphonuclear leukocyte chemiluminescence (PMN CL) assays of opsonophagocytosis was developed for a microtitre-plate luminometer. The complete procedure was performed in a single microtitre plate and was simpler and more efficient than previous protocols. The kinetics of the PMN CL response were best when microtitre plates were incubated on a shaking incubator between readings. The new protocol was used in a study of the pathogenicity of Corynebacterium jeikeium, an organism found in association with infection in the immunocompromised. No differences were found when PMN CL induction by 15 strains of C. jeikeium were compared with 15 isolates of other corynebacteria. Both groups of organisms required complement for efficient opsonophagocytosis; C. jeikeium strains showed no requirement for specific antibody. Resistance to opsonophagocytosis does not appear to be an explanation for the increased pathogenicity of C. jeikeium. Microtitre-plate luminometers are particularly well suited to bacterial opsonization studies where large numbers of strains often need to be assessed.
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  • 69
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 479-483 
    ISSN: 0884-3996
    Keywords: Quenched CL ; peroxyoxalate ; mechanism ; detection ; HPLC ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Several analytes such as the inorganic anions bromide, iodide, sulphite and nitrite and organic compounds as substituted anilines and sulphur compounds cause quenching of peroxyoxalate chemiluminescence. A detection method for liquid chromatography based on the quenching phenomenon has been developed. It makes use of an immobilized luminophore, i.e. 3-aminofluoranthene covalently bound via an alkyl-spacer on controlled pore glass, packed in the detector cell.The mechanism behind the quenching has been elucidated by investigating the roles of luminophores (both in the liquid and in solid state) and oxalates in peroxylate CL with respect to quenchers. Most probably the quencher destroys the radical ion pair produced after electron transfer in the last stage of the CIEEL reaction scheme, thus preventing the formation of electronically excited luminophore.
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 159-166 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 71
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 155-158 
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    Keywords: Bioluminescence ; chemiluminescence ; acridinium ester ; firefly luciferase ; kinetics ; photographic film ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Frozen assay reagents have been used to reduce the rate of light emission from the rapid chemiluminescent acridinium ester and the bioluminescent firefly luciferase reactions. Melting of the assay reagent delays the initiation of the light emission, thus eliminating the need to initiate these rapid reactions by injection of the assay reagents in front of the photodetector.
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    Journal of Bioluminescence and Chemiluminescence 3 (1989) 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 73
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 74
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 169-174 
    ISSN: 0884-3996
    Keywords: Diabetes ; albuminuria ; chemiluminescent immunoassay ; acridinium ester ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A simple chemiluminescent immunoassay (CLIA) for urinary albumin has been developed based on the use of a chemiluminescent acridinium ester-labelled human albumin and a commercially available antiserum. It includes two incubation steps and a second polyethylene glycol-assisted antibody separation. The sensitivity of detection is 0.016 mg/l, the assay working range is 0.1-5 mg/l, and the inter-assay CVs are ≤ 15%. Using 10- and 50-fold sample dilutions in assay buffer, a wide working range (1-250 mg/l) is obtained covering normal and pathological conditions. Timed overnight urine samples (bed rest conditions) were collected on three consecutive days for each patient. Albumin excretion rate (AER) was 4.7 ± 2.7 μg/min (x ± SD), range 1-15.9 μg/min in 36 healthy subjects (17♂, 19♀, ages 4-56 years), with day-to-day variations of 28.5 ± 20% (x ± SD), range 3.3-76.1%. The use of an acridinium ester as a chemiluminescent (CL) label overcomes the disadvantages of short shelf-life and health and safety hazards associated with radioisotopes. Results compare favourably with those obtained using a commercially available RIA kit.
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  • 75
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 193-199 
    ISSN: 0884-3996
    Keywords: Gene regulation ; bacterial bioluminescence ; Vibrio fischeri ; Vibrio harveyi ; lux genes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Comparison of the nucleotide sequences and gene organization of the lux systems from Vibrio harveyi and Vibrio fischeri has demonstrated that the location and order of the lux structural genes are highly conserved whereas considerable divergence has arisen in the location and/or presence of lux regulatory genes in the two marine bacteria. The order of the lux structural genes (luxCDABE) are identical in the two bacteria with the three fatty acid reductase genes (luxCDE) required for aldehyde biosynthesis flanking the luciferase genes (luxAB). Complementation in trans of the upstream V. fischeri DNA containing the luxC and luxD structural genes with the downstream luxA, B and E genes of V. harveyi resulted in luminescent Escherichia coli that did not require any exogenous aldehyde. These results indicate that the light-emitting systems are very similar in the two bacteria.However, the lux regulatory systems in these two bacteria appear to have clearly diverged. In V. harveyi, an open reading frame of 〉40 codons does not exist within 600 bp of the start of luxC in contrast to the luxl regulatory gene present in the V. fischeri lux system. An open reading frame of 615 bp is present farther upstream of luxC with the same direction of transcription and approximate location as the luxR regulatory gene of V. fischeri; however, apparent homologies do not exist between the two genes. The similarities in organization of the lux structural genes and the differences in the existence and/or location of lux regulatory genes in the two Vibrio species raises the question of how these marine bacteria can have a similar growth-dependent regulation of luminescence expression.
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  • 76
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 213-216 
    ISSN: 0884-3996
    Keywords: ATP ; monuments ; stone ; biodeterioration ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The examination of the state of conservation of works of art in stone includes the assessment of the presence of microbiological agents on the surface of the decayed monuments. These microorganisms can accelerate, via their metabolic activity, the decay process of the stone surface. At present this assessment is made with the traditional techniques for the microbiological examination of the soil, provides results only after a delay of 30 days. A bioluminescent ATP assay should provide rapid quantitation of actively growing organisms on the surface of a stone monument, and the applicability of this technique was verified on some samples of sandstone (Pietraforte) collected from a historic building (the Strozzi Palace) in Florence. These samples were evaluated for the amount of the ATP and the total number of microorganisms.The results obtained suggest that the bioluminescent assay could be suitable for detecting and quantitating the presence of microorganisms in a sample of stone.
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 29-30 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 51-58 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 79
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 40-48 
    ISSN: 0884-3996
    Keywords: Purification ; site-directed mutagenesis ; mutant enzymes ; lux genes ; aldehyde substrate inhibition ; FMNH2 binding ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: It has been appreciated for many years that the luciferase from the luminous marine bacterium Vibrio harveyi has a highly reactive cysteinyl residue which is protected from alkylation by binding of flavin. Alkylation of the reactive thiol, which resides in a hydrophobic pocket, leads to inactivation of the enzyme. To determine conclusively whether the reactive thiol is required for the catalytic mechanism, we have constructed a mutant by oligonucleotide directed site-specific mutagenesis in which the reactive cysteinyl residue, which resides at position 106 of the α subunit, has been replaced with a seryl residue. The resulting α106Ser luciferase retains full activity in the bioluminescence reaction, although the mutant enzyme has a ca 100-fold increase in the FMNH2 dissociation constant. The α106Ser luciferase is still inactivated by N-ethylmaleimide, albeit at about 1/10 the rate of the wild-type (α106Cys) enzyme, demonstrating the existence of a second, less reactive, cysteinyl residue that was obscured in the wild-type enzyme by the highly reactive cysteinyl residue at position α106. An α106Ala variant luciferase was also active, but the α106Val mutant enzyme was about 50-fold less active than the wild type. All three variants (Ser, Ala and Val) appeared to have somewhat reduced affinities for the aldehyde substrate, the valine mutant being the most affected.It is interesting to note that the α106 mutant luciferases are much less subject to aldehyde substrate inhibition than is the wild-type V. harveyi luciferase, suggesting that the molecular mechanism of aldehyde substrate inhibition involves the Cys at α106.
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  • 80
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 59-78 
    ISSN: 0884-3996
    Keywords: Ultrasensitive immunoassay ; fluorescent microspot immunoassay ; confocal microscopy ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The sensitivities of immunoassays relying on conventional radioisotopic labels (i.e. radioimmunoassay (RIA) and immunoradiometric assay (IRMA)) permit the measurement of analyte concentrations above ca 107 molecules/ml. This limitation primarily derives, in the case of ‘competitive’ or ‘limited reagent’ assays, from the manipulation errors arising in the system combined with the physicochemical characteristics of the particular antibody used; however, in the case of ‘non-competitive’ systems, the specific activity of the label may play a more important constraining role. It is theoretically demonstrable that the development of assay techniques yielding detection limits significantly lower than 107 molecules/ml depends on: 1the adoption of ‘non-competitive’ assays designs;2the use of labels of higher specific activity than radioisotopes;3highly efficient discrimination between the products of the immunological reactions involved.Chemiluminescent and fluorescent substances are capable of yielding higher specific activities than commonly used radioisotopes when used as direct reagent labels in this context, and both thus provide a basis for the development of ‘ultra-sensitive’, non-competitive, immunoassay methodologies. Enzymes catalysing chemiluminescent reactions or yielding fluorescent reaction products can likewise be used as labels yielding high effective specific activities and hence enhanced assay sensitivities.A particular advantage of fluorescent labels (albeit one not necessarily confined to them) lies in the possibility they offer of revealing immunological reactions localized in ‘microspots’ distributed on an inert solid support. This opens the way to the development of an entirely new generatio of ‘ambient analyte’ microspot immunoassays perrnitting the simultaneous measurement of tens or even hundreds of different analytes in the same small sample, using (for example) laser scanning techniques. Early experience suggests that microspot assays with sensitivities surpassing that of isotopically based methodologies can readily be developed.
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 112-118 
    ISSN: 0884-3996
    Keywords: Streptavidin ; biotin ; chemiluminescence ; universal marker ; immunoassay ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The tetrameric structure of streptavidin and its exceptionally strong affinity to biotin (Ka = 1015M-1) can be exploited to achieve an amplification of the signal in immunoassays. In the approach described here streptavidin (STAV) labelled with aminobutylethyl-isoluminol (ABEI) served as a universal marker in immunoassays for both haptens and big antigens. The advantageous features of streptavidin can be applied to any immunoassay using biotinylated antibodies as the primary probe.In two-site immunometric assays for larger antigens the liquid phase ‘tracer’ antibody is biotinylated. In hapten assays the solid phase antigen technique (Wood et al., 1982) is employed, in which sample-antigen and solid phase-antigen compete for a biotinylated antibody. In this paper we demonstrate the use of STAV-ABEI as a universal chemiluminescent label in steroid assays and in an immunometric assay using human growth hormone (hGH) as an example.
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  • 82
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 129-135 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; enzyme ; xanthine oxidase ; long-term signal ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A new enzyme label system is described which is superior to all existing chemiluminescence labels used in immunoassays. The system consists of the enzyme xanthine oxidase with hypoxanthine as substrate. The signal reagent contains perborate, an Fe-EDTA complex and luminol. The enzyme preparation and the signal reagent are very stable upon storage. The main features of the system are a long duration of the chemiluminescent signal (half-life time of 30 hours) and a very low limit of detection (about 3 amol). Possibilities and implications for the use of various measuring system are discussed.
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  • 83
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 140-148 
    ISSN: 0884-3996
    Keywords: NADH ; NADPH ; chemiluminescence ; chemiluminescent EIA ; enzyme activity ; DNA hybridization ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A highly sensitive chemiluminescent assay for NAD(P)H have been developed. The principle of the method is as follows; NAD(P)H reduces molecular oxygen to superoxide anion (O2-) and hydrogen peroxide (H2O2) in the presence of 1-methoxy-5-methylphenazinium methyl sulphate (1-MPMS) as electron mediator. The produced O2- and H2O2 can be measured by chemiluminescent reaction using isoluminol (IL) and microperoxidase (m-POD). A linear relationship between chemiluminescence intensity and NAD(P)H concentration (log/log) was obtained ranged from 10-9 mol/I to 10-5 mol/I. This chemiluminescent reaction has been coupled to the assay of glucose-6-phosphate dehydrogenase (G6PDH), β-D-galactosidase (β-Gal) and alkaline phosphatase (ALP). The detection limits of G6PDH, β-Gal and ALP were 10-18 mol, 10-20 mol and 10-18 mol per assay, respectively. The chemiluminescent assay of these enzymes applied to chemiluminescent enzyme immunoassay for 17α-hydroxy-progesterone and DNA hybridization assay using these enzymes as label.
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  • 84
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 154-158 
    ISSN: 0884-3996
    Keywords: Chemiluminescence: isoluminol ; immunoassay ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Isoluminol derivatives (ID) were early employed in the preparation of tracers for immunoassay. Their wide use was mainly due to their high quantum efficiency, low molecular weight, well-known chemical structure, low cost and high stability. Moreover the light efficiency of some ID may be modified by specific binding to the antibody, thus allowing the development of homogenous immunoassays requiring no bound/free separation step. Some ID are commercially available under both the amino terminal and carboxyl terminal form for conjugation to respectively carboxy derivative of steroid and amino residues of protein. The linking reaction can be commonly carried on via active esters chemistry and can be easily accomplished within one day. Steroid conjugates can then be rapidly purified by silica gel thin layer chromatography, and protein conjugates by gel filtration on a short disposable column. In the field of steroid studies, isoluminol derivative conjugates were prepared for the immunoassay of almost all compounds of clinical interest. When dealing with protein, both antigens and antibody were labelled in this way, resulting in highly specific activity tracers for competitive and non-competitive immunoassays. Recently the labelling of streptavidin with amino-buty-ethyl-isoluminol allowed the development of very sensitive immunoassay methods which take advantage of the biotin-avidin system.
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  • 85
    ISSN: 0884-3996
    Keywords: Nucleotide sequence ; genetic regulation ; bacterial luciferase ; amino acid sequence ; luxR ; autoinducer ; luxN ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We have determined the complete nucleotide sequence of a 7622 base pair fragment of DNA from Vibrio fischeri strain ATCC7744 that contains all the information required to confer plasmid-borne, regulated bioluminescence upon strains of Escherichia coli. The lux regulon from V. fischeri consists of two divergently transcribed operons, L (left) and R (right), and at least seven genes, luxR (L operon) and luxICDABE (R operon) and the intervening control region. The luxA and luxB genes encode respectively the α and β subunits of luciferase. The gene order luxCDABE seen in V. fischeri is the same as for V. harveyi. We have determined the sequence of the luxAB and flanking regions from Photobacterium leiognathi and have found upstream sequences homologous with luxC from the Vibrio species, but between luxB and luxE, there is an open reading frame encoding a protein of 227 amino acids (26,229 molecular weight) that is not found in this location in the Vibrio species. The amino terminal amino acid sequence of the encoded protein is nearly identical to that determined by O'Kane and Lee (University of Georgia) for the non-fluorescent flavoprotein from a closely related Photobacterium species (Dr Dennis O'Kane, personal communication). We have therefore designated this gene luxN.There is a 20-base inverted repeat ACCTGTAGGA×TCGTACAGGT, centred between bases 927 and 928 in the region between the two operons of V. fischeri. This region appears to fulfil two functions: it is critical for the LuxR protein to exert its effect and it is a consensus binding site for the E. coli LexA protein, a negative regulatory protein involved with the SOS response. There are sequences within the luxR coding region that appear to function in a cis-acting fashion to repress transcription from both the leftward and rightward promoters in the absence of the respective transcriptional activator proteins, thereby resulting in low basal levels of transcription. It now appears clear that there are multiple levels of control on the lux system allowing for a modulation of the intensity of bioluminescence of over four orders of magnitude.
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  • 86
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 611-614 
    ISSN: 0884-3996
    Keywords: Chemiluminescence immunoassay ; acridinium ester ; free T4 ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Chemiluminescence immunoassays have now achieved a recognized place in the diagnostic laboratory. The advantages of this non-isotopic technology derive from the use of acridinium esters which can be used to label antigens and antibodies to high specific activities, as well as from optimized immunochemistry. The availability of simple, reliable instrumentation for chemiluminescence measurement together with a range of assay kits offers a logical alternative to traditional radioimmunoassay.
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  • 87
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 615-619 
    ISSN: 0884-3996
    Keywords: Magic lite ; two-point adjustment ; immunoassay ; stability ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Ciba Corning Diagnostics has developed a two-point adjustment algorithm for use with the Magic Lite System which allows for extended stability of a full 7-10 point calibration curve over the life of a kit. This adjustment is accomplished by using a master calibration curve established during manufacturing along with two calibrators for each assay.Most conventional non-automated immunoassays contain anywhere from 6 to 10 calibrators which are included with each run of an assay.Eliminating the need to run full standard curves and using the two-point adjustment algorithm results in significant savings in both labour and reagent usage.
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  • 88
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 620-626 
    ISSN: 0884-3996
    Keywords: Immunochemiluminometric assay ; thyrotropin ; monoclonal antibody ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A recently available immunochemiluminometric assay (ICMA) for TSH developed by Ciba Corning Corp. has been evaluated. This system (Magic Lite) uses an acridinium-ester-labelled antibody and magnetizable particle for bound-free separation. In each assay only two calibrators are carried out and used to re-scale a manufacturer-generated curve stored in the memory of the luminometer. The precision of the response (RLU) estimated by all duplicates of 14 runs was about 4% for responses 〉12,000 RLU (corresponding to a concentration interval 0.7-113 μlU/ml) and worsened in the lower range (up to 10% CV); the sensitivity, computed from the mean within-assay precision profile, was 0.028 μlU/ml; the between-assay precision ranged from 4.6 to 13.1 CV%. Regression analysis of ICMA results (y) against consensus values of Behring IRMA (x) on 15 QC sera assayed in an inter-laboratory survey (concentration range 1-30 μlU/ml) gave y = -0.003 + 0.98x indicating a good agreement of the two techniques. Similar conclusions have been derived from the comparison of the ICMA results (y) in the low TSH concentration range (〈 1 μlU/ml) against the IRMA Boots Celltech (x) on 80 patient samples (y = 0.04 + 1.04x).
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  • 89
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 627-634 
    ISSN: 0884-3996
    Keywords: Free thyroxine ; chemiluminescence ; equilibrium dialysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A study of 239 patients compared free thyroxine (FT4) measurements made by equilibrium dialysis (ED) with measurements made using the Magic Lite FT4 chemiluminescence (Cl) immunoassay (Ciba Corning Immunodiagnostics). Patient groups: 41 normals; 27 hyperthyroid; 29 hypothyroid; 37 sick euthyroid; 10 chronic renal failure (CRF) and 25 pregnant patients; 13 oestrogen; 10 heparin; 12 salicylate; and 9 dilantin-treated patients; 3 lipaemic; 5 haemolysed; 6 hyperbilirubinaemic patients; 6 low thyroid binding protein (TBP) and 6 high TBP level patients. The two assays gave comparable results in most groups. Both assays tended to give elevated values in heparinized patients but FT4-ED results were more obviously affected. Pregnant patients and women on oral oestrogen had higher mean values with FT4-ED. In both assays the sick euthyroid and CRF patients had mean FT4 values similar to healthy euthyroid patients; the range of values in sick euthyroid and CRF patients was similar in both assays but wider than in healthy euthyroid patients. A supplemental study of 81 unselected acutely ill patients using FT4-Cl alone confirmed the wider range of values to be anticipated in sick euthyroid patients.
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  • 90
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 89-92 
    ISSN: 0884-3996
    Keywords: Spontaneous bacterial infection ; ascitic fluid ; alcoholic cirrhosis ; peritoneal carcinoma ; opsonic activity ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Cirrhotic ascites are highly susceptible to spontaneous bacterial infection, whereas carcinogenic ascites are seldom infected. This difference may be explained by differences in their chemotactic, bactericidal and/or opsomic activities. We measured the chemotactic and opsonic activity of ascitic fluids from 35 alcoholic cirrhotic ascites and of 12 peritoneal carcinogenic ascites. Chemotactic activity was measured by the under-agarose technique and opsonic activity by a luminol-enhanced method. Ascitic fluids from alcoholic cirrhosis had low chemotactic (62 ± 24.5% that of N-formylated peptide) and opsonic (67 ± 50% of normal serum) activities on normal human neutrophils. In contrast, ascitic fluids from peritoneal carcinoma were found to possess high opsonic activity (114 ± 49% of normal serum) and chemotactic activity similar to that of N-formylated peptide. During a 3-month follow-up, 11 spontaneous bacterial infections were observed among the first group against none in the carcinogenic group.
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  • 91
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 177-184 
    ISSN: 0884-3996
    Keywords: Time-resolved fluorescence immunoassay ; non-separation or homogeneous assay ; urinary steroid ; drug metabolites ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: 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.
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  • 92
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 185-190 
    ISSN: 0884-3996
    Keywords: Luminescence immunoassay ; thyrotropin ; coated tube ; monoclonal antibody ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A recently available chemiluminescence immunoassay (LIA-MAT) for TSH, set up by Byk-Sangtec Diagnostica, has been evaluated and compared with IRMA methods. The system LIA-MAT uses two monoclonal antibodies: one coated on tubes and the other labelled with isoluminol. To compute the within-assay precision profile, we estimated the response error relationship from all the duplicates (420) of eleven experiments. The CV of the response was 4-5% from the maximal response until 10,000 RLU (corresponding to about 1 μlU/ml); in the lower response range the CV worsened up to 8%. The sensitivity, derived from the precision profile, was 0.052 μlU/ml similar to that found in IRMA-MAT (0.044 μlU/ml); the working range extended from 0.33 to 100 μlU/ml. Results from LIA-MAT in the concentration range 1-30 μlU/ml were compared with the consensus mean produced by users of IRMA methods (IRMA-MAT, IRMA-Behring, Maiaclone Serono) participating in an inter-laboratory survey; a good correlation with IRMA techniques was found. The distribution of TSH determinations produced by LIA-MAT on 62 low concentration sample (〈0.3 μlU/ml) from patients unresponsive to TRH test, was found similar to that observed for the kit IRMA Boots-Celltech assumed as reference.
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  • 93
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 219-225 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; excited quinone ; semiquinone ; electron transfer reactions ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The reaction between p-benzoquinone and H2O2 in slightly alkaline solutions yields three major quinoid products that accumulate in the reaction mixture: (a) 2,3-epoxy-p-benzoquinone, (b) 2-hydroxy-p-benzoquinone and (c) p-benzohydroquinone. The reaction is accompanied by photoemission, probably originating from excited triplet 2-hydroxy-p-benzoquinone. These products originate from hydrogen peroxide and hydroxide nucleophilic addition to the C2=C3 double bond, as well as secondary redox interactions. The hydroxy substituent and the epoxide ring exert a substantial influence on the electronic distribution in the p-benzoquinone molecule leading to a decrease in the half-wave potential, as compared to the parent p-benzoquinone.The generation of electronically excited states is the result of reactions secondary to the nucleophilic additions involving 2-hydroxy-p-benzosemiquinone, H2O2 and hydroxyl radical. The process involves the primary oxidation of 2-hydroxy-p-benzosemiquinone by hydrogen peroxide, followed by oxidation of the semiquinone by hydroxyl radical leading to the formation of the electronically excited quinone. The decay of the excited triplet to the ground state is accompanied by photoemission with maximal intensity at 485-530 nm. Thermodynamic calculations along with an observed increase of photoemission intensity in anaerobiosis point to the triplet (n, π*) multiplicity of the excited state. The efficiency of chemiluminescence could be calculated as 10-8 photons/2-hydroxy-p-benzoquinone molecule formed.Photoemission arising from the p-benzoquinone/H2O2 reaction was inhibited efficiently by addition of GSH to the reaction mixture. This may be due to deactivation of the triplet quinone by a 2-glutathionyl-p-benzohydroquinone adduct, involving thioether α-hydrogen atom-transfer to the triplet ketone.
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  • 94
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 231-240 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; lipid peroxidation ; peroxidase-catalysed reaction ; chlorpromazine oxidation ; fertilization of sea urchin eggs ; activated leukocytes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Low-level chemiluminescence during lipid peroxidation and enzymatic reaction have been analysed by a filter type spectrometer. Tyrosine and tryprophan residues in proteins were found to be emitters in the visible region during their enzymatic oxidation. The natural chemiluminescence from fertilization of sea urchin eggs was found to have originated from tyrosine - cation radical mediated reaction in ovo-peroxidase - membrane protein - H2O2 system.
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  • 95
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 241-244 
    ISSN: 0884-3996
    Keywords: Oxidative stress ; anti-inflammatory drugs ; rat heart eicosanoids ; ischaemia ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Detection of ultraweak chemiluminescence (CL) emission from the surface of the organ is a sensitive and non-disruptive tool to evaluate the oxidative stress in rat heart. Indeed, an increased photon emission rate can be observed when cellular antioxidants such as glutathione or vitamin E are depleted, or when organic hydroperoxides are infused. We used CL recording to demonstrate in rat heart that: (i) different diets may lead to different heart sensitivity to an oxidative stress; and (ii) post-ischaemic reoxygenation induces an oxidative stress. CL emission induced by an oxidative stress is accompanied by an increased release of eicosanoids. However, while non-steroid anti-inflammatory drugs (aspirin, indomethacin and ibuprofen) prevented eicosanoid release, these compounds dramatically enhanced hydroperoxide-dependent CL. The nature of this phenomenon is still obscure, but the increase of steady-state concentration of excited species caused by anti-inflammatory drugs seems to be pathophysiologically relevant, since in all our experimental conditions tissue damage was proportional to CL emission rate.
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  • 96
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 263-266 
    ISSN: 0884-3996
    Keywords: Leukocytes ; respiratory burst ; granule ; calcium ionophore ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The granulocyte luminol-dependent chemiluminescence (CL) reaction is linked to the enzyme myeloperoxidase reacting with products of the respiratory burst activation. The results presented in this paper, show that the light generated in granulocytes originate both from intracellular and extracellular reactions; however, depending on the stimulus used the one or the other will dominate the activity measured. Furthermore, lysosomal fusion is proposed to be required for the intracellular CL reaction.
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  • 97
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 279-286 
    ISSN: 0884-3996
    Keywords: Chemiluminescence ; phagocytosis ; alveolar macrophages ; asthma ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Human alveolar macrophages (AM) were demonstrated to generate reactive toxic derivatives of oxygen in many pulmonary disorders. These cells are involved in local inflammation which characterizes bronchial asthma. In the present work, we studied the ability of stimulated macrophages from healthy volunteers, and asthmatic patients to generate oxygen species in vitro. AM obtained by bronchoalveolar lavage were purified by adherence. The production of oxygen species was measured by luminol-enhanced chemiluminescence (CL) after challenge with opsonized zymosan. The maximal values were significantly (p 〈 0.03 and p 〈 0.01) higher in AM from asthmatics than in AM from healthy subjects. A significant correlation (p 〈 0.01) was observed between maximal value of CL and the severity of asthma as assessed by the clinical score. But, no difference was observed between AM from asthmatics in a stable state and healthy subjects. On the other hand, assays for superoxide anion generation emphasized the activation state of these macrophages stimulated by formyl-peptides.
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  • 98
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    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 317-325 
    ISSN: 0884-3996
    Keywords: Bacterial luminescence ; LexA ; Sigma 32 ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We have recently shown that the transcription of the PR lux operon for Vibrio fischeri luminescence is positively controlled by the htpR (σ32) protein. It was suggested that the LexA protein might negatively control the lux genes. This paper extends these findings. It was found that Escherichia coli cells that contain the entire lux operon (pChv1) in RecA or LexA mutants which are unable to remove the LexA protein are considerable dimmer than the wild-type strain. Mutants that do not make LexA or from a weakly bound LexA are very bright. The role of σ32 protein was studied on luxR-luxl genes that are fused to β-galactosidase. The addition of V. fischeri inducer brings about the formation of β-galactosidase activity in htpR+ but not in htpR- strains of E. coli/pMJ3. Similar to the effect of starvation on the induction of luminescence in marine bacteria and in E. coli/pChv1 cells, β-galactrosidase activity in such constructs is preferentially induced by low nutrient concentrations. A new model for the regulatory control of the V. fischeri luminescence system is discussed.
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  • 99
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    Journal of Bioluminescence and Chemiluminescence 3 (1989) 
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
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  • 100
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    Journal of Bioluminescence and Chemiluminescence 3 (1989), S. 1-5 
    ISSN: 0884-3996
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The untimely death of Marlene DeLuca in 1987 has deprived the scientific community of an outstanding expert on bioluminescence. Earlier in that year she was honoured as thethirty-ninth recipient of the Otto Mitchell Smith Lectureship Award at Oklahoma State University, Stillwater, Oklahoma.On 20 March 1987 Dr DeLuca presented a scientific lecture entitled ‘Firefly Luciferase-Mechanism of Action, Cloning, and Expression of the Active Enzyme’ and a popular lecture at the banquet that evening entitled ‘Light and Life’. She was selected for her excellence in research, her oral presentation ability, and her personableness. Marlene was the first woman so honoured.To honour Dr Otto M. Smith the Alpha Delta Chapter of Phi Lambda Upsilon, a national chemistry honorary organization, inaugurated The Otto Mitchell Smith Lectureship in 1948 at Oklahoma State University. Former awardees include Nobel Laureates H. C. Brown, Stanford Moore, and Arthur Kornberg and the following prominent biochemists/molecular biologists: Robert A. Alberty, University of Wisconsin; Daniel E. Koshland, Brookhaven National Laboratory; Sol Spiegelman, University of Illinois; Carl Djerassi, Stanford University; and John T. Edsall, Harvard University. The lectureship honours Dr O. M. Smith, who was Director of the Research Foundation, professor, and Head of the Departments of Chemistry and Chemical Engineering.As a tribute to Dr DeLuca's outstanding contibution to bioluminescence we reproduce here the edited text of her Otto Mitchell Smith Lectureship and a selected bibliography of her work on firefly bioluminescence.
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