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Kinetics of luminol-enhanced chemiluminescence induced in murine splenocytes and bone marrow cells by various stimulating agents.

The luminol-amplified chemiluminescence (CL) activity of murine splenocytes and bone marrow cells was investigated and compared. Phagocytosis was activated by adding starch grains, opsonized zymosan particles (OZP), or phorbol myristate acetate (PMA) to cell suspensions. CL was studied within a concentration range of 3.13 x 10(6)-1.25 x 10(8) splenocytes/ml, and 1.56 x 10(6)-6.25 x 10(7) bone marrow cells/ml. The CL reaction intensity was increasing with rising cell concentration throughout the whole range studied. Starch grains and OZP elicited a unimodal kinetics of the CL response with a single peak after 10 min for splenocytes and 13-16 min for bone marrow cells. PMA activation induced a bimodal reaction with the first peak appearing after 3 min for either of the cell populations, and the second maximal peak being recorded after 6-7 min for splenocytes and 9-10 min for bone marrow cells.

Animals↗

[The action of gamma irradiation on the luminol-dependent chemiluminescence of activated thymocytes].

The method of luminol-dependent chemiluminescence was used to study the affect of gamma radiation on activation of cells of thymus and bursa of Fabricius upon their adhesion on kapron. Thymocyte activation by adhesion on glass or kapron provoked chemiluminescence which was connected with the development in cells of active oxygen forms. The chemiluminescence amplitude was shown to increase with adhesion of thymocytes exposed to 1-5 Gy radiation and to decrease after a dose of 10 Gy. In cells of bursa of Fabricius, the analogous response to activation or irradiation was not observed.

Animals↗

Luminol-chemiluminescence in free peritoneal cells in hemorrhagic shock in rats treated with PAF-receptor-antagonist BN 52021.

The activity of rat peritoneal cells were assessed by the phorbol mirystinian acetate (PMA)-induced luminol chemiluminescence (LCL). Results in control groups (0 - no manipulation, and I - carotid artery cannulation) were compared with those in the untreated hemorrhagic shock (group II), in the shock treated with the standard polyelectrolyte solution (PES) (group III), and in shock treated with PAF receptor-antagonist BN 52021 + PES (group IV). The maximal and the most rapid LCL was observed in the group treated with BN 52021 (group IV), while chemiluminescent response in the the untreated shock (group II) and in shock treated with PES was minimally expressed and late. The findings indicate for a rapid activation of peritoneal cells during ca 1 hour of hemorrhagic shock. This leads to exhausting their ability to the superoxide anion generation 15 minutes later. Peritoneal cells obtained from the group treated with the BN 52021 revealed a preserved ability to the respiratory burst. It can be concluded that BN 52021 effectively inhibits activation of the PC during hemorrhagic shock.

Analysis of Variance↗

[The effect of norfloxacin on the luminol-dependent chemiluminescence and adhesion of leukocytes].

The in vitro effect of norfloxacin on the luminol-dependent chemiluminescence and adherence of polymorphonuclear leukocytes of guinea pigs was studied with the use of a wide range of the drug concentrations from 0.1 to 100 micrograms/ml under two different conditions, i.e. during the cell incubation in the presence of norfloxacin various concentrations and after the cell washing to remove the drug. In a concentration of 1 microgram/ml norfloxacin stimulated the zymosan-induced chemiluminescence. When the drug was used in a concentration of 10 micrograms/ml, the stimulating effect was less pronounced. In a concentration of 100 mu/ml norfloxacin significantly inhibited the leukocyte chemiluminescence. In concentrations of 0.1 to 100 micrograms/ml norfloxacin had no effect on the leukocyte adherence to the plastic surface covered by albumin.

Animals↗

Modulation of luminol-dependent chemiluminescence of murine macrophages by flavone and its synthetic derivatives.

The effect of flavone (CAS 525-82-6, 2-phenylbenzopyran-4-one, 1), flavone-8-acetic acid (CSA 87626-55-9, FAA, 2) and 10 substituted flavones on the luminol-dependent chemiluminescence of murine macrophages was studied in vitro. The synthetic derivatives were variously substituted with halo, nitro, amino, hydroxy and methoxy substituents in the 3' and 4' positions. Chemiluminescence was used in this study as an indicator for the production of reactive oxygen species by macrophages, stimulated in vitro by phorbol myristate acetate (PMA). All flavones except FAA (2) showed more than 20% inhibition at 10 mumol/l or 100 mumol/l. 3'-Amino-4'-hydroxyflavone (8) was the most potent inhibitor. The IC50s for inhibition of chemiluminescence were 4.2 +/- 1.1 mumol/l, 5.0 +/- 1.0 mumol/l and 3.3 +/- 1.4 mumol/l for resident, elicited and LPS-Poly I:C-primed macrophages, respectively. Small but statistically significant enhancements of chemiluminescence were caused by low concentrations of flavone (1), FAA (2) and 4'-methoxyflavone (6). These results suggest that modulation of the chemiluminescent capacity of macrophages depends on the nature of the substituents and the concentration of the flavones.

Animals↗

[Effect of lomefloxacin on luminol-dependent chemiluminescence and adhesion of leukocytes].

The in vitro effect of lomefloxacin within the wide dose ranges from 0.1 to 100 micrograms/ml on luminol-dependent chemiluminescence and adherence of polymorphonuclear leukocytes of guinea pigs was studied under two different conditions: during the cell incubation in the presence of various concentrations of the drug and after the cell washing to remove lomefloxacin. In concentrations of 10 and 100 micrograms/ml the drug inhibited the response under the both conditions while in a concentration of 1 microgram/ml it stimulated the chemiluminescence. In concentrations of 0.1 to 100 micrograms/ml lomefloxacin stimulated the leukocyte adherence. The highest stimulation was observed with the use of the drug in a concentration of 1 microgram/ml.

Animals↗

Phenomenon of specific inhibition of luminol-dependent chemiluminescence of polymorphonuclear leukocytes under the influence of allergen.

The purpose of our investigation was to establish how the intensity of induced luminol-dependent chemiluminescence of polymorphonuclear leukocytes is affected by the cultivation of allergen with the blood of allergic patients (pollinosis, bronchial asthma and penicillin allergy) and animals with allergic processes (Arthus' phenomenon, guinea pigs sensitized by horse serum). The results of all assays were uniform. In all cases three phases of changes in induced chemiluminescence were established. In the first phase with the minimum doses of specific and unspecific allergens, either no changes in chemiluminescence were found or chemiluminescence even increased. With higher doses of allergen came the second phase - inhibition of stimulated chemiluminescence when specific allergen had been added and the absence of changes in the tests with unspecific allergen. We defined this phase as the phase of specific chemiluminescence inhibition. With a further increase in the concentration of allergen came the third phase. It expressed itself in the inhibition of stimulated chemiluminescence under the influence of both specific and unspecific allergen. We viewed this phase as a manifestation of the toxic influence of allergen on polymorphonuclear leukocytes. We suggested using the second phase for diagnosing sensitization.

Allergens↗

[The measuring of blood plasma antioxidant activity by the hemoglobin-hydrogen peroxide-luminol system].

The method of valuation of the blood plasma antioxidant activity (AOA) by the hemoglobin-hydrogen peroxide-luminol chemiluminescence system has been proposed. The method is based on the measuring the induction time of chemiluminescence that is directly proportional to the added volume of plasma or concentration of standard antioxidant. The ascorbate was taken as a standard. Blood plasma AOA has been expressed through the concentration of equivalent ascorbate solution (ascorbate equivalent). The influence of hemolysis and storage conditions of plasma on its detected AOA was investigated. The change of human blood plasma AOA after single administration of 2 g the ascorbate and its dynamics in patients with acute pancreatitis was studied.

Acute Disease↗

[Effect of metronidazole on luminol dependent chemiluminescence of neutrophils in whole blood of patients with chlamydial infection].

In vivo effect of metronidazole on the active oxygen production by neutrophils in the peripheral blood of patients with urogenital chlamydial infection (9 females and 7 males) was studied with the use of luminol dependent chemiluminescence. Metronidazole was administered orally in a dose of 250 mg twice a day for 10 days. Blood for the specimens was collected prior to the treatment and on the next day of the treatment course completion. No effect on the spontaneous and zymozan or phorbol-12-myristate-13-acetate stimulated chemiluminescence was observed, although a significant increase in the chemiluminescence indices was stated in 3 females and 1 male. It was suggested that the changes in the individual values of the neutrophil chemiluminescence after the treatment were associated with the influence of metronidazole and/or its metabolites on the blood cells and with different activity of the host enzymes participating in the drug biotransformation.

Administration, Oral↗

[The effect of ionizing radiation and radiosensitizing agent AK-2123 on luminol-dependent chemiluminescence of human neutrophils].

The method of luminol-dependent chemiluminescence (CL) was used to study the influence of gamma- and fast electrons-irradiation on secretion of reactive oxygen species of neutrophils in enrich suspension and in presence of all blood components. In vitro gamma-irradiation (from 5 to 25 Gy) of the neutrophil suspension inhibited spontaneous CL and activated stimulated by phorbol-12-myristate-13-acetate (PMA) (100 nM) in the period 30-60 min after radiation. The time of maximum PMA-stimulated CL amplitude was decreased with the dose range from 2(5) to 25(30) Gy for gamma- and fast electrons-irradiation. The addition of radiosensitizer AK-2123 (0.05% or 2.2 mM) depressed the early PMA-stimulated CL response on gamma- and fast electrons-irradiation. The obtained results suggest that this effect is connected with influence of AK-2123 on ion canals of neutrophils.

Gamma Rays↗

Inhibition of monocyte luminol-dependent chemiluminescence by tetrahydrobiopterin, and the free radical oxidation of tetrahydrobiopterin, dihydrobiopterin and dihydroneopterin.

Luminol-dependent chemiluminescence of normal human monocytes activated by zymosan is demonstrated to be inhibited by tetrahydrobiopterin in a concentration-dependent manner. The reduced pterins tetrahydrobiopterin, dihydrobiopterin, and dihydroneopterin are all shown to be readily oxidized by the hydroxyl radical. The susceptibility of reduced pterins to free radical attack may explain the inhibition of chemiluminescence observed and an additional role of reduced pterins as free radical scavengers in tissues is considered.

Ascorbic Acid↗

[Modification of phagocytic microbicide function by antifungal agents--measuring luminol enhanced chemoluminescence in full blood].

Fungus infections are becoming more important in surgical intensive medicine, and various preparations are now available to treat them. The goal of our investigations was to determine the influence of the antimycotics in current use on the microbicide phagocytic function by measuring chemiluminescence. To this end the luminol-enhanced chemiluminescence in whole blood samples from a total of 29 healthy donors was measured with a LKB 1251 Luminometer after stimulation with zymosan or a Canadida albicans preparation. We tested the substances amphotericin B, flucytosin (Ancotil), fluconazol (Diflucan) and itraconazol (Sempera), each in three different concentrations within the recommended dose range and compared the results with those in an untreated sample. For the insertions of amphotericin B or intraconazol after zymosan stimulation no significant differences in the measurements were found (50.84 vs 47.99 mV and 46.10 vs 47.89 mV) compared with the blank test. Similar situations revealed by the tests with C. albicans administration (15.21 vs 12.35 mV and 11.16 vs 11.91 mV). However, the tests with flucytosin in the higher concentration range, after stimulation with either zymosan or C. albicans, evidenced a significant reduction in the measurements (34.70 vs 52.74 mV, P<0.005, and 10.98 vs 14.57 mV, P<0.01). The tests with fluconazol showed a decrease of the chemiluminescence exclusively for the highest concentration in the C. albicans group (14.36 vs 17.20 mV, P<0.005). Our results indicate a negative influence of the phagocytes on the oxidative metabolism especially with flucytosin in the higher concentrations. This emphatically confirms demands for exact indications and dosage of antimycotics and their correct administration.

Antifungal Agents↗

Decreased luminol-dependent chemiluminescence response of neutrophils to recombinant human tumour necrosis factor in patients with gastric cancer.

Production of reactive oxygen species by neutrophils was analyzed by a chemiluminometric method in the presence of luminol in ten healthy donors, eight patients with gastric cancer and ten patients with gastric precancer. The neutrophil chemiluminominescence response to recombinant tumour necrosis factor (rTNF) was almost five times lower in the gastric patients when compared with healthy donors and precancer patients. The chemiluminescence response to zymosan was decreased only in the gastric cancer patients with chemiluminescence index activation (CTNF) less than 1. The observed changes of neutrophil functions are thought to result in a decrease of neutrophil cytotoxic activity in gastric cancer patients.

Adult↗

Determination of proteins at nanogram levels by their quenching effect on the chemiluminescence reaction between luminol and hydrogen peroxide with manganese-tetrasulfonatophthalocyanine as a new catalyst.

The manganese-tetrasulfonatophthalocyanine (MnTSPc) catalyzed luminol-hydrogen peroxide chemiluminescence (CL) systems can be quenched in the presence of proteins. A highly sensitive CL quenching method has been developed for the determination of proteins. Under optimum conditions, the linear ranges of the calibration curves were 0.1-20 microg/mL for human serum albumin (HSA), 0.2-20 microg/mL for human gamma-IgG, and 0.5-50 microg/mL for the bovine serum albumin (BSA) with the corresponding detection limits were 1.9 ng/mL, 2.7 ng/mL, and 3.4 ng/mL. The method has been applied to the analysis of total proteins in human serum samples and the results were in good agreement with clinical data provided.

Animals↗

Determination of ATP via the photochemical generation of hydrogen peroxide using flow injection luminol chemiluminescence detection.

The determination of ATP using the coupling between a photochemical reaction and a chemiluminescence reaction in a flow injection (FI) system is described. The method is based on the reaction of glucose with ATP catalyzed by hexokinase and Mg(2+) ions. The glucose that is not consumed by ATP is subsequently photooxidized using 9,10-anthraquinone-2,6-disulfonate as a sensitizer. The hydrogen peroxide produced in the photochemical reaction is monitored through the chemiluminescence reaction with luminol catalyzed by hematine. There is a linear relationship between the decrease in the chemiluminescence response and the ATP concentration at a constant concentration of glucose. Under the optimum conditions, the calibration graph is linear in the range 0.20-50.5 mg L(-1) with a throughput of 25 samples per hour and relative standard deviations between +/-0.62 and +/-1.42%. The limit of detection is 0.07 mg L(-1). The method was used for the determination of ATP in pharmaceuticals, milk, and soils.

Journal Article↗

Fluorimetry studies of oscillating chemilumnescence in the luminol-H2O2-KSCN-CuSO4-TMAOH system.

Oscillating chemical reactions are complex systems, involving a large number of chemical species. In oscillating chemical reaction, some species, usually a reaction intermediate, exhibit fluctuation in its concentration. In this report, visible oscillating chemiluminescence produced by the addition of luminol (3-aminophthalhydrazide) to the oscillating system of H2O2-KSCN-CuSO4-TMAOH was investigated using spectrofluorimetry method. The effects of ingredient concentration of the oscillating system and complexing agents like citric acid and cysteine on the behavior of the oscillating system were investigated. Moreover, the influence of nonaqueous solvents such as ethanol and ethylene glycol has been studied.

Journal Article↗

Development and optimization of an analytical method for the determination of Sudan dyes in hot chilli pepper by high-performance liquid chromatography with on-line electrogenerated BrO- -luminol chemiluminescence detection.

The determination of four Sudan dyes by means of high-performance liquid chromatography (HPLC) with chemiluminescence (CL) detection was proposed. The method was based on the enhancement effect of Sudan dyes on the chemiluminescence reaction between luminol and BrO-, which was on-line electrogenerated by constant current electrolysis. The separation was carried out on Nucleosil RP-C18 column (250 mm x 4.6 mm i.d., 5 microm, pore size, 100 A) at 35 degrees C. The mobile phase consisted of a V (methanol): V (0.2% aqueous formic acid) = 90:10 solution. At a flow-rate of 1.0 ml min(-1), the total run time was 25 min. The effects of several parameters on the HPLC resolution and CL emission were studied systematically. For the four Sudan dyes, the limits of detection (LOD) at a signal-to-noise of 3 ranged from 4 to 8 microg kg(-1) and the limits of quantification (LOQ) at a signal-to-noise of 10 ranged from 13 to 27 microg kg(-1). The relative standard deviations (RSD) of intra-and inter-day precision were below 4.4%. The average recoveries for all four Sudan dyes (spiked at the levels of 1.0 and 1.5 mg kg(-1)) in chilli tomato sauce and hot chilli pepper ranged from 94% to 105%, and the relative standard deviations of the quantitative results were from 2.5 to 4.2%. The proposed method had been successfully applied to the determination of four Sudan dyes in hot chilli products.

Azo Compounds↗

Trace analysis of phosphorus in water by sorption preconcentration and luminol chemiluminescence

A new, highly sensitive chemiluminescence method for the determination of sub-ppb quantities of phosphorus in water is described. The method is based on sorption preconcentration of phosphorus as a yellow vanadomolybdophosphoric heteropoly acid (HPA) in the presence or absence of a cationic surfactant on a paper filter, followed by direct chemiluminescence detection of the phosphorus concentrate via reaction with an alkaline luminol solution. The molar ratio of cationic surfactant to HPA in the ion associate sorbed on the filter is 4:1. The detection limits for phosphorus are 0.02 microgram of P L-1 in the presence of surfactant and 0.1 microgram of P L-1 in the absence of surfactant for a sample volume of 150 mL. The calibration plot is linear from 0.06 to 1.7 micrograms of P L-1 in the presence of a surfactant, and the time required for analysis is 25 min. In the absence of surfactant, the selectivities against Si4+ and As5+ are 5 and 40 times greater than those for the standard colorimetric method based on the formation of the blue molybdophosphoric HPA. Applications of the method to the analyses of river water, seawater, and the turbine vapor condensate from a coal-fired power plant are described. It is demonstrated that the sensitivity advantage of the chemiluminescence technique can be combined with the magnesium-induced coprecipitation (MAGIC) method for a more selective measurement of soluble reactive phosphorus.

Journal Article↗