Nitric oxide donor-mediated killing of bioluminescent Escherichia coli

Virta, Marko; Karp, Matti; Vuorinen, Pauli

doi:10.1128/aac.38.12.2775

Cited by 30 publications

(21 citation statements)

References 28 publications

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“…Variation between the measurements was less than 10%, in agreement with previous studies (29,31). RLU, relative light units.…”

Section: Discussionsupporting

confidence: 78%

“…Therefore, many applications for luminescent cells have been created during the last few years, for instance, an assay for the extremely sensitive detection of the drug susceptibility of Mycobacterium tuberculosis (9). We have used bacteria cloned either with bacterial luciferase (lux) genes or with insect luciferase (luc) genes as general detectors of toxic substances (17), in studies concerning the killing of E. coli cells with nitric oxide-donating chemicals (29), and for the specific detection of heavy metals such as mercuric ions (30).…”

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confidence: 99%

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Antimicrobial Compounds

2013

Freshwater Phytopharmaceutical Compounds

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The oral bacterium Streptococcus mutans was transformed by electroporation with a shuttle vector (pCSS945) containing insect luciferase gene from a click beetle (Pyrophorus plagiophthalamus) resulting in a bioluminescent phenotype. This S. mutans strain was used in experiments in which light emission was used as a rapid and, compared to conventional CFU counting, more convenient means of estimating the effects of various antimicrobial treatments. The basic parameters affecting in vivo light production by the strain were studied. It was found that pH 6.0 was optimal for incorporation of the substrate D-luciferin for the luciferase reaction. The optimum concentration of D-luciferin was approximately 150 M at room temperature. Under optimum conditions the light emission in vivo increased rapidly to a constant level and thereafter had a decay of 0.6%/min when logarithmic-growth-phase cells were used. The light emission closely paralleled the numbers of CFU, giving a detectable signal from 30,000 cells and having a dynamic measurement range over 4 log CFU/relative light unit. The cells were treated with various antimicrobial agents, and the emitted bioluminescence was measured. With the bioluminescent measurements, the results were obtained within hours compared to the days required for agar plates, and also, the kinetics of the antibacterial actions could be followed. Thus, the light emission was found to be a reliable, sensitive, and real-time indicator of the bacteriostatic actions of the antimicrobial agents tested.

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“…Variation between the measurements was less than 10%, in agreement with previous studies (29,31). RLU, relative light units.…”

Section: Discussionsupporting

confidence: 78%

mentioning

confidence: 99%

Antimicrobial Compounds

2013

Freshwater Phytopharmaceutical Compounds

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“…These compounds have been shown to inhibit neutrophil functions (Moilanen et al, 1993;, suppress tumour-cell growth (Vilpo et al, 1994), and to have antibacterial (Virta et al, 1994), vasodilator, antiplatelet and fibrinolytic (Corell et al, 1994) and low density lipoprotein oxidation inhibitory activities . In guinea-pig trachea and human neutrophils the biological activity was associated with an increase in the production of cyclic GMP (Moilanen et al, 1993;Corell et al, 1994).…”

Section: Introductionmentioning

confidence: 99%

Nitric oxide‐donating properties of mesoionic 3‐aryl substituted oxatriazole‐5‐imine derivatives

Kankaanranta

Rydell

Petersson

et al. 1996

British J Pharmacology

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1 The nitric oxide (NO)-releasing properties of two new mesoionic 3-aryl substituted oxatriazole-5-imine derivatives (GEA 3162 and GEA 3175) were characterized and compared with the known NOdonors 3-morpholino-sydnonimine (SIN-1) and S-nitroso-N-acetylpenicillamine (SNAP). 2 GEA 3162, GEA 3175, SIN-i and SNAP inhibited adenosine 5'-diphosphate-induced platelet aggregation (IC50 values 0.18, 0.39, 3.73 and 2.12 gM, respectively). All four compounds induced a dosedependent and more than 4 fold increase in cyclic GMP in platelets. The increase in cyclic GMP concentration was potentiated more than 1.5 fold by a phosphodiesterase inhibitor, zaprinast (10 gM) and inhibited 38-97% by oxyhaemoglobin (10-45 gM).3 All of the four compounds studied converted oxyhaemoglobin to methaemoglobin and formed a paramagnetic NO-haemoglobin complex. All but GEA 3175 formed nitrite and nitrate in phosphate buffer. During a 40 min incubation, GEA 3162, SIN-I and SNAP (100 pM) produced 50-70 gM NO2-+ NO3-as determined by high performance liquid chromatography. The release of NO and NO2 by GEA 3175 was increased 140 fold in the presence of human plasma (0.14 and 19.7 ppb in the absence and presence of 1% human plasma, respectively) as analyzed by ozone chemiluminescence. 4 The results suggest that the mesoionic 3-aryl substituted oxatriazole-5-imine derivatives GEA 3162 and GEA 3175 as well as SIN-l and SNAP release nitric oxide.

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“…Green fluorescent protein (GFP) is a highly fluorescent, extremely stable protein that accumulates in cells during growth; therefore, the measured fluorescence signal is proportional to the total number of cells [28]. The luciferase reaction is dependent on ATP produced by living cells [29], so that the bioluminescence level is a direct measure of viable cells [22]. Figs.…”

Section: Resultsmentioning

confidence: 99%

“…There are several different methods used for these purposes, and in general each method is most suitable for a given application. Plate counting is traditionally used for measuring bacterial viability [22], whilst the conventional technique for measuring the susceptibility of bacterial cells to various antimicrobial agents is the microbroth dilution method [23]. The checkerboard method is an in vitro method most frequently used to evaluate the synergistic/antagonistic action of two drugs at one time against a single microorganism [24].…”

Section: Introductionmentioning

confidence: 99%