Bioluminescent methods are gaining more and more attention among scientists due to their sensitivity, selectivity and simplicity; coupled with the fact that the bioluminescence can be monitored both in vitro and in vivo. Since the discovery of bioluminescence in the 19th century, enzymes involved in the bioluminescent process have been isolated and cloned. The bioluminescent reactions in several different organisms have also been fully characterized and used as reporters in a wide variety of biochemical assays. From the 1990s it became clear that bioluminescence can be detected and quantified directly from inside a living cell. This gave rise to numerous possibilities for the in vivo monitoring of intracellular processes non-invasively using bioluminescent molecules as reporters. This review describes recent developments in the area of bioluminescent imaging for cell biology. Newly developed imaging methods allow transcriptional/translational regulation, signal transduction, protein–protein interaction, oncogenic transformation, cell and protein trafficking, and target drug action to be monitored in vivo in real-time with high temporal and spatial resolution; thus providing researchers with priceless information on cellular functions. Advantages and limitations of these novel bioluminescent methods are discussed and possible future developments identified.

Bioluminescence imaging

10.3184/003685007x215986

Cite

Citations (11)

Characterization of immune-active peptides obtained from milk fermented byLactobacillus helveticus

Journal of Dairy Research (2010)

Angela M. Tellez Milena Corredig Lubov Brovko Mansel W. Griffiths

The objectives of this research were to confirm the effect of compounds derived from milk fermented by Lactobacillus helveticus (LH-2) on the nonspecific host defence system, and isolate and characterize the active peptides that mediate the immune response. The cell-free supernatant obtained from the fermented milk and its fractions were tested in vitro for immuno-modulating activity using murine macrophages (RAW 264·7 cell line). Cytokine production (Interleukin-6 (IL-6), Tumor Necrosis Factor-α (TNF-α), and Interleukin-1β (IL1-β)), nitric oxide (NO) production and phagocytosis were used as biomarkers. Macrophages stimulated with cell-free supernatant of fermented milk showed higher production of cytokines and NO compared with macrophages stimulated with LPS (Lipopolysaccharide) and a commercial immunomodulator derived from β-casein (f54-59). Phagocytosis was observed by macrophages stimulated with the supernatant. Two of nine fractions collected from the supernatant using size exclusion chromatography produced the highest response when used to stimulate macrophages. The results of the dose-response study of the effect of the fraction with the highest stimulation effect on the production of TNF-α showed a direct correlation between protein concentration and TNF-α release. The fraction contained four novel peptides, three derived from the hydrolysis of β-casein and one from the hydrolysis of α-lactalbumin. These results confirm that fermentation of milk by Lactobacillus helveticus (LH-2) results in the production of specific peptides capable of modulating macrophage activity.

Lactobacillus helveticus

10.1017/s002202990999046x

Cite

Citations (66)

Luciferase intramolecular dynamics studied by time-resolved fluorescence spectroscopy

Proceedings of SPIE, the International Society for Optical Engineering/Proceedings of SPIE (1993)

Lubov Brovko Andrey Y. Chikishev Olga Gandelman A. P. Shkurinov

Earlier a comprehensive study of reaction substrate, product and their complexes with the enzyme has been carried out by means of stationary fluorescence spectroscopy. A hypothesis has been suggested according to which protein changes its conformation during the reaction. In this respect it is quite interesting to study both static and dynamic properties of the enzyme molecule. Luckily, according to genetic engineering data, luciferase molecule contains only one tryptophan residue, which makes it a very convenient object for the studies by means of time-resolved fluorescence spectroscopy. The scope of this paper is the study of luciferase tryptophan fluorescence. The methods applied were fluorescence spectrochronography and anisotropy decay analysis.

Fluorescence Correlation Spectroscopy

Molecular biophysics

10.1117/12.146114

Cite

Citations (0)

Bioluminescence of free and poly(vinyl alcohol) cryogel-entrapped recombinant Escherichia coli cells expressing firefly luciferase.

PubMed (1998)

Nadya Romanova Lubov Brovko T. Makhlis E M Sinodova Н.Н. Угарова

Vinyl alcohol

Membrane permeability

Source

Cite

Citations (1)

Bioluminescence lights the way to food safety

Proceedings of SPIE, the International Society for Optical Engineering/Proceedings of SPIE (2003)

Lubov Brovko Mansel W. Griffiths

The food industry is increasingly adopting food safety and quality management systems that are more proactive and preventive than those used in the past which have tended to rely on end product testing and visual inspection. The regulatory agencies in many countries are promoting one such management tool, Hazard Analysis Critical Control Point (HACCP), as a way to achieve a safer food supply and as a basis for harmonization of trading standards. Verification that the process is safe must involve microbiological testing but the results need not be generated in real-time. Of all the rapid microbiological tests currently available, the only ones that come close to offering real-time results are bioluminescence-based methods. Recent developments in application of bioluminescence for food safety issues are presented in the paper. These include the use of genetically engineered microorganisms with bioluminescent and fluorescent phenotypes as a real time indicator of physiological state and survival of food-borne pathogens in food and food processing environments as well as novel bioluminescent-based methods for rapid detection of pathogens in food and environmental samples. Advantages and pitfalls of the methods are discussed.

SAFER

Harmonization

Food safety management

10.1117/12.477888

Cite

Citations (1)

Bioluminescent Analysis of β-Galactosidase and Its Application for Immunoassay and DNA-Probes

Analytical Letters (1994)

Lubov Brovko Ekaterina Dementieva E. A. Aronova P. V. Yevseev Н.Н. Угарова

Abstract Bioluminescent method for β-galactosidase assay is proposed with sensitivity as high as 0.25 fmol/1 with the help of bioluminogenic substrate – firefly luciferin derivative. Method was applied to immunoassay of foot-and-mouth disease viral antigen and biotinylated DNA-probe analysis with the help of β-galactoside labelled conjugates. Methods proved to be very sensitive, rapid and simple in performance. Photographic detection and reprobing of the samples are possible. Key Words: ImmunoassayDNA ProbesB-galactosidaseBioluminescent

Luminescent Measurements

10.1080/00032719408000303

Cite

Citations (0)