34 research outputs found
Evaluating the Effect of Substrates on the Stability of the Bacterial Luminescent System during Co-Immobilization in the Starch Gel
This study presents the evaluation of the effect of the substrates: tetradecanal and NADH on the activity of the coupled enzyme system NAD(P)H: FMN oxidoreductase β luciferase during co-immobilization and long-term storage.Π£ΡΠ°ΡΡΠΈΠ΅ Π² IX ΠΠ΅ΠΆΠ΄ΡΠ½Π°ΡΠΎΠ΄Π½ΠΎΠΉ ΠΌΠΎΠ»ΠΎΠ΄Π΅ΠΆΠ½ΠΎΠΉ Π½Π°ΡΡΠ½ΠΎΠΉ ΠΊΠΎΠ½ΡΠ΅ΡΠ΅Π½ΡΠΈΠΈ Β«Π€ΠΈΠ·ΠΈΠΊΠ°. Π’Π΅Ρ
Π½ΠΎ-Π»ΠΎΠ³ΠΈΠΈ. ΠΠ½Π½ΠΎΠ²Π°ΡΠΈΠΈ. Π€Π’Π-2022Β» ΠΏΠΎΠ΄Π΄Π΅ΡΠΆΠ°Π½ΠΎ ΠΡΠ°ΡΠ½ΠΎΡΡΡΠΊΠΈΠΌ ΠΊΡΠ°Π΅Π²ΡΠΌ ΡΠΎΠ½Π΄ΠΎΠΌ Π½Π°ΡΠΊΠΈ
ΠΡΠ΅Π½ΠΊΠ° ΠΏΡΠΎΡΠΈΠ²ΠΎΠΎΠΏΡΡ ΠΎΠ»Π΅Π²ΡΡ , ΡΠΎΠΊΡΠΈΡΠ΅ΡΠΊΠΈΡ ΡΡΡΠ΅ΠΊΡΠΎΠ² ΠΈ Ρ Π°ΡΠ°ΠΊΡΠ΅ΡΠ° ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ Π³Π΅Π½ΠΎΠ²-ΠΌΠΈΡΠ΅Π½Π΅ΠΉ miR-204-5p ΠΏΡΠΈ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠΈ Π΅Π΅ ΠΈΠΌΠΈΡΠ°ΡΠΎΡΠ° Π½Π° ΠΌΠΎΠ΄Π΅Π»ΠΈ ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ Π-16 in vivo
Objective. To evaluate anti-tumor, toxic effect of miR-204-5p mimic applicaton on melanoma B-16-bearing mice followed by miR-204-5p target gene expression estimation in melanoma tumor and distant organs. Material and Methods. C57Bl/6 melanoma B-16-bearing mice were used. The animals of the experimental group were intraperitoneally injected with a 5 nM miR-204-5p miRNA simulator (mimic) on the 8th, 10th, and 12th days after melanoma B-16 cell transplantation. Based on the results of bioinformatic analysis, miR-204-5p target genes BCL2 and SIRT1 expression levels were determined by quantitative real-time PCR. The toxic effect of miR-204-5p mimic was estimated by the evaluation of body weight, mass of the internal organs, and motor activity. Results. On the 13-14th days of the experiment, the motor activity of animals in the control groups decreased signifcantly compared to the group of animals treated by miR-204-5p. Target gene BCL2 showed increased expression in the lungs and kidneys and SIRT1 levels were increased in the lungs of miR-204-5p mimic treated animals (pΛ0.05). Tumor mass tended to decrease in the animals treated by miR-204-5p mimic. Conclusion. Modulation of the level of miR-204-5p microRNA led to changes in the expression of SIRT1 and BCL2 in the lungs of animals, and changes in the expression of BCL2in the kidneys. MiR-204-5p mimic application did not have toxic effect on animals treated. Further studies are necessary to clarify miR-204-5p implication in melanoma cell proliferation regulation as well as itβs biodistibution in the tumor tissue.Π¦Π΅Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ β ΠΎΡΠ΅Π½ΠΊΠ° Π²Π»ΠΈΡΠ½ΠΈΡ ΠΈΠΌΠΈΡΠ°ΡΠΎΡΠ° miR-204-5p Π½Π° ΡΠΎΡΡ ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ Π-16 in vivo ΠΏΡΠΈ Π²Π½ΡΡΡΠΈΠ±ΡΡΡΠΈΠ½Π½ΠΎΠΌ ΡΡΠ΅Ρ
ΠΊΡΠ°ΡΠ½ΠΎΠΌ Π΅Π³ΠΎ Π²Π²Π΅Π΄Π΅Π½ΠΈΠΈ, ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΠ΅ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΠΏΡΠΈ ΡΡΠΎΠΌ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ Π³Π΅Π½ΠΎΠ²-ΠΌΠΈΡΠ΅Π½Π΅ΠΉ miR-204-5p Π² ΠΎΠΏΡΡ
ΠΎΠ»ΠΈ ΠΈ Π΄ΠΈΡΡΠ°Π½ΡΠ½ΡΡ
ΠΎΡΠ³Π°Π½Π°Ρ
, Π° ΡΠ°ΠΊΠΆΠ΅ Π²ΡΡΠ°ΠΆΠ΅Π½Π½ΠΎΡΡΠΈ ΡΠΎΠΊΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΡΠ΅Π°ΠΊΡΠΈΠΉ.ΠΠ°ΡΠ΅ΡΠΈΠ°Π» ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ. ΠΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΠ»ΠΈ Π½Π° ΠΌΡΡΠ°Ρ
C57Bl/6 Ρ ΠΏΠΎΠ΄ΠΊΠΎΠΆΠ½ΠΎ ΠΏΠ΅ΡΠ΅Π²ΠΈΡΠΎΠΉ ΠΌΠ΅Π»Π°Π½ΠΎΠΌΠΎΠΉ B-16. ΠΠΈΠ²ΠΎΡΠ½ΡΠΌ ΠΎΠΏΡΡΠ½ΠΎΠΉ Π³ΡΡΠΏΠΏΡ Π²Π½ΡΡΡΠΈΠ±ΡΡΡΠΈΠ½Π½ΠΎ Π²Π²ΠΎΠ΄ΠΈΠ»ΠΈ ΠΈΠΌΠΈΡΠ°ΡΠΎΡ ΠΌΠΈΠΊΡΠΎΠ ΠΠ miR-204-5p (5Π½ΠΠΎΠ»Ρ) Π½Π° 8, 10, 12-Π΅ ΡΡΡ ΠΏΠΎΡΠ»Π΅ ΡΡΠ°Π½ΡΠΏΠ»Π°Π½ΡΠ°ΡΠΈΠΈ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ. Π‘ΠΎΠ³Π»Π°ΡΠ½ΠΎ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠ°ΠΌ Π±ΠΈΠΎΠΈΠ½ΡΠΎΡΠΌΠ°ΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π°Π½Π°Π»ΠΈΠ·Π° ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ ΡΡΠΎΠ²Π΅Π½Ρ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ Π³Π΅Π½ΠΎΠ²-ΠΌΠΈΡΠ΅Π½Π΅ΠΉ ΠΌΠΈΠΊΡΠΎΠ ΠΠ BCL2 ΠΈ SIRT1 ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΠ¦Π Π² ΡΠ΅Π°Π»ΡΠ½ΠΎΠΌ Π²ΡΠ΅ΠΌΠ΅Π½ΠΈ. ΠΠΏΡΠ΅Π΄Π΅Π»ΡΠ»ΠΈ ΡΠΎΠΊΡΠΈΡΠ΅ΡΠΊΠΈΠΉ ΡΡΡΠ΅ΠΊΡ Π²ΠΎΠ·Π΄Π΅ΠΉΡΡΠ²ΠΈΡ ΠΈΠΌΠΈΡΠ°ΡΠΎΡΠ° ΠΏΠΎ Π΄ΠΈΠ½Π°ΠΌΠΈΠΊΠ΅ ΠΌΠ°ΡΡΡ ΡΠ΅Π»Π° ΠΈ ΠΎΡΠ³Π°Π½ΠΎΠ², ΠΎΠ±ΡΠ΅ΠΌΡ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΠΎΠ³ΠΎ ΡΠ·Π»Π°, ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ Π΄Π²ΠΈΠ³Π°ΡΠ΅Π»ΡΠ½ΠΎΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΈ Π²Π½Π΅ΡΠ½Π΅Π³ΠΎ Π²ΠΈΠ΄Π° ΠΆΠΈΠ²ΠΎΡΠ½ΡΡ
Π² ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ. ΠΡΠ΅Π½ΠΊΠ° Π²Π½Π΅ΡΠ½ΠΈΡ
ΠΏΡΠΈΠ·Π½Π°ΠΊΠΎΠ² ΠΈ Π΄ΠΈΠ½Π°ΠΌΠΈΠΊΠΈ Π΄Π²ΠΈΠ³Π°ΡΠ΅Π»ΡΠ½ΠΎΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΆΠΈΠ²ΠΎΡΠ½ΡΡ
, Π° ΡΠ°ΠΊΠΆΠ΅ Π΄ΠΈΠ½Π°ΠΌΠΈΠΊΠΈ ΠΈΡ
ΠΌΠ°ΡΡΡ ΠΈ ΠΌΠ°ΡΡΡ ΠΎΡΠ³Π°Π½ΠΎΠ² ΠΏΡΠΈ Π²ΡΠΊΡΡΡΠΈΠΈ ΡΠ²ΠΈΠ΄Π΅ΡΠ΅Π»ΡΡΡΠ²ΡΠ΅Ρ ΠΎΠ± ΠΎΡΡΡΡΡΡΠ²ΠΈΠΈ ΡΠΎΠΊΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΡΡΡΠ΅ΠΊΡΠ° ΠΈΠΌΠΈΡΠ°ΡΠΎΡΠ° miR-204-5p. Π 13β14-ΠΌΡ Π΄Π½Ρ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ° Π΄Π²ΠΈΠ³Π°ΡΠ΅Π»ΡΠ½Π°Ρ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ Π² ΠΊΠΎΠ½ΡΡΠΎΠ»ΡΠ½ΡΡ
Π³ΡΡΠΏΠΏΠ°Ρ
ΠΆΠΈΠ²ΠΎΡΠ½ΡΡ
ΡΡΠ°ΡΠΈΡΡΠΈΡΠ΅ΡΠΊΠΈ Π·Π½Π°ΡΠΈΠΌΠΎ ΡΠ½ΠΈΠ·ΠΈΠ»Π°ΡΡ ΠΏΠΎ ΡΡΠ°Π²Π½Π΅Π½ΠΈΡ Ρ Π³ΡΡΠΏΠΏΠΎΠΉ ΠΆΠΈΠ²ΠΎΡΠ½ΡΡ
, ΠΊΠΎΡΠΎΡΡΠΌ Π²Π²ΠΎΠ΄ΠΈΠ»ΡΡ ΠΈΠΌΠΈΡΠ°ΡΠΎΡ miR-204-5p (Ρ=0,011) ΠΡΠΌΠ΅ΡΠ΅Π½ΠΎ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΠ΅ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ BCL2 Π² Π»Π΅Π³ΠΊΠΈΡ
ΠΈ ΠΏΠΎΡΠΊΠ°Ρ
ΠΌΡΡΠ΅ΠΉ ΠΈ SIRT1 β Π² Π»Π΅Π³ΠΊΠΈΡ
ΠΌΡΡΠ΅ΠΉ (pΛ0,05). ΠΡΠΌΠ΅ΡΠ°Π»Π°ΡΡ ΡΠ΅Π½Π΄Π΅Π½ΡΠΈΡ ΠΊ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΡ ΠΌΠ°ΡΡΡ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΠΎΠ³ΠΎ ΡΠ·Π»Π° ΠΊ 14-ΠΌΡ Π΄Π½Ρ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°.ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅. ΠΠΎΠ΄ΡΠ»ΡΡΠΈΡ ΡΡΠΎΠ²Π½Ρ ΠΌΠΈΠΊΡΠΎΠ ΠΠ miR-204-5p ΠΏΡΠΈΠ²ΠΎΠ΄ΠΈΡ ΠΊ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ Π³Π΅Π½ΠΎΠ²-ΠΌΠΈΡΠ΅Π½Π΅ΠΉ β SIRT1 ΠΈ BCL2 Π² Π»Π΅Π³ΠΊΠΈΡ
ΠΆΠΈΠ²ΠΎΡΠ½ΡΡ
, BCL2 β Π² ΠΏΠΎΡΠΊΠ°Ρ
. ΠΠ²Π΅Π΄Π΅Π½ΠΈΠ΅ ΠΈΠΌΠΈΡΠ°ΡΠΎΡΠ° ΠΌΠΈΠΊΡΠΎΠ ΠΠ Π½Π΅ Π²ΡΠ·ΡΠ²Π°Π΅Ρ Π½Π°ΡΡΡΠ΅Π½ΠΈΠΉ Π΄Π²ΠΈΠ³Π°ΡΠ΅Π»ΡΠ½ΠΎΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ ΠΆΠΈΠ²ΠΎΡΠ½ΡΡ
, ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΠΌΠ°ΡΡΡ Π²Π½ΡΡΡΠ΅Π½Π½ΠΈΡ
ΠΎΡΠ³Π°Π½ΠΎΠ², ΡΡΠΎ ΠΌΠΎΠΆΠ΅Ρ ΡΠ²ΠΈΠ΄Π΅ΡΠ΅Π»ΡΡΡΠ²ΠΎΠ²Π°ΡΡ ΠΎΠ± ΠΎΡΡΡΡΡΡΠ²ΠΈΠΈ ΡΠ°Π·Π²ΠΈΡΠΈΡ ΡΠΎΠΊΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΡΡΡΠ΅ΠΊΡΠ°. ΠΠ°Π»ΡΠ½Π΅ΠΉΡΠ΅Π΅ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΠ΅ ΡΡΠ΅Π±ΡΠ΅ΡΡΡ Π΄Π»Ρ ΡΠ°Π·ΡΡΡΠ½Π΅Π½ΠΈΡ Π±ΠΈΠΎΠ΄ΠΎΡΡΡΠΏΠ½ΠΎΡΡΠΈ ΠΌΠΎΠ΄ΡΠ»ΡΡΠΎΡΠΎΠ² ΠΌΠΈΠΊΡΠΎΠ ΠΠ Π² ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΡΡ ΡΠΊΠ°Π½Ρ, Π° ΡΠ°ΠΊΠΆΠ΅ Π²Π»ΠΈΡΠ½ΠΈΡ ΠΈΠΌΠΈΡΠ°ΡΠΎΡΠ° miR-204-5p Π½Π° ΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΡ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ in vivo
Applications of luminous bacteria enzymes in toxicology
This review describes the principle and applications of bioluminescent enzymatic toxicity bioassays. This type of assays uses bacterial coupled enzyme systems: NADH:FMN-oxidoreductase and luciferase to replace living organisms in developing cost-competitive biosensors for environmental, medical and industrial applications. These biosensors instantly signal chemical and
biological hazards and allow for detecting a great amount of toxic compounds with advantages associated with fast results, high sensitivity, simplicity, low cost and safety of the procedure
Applications of luminous bacteria enzymes in toxicology
This review describes the principle and applications of bioluminescent enzymatic toxicity bioassays. This type of assays uses bacterial coupled enzyme systems: NADH:FMN-oxidoreductase and luciferase to replace living organisms in developing cost-competitive biosensors for environmental, medical and industrial applications. These biosensors instantly signal chemical and
biological hazards and allow for detecting a great amount of toxic compounds with advantages associated with fast results, high sensitivity, simplicity, low cost and safety of the procedure
NAD(P)H:FMN-oxidoreductase functioning under macromolecular crowding: in vitro modeling
Π’Π΅ΠΊΡΡ ΡΡΠ°ΡΡΠΈ Π½Π΅ ΠΏΡΠ±Π»ΠΈΠΊΡΠ΅ΡΡΡ Π² ΠΎΡΠΊΡΡΡΠΎΠΌ Π΄ΠΎΡΡΡΠΏΠ΅ Π² ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΠΈΠΈ Ρ ΠΏΠΎΠ»ΠΈΡΠΈΠΊΠΎΠΉ ΠΆΡΡΠ½Π°Π»Π°
Rapid biosensing tools for cancer biomarkers
Π’Π΅ΠΊΡΡ ΡΡΠ°ΡΡΠΈ Π½Π΅ ΠΏΡΠ±Π»ΠΈΠΊΡΠ΅ΡΡΡ Π² ΠΎΡΠΊΡΡΡΠΎΠΌ Π΄ΠΎΡΡΡΠΏΠ΅ Π² ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΠΈΠΈ Ρ ΠΏΠΎΠ»ΠΈΡΠΈΠΊΠΎΠΉ ΠΆΡΡΠ½Π°Π»Π°.The present review critically discusses the latest developments in the field of smart diagnostic systems for cancer biomarkers. A wide coverage of recent biosensing approaches involving aptamers, enzymes, DNA probes, fluorescent probes, interacting proteins and antibodies in vicinity to transducers such as electrochemical, optical and piezoelectric is presented. Recent advanced developments in biosensing approaches for cancer biomarker owes much credit to functionalized nanomaterials due to their unique opto-electronic properties and enhanced surface to volume ratio. Biosensing methods for a plenty of cancer biomarkers has been summarized emphasizing the key principles involved
Rapid biosensing tools for cancer biomarkers
Π’Π΅ΠΊΡΡ ΡΡΠ°ΡΡΠΈ Π½Π΅ ΠΏΡΠ±Π»ΠΈΠΊΡΠ΅ΡΡΡ Π² ΠΎΡΠΊΡΡΡΠΎΠΌ Π΄ΠΎΡΡΡΠΏΠ΅ Π² ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΠΈΠΈ Ρ ΠΏΠΎΠ»ΠΈΡΠΈΠΊΠΎΠΉ ΠΆΡΡΠ½Π°Π»Π°.The present review critically discusses the latest developments in the field of smart diagnostic systems for cancer biomarkers. A wide coverage of recent biosensing approaches involving aptamers, enzymes, DNA probes, fluorescent probes, interacting proteins and antibodies in vicinity to transducers such as electrochemical, optical and piezoelectric is presented. Recent advanced developments in biosensing approaches for cancer biomarker owes much credit to functionalized nanomaterials due to their unique opto-electronic properties and enhanced surface to volume ratio. Biosensing methods for a plenty of cancer biomarkers has been summarized emphasizing the key principles involved
Impact of enzyme stabilizers on the characteristics of biomodules for bioluminescent biosensors
Π’Π΅ΠΊΡΡ ΡΡΠ°ΡΡΠΈ Π½Π΅ ΠΏΡΠ±Π»ΠΈΠΊΡΠ΅ΡΡΡ Π² ΠΎΡΠΊΡΡΡΠΎΠΌ Π΄ΠΎΡΡΡΠΏΠ΅ Π² ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΠΈΠΈ Ρ ΠΏΠΎΠ»ΠΈΡΠΈΠΊΠΎΠΉ ΠΆΡΡΠ½Π°Π»Π°.The biomodule of bioluminescent biosensor based on a coupled enzyme system NADH:FMN-oxidoreductase and luciferase, co-immobilized with substrates in dried starch or gelatin gels, has been developed. We studied the impact of several stabilizers-dithiothreitol (DTT), bovine serum albumin (BSA) and mercaptoethanol (ME) on the biomodule's activity, storage stability and sensitivity to toxic substances. The inclusion of stabilizers increases the activity of the biological module by more than 150%. To achieve the combination of high activity, prolonged storage time and acute sensitivity to toxic substances within maximum permissible concentration we used starch gel as a carrier adding 100 mu M DTT to the immobilized preparation. The gelatin-based biological module had greater storage stability than the starch-based one but demonstrated less sensitivity to toxic substances. (C) 2015 Elsevier B.V. All rights reserved
Gelatin and starch: What better stabilizes the enzyme activity?
The regularities of the functioning of a number of enzymes in a viscous environment created by natural polymers, starch and gelatin are examined. Based on the analysis of kinetic curves of thermal inactivation, mechanisms of thermal inactivation of enzymes in a viscous microenvironment are proposed. Using the example of butyrylcholinesterase, NAD(P)H:FMN oxidoreductase, and coupled system of the luminous bacteria (NAD(P)H:FMN oxidoreductase + luciferase), the conditions, under which starch and gelatin have a stabilizing effect on enzyme activity during storage and exposure to various physical and chemical environmental factors, were found. A significant increase in the stabilizing effect is achieved by eliminating water during drying the enzyme preparations immobilized in starch and gelatin polymer gels