64 research outputs found
Red fluorescent genetically encoded indicator for intracellular hydrogen peroxide
Reactive oxygen species (ROS) are conserved regulators of numerous cellular functions, and overproduction of ROS is a hallmark of various pathological processes. Genetically encoded fluorescent probes are unique tools to study ROS production in living systems of different scale and complexity. However, the currently available recombinant redox sensors have green emission, which overlaps with the spectra of many other probes. Expanding the spectral range of recombinant in vivo ROS probes would enable multiparametric in vivo ROS detection. Here we present the first genetically encoded red fluorescent sensor for hydrogen peroxide detection, HyPerRed. The performance of this sensor is similar to its green analogues. We demonstrate the utility of the sensor by tracing low concentrations of H2O2 produced in the cytoplasm of cultured cells upon growth factor stimulation. Moreover, using HyPerRed we detect local and transient H2O2 production in the mitochondrial matrix upon inhibition of the endoplasmic reticulum Ca(2+) uptake
Π ΠΠΠ¬ ΠΠΠΠ’ΠΠΠΠΠΠ¬ΠΠ-ΠΠΠΠΠΠ₯ΠΠΠΠΠ¬ΠΠΠΠ ΠΠΠ ΠΠ₯ΠΠΠ Π ΠΠ£Π’ΠΠ€ΠΠΠΠ Π ΠΠ ΠΠ’ΠΠΠΠΠΠ£Π₯ΠΠΠΠΠΠ ΠΠ’ΠΠΠ’Π ΠΠΠΠ’ΠΠ§ΠΠ«Π₯ ΠΠΠΠΠ ΠΠΠΠΠΠΠΠ« ΠΠ Π’ΠΠ ΠΠΠ’ΠΠΠ ΠΠΠΠΠΠΠ ΠΠΠΠΠΠ MEK ΠΈ mTOR ΠΠΠΠΠ
Introduction. Cutaneous melanoma is a challenge to treat due to rapid progression of disease and acquired resistance to therapy. Autophagy and the epithelial-to-mesenchymal transition (EMT) are closely interrelated and play a key role in tumor progression. Targeted co-inhibition of MEK and mTOR kinases is a potential target for melanoma therapy by downregulatoin of the EMT.Objective: to study the effect of MEK and mTOR co-inhibition on cell viability, ability to form 3D-spheroids and migratory capacity of melanoma cell lines, and correlation of these changes with EMTand autophagy-related markers.Material and Methods. Melanoma cell lines Mel Z and Mel MTP were derived from patients, who were treated at the N.N. Blokhin National Medical Research Center of Oncology. The antiproliferative effect of binimetinib and/or rapamycin was studied by the MTT -test. 3D spheroids were formed using RGD peptides. Cell migration and invasion were assessed by a Boyden chamber migration assay. The expression levels of autophagy and EMT markers were investigated by immunocytochemistry or immunoblotting.Results. Rapamycin increased cytotoxicity of binimetinib in both 2D and 3D melanoma cell line cultures. At the same time, binimetinib and rapamycin reduced invasion, but not migration capacity of melanoma cells in vitro. The effectiveness of the combination was associated with a decrease in the EMT markers (N-cadherin and Ξ²-catenin) and autophagy markers (Beclin 1, p62/SQST M1 and LC3BII ) in melanoma cells.Conclusion. Inactivation of autophagy and EMT leads to overcoming the resistance to current anti-melanoma therapy and can be considered as a promising target for the treatment of melanoma.ΠΠ²Π΅Π΄Π΅Π½ΠΈΠ΅. ΠΠΎΠ·Π½ΠΈΠΊΠ½ΠΎΠ²Π΅Π½ΠΈΠ΅ ΡΠ΅Π·ΠΈΡΡΠ΅Π½ΡΠ½ΠΎΡΡΠΈ ΠΈ Π΄Π°Π»ΡΠ½Π΅ΠΉΡΠ°Ρ ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²Π°Ρ ΠΏΡΠΎΠ³ΡΠ΅ΡΡΠΈΡ ΠΎΡΡΠ°ΡΡΡΡ Π°ΠΊΡΡΠ°Π»ΡΠ½ΠΎΠΉ ΠΏΡΠΎΠ±Π»Π΅ΠΌΠΎΠΉ Π² Π»Π΅ΡΠ΅Π½ΠΈΠΈ ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ ΠΊΠΎΠΆΠΈ. ΠΡΠΎΡΠ΅ΡΡ Π°ΡΡΠΎΡΠ°Π³ΠΈΠΈ ΠΈ ΡΠΏΠΈΡΠ΅Π»ΠΈΠ°Π»ΡΠ½ΠΎ-ΠΌΠ΅Π·Π΅Π½Ρ
ΠΈΠΌΠ°Π»ΡΠ½ΡΠΉ ΠΏΠ΅ΡΠ΅Ρ
ΠΎΠ΄ (ΠΠΠ) ΡΠ΅ΡΠ½ΠΎ ΡΠ²ΡΠ·Π°Π½Ρ ΠΌΠ΅ΠΆΠ΄Ρ ΡΠΎΠ±ΠΎΠΉ ΠΈ ΠΈΠ³ΡΠ°ΡΡ ΠΊΠ»ΡΡΠ΅Π²ΡΡ ΡΠΎΠ»Ρ Π² ΠΎΠΏΡΡ
ΠΎΠ»Π΅Π²ΠΎΠΉ ΠΏΡΠΎΠ³ΡΠ΅ΡΡΠΈΠΈ. Π’Π°ΡΠ³Π΅ΡΠ½ΠΎΠ΅ ΠΊΠΎΠΈΠ½Π³ΠΈΠ±ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΠΠ ΠΈ mTOR ΠΊΠΈΠ½Π°Π· ΡΠ²Π»ΡΠ΅ΡΡΡ ΠΏΠΎΡΠ΅Π½ΡΠΈΠ°Π»ΡΠ½ΠΎΠΉ ΠΌΠΈΡΠ΅Π½ΡΡ Π΄Π»Ρ ΡΠ΅ΡΠ°ΠΏΠΈΠΈ ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ, Π½Π°ΡΠ΅Π»Π΅Π½Π½ΠΎΠΉ Π½Π° Π±Π»ΠΎΠΊΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΠΠ.Π¦Π΅Π»Ρ ΡΠ°Π±ΠΎΡΡ β ΠΈΠ·ΡΡΠ΅Π½ΠΈΠ΅ Π²Π»ΠΈΡΠ½ΠΈΡ ΠΊΠΎ-ΠΈΠ½Π³ΠΈΠ±ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΠΠ ΠΈ mTOR ΠΊΠΈΠ½Π°Π· Π½Π° Π²ΡΠΆΠΈΠ²Π°Π΅ΠΌΠΎΡΡΡ, Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΡ ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ 3D-ΡΡΠ΅ΡΠΎΠΈΠ΄ΠΎΠ² ΠΈ ΠΌΠΈΠ³ΡΠ°ΡΠΈΠΎΠ½Π½ΡΠ΅ ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΡΡΠΈ ΠΊΠ»Π΅ΡΠΎΡΠ½ΡΡ
Π»ΠΈΠ½ΠΈΠΉ ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ, Π° ΡΠ°ΠΊΠΆΠ΅ Π²Π·Π°ΠΈΠΌΠΎΡΠ²ΡΠ·Ρ ΡΡΠΈΡ
ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΠΉ Ρ ΠΌΠ°ΡΠΊΠ΅ΡΠ°ΠΌΠΈ ΠΠΠ ΠΈ Π°ΡΡΠΎΡΠ°Π³ΠΈΠΈ.ΠΠ°ΡΠ΅ΡΠΈΠ°Π» ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ. Π Π°Π±ΠΎΡΠ° ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½Π° Π½Π° ΠΊΠ»Π΅ΡΠΎΡΠ½ΡΡ
Π»ΠΈΠ½ΠΈΡΡ
ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ Mel Z ΠΈ Mel MTP, ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΡΡ
ΠΎΡ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ², ΠΏΡΠΎΡ
ΠΎΠ΄ΠΈΠ²ΡΠΈΡ
Π»Π΅ΡΠ΅Π½ΠΈΠ΅ Π² ΠΠΠΠ¦ ΠΎΠ½ΠΊΠΎΠ»ΠΎΠ³ΠΈΠΈ ΠΈΠΌ. Π.Π. ΠΠ»ΠΎΡ
ΠΈΠ½Π°. ΠΡΠ΅Π½ΠΊΡ Π°Π½ΡΠΈΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΠΎΠΉ Π°ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ Π±ΠΈΠ½ΠΈΠΌΠ΅ΡΠΈΠ½ΠΈΠ±Π° ΠΈ/ΠΈΠ»ΠΈ ΡΠ°ΠΏΠ°ΠΌΠΈΡΠΈΠ½Π° ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π»ΠΈ ΠΠ’Π’-ΡΠ΅ΡΡΠΎΠΌ. 3D-ΡΡΠ΅ΡΠΎΠΈΠ΄Ρ ΠΏΠΎΠ»ΡΡΠ°Π»ΠΈ Π½Π° ΠΎΡΠ½ΠΎΠ²Π΅ RGDΠΏΠ΅ΠΏΡΠΈΠ΄ΠΎΠ², ΠΌΠΈΠ³ΡΠ°ΡΠΈΠΎΠ½Π½ΡΡ ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΡΡΡ ΠΈ ΠΈΠ½Π²Π°Π·ΠΈΠ²Π½ΠΎΡΡΡ ΠΎΡΠ΅Π½ΠΈΠ²Π°Π»ΠΈ ΠΊ ΠΊΠ°ΠΌΠ΅ΡΠ΅ ΠΠΎΠΉΠ΄Π΅Π½Π° ΠΈ Π±Π°Π·Π°Π»ΡΠ½ΠΎΠΌ ΠΌΠ°ΡΡΠΈΠΊΡΠ΅. ΠΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ ΠΌΠ°ΡΠΊΠ΅ΡΠΎΠ² Π°ΡΡΠΎΡΠ°Π³ΠΈΠΈ ΠΈ ΠΠΠ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½Ρ ΠΈΠΌΠΌΡΠ½ΠΎΡΠΈΡΠΎΡ
ΠΈΠΌΠΈΡΠ΅ΡΠΊΠΈ ΠΈΠ»ΠΈ ΠΈΠΌΠΌΡΠ½ΠΎΠ±Π»ΠΎΡΡΠΈΠ½Π³ΠΎΠΌ.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ. Π Π°ΠΏΠ°ΠΌΠΈΡΠΈΠ½ ΡΡΠΈΠ»ΠΈΠ²Π°Π» ΡΠΈΡΠΎΡΠΎΠΊΡΠΈΡΠ½ΠΎΡΡΡ Π±ΠΈΠ½ΠΈΠΌΠ΅ΡΠΈΠ½ΠΈΠ±Π° ΠΊΠ°ΠΊ Π² 2D-, ΡΠ°ΠΊ ΠΈ Π² 3D-ΠΊΡΠ»ΡΡΡΡΠ°Ρ
ΠΊΠ»Π΅ΡΠΎΡΠ½ΡΡ
Π»ΠΈΠ½ΠΈΠΉ ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ. ΠΡΠΈ ΡΡΠΎΠΌ Π±ΠΈΠ½ΠΈΠΌΠ΅ΡΠΈΠ½ΠΈΠ± ΠΈ ΡΠ°ΠΏΠ°ΠΌΠΈΡΠΈΠ½ ΡΠ½ΠΈΠΆΠ°Π»ΠΈ ΠΈΠ½Π²Π°Π·ΠΈΡ, Π½ΠΎ Π½Π΅ ΠΌΠΈΠ³ΡΠ°ΡΠΈΡ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ in vitro. ΠΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ ΠΊΠΎΠΌΠ±ΠΈΠ½Π°ΡΠΈΠΈ ΡΠ²ΡΠ·Π°Π½Π° ΡΠΎ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΠ΅ΠΌ ΠΌΠ°ΡΠΊΠ΅ΡΠΎΠ² ΠΠΠ N-ΠΊΠ°Π΄Ρ
Π΅ΡΠΈΠ½Π° ΠΈ Ξ²-ΠΊΠ°ΡΠ΅Π½ΠΈΠ½Π° ΠΈ Π°ΡΡΠΎΡΠ°Π³ΠΈΠΈ Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ
ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ β ΠΠ΅ΠΊΠ»ΠΈΠ½ 1, Ρ62/SQST M1 ΠΈ LC3BII .ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅. ΠΠ½Π°ΠΊΡΠΈΠ²Π°ΡΠΈΡ Π°ΡΡΠΎΡΠ°Π³ΠΈΠΈ ΠΈ ΠΠΠ ΠΏΠΎΠ·Π²ΠΎΠ»ΡΠ΅Ρ ΠΏΡΠ΅ΠΎΠ΄ΠΎΠ»Π΅Π²Π°ΡΡ ΡΠ΅Π·ΠΈΡΡΠ΅Π½ΡΠ½ΠΎΡΡΡ ΠΊ ΡΡΡΠ΅ΡΡΠ²ΡΡΡΠ΅ΠΉ ΡΠ΅ΡΠ°ΠΏΠΈΠΈ ΠΈ ΠΌΠΎΠΆΠ΅Ρ Π±ΡΡΡ ΡΠ°ΡΡΠΌΠΎΡΡΠ΅Π½Π° ΠΊΠ°ΠΊ ΠΏΠ΅ΡΡΠΏΠ΅ΠΊΡΠΈΠ²Π½Π°Ρ ΠΌΠΈΡΠ΅Π½Ρ Π΄Π»Ρ ΡΠ΅ΡΠ°ΠΏΠΈΠΈ ΠΌΠ΅Π»Π°Π½ΠΎΠΌΡ
Optimization of the encapsulation of Thrombin Receptor Agonist Peptide in PLGA microbeads
Optimization of the encapsulation of Thrombin Receptor Agonist Peptide in PLGA microbeads
Alginate-microparticles coated with chitosan and loaded with synthetic ligands for specific removal of proteins
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