22 research outputs found
ΠΡΠ΅Π΄ΠΏΠΎΡΡΠ»ΠΊΠΈ ΠΈ ΡΠΌΡΡΠ»ΠΎΠ²ΠΎΠΉ Π³ΠΎΡΠΈΠ·ΠΎΠ½Ρ ΠΏΠΎΠ½ΡΡΠΈΡ Β«Π²ΠΎΠ²Π»Π΅ΡΠ΅Π½ΠΈΠ΅Β» Π² ΡΠΈΠ»ΠΎΡΠΎΡΠΈΠΈ Π. ΠΡΠ½ΡΠ΅
This text describes the concept of engagement by the French philosopher E. Mounier. It examines the role of engagement in 20th century philosophy. The concept of engagement was brougtn in the personalisme by a german philosopher Landsberg. Mounier found it the key to undestarding the problem of personal engagement in history. For Mounier, personality exists to cross the engagement. The engagement according to Mounier is an intimasse act and that is why it is sacred. This character of engagement gave birth to the personality that is capable to take responsibility for humanity. By the French philosopher, the necessity of engagement was the principal vocation of her generation.Π ΡΠΈΠ»ΠΎΡΠΎΡΠΈΠΈ ΠΏΠ΅ΡΡΠΎΠ½Π°Π»ΠΈΠ·ΠΌΠ° ΠΏΠΎΠ½ΡΡΠΈΠ΅ Β«Π²ΠΎΠ²Π»Π΅ΡΠ΅Π½ΠΈΡΒ» (Β«engagementΒ») ΡΠ²Π»ΡΠ΅ΡΡΡ ΠΊΠ»ΡΡΠ΅Π²ΡΠΌ ΠΈ Π²ΠΌΠ΅ΡΡΠ΅ Ρ ΡΠ΅ΠΌ Π½Π°ΠΈΠ±ΠΎΠ»Π΅Π΅ ΡΡΡΠ΄Π½ΡΠΌ Π΄Π»Ρ ΠΏΠΎΠ½ΠΈΠΌΠ°Π½ΠΈΡ ΠΈ ΠΈΡΡΠΎΡΠΈΠΊΠΎ-ΡΠΈΠ»ΠΎΡΠΎΡΡΠΊΠΎΠΉ ΠΈΠ½ΡΠ΅ΡΠΏΡΠ΅ΡΠ°ΡΠΈΠΈ. ΠΠ°ΠΊ ΠΏΠΎΠ»Π°Π³Π°Π» ΠΎΠ΄ΠΈΠ½ ΠΈΠ· ΠΎΡΠ½ΠΎΠ²Π°ΡΠ΅Π»Π΅ΠΉ ΠΈ Π²Π΅Π΄ΡΡΠΈΡ
ΠΏΡΠ΅Π΄ΡΡΠ°Π²ΠΈΡΠ΅Π»Π΅ΠΉ ΡΡΠ°Π½ΡΡΠ·ΡΠΊΠΎΠ³ΠΎ ΠΏΠ΅ΡΡΠΎΠ½Π°Π»ΠΈΠ·ΠΌΠ° Π. ΠΡΠ½ΡΠ΅, ΠΊΠ»ΡΡΠ΅Π²ΡΠΌ Π΄Π»Ρ ΠΏΠΎΠ½ΠΈΠΌΠ°Π½ΠΈΡ Β«engagementΒ» ΡΠ²Π»ΡΠ΅ΡΡΡ ΠΊΠΎΠ½ΡΠ΅ΠΏΡΠΈΡ Π»ΠΈΡΠ½ΠΎΡΡΠΈ. ΠΠΈΡΠ½ΠΎΡΡΡ, ΠΏΠΎ Π΅Π³ΠΎ ΠΌΠ½Π΅Π½ΠΈΡ, ΠΏΠΎΠ΄Π΄Π΅ΡΠΆΠΈΠ²Π°Π΅Ρ ΡΠ²ΠΎΡ Β«ΡΡΠ±ΡΡΠ°Π½ΡΠΈΡ ΡΠ΅ΡΠ΅Π· ΡΠ²ΡΠ·Ρ Ρ ΠΈΠ΅ΡΠ°ΡΡ
ΠΈΠ΅ΠΉ ΡΠ΅Π½Π½ΠΎΡΡΠ΅ΠΉ, ΡΠ²ΠΎΠ±ΠΎΠ΄Π½ΠΎ Π°Π΄Π°ΠΏΡΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
, Π°ΡΡΠΈΠΌΠΈΠ»ΠΈΡΠΎΠ²Π°Π½Π½ΡΡ
ΠΈ ΠΏΠ΅ΡΠ΅ΠΆΠΈΡΡΡ
ΠΎΡΠ²Π΅ΡΡΡΠ²Π΅Π½Π½ΡΠΌ Π²ΠΎΠ²Π»Π΅ΡΠ΅Π½ΠΈΠ΅ΠΌ (engagement) ΠΈ ΠΏΠΎΡΡΠΎΡΠ½Π½ΡΠΌ ΠΎΠ±ΡΠ°ΡΠ΅Π½ΠΈΠ΅ΠΌ (conversion). ΠΠ½Π° ΡΠ°ΠΊΠΆΠ΅ ΠΎΠ±ΡΠ΅Π΄ΠΈΠ½ΡΠ΅Ρ Π²ΡΡ ΡΠ²ΠΎΡ Π΄Π΅ΡΡΠ΅Π»ΡΠ½ΠΎΡΡΡ Π² ΡΠ²ΠΎΠ±ΠΎΠ΄Π΅ ΠΈ, ΠΊΡΠΎΠΌΠ΅ ΡΠΎΠ³ΠΎ, ΡΠ°Π·Π²ΠΈΠ²Π°Π΅Ρ Π΅Π΄ΠΈΠ½ΠΈΡΠ½ΠΎΡΡΡ ΡΠ²ΠΎΠ΅Π³ΠΎ ΠΏΡΠΈΠ·Π²Π°Π½ΠΈΡΒ» [14. Π . 523]. ΠΠΎΡΡΠΎΠΌΡ Π²ΠΎΠ²Π»Π΅ΡΠ΅Π½ΠΈΠ΅ ΡΠ»Π΅Π΄ΡΠ΅Ρ ΡΠ°ΡΡΠΌΠ°ΡΡΠΈΠ²Π°ΡΡ ΠΊΠ°ΠΊ ΡΡΠ»ΠΎΠ²ΠΈΠ΅ ΠΏΠ΅ΡΡΠΎΠ½Π°Π»ΡΠ½ΠΎΠ³ΠΎ Π±ΡΡΠΈΡ
The resultant on compact Riemann surfaces
We introduce a notion of resultant of two meromorphic functions on a compact
Riemann surface and demonstrate its usefulness in several respects. For
example, we exhibit several integral formulas for the resultant, relate it to
potential theory and give explicit formulas for the algebraic dependence
between two meromorphic functions on a compact Riemann surface. As a particular
application, the exponential transform of a quadrature domain in the complex
plane is expressed in terms of the resultant of two meromorphic functions on
the Schottky double of the domain.Comment: 44 page
Efficiency of RNA Hydrolysis by Binase from Bacillus pumilus: The Impact of Substrate Structure, Metal Ions, and Low Molecular Weight Nucleotide Compounds
Binase is an extracellular guanyl-preferring ribonuclease from Bacillus pumilus. The main biological function of binase is RNA degradation with the formation of guanosine-2',3'-cyclic phosphate and its subsequent hydrolysis to 3'-phosphate. Extracellular RNases are believed to be key agents that affect the functional activity of the body, as they directly interact with epithelial and immune cells. The biological effects of the enzyme may consist of both direct RNA degradation, and the accumulation of 2',3'-cGMP in the human body. In this work, we have performed a comparative analysis of the cleavage efficiency of model RNA substrates, i.e., short hairpin structures that contain guanosine at various positions. It has been shown that the hydrolysis efficiency of the model RNA substrates depends on the position of guanosine. We have also demonstrated the influence of various divalent metal ions and low molecular weight nucleotide compounds on the binase-catalyzed endoribonucleolytic reaction
Efficiency of RNA Hydrolysis by Binase from Bacillus pumilus: The Impact of Substrate Structure, Metal Ions, and Low Molecular Weight Nucleotide Compounds
Β© 2020, Pleiades Publishing, Inc. Abstract: Binase is an extracellular guanyl-preferring ribonuclease from Bacillus pumilus. The main biological function of binase is RNA degradation with the formation of guanosine-2',3'-cyclic phosphate and its subsequent hydrolysis to 3'-phosphate. Extracellular RNases are believed to be key agents that affect the functional activity of the body, as they directly interact with epithelial and immune cells. The biological effects of the enzyme may consist of both direct RNA degradation, and the accumulation of 2',3'-cGMP in the human body. In this work, we have performed a comparative analysis of the cleavage efficiency of model RNA substrates, i.e., short hairpin structures that contain guanosine at various positions. It has been shown that the hydrolysis efficiency of the model RNA substrates depends on the position of guanosine. We have also demonstrated the influence of various divalent metal ions and low molecular weight nucleotide compounds on the binase-catalyzed endoribonucleolytic reaction
Π ΠΎΠ»Ρ Π°Π΄ΡΠ΅Π½ΠΎΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠΎΠ² Π² ΡΠ΅Π°Π»ΠΈΠ·Π°ΡΠΈΠΈ ΡΡΠΈΠΌΡΠ»ΡΡΠΎΡΠ½ΠΎΠ³ΠΎ Π΄Π΅ΠΉΡΡΠ²ΠΈΡ ΡΠ΅ΡΠΎΡΠΎΠ½ΠΈΠ½Π° Π½Π° ΠΌΠΎΡΠΎΡΠΈΠΊΡ ΠΆΠ΅Π»ΡΠ΄ΠΊΠ°
Purpose of the study - study of the role of adrenoreceptors in the development of the stimulatory action of serotonin on the gastric motor activity. Materials and methods. The experiments were performed on rats (27) of the Wistar line in the surgical stage of anesthesia. Electromyogram and hydrostatic pressure in the stomach cavity were recorded using a BioAmp ML132 amplifier (Adinstruments, Australia), an Maclab 8e analog-to-digital converter (Adinstruments, Australia), a Macintosh Performa 6400/180 computer, and Chart 4.2.3. program. Serotonin injected into the body to intact animals and against the background of separate and joint blockade of Ξ±- and Ξ²-adrenoreceptors. The results of the study. In experiments on rats established that the preliminary simultaneous blockade of Ξ±- and Ξ²- adrenoreceptors leads to an increase in the stimulatory effect of the stomach with the introduction of serotonin by 58%, blockade of Ξ±-adrenoreceptors only - by 62%, Ξ²-adrenoreceptor blockade - by 89%. In intact animals, the stimulatory effect of serotonin is only + 26%. Simultaneous blockade of Ξ±- and Ξ²-adrenoreceptors and blockade of Ξ±-adrenoreceptors only (without serotonin administration) did not affect the gastric motor activity of intact animals. Blockade only Ξ²-adrenoreceptors will lead to an increase in gastric contractions by 34%. Conclusion. Intact Ξ±- and Ξ²-adrenoreceptors inhibit the stimulatory effect of serotonin on gastric motor activity.Π¦Π΅Π»Ρ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ - ΠΈΠ·ΡΡΠ΅Π½ΠΈΠ΅ ΡΠΎΠ»ΠΈ Π°Π΄ΡΠ΅Π½ΠΎΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠΎΠ² Π² ΡΠ°Π·Π²ΠΈΡΠΈΠΈ ΡΡΠΈΠΌΡΠ»ΡΡΠΎΡΠ½ΠΎΠ³ΠΎ Π΄Π΅ΠΉΡΡΠ²ΠΈΡ ΡΠ΅ΡΠΎΡΠΎΠ½ΠΈΠ½Π° Π½Π° ΠΌΠΎΡΠΎΡΠΈΠΊΡ ΠΆΠ΅Π»ΡΠ΄ΠΊΠ°. ΠΠ°ΡΠ΅ΡΠΈΠ°Π»Ρ ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ. ΠΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΡ Π²ΡΠΏΠΎΠ»Π½Π΅Π½Ρ Π½Π° ΠΊΡΡΡΠ°Ρ
(27) Π»ΠΈΠ½ΠΈΠΈ ΠΠΈΡΡΠ°Ρ Π² ΡΡΠ»ΠΎΠ²ΠΈΡΡ
Ρ
ΠΈΡΡΡΠ³ΠΈΡΠ΅ΡΠΊΠΎΠΉ ΡΡΠ°Π΄ΠΈΠΈ Π½Π°ΡΠΊΠΎΠ·Π°. Π Π΅Π³ΠΈΡΡΡΠΈΡΠΎΠ²Π°Π»ΠΈ ΡΠ»Π΅ΠΊΡΡΠΎΠΌΠΈΠΎΠ³ΡΠ°ΠΌΠΌΡ ΠΈ Π³ΠΈΠ΄ΡΠΎΡΡΠ°ΡΠΈΡΠ΅ΡΠΊΠΎΠ΅ Π΄Π°Π²Π»Π΅Π½ΠΈΠ΅ Π² ΠΏΠΎΠ»ΠΎΡΡΠΈ ΠΆΠ΅Π»ΡΠ΄ΠΊΠ° Ρ ΠΏΠΎΠΌΠΎΡΡΡ ΡΡΠΈΠ»ΠΈΡΠ΅Π»Ρ BioAmp ML132 (Adinstruments, ΠΠ²ΡΡΡΠ°Π»ΠΈΡ), Π°Π½Π°Π»ΠΎΠ³ΠΎ-ΡΠΈΡΡΠΎΠ²ΠΎΠ³ΠΎ ΠΏΡΠ΅ΠΎΠ±ΡΠ°Π·ΠΎΠ²Π°ΡΠ΅Π»Ρ Maclab 8e (Adinstruments, ΠΠ²ΡΡΡΠ°Π»ΠΈΡ), ΠΊΠΎΠΌΠΏΡΡΡΠ΅ΡΠ° ΠΠ°Ρintosh Performa 6400/180 ΠΈ ΠΏΡΠΎΠ³ΡΠ°ΠΌΠΌΡ Chart 4.2.3. Π‘Π΅ΡΠΎΡΠΎΠ½ΠΈΠ½ Π²Π²ΠΎΠ΄ΠΈΠ»ΠΈ Π² ΠΎΡΠ³Π°Π½ΠΈΠ·ΠΌ ΠΈΠ½ΡΠ°ΠΊΡΠ½ΡΠΌ ΠΆΠΈΠ²ΠΎΡΠ½ΡΠΌ ΠΈ Π½Π° ΡΠΎΠ½Π΅ ΡΠ°Π·Π΄Π΅Π»ΡΠ½ΠΎΠΉ ΠΈ ΡΠΎΠ²ΠΌΠ΅ΡΡΠ½ΠΎΠΉ Π±Π»ΠΎΠΊΠ°Π΄Ρ Ξ±- ΠΈ Ξ²-Π°Π΄ΡΠ΅Π½ΠΎΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠΎΠ². Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ. Π ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°Ρ
Π½Π° ΠΊΡΡΡΠ°Ρ
ΡΡΡΠ°Π½ΠΎΠ²ΠΈΠ»ΠΈ, ΡΡΠΎ ΠΏΡΠ΅Π΄Π²Π°ΡΠΈΡΠ΅Π»ΡΠ½Π°Ρ ΠΎΠ΄Π½ΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½Π°Ρ Π±Π»ΠΎΠΊΠ°Π΄Π° Ξ±-ΠΈ Ξ²- Π°Π΄ΡΠ΅Π½ΠΎΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠΎΠ² Π²Π΅Π΄Π΅Ρ ΠΊ ΡΠ²Π΅Π»ΠΈΡΠ΅Π½ΠΈΡ ΡΡΠΈΠΌΡΠ»ΡΡΠΎΡΠ½ΠΎΠ³ΠΎ ΡΡΡΠ΅ΠΊΡΠ° ΠΆΠ΅Π»ΡΠ΄ΠΊΠ° ΠΏΡΠΈ Π²Π²Π΅Π΄Π΅Π½ΠΈΠΈ ΡΠ΅ΡΠΎΡΠΎΠ½ΠΈΠ½Π° Π½Π° 58%, Π±Π»ΠΎΠΊΠ°Π΄Π° ΡΠΎΠ»ΡΠΊΠΎ Ξ±-Π°Π΄ΡΠ΅Π½ΠΎΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠΎΠ²-Π½Π° 62%, Π±Π»ΠΎΠΊΠ°Π΄Π° Ξ²- Π°Π΄ΡΠ΅Π½ΠΎΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠΎΠ²-Π½Π° 89%. Π£ ΠΈΠ½ΡΠ°ΠΊΡΠ½ΡΡ
ΠΆΠΈΠ²ΠΎΡΠ½ΡΡ
ΡΡΠΈΠΌΡΠ»ΡΡΠΎΡΠ½ΡΠΉ ΡΡΡΠ΅ΠΊΡ ΡΠ΅ΡΠΎΡΠΎΠ½ΠΈΠ½Π° ΡΠΎΡΡΠ°Π²Π»ΡΠ΅Ρ Π»ΠΈΡΡ + 26%. ΠΠ΄Π½ΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½Π°Ρ Π±Π»ΠΎΠΊΠ°Π΄Π° Ξ±- ΠΈ Ξ²- Π°Π΄ΡΠ΅Π½ΠΎΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠΎΠ² ΠΈ Π±Π»ΠΎΠΊΠ°Π΄Π° ΡΠΎΠ»ΡΠΊΠΎ Ξ±-Π°Π΄ΡΠ΅Π½ΠΎΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠΎΠ² (Π±Π΅Π· Π²Π²Π΅Π΄Π΅Π½ΠΈΡ ΡΠ΅ΡΠΎΡΠΎΠ½ΠΈΠ½Π°) Π½Π΅ Π²Π»ΠΈΡΠ»ΠΈ Π½Π° ΠΌΠΎΡΠΎΡΠΈΠΊΡ ΠΆΠ΅Π»ΡΠ΄ΠΊΠ° ΠΈΠ½ΡΠ°ΠΊΡΠ½ΡΡ
ΠΆΠΈΠ²ΠΎΡΠ½ΡΡ
. ΠΠ»ΠΎΠΊΠ°Π΄Π° ΠΆΠ΅ ΡΠΎΠ»ΡΠΊΠΎ Ξ²- Π°Π΄ΡΠ΅Π½ΠΎΡΠ΅ΡΠ΅ΠΏΡΠΎΡΠΎΠ² Π²Π΅Π΄Π΅Ρ ΠΊ ΡΡΠΈΠ»Π΅Π½ΠΈΡ ΡΠΎΠΊΡΠ°ΡΠ΅Π½ΠΈΠΉ ΠΆΠ΅Π»ΡΠ΄ΠΊΠ° Π½Π° 34%. ΠΡΠ²ΠΎΠ΄. ΠΠ½ΡΠ°ΠΊΡΠ½ΡΠ΅ a- ΠΈ b- Π°Π΄ΡΠ΅Π½ΠΎΡΠ΅ΡΠ΅ΠΏΡΠΎΡΡ ΠΏΡΠ΅ΠΏΡΡΡΡΠ²ΡΡΡ ΡΡΠΈΠΌΡΠ»ΡΡΠΎΡΠ½ΠΎΠΌΡ Π΄Π΅ΠΉΡΡΠ²ΠΈΡ ΡΠ΅ΡΠΎΡΠΎΠ½ΠΈΠ½Π° Π½Π° ΠΌΠΎΡΠΎΡΠΈΠΊΡ ΠΆΠ΅Π»ΡΠ΄ΠΊΠ°
ΠΡΠΈΡΠ΅ΡΠΈΠΈ Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΠΊΠΈ Π»ΠΈΠΌΡΠΎΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΡΡ Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΠΉ ΠΏΠΎ ΠΏΠ΅ΡΠΈΡΠ΅ΡΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΊΡΠΎΠ²ΠΈ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ² ΠΏΡΠΈ ΠΏΠΎΠΌΠΎΡΠΈ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠ³ΠΎ Π±ΠΈΠΎΡΠΈΠΏΠ°
Background: At present, the diagnosis of lymphoproliferative disorders is based on the combination of blood or bone marrow smear morphology and immunophenotyping by flow cytometry. Immunophenotypic testing by flow cytometry technique is available only in big medical centers, which is not always convenient for a Β patient. Therefore, development of an available method for preliminary diagnosis of lymphoproliferative diseases not requiring special equipment seems relevant.Materials and methods: Peripheral blood mononuclear cells from 17 Β patients admitted to the hospital with suspicion of a Β lymphoproliferative disorder, and 17 Β healthy donors were studied on a cell biochip for determination of proportions of cells positive for various surface CD antigens. The diagnosis was verified by flow cytometry.Results: Compared to healthy controls and patients with T-cell lymphoproliferative disorders (TCLPD), the patients with B-cell lymphoproliferative disorders (BCLPD) had significantly lower proportion of CD7+ cells (medians, 7% and 73% respectively, p=2Γ10-6 for comparison with healthy controls; median Β 7% and 93% for comparison with TCLPD, p=0.032). In addition, the patients with BCLPD had higher proportion of peripheral Π‘D19+ mononuclear cells, compared to that in the patients with TCLPD and healthy donors (medians 84% and 13% for comparison between BCLPD and healthy control, p=2Γ10-5; 84% and 3% for comparison of BCLPD and TCLPD, p=0.033). The patients with B-cell chronic lymphocytic leukemia had significantly higher CD5+ cells in the cell biochip compared to the patients with other BCLPD (medians 72% and 9%, p=0.024). The patients with TCLPD had significantly lower proportion of CD19+ cells than the healthy controls (medians, 3% and 13%, respectively, Ρ=0.042).Conclusion: The study has demonstrated the potential to use the previously developed cell biochip for diagnosis of lymphoproliferative diseases. The biochip makes it possible to sort out white blood cells according to their surface differentiation antigen for their further morphological analysis. The cell biochip allows for the differential diagnosis between BCLPD and TCLPD and determination the lymphocyte clones based on the expression of immunoglobulin light chains.ΠΠΊΡΡΠ°Π»ΡΠ½ΠΎΡΡΡ. Π Β Π½Π°ΡΡΠΎΡΡΠ΅Π΅ Π²ΡΠ΅ΠΌΡ Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΠΊΠ° Π»ΠΈΠΌΡΠΎΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΡΡ
Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΠΉ Π±Π°Π·ΠΈΡΡΠ΅ΡΡΡ Π½Π° ΡΠΎΡΠ΅ΡΠ°Π½ΠΈΠΈ ΠΌΠΎΡΡΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΠΌΠ°Π·ΠΊΠΎΠ² ΠΊΡΠΎΠ²ΠΈ ΠΈΠ»ΠΈ ΠΊΠΎΡΡΠ½ΠΎΠ³ΠΎ ΠΌΠΎΠ·Π³Π° Ρ Β ΠΈΠΌΠΌΡΠ½ΠΎΡΠ΅Π½ΠΎΡΠΈΠΏΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΏΡΠΎΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΠΎΠΌΠ΅ΡΡΠΈΠΈ. ΠΡΠΎΠ²Π΅Π΄Π΅Π½ΠΈΠ΅ ΠΈΠΌΠΌΡΠ½ΠΎΡΠ΅Π½ΠΎΡΠΈΠΏΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΈΡ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ ΠΏΡΠΎΡΠΎΡΠ½ΠΎΠΉ ΡΠΈΡΠΎΠΌΠ΅ΡΡΠΈΠΈ Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎ ΡΠΎΠ»ΡΠΊΠΎ Π² Β ΠΊΡΡΠΏΠ½ΡΡ
ΠΌΠ΅Π΄ΠΈΡΠΈΠ½ΡΠΊΠΈΡ
ΡΠ΅Π½ΡΡΠ°Ρ
, ΡΡΠΎ Π½Π΅ Π²ΡΠ΅Π³Π΄Π° ΡΠ΄ΠΎΠ±Π½ΠΎ Π΄Π»Ρ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠ°. Π Β ΡΡΠΎΠΉ ΡΠ²ΡΠ·ΠΈ ΡΠΎΠ·Π΄Π°Π½ΠΈΠ΅ Π΄ΠΎΡΡΡΠΏΠ½ΠΎΠ³ΠΎ ΠΌΠ΅ΡΠΎΠ΄Π° ΠΏΡΠ΅Π΄Π²Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎΠΉ Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΠΊΠΈ Π»ΠΈΠΌΡΠΎΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΡΡ
Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΠΉ, ΠΊΠΎΡΠΎΡΡΠΉ Π½Π΅ ΡΡΠ΅Π±ΡΠ΅Ρ ΡΠΏΠ΅ΡΠΈΠ°Π»ΡΠ½ΠΎΠ³ΠΎ ΠΎΠ±ΠΎΡΡΠ΄ΠΎΠ²Π°Π½ΠΈΡ, ΠΏΡΠ΅Π΄ΡΡΠ°Π²Π»ΡΠ΅ΡΡΡ Π°ΠΊΡΡΠ°Π»ΡΠ½ΠΎΠΉ Π·Π°Π΄Π°ΡΠ΅ΠΉ.ΠΠ°ΡΠ΅ΡΠΈΠ°Π» ΠΈ ΠΌΠ΅ΡΠΎΠ΄Ρ. ΠΠΎΠ½ΠΎΠ½ΡΠΊΠ»Π΅Π°ΡΡ ΠΏΠ΅ΡΠΈΡΠ΅ΡΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΊΡΠΎΠ²ΠΈ 17 Β ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ², ΠΏΠΎΡΡΡΠΏΠΈΠ²ΡΠΈΡ
Π² Β ΡΡΠ°ΡΠΈΠΎΠ½Π°Ρ Ρ Β ΠΏΠΎΠ΄ΠΎΠ·ΡΠ΅Π½ΠΈΠ΅ΠΌ Π½Π° Π»ΠΈΠΌΡΠΎΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΠΎΠ΅ Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΠ΅, ΠΈ 17 Π·Π΄ΠΎΡΠΎΠ²ΡΡ
Π΄ΠΎΠ½ΠΎΡΠΎΠ² ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π»ΠΈΡΡ Π½Π° ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΌ Π±ΠΈΠΎΡΠΈΠΏΠ΅ Π΄Π»Ρ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½ΠΈΡ Π΄ΠΎΠ»ΠΈ ΠΊΠ»Π΅ΡΠΎΠΊ, ΠΏΠΎΠ»ΠΎΠΆΠΈΡΠ΅Π»ΡΠ½ΡΡ
ΠΏΠΎ ΡΠ°Π·Π»ΠΈΡΠ½ΡΠΌ ΠΏΠΎΠ²Π΅ΡΡ
Π½ΠΎΡΡΠ½ΡΠΌ CD-Π°Π½ΡΠΈΠ³Π΅Π½Π°ΠΌ. ΠΠ»Ρ Π²Π΅ΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ Π΄ΠΈΠ°Π³Π½ΠΎΠ·Π° ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π»ΠΈ ΠΏΡΠΎΡΠΎΡΠ½ΡΡ ΡΠΈΡΠΎΠΌΠ΅ΡΡΠΈΡ.Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ. ΠΠ°ΡΠΈΠ΅Π½ΡΡ Ρ Β Π-ΠΊΠ»Π΅ΡΠΎΡΠ½ΡΠΌΠΈ Π»ΠΈΠΌΡΠΎΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΡΠΌΠΈ Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΡΠΌΠΈ (Π-ΠΠΠ) ΡΡΠ°ΡΠΈΡΡΠΈΡΠ΅ΡΠΊΠΈ Π·Π½Π°ΡΠΈΠΌΠΎ ΠΎΡΠ»ΠΈΡΠ°Π»ΠΈΡΡ ΠΎΡ Π·Π΄ΠΎΡΠΎΠ²ΡΡ
Π΄ΠΎΠ½ΠΎΡΠΎΠ² Π±ΠΎΠ»Π΅Π΅ Π½ΠΈΠ·ΠΊΠΎΠΉ Π΄ΠΎΠ»Π΅ΠΉ CD7+ (ΠΌΠ΅Π΄ΠΈΠ°Π½Ρ Β 7 ΠΈ Β 73% ΠΏΡΠΈ ΡΡΠ°Π²Π½Π΅Π½ΠΈΠΈ Π-ΠΠΠ ΠΈ Β ΠΊΠΎΠ½ΡΡΠΎΠ»Ρ, p=2Γ10-6; ΠΌΠ΅Π΄ΠΈΠ°Π½Ρ 93 ΠΈ 7% ΠΏΡΠΈ ΡΡΠ°Π²Π½Π΅Π½ΠΈΠΈ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ² Ρ Β T-ΠΊΠ»Π΅ΡΠΎΡΠ½ΡΠΌΠΈ Π»ΠΈΠΌΡΠΎΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΡΠΌΠΈ Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΡΠΌΠΈ (Π’-ΠΠΠ) ΠΈ Π-ΠΠΠ, p=0,032) ΠΈ Π±ΠΎΠ»Π΅Π΅ Π²ΡΡΠΎΠΊΠΎΠΉ Π΄ΠΎΠ»Π΅ΠΉ Π‘D19+ ΠΌΠΎΠ½ΠΎΠ½ΡΠΊΠ»Π΅Π°ΡΠ½ΡΡ
ΠΊΠ»Π΅ΡΠΎΠΊ ΠΏΠ΅ΡΠΈΡΠ΅ΡΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΊΡΠΎΠ²ΠΈ ΠΏΠΎ ΡΡΠ°Π²Π½Π΅Π½ΠΈΡ Ρ Β ΠΏΠ°ΡΠΈΠ΅Π½ΡΠ°ΠΌΠΈ Ρ Β Π’-ΠΠΠ ΠΈ Β Π·Π΄ΠΎΡΠΎΠ²ΡΠΌΠΈ Π΄ΠΎΠ½ΠΎΡΠ°ΠΌΠΈ (ΠΌΠ΅Π΄ΠΈΠ°Π½Ρ Β 84 ΠΈ Β 13% ΠΏΡΠΈ ΡΡΠ°Π²Π½Π΅Π½ΠΈΠΈ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ² Ρ Β Π-ΠΠΠ ΠΈ Β ΠΊΠΎΠ½ΡΡΠΎΠ»Ρ, p=2Γ10-5; 84 ΠΈ Β 3% ΠΏΡΠΈ ΡΡΠ°Π²Π½Π΅Π½ΠΈΠΈ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ² Ρ B-ΠΠΠ ΠΈ T-ΠΠΠ, p=0,033). ΠΠ°ΡΠΈΠ΅Π½ΡΡ Ρ Β Π-ΠΊΠ»Π΅ΡΠΎΡΠ½ΡΠΌ Ρ
ΡΠΎΠ½ΠΈΡΠ΅ΡΠΊΠΈΠΌ Π»ΠΈΠΌΡΠΎΠ»Π΅ΠΉΠΊΠΎΠ·ΠΎΠΌ (Π-Π₯ΠΠ) ΡΡΠ°ΡΠΈΡΡΠΈΡΠ΅ΡΠΊΠΈ Π·Π½Π°ΡΠΈΠΌΠΎ ΠΎΡΠ»ΠΈΡΠ°Π»ΠΈΡΡ ΠΎΡ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ² Ρ Π΄ΡΡΠ³ΠΈΠΌΠΈ Π-ΠΠΠ Π±ΠΎΠ»Π΅Π΅ Π²ΡΡΠΎΠΊΠΎΠΉ Π΄ΠΎΠ»Π΅ΠΉ Π‘D5+ ΠΊΠ»Π΅ΡΠΎΠΊ Π½Π° ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΌ Π±ΠΈΠΎΡΠΈΠΏΠ΅ (ΠΌΠ΅Π΄ΠΈΠ°Π½Ρ Β 72 ΠΈ Β 9% ΠΏΡΠΈ ΡΡΠ°Π²Π½Π΅Π½ΠΈΠΈ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ² Ρ Π-Π₯ΠΠ ΠΈ Π-ΠΠΠ, p=0,024). ΠΠ°ΡΠΈΠ΅Π½ΡΡ Ρ Π’-ΠΠΠ ΡΡΠ°ΡΠΈΡΡΠΈΡΠ΅ΡΠΊΠΈ Π·Π½Π°ΡΠΈΠΌΠΎ ΠΎΡΠ»ΠΈΡΠ°Π»ΠΈΡΡ ΠΎΡ Π·Π΄ΠΎΡΠΎΠ²ΡΡ
Π΄ΠΎΠ½ΠΎΡΠΎΠ² Π±ΠΎΠ»Π΅Π΅ Π½ΠΈΠ·ΠΊΠΎΠΉ Π΄ΠΎΠ»Π΅ΠΉ Π‘D19+ ΠΊΠ»Π΅ΡΠΎΠΊ (ΠΌΠ΅Π΄ΠΈΠ°Π½Ρ 3 ΠΈ 13% ΠΏΡΠΈ ΡΡΠ°Π²Π½Π΅Π½ΠΈΠΈ ΠΏΠ°ΡΠΈΠ΅Π½ΡΠΎΠ² Ρ Π’-ΠΠΠ ΠΈ ΠΊΠΎΠ½ΡΡΠΎΠ»Ρ, Ρ=0,042).ΠΠ°ΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅. ΠΠΎΠΊΠ°Π·Π°Π½Π° Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΡ Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΠΊΠΈ Π»ΠΈΠΌΡΠΎΠΏΡΠΎΠ»ΠΈΡΠ΅ΡΠ°ΡΠΈΠ²Π½ΡΡ
Π·Π°Π±ΠΎΠ»Π΅Π²Π°Π½ΠΈΠΉ Ρ Β ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΠ°Π·ΡΠ°Π±ΠΎΡΠ°Π½Π½ΠΎΠ³ΠΎ ΡΠ°Π½Π΅Π΅ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠ³ΠΎ Π±ΠΈΠΎΡΠΈΠΏΠ°. Π‘ Β Π΅Π³ΠΎ ΠΏΠΎΠΌΠΎΡΡΡ ΠΌΠΎΠΆΠ½ΠΎ ΡΠ°ΡΡΠΎΡΡΠΈΡΠΎΠ²Π°ΡΡ Π»Π΅ΠΉΠΊΠΎΡΠΈΡΡ Π² Β ΠΏΡΠΎΡΡΡΠ°Π½ΡΡΠ²Π΅ ΠΏΠΎ ΠΈΡ
ΠΏΠΎΠ²Π΅ΡΡ
Π½ΠΎΡΡΠ½ΡΠΌ Π΄ΠΈΡΡΠ΅ΡΠ΅Π½ΡΠΈΡΠΎΠ²ΠΎΡΠ½ΡΠΌ Π°Π½ΡΠΈΠ³Π΅Π½Π°ΠΌ Π΄Π»Ρ Π΄Π°Π»ΡΠ½Π΅ΠΉΡΠ΅Π³ΠΎ ΠΌΠΎΡΡΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π°Π½Π°Π»ΠΈΠ·Π°. ΠΠ»Π΅ΡΠΎΡΠ½ΡΠΉ Π±ΠΈΠΎΡΠΈΠΏ ΠΏΠΎΠ·Π²ΠΎΠ»ΡΠ΅Ρ ΠΏΡΠΎΠ²ΠΎΠ΄ΠΈΡΡ Π΄ΠΈΡΡΠ΅ΡΠ΅Π½ΡΠΈΠ°Π»ΡΠ½ΡΡ Π΄ΠΈΠ°Π³Π½ΠΎΡΡΠΈΠΊΡ ΠΌΠ΅ΠΆΠ΄Ρ Π- ΠΈ Β Π’-ΠΠΠ ΠΈ Β ΠΎΠΏΡΠ΅Π΄Π΅Π»ΡΡΡ ΠΊΠ»ΠΎΠ½Π°Π»ΡΠ½ΠΎΡΡΡ Π-Π»ΠΈΠΌΡΠΎΡΠΈΡΠΎΠ² Π½Π° ΠΎΡΠ½ΠΎΠ²Π΅ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ Π»Π΅Π³ΠΊΠΈΡ
ΡΠ΅ΠΏΠ΅ΠΉ ΠΈΠΌΠΌΡΠ½ΠΎΠ³Π»ΠΎΠ±ΡΠ»ΠΈΠ½ΠΎΠ²
The formation of catalytically competent enzymeβsubstrate complex is not a bottleneck in lesion excision by human alkyladenine DNA glycosylase
<p>Human alkyladenine DNA glycosylase (AAG) protects DNA from alkylated and deaminated purine lesions. AAG flips out the damaged nucleotide from the double helix of DNA and catalyzes the hydrolysis of the N-glycosidic bond to release the damaged base. To understand better, how the step of nucleotide eversion influences the overall catalytic process, we performed a pre-steady-state kinetic analysis of AAG interaction with specific DNA-substrates, 13-base pair duplexes containing in the 7th position 1-N6-ethenoadenine (Ξ΅A), hypoxanthine (Hx), and the stable product analogue tetrahydrofuran (F). The combination of the fluorescence of tryptophan, 2-aminopurine, and 1-N6-ethenoadenine was used to record conformational changes of the enzyme and DNA during the processes of DNA lesion recognition, damaged base eversion, excision of the N-glycosidic bond, and product release. The thermal stability of the duplexes characterized by the temperature of melting, <i>T</i><sub>m</sub>, and the rates of spontaneous opening of individual nucleotide base pairs were determined by NMR spectroscopy. The data show that the relative thermal stability of duplexes containing a particular base pair in position 7, (<i>T</i><sub>m</sub>(F/T)β<β<i>T</i><sub>m</sub>(Ξ΅A/T)β<β<i>T</i><sub>m</sub>(Hx/T)β<β<i>T</i><sub>m</sub>(A/T)) correlates with the rate of reversible spontaneous opening of the base pair. However, in contrast to that, the catalytic lesion excision rate is two orders of magnitude higher for Hx-containing substrates than for substrates containing Ξ΅A, proving that catalytic activity is not correlated with the stability of the damaged base pair. Our study reveals that the formation of the catalytically competent enzymeβsubstrate complex is not the bottleneck controlling the catalytic activity of AAG.</p