32 research outputs found
Anomaly Detection Algorithms and Techniques for Network Intrusion Detection Systems
In recent years, many deep learning-based models have been proposed for anomaly detection. This thesis presents a comparison of selected deep autoencoding models and classical anomaly detection methods on three modern network intrusion detection datasets. We experiment with different configurations and architectures of the selected models, as well as aggregation techniques for input preprocessing and output postprocessing. We propose a methodology for creating benchmark datasets for the evaluation of the methods in different settings. We provide a statistical comparison of the performance of the selected techniques. We conclude that the deep autoencoding models, in particular AE and VAE, systematically outperform the classic methods. Furthermore, we show that aggregating input network flow data improves the overall performance. In general, the tested techniques are promising regarding their application in network intrusion detection systems. However, secondary techniques must be employed to reduce the high numbers of generated false alarms
Molecular Dynamics Study of Self-Diffusion in Zr
We employed a recently developed semi-empirical Zr potential to determine the
diffusivities in the hcp and bcc Zr via molecular dynamics simulation. The
point defect concentration was determined directly from MD simulation rather
than from theoretical methods using T=0 calculations. We found that the
diffusion proceeds via the interstitial mechanism in the hcp Zr and both the
vacancy and interstitial mechanisms give contribution in diffusivity in the bcc
Zr. The agreement with the experimental data is excellent for the hcp Zr and
for the bcc Zr it is rather good at high temperatures but there is a
considerable disagreement at low temperatures
Numerical and experimental study of the weld joints formation in welding foam materials
A numerical analysis of fusion welding of steel- and aluminum-based foam materials is carried out. The schemes of the structured and stochastic pore distribution are considered. The research results were used to conduct the experiments which confirmed the reliability of the numerical calculations
Endless Bridge
1 volume, 80 pages. A visual retelling of creation, Mikhail Mishin\u27s Endless Bridge utilizes juxtapositions and stark comparisons in a narrative that spans from birth to death and back again. iPhone photos, newspaper clippings and other found materials are the vocabulary used in this book in which light is separated from dark, and order emerges from chaos, only to revert back into nothingness. Images in this book have the quality of archaeological artifacts, where the transient qualities of an inflated tarp, a light reflection on the pavement or footprint in the concrete are highlighted. We know what comes after, we see what came before. Yet these fleeting moments hold great weight in the larger context of this cycle. --Publisher\u27s website, viewed April 25, 2019. Limited edition of 100 copies. Black on black illustration on paper cover. Perfect binding.https://digitalcommons.risd.edu/specialcollections_artistsbooks/1155/thumbnail.jp
Deep Laser Cooling of Thulium Atoms to Sub-K Temperatures in Magneto-Optical Trap
Deep laser cooling of atoms, ions, and molecules facilitates the study of
fundamental physics as well as applied research. In this work, we report on the
narrow-line laser cooling of thulium atoms at the wavelength of
with the natural linewidth of , which
widens the limits of atomic cloud parameters control. Temperatures of about
, phase-space density of up to and
number of trapped atoms were achieved. We have also demonstrated
formation of double cloud structure in an optical lattice by adjusting
parameters of the magneto-optical trap. These results can
be used to improve experiments with BEC, atomic interferometers, and optical
clocks.Comment: 12 pages, 6 figure
Structural insights into thrombolytic activity of destabilase from medicinal leech
Destabilase from the medical leech Hirudo medicinalis belongs to the family of i-type lysozymes. It has two different enzymatic activities: microbial cell walls destruction (muramidase activity), and dissolution of the stabilized fibrin (isopeptidase activity). Both activities are known to be inhibited by sodium chloride at near physiological concentrations, but the structural basis remains unknown. Here we present two crystal structures of destabilase, including a 1.1Β Γ
-resolution structure in complex with sodium ion. Our structures reveal the location of sodium ion between Glu34/Asp46 residues, which were previously recognized as a glycosidase active site. While sodium coordination with these amino acids may explain inhibition of the muramidase activity, its influence on previously suggested Ser49/Lys58 isopeptidase activity dyad is unclear. We revise the Ser49/Lys58 hypothesis and compare sequences of i-type lysozymes with confirmed destabilase activity. We suggest that the general base for the isopeptidase activity is His112 rather than Lys58. pKa calculations of these amino acids, assessed through the 1Β ΞΌs molecular dynamics simulation, confirm the hypothesis. Our findings highlight the ambiguity of destabilase catalytic residues identification and build foundations for further research of structureβactivity relationship of isopeptidase activity as well as structure-based protein design for potential anticoagulant drug development.</p
Biosynthesis lipase from the fungus
The article provides a method of isolating a lipase producer and identification methods by classical methods in microbiology of an isolated micromycete producing lipase. The species affiliation of Penicillium hordei has been determined. The temperature of micromycete cultivation was studied, as well as the dynamics of Penicillium hordei lipase biosynthesis. Screening plans were also carried out (Plackett-Berman Plan), then optimization according to the steep climb plan and an experiment based on the plan of a full-factor experiment (three-level CFE) was also conducted to determine the optimal concentrations that are significant for lipase biosynthesis of components. A mathematical model is constructed that describes the relationship between the influence of two significant factors on the volumetric activity of P. Hordei culture fluid. The most optimal composition for the culture fluid for lipase biosynthesis, Penicillium hordei micromycete, was compiled
Π ΠΎΠ·ΡΠΎΠ±ΠΊΠ° ΡΠΎΠ·ΠΌΡΡΠΎΡΡΠ°Π±ΡΠ»ΡΠ½ΠΈΡ ΡΡ Π΅ΠΌ Π±Π°Π³Π°ΡΠΎΡΠ°ΡΠΎΠ²ΠΈΡ ΡΡΡΠ±ΠΎΠΏΡΠΎΠ²ΠΎΠ΄ΡΠ² Ρ ΡΠΈΠ»ΡΠ½Π΄ΡΠΈΡΠ½ΠΈΡ ΠΏΠΎΡΡΠ΄ΠΈΠ½ ΡΠΈΡΠΊΡ Π· Π²ΡΠ³Π»Π΅ΠΏΠ»Π°ΡΡΠΈΠΊΡΠ²
In the framework of the momentless theory of cylindrical thin shells, the elastic deformation of multilayer pipes and pressure vessels is investigated. It is assumed that the pipes and pressure vessels are made by two-way spiral winding of carbon fiber reinforced plastic tape on a metal mandrel.The analysis of the dependences of elastic deformations on the reinforcement angles is performed. The relations for axial and circumferential deformations of the wall, depending on the structure of the layer package, reinforcement angles under static loading are obtained. The separate and combined effect of internal pressure and temperature is considered. For the separate effect of loads, the graphs of deformations against the winding angle are plotted.Composite pipes made of KMU-4L carbon fiber reinforced plastic, as well as composite metal-composite pipes, are investigated. The results obtained for thermal loads are in good agreement with the data of the known experiment and solution. Depending on the load parameters, composite and metal-composite structures with dimensionally stable properties are determined.It is shown that dimensionally stable structures can be used to solve the problem of compensation of elastic deformations of pipelines. For this purpose, using the ASCP software package, the variant analysis of model structures is performed. By the comparative analysis of the three versions of the structure, layer package structures and reinforcement schemes, ensuring a significant reduction of loads on the supporting elements are obtained. On the example of a pipeline with a flowing fluid, it is shown that the use of dimensionally stable multilayer pipes makes it possible to eliminate bending deformations and significantly reduce the level of working forces and stresses.Dimensionally stable composite multilayer pipes open up new approaches to the design of pipelines and pressure vessels. It is possible to create structures with predetermined (not necessarily zero) displacement fields, consistent with the fields of the initial technological displacements, as well as with the displacements of conjugate elastic elements and equipment when the operating mode changes. The scope of such structures is not limited to "hot" pipes. The results can be used in cryogenic engineeringΠ ΡΠ°ΠΌΠΊΠ°Ρ
Π±Π΅Π·ΠΌΠΎΠΌΠ΅Π½ΡΠ½ΠΎΠΉ ΡΠ΅ΠΎΡΠΈΠΈ ΡΠΈΠ»ΠΈΠ½Π΄ΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΡΠΎΠ½ΠΊΠΈΡ
ΠΎΠ±ΠΎΠ»ΠΎΡΠ΅ΠΊ ΠΈΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½ΠΎ ΡΠΏΡΡΠ³ΠΎΠ΅ Π΄Π΅ΡΠΎΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΌΠ½ΠΎΠ³ΠΎΡΠ»ΠΎΠΉΠ½ΡΡ
ΡΡΡΠ± ΠΈ ΡΠΎΡΡΠ΄ΠΎΠ² Π΄Π°Π²Π»Π΅Π½ΠΈΡ. ΠΡΠ΅Π΄ΠΏΠΎΠ»Π°Π³Π°Π΅ΡΡΡ, ΡΡΠΎ ΡΡΡΠ±Ρ ΠΈ ΡΠΎΡΡΠ΄Ρ Π΄Π°Π²Π»Π΅Π½ΠΈΡ Π²ΡΠΏΠΎΠ»Π½Π΅Π½Ρ ΠΏΠ΅ΡΠ΅ΠΊΡΠ΅ΡΡΠ½ΠΎΠΉ ΡΠΏΠΈΡΠ°Π»ΡΠ½ΠΎΠΉ Π½Π°ΠΌΠΎΡΠΊΠΎΠΉ Π°ΡΠΌΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠΉ Π»Π΅Π½ΡΡ ΠΈΠ· ΡΠ³Π»Π΅ΠΏΠ»Π°ΡΡΠΈΠΊΠ° Π½Π° ΠΌΠ΅ΡΠ°Π»Π»ΠΈΡΠ΅ΡΠΊΡΡ ΠΎΠΏΡΠ°Π²ΠΊΡ.ΠΡΠΏΠΎΠ»Π½Π΅Π½ Π°Π½Π°Π»ΠΈΠ· Π·Π°Π²ΠΈΡΠΈΠΌΠΎΡΡΠ΅ΠΉ ΡΠΏΡΡΠ³ΠΈΡ
Π΄Π΅ΡΠΎΡΠΌΠ°ΡΠΈΠΉ ΠΎΡ ΡΠ³Π»ΠΎΠ² Π°ΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ. ΠΠΎΠ»ΡΡΠ΅Π½Ρ ΡΠΎΠΎΡΠ½ΠΎΡΠ΅Π½ΠΈΡ Π΄Π»Ρ ΠΎΡΠ΅Π²ΡΡ
ΠΈ ΠΎΠΊΡΡΠΆΠ½ΡΡ
Π΄Π΅ΡΠΎΡΠΌΠ°ΡΠΈΠΉ ΡΡΠ΅Π½ΠΊΠΈ Π² Π·Π°Π²ΠΈΡΠΈΠΌΠΎΡΡΠΈ ΠΎΡ ΡΡΡΡΠΊΡΡΡΡ ΠΏΠ°ΠΊΠ΅ΡΠ° ΡΠ»ΠΎΠ΅Π², ΡΠ³Π»ΠΎΠ² Π°ΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΏΡΠΈ ΡΡΠ°ΡΠΈΡΠ΅ΡΠΊΠΎΠΌ Π½Π°Π³ΡΡΠΆΠ΅Π½ΠΈΠΈ. Π Π°ΡΡΠΌΠΎΡΡΠ΅Π½ΠΎ ΠΎΠ±ΠΎΡΠΎΠ±Π»Π΅Π½Π½ΠΎΠ΅ ΠΈ ΠΊΠΎΠΌΠ±ΠΈΠ½ΠΈΡΠΎΠ²Π°Π½Π½ΠΎΠ΅ Π΄Π΅ΠΉΡΡΠ²ΠΈΠ΅ Π²Π½ΡΡΡΠ΅Π½Π½Π΅Π³ΠΎ Π΄Π°Π²Π»Π΅Π½ΠΈΡ ΠΈ ΡΠ΅ΠΌΠΏΠ΅ΡΠ°ΡΡΡΡ. ΠΠ»Ρ ΠΎΠ±ΠΎΡΠΎΠ±Π»Π΅Π½Π½ΠΎΠ³ΠΎ Π΄Π΅ΠΉΡΡΠ²ΠΈΡ Π½Π°Π³ΡΡΠ·ΠΎΠΊ ΠΏΠΎΡΡΡΠΎΠ΅Π½Ρ Π³ΡΠ°ΡΠΈΠΊΠΈ Π·Π°Π²ΠΈΡΠΈΠΌΠΎΡΡΠ΅ΠΉ Π΄Π΅ΡΠΎΡΠΌΠ°ΡΠΈΠΉ ΠΎΡ ΡΠ³Π»Π° Π½Π°ΠΌΠΎΡΠΊΠΈ.ΠΡΡΠ»Π΅Π΄ΠΎΠ²Π°Π½Ρ ΠΊΠΎΠΌΠΏΠΎΠ·ΠΈΡΠ½ΡΠ΅ ΡΡΡΠ±Ρ, ΠΈΠ·Π³ΠΎΡΠΎΠ²Π»Π΅Π½Π½ΡΠ΅ ΠΈΠ· ΡΠ³Π»Π΅ΠΏΠ»Π°ΡΡΠΈΠΊΠ°[s1]Β KΠΠ£-4Π, Π° ΡΠ°ΠΊΠΆΠ΅ ΡΠΎΡΡΠ°Π²Π½ΡΠ΅ ΠΌΠ΅ΡΠ°Π»Π»ΠΎ-ΠΊΠΎΠΌΠΏΠΎΠ·ΠΈΡΠ½ΡΠ΅ ΡΡΡΠ±Ρ. Π Π΅Π·ΡΠ»ΡΡΠ°ΡΡ, ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΡΠ΅ Π΄Π»Ρ ΡΠ΅ΠΏΠ»ΠΎΠ²ΡΡ
Π½Π°Π³ΡΡΠ·ΠΎΠΊ, Ρ
ΠΎΡΠΎΡΠΎ ΡΠΎΠ³Π»Π°ΡΡΡΡΡΡ Ρ Π΄Π°Π½Π½ΡΠΌΠΈ ΠΈΠ·Π²Π΅ΡΡΠ½ΠΎΠ³ΠΎ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ° ΠΈ ΡΠ΅ΡΠ΅Π½ΠΈΡ. Π Π·Π°Π²ΠΈΡΠΈΠΌΠΎΡΡΠΈ ΠΎΡ ΠΏΠ°ΡΠ°ΠΌΠ΅ΡΡΠΎΠ² Π½Π°Π³ΡΡΠ·ΠΎΠΊ ΠΎΠΏΡΠ΅Π΄Π΅Π»Π΅Π½Ρ ΠΊΠΎΠΌΠΏΠΎΠ·ΠΈΡΠ½ΡΠ΅ ΠΈ ΠΌΠ΅ΡΠ°Π»Π»ΠΎ-ΠΊΠΎΠΌΠΏΠΎΠ·ΠΈΡΠ½ΡΠ΅ ΡΡΡΡΠΊΡΡΡΡ Ρ ΡΠ°Π·ΠΌΠ΅ΡΠΎΡΡΠ°Π±ΠΈΠ»ΡΠ½ΡΠΌΠΈ ΡΠ²ΠΎΠΉΡΡΠ²Π°ΠΌΠΈ.ΠΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ ΡΠ°Π·ΠΌΠ΅ΡΠΎΡΡΠ°Π±ΠΈΠ»ΡΠ½ΡΠ΅ ΡΡΡΡΠΊΡΡΡΡ ΠΌΠΎΠ³ΡΡ Π±ΡΡΡ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½Ρ Π΄Π»Ρ ΡΠ΅ΡΠ΅Π½ΠΈΡ ΠΏΡΠΎΠ±Π»Π΅ΠΌΡ ΠΊΠΎΠΌΠΏΠ΅Π½ΡΠ°ΡΠΈΠΈ ΡΠΏΡΡΠ³ΠΈΡ
Π΄Π΅ΡΠΎΡΠΌΠ°ΡΠΈΠΉ ΡΡΡΠ±ΠΎΠΏΡΠΎΠ²ΠΎΠ΄ΠΎΠ². Π‘ ΡΡΠΎΠΉ ΡΠ΅Π»ΡΡ ΠΏΡΠΈ ΠΏΠΎΠΌΠΎΡΠΈ ΠΏΡΠΎΠ³ΡΠ°ΠΌΠΌΠ½ΠΎΠ³ΠΎ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠ° ASCP Π²ΡΠΏΠΎΠ»Π½Π΅Π½ Π²Π°ΡΠΈΠ°Π½ΡΠ½ΡΠΉ Π°Π½Π°Π»ΠΈΠ· ΠΌΠΎΠ΄Π΅Π»ΡΠ½ΠΎΠΉ ΠΊΠΎΠ½ΡΡΡΡΠΊΡΠΈΠΉ. ΠΡΡΠ΅ΠΌ ΡΠΎΠΏΠΎΡΡΠ°Π²ΠΈΡΠ΅Π»ΡΠ½ΠΎΠ³ΠΎ Π°Π½Π°Π»ΠΈΠ·Π° ΡΡΠ΅Ρ
Π²Π°ΡΠΈΠ°Π½ΡΠΎΠ² ΠΊΠΎΠ½ΡΡΡΡΠΊΡΠΈΠΈ ΠΏΠΎΠ»ΡΡΠ΅Π½Ρ ΡΡΡΡΠΊΡΡΡΡ ΠΏΠ°ΠΊΠ΅ΡΠΎΠ² ΡΠ»ΠΎΠ΅Π² ΠΈ ΡΡ
Π΅ΠΌΡ Π°ΡΠΌΠΈΡΠΎΠ²Π°Π½ΠΈΡ, ΠΎΠ±Π΅ΡΠΏΠ΅ΡΠΈΠ²Π°ΡΡΠΈΠ΅ Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎΠ΅ ΡΠ½ΠΈΠΆΠ΅Π½ΠΈΠ΅ Π½Π°Π³ΡΡΠ·ΠΎΠΊ Π½Π° ΠΎΠΏΠΎΡΠ½ΡΠ΅ ΡΠ»Π΅ΠΌΠ΅Π½ΡΡ. ΠΠ° ΠΏΡΠΈΠΌΠ΅ΡΠ΅ ΡΡΡΠ±ΠΎΠΏΡΠΎΠ²ΠΎΠ΄Π° Ρ ΠΏΡΠΎΡΠ΅ΠΊΠ°ΡΡΠ΅ΠΉ ΠΆΠΈΠ΄ΠΊΠΎΡΡΡΡ ΠΏΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ ΡΠ°Π·ΠΌΠ΅ΡΠΎΡΡΠ°Π±ΠΈΠ»ΡΠ½ΡΡ
ΠΌΠ½ΠΎΠ³ΠΎΡΠ»ΠΎΠΉΠ½ΡΡ
ΡΡΡΠ± ΠΏΠΎΠ·Π²ΠΎΠ»ΡΠ΅Ρ ΠΈΡΠΊΠ»ΡΡΠΈΡΡ Π΄Π΅ΡΠΎΡΠΌΠ°ΡΠΈΠΈ ΠΈΠ·Π³ΠΈΠ±Π° ΠΈ Π·Π°ΠΌΠ΅ΡΠ½ΠΎ ΠΏΠΎΠ½ΠΈΠ·ΠΈΡΡ ΡΡΠΎΠ²Π΅Π½Ρ ΡΠ°Π±ΠΎΡΠΈΡ
ΡΡΠΈΠ»ΠΈΠΉ ΠΈ Π½Π°ΠΏΡΡΠΆΠ΅Π½ΠΈΠΉ.Π Π°Π·ΠΌΠ΅ΡΠΎΡΡΠ°Π±ΠΈΠ»ΡΠ½ΡΠ΅ ΠΌΠ½ΠΎΠ³ΠΎΡΠ»ΠΎΠΉΠ½ΡΠ΅ ΡΡΡΠ±Ρ ΠΈΠ· ΠΊΠΎΠΌΠΏΠΎΠ·ΠΈΡΠΎΠ² ΠΎΡΠΊΡΡΠ²Π°ΡΡ Π½ΠΎΠ²ΡΠ΅ ΠΏΠΎΠ΄Ρ
ΠΎΠ΄Ρ ΠΊ ΠΏΡΠΎΠ΅ΠΊΡΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΡΠ±ΠΎΠΏΡΠΎΠ²ΠΎΠ΄ΠΎΠ² ΠΈ ΡΠΎΡΡΠ΄ΠΎΠ² ΠΏΠΎΠ΄ Π΄Π°Π²Π»Π΅Π½ΠΈΠ΅ΠΌ. ΠΠΎΡΠ²Π»ΡΡΡΡΡ Π²ΠΎΠ·ΠΌΠΎΠΆΠ½ΠΎΡΡΠΈ ΡΠΎΠ·Π΄Π°Π½ΠΈΡ ΠΊΠΎΠ½ΡΡΡΡΠΊΡΠΈΠΉ Ρ Π½Π°ΠΏΠ΅ΡΠ΅Π΄ Π·Π°Π΄Π°Π½Π½ΡΠΌΠΈ (Π½Π΅ ΠΎΠ±ΡΠ·Π°ΡΠ΅Π»ΡΠ½ΠΎ Π½ΡΠ»Π΅Π²ΡΠΌΠΈ) ΠΏΠΎΠ»ΡΠΌΠΈ ΠΏΠ΅ΡΠ΅ΠΌΠ΅ΡΠ΅Π½ΠΈΠΉ, ΡΠΎΠ³Π»Π°ΡΠΎΠ²Π°Π½Π½ΡΠΌΠΈ Ρ ΠΏΠΎΠ»ΡΠΌΠΈ Π½Π°ΡΠ°Π»ΡΠ½ΡΡ
ΡΠ΅Ρ
Π½ΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΏΠ΅ΡΠ΅ΠΌΠ΅ΡΠ΅Π½ΠΈΠΉ, Π° ΡΠ°ΠΊΠΆΠ΅ Ρ ΠΏΠ΅ΡΠ΅ΠΌΠ΅ΡΠ΅Π½ΠΈΡΠΌΠΈ ΡΠΎΠΏΡΡΠΆΠ΅Π½Π½ΡΡ
ΡΠΏΡΡΠ³ΠΈΡ
ΡΠ»Π΅ΠΌΠ΅Π½ΡΠΎΠ² ΠΈ ΠΎΠ±ΠΎΡΡΠ΄ΠΎΠ²Π°Π½ΠΈΡ ΠΏΡΠΈ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΠΈ ΡΠ΅ΠΆΠΈΠΌΠ° ΡΠ°Π±ΠΎΡΡ. ΠΠ±Π»Π°ΡΡΡ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΡ ΠΏΠΎΠ΄ΠΎΠ±Π½ΡΡ
ΠΊΠΎΠ½ΡΡΡΡΠΊΡΠΈΠΉ Π½Π΅ ΠΎΠ³ΡΠ°Π½ΠΈΡΠΈΠ²Π°Π΅ΡΡΡ "Π³ΠΎΡΡΡΠΈΠΌΠΈ" ΡΡΡΠ±Π°ΠΌΠΈ. ΠΠΎΠ»ΡΡΠ΅Π½Π½ΡΠ΅ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΡ ΠΌΠΎΠ³ΡΡ Π½Π°ΠΉΡΠΈ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ Π² ΠΊΡΠΈΠΎΠ³Π΅Π½Π½ΠΎΠΉ ΡΠ΅Ρ
Π½ΠΈΠΊΠ΅Β Π ΡΠ°ΠΌΠΊΠ°Ρ
Π±Π΅Π·ΠΌΠΎΠΌΠ΅Π½ΡΠ½ΠΎΡ ΡΠ΅ΠΎΡΡΡ ΡΠΈΠ»ΡΠ½Π΄ΡΠΈΡΠ½ΠΈΡ
ΡΠΎΠ½ΠΊΠΈΡ
ΠΎΠ±ΠΎΠ»ΠΎΠ½ΠΎΠΊ Π΄ΠΎΡΠ»ΡΠ΄ΠΆΠ΅Π½ΠΎ ΠΏΡΡΠΆΠ½Π΅ Π΄Π΅ΡΠΎΡΠΌΡΠ²Π°Π½Π½Ρ Π±Π°Π³Π°ΡΠΎΡΠ°ΡΠΎΠ²ΠΈΡ
ΡΡΡΠ± Ρ ΠΏΠΎΡΡΠ΄ΠΈΠ½ ΡΠΈΡΠΊΡ. ΠΠ΅ΡΠ΅Π΄Π±Π°ΡΠ°ΡΡΡΡΡ, ΡΠΎ ΡΡΡΠ±ΠΈ Ρ ΠΏΠΎΡΡΠ΄ΠΈΠ½ΠΈ ΡΠΈΡΠΊΡ Π²ΠΈΠΊΠΎΠ½Π°Π½Ρ ΠΏΠ΅ΡΠ΅Ρ
ΡΠ΅ΡΠ½ΠΎΡ ΡΠΏΡΡΠ°Π»ΡΠ½ΠΎΡ Π½Π°ΠΌΠΎΡΡΠ²Π°Π½Π½ΡΠΌ Π°ΡΠΌΠΎΠ²Π°Π½ΠΎΡ ΡΡΡΡΡΠΊΠΈ Π· Π²ΡΠ³Π»Π΅ΠΏΠ»Π°ΡΡΠΈΠΊΠ° Π½Π° ΠΌΠ΅ΡΠ°Π»Π΅Π²Ρ ΠΎΠΏΡΠ°Π²Π»Π΅Π½Π½Ρ.ΠΠΈΠΊΠΎΠ½Π°Π½ΠΎ Π°Π½Π°Π»ΡΠ· Π·Π°Π»Π΅ΠΆΠ½ΠΎΡΡΠ΅ΠΉ ΠΏΡΡΠΆΠ½ΠΈΡ
Π΄Π΅ΡΠΎΡΠΌΠ°ΡΡΠΉ Π²ΡΠ΄ ΠΊΡΡΡΠ² Π°ΡΠΌΡΠ²Π°Π½Π½Ρ. ΠΡΡΠΈΠΌΠ°Π½ΠΎ ΡΠΏΡΠ²Π²ΡΠ΄Π½ΠΎΡΠ΅Π½Π½Ρ Π΄Π»Ρ ΠΎΡΡΠΎΠ²ΠΈΡ
Ρ ΠΎΠΊΡΡΠΆΠ½ΠΈΡ
Π΄Π΅ΡΠΎΡΠΌΠ°ΡΡΠΉ ΡΡΡΠ½ΠΊΠΈ Π² Π·Π°Π»Π΅ΠΆΠ½ΠΎΡΡΡ Π²ΡΠ΄ ΡΡΡΡΠΊΡΡΡΠΈ ΠΏΠ°ΠΊΠ΅ΡΠ° ΡΠ°ΡΡΠ², ΠΊΡΡΡΠ² Π°ΡΠΌΡΠ²Π°Π½Π½Ρ ΠΏΡΠΈ ΡΡΠ°ΡΠΈΡΠ½ΠΎΠΌΡ Π½Π°Π²Π°Π½ΡΠ°ΠΆΠ΅Π½Π½Ρ. Π ΠΎΠ·Π³Π»ΡΠ½ΡΡΠΎ Π²ΡΠ΄ΠΎΠΊΡΠ΅ΠΌΠ»Π΅Π½Π° Ρ ΠΊΠΎΠΌΠ±ΡΠ½ΠΎΠ²Π°Π½Π° Π΄ΡΡ Π²Π½ΡΡΡΡΡΠ½ΡΠΎΠ³ΠΎ ΡΠΈΡΠΊΡ Ρ ΡΠ΅ΠΌΠΏΠ΅ΡΠ°ΡΡΡΠΈ. ΠΠ»Ρ Π²ΡΠ΄ΠΎΠΊΡΠ΅ΠΌΠ»Π΅Π½ΠΎΡ Π΄ΡΡ Π½Π°Π²Π°Π½ΡΠ°ΠΆΠ΅Π½Ρ ΠΏΠΎΠ±ΡΠ΄ΠΎΠ²Π°Π½Ρ Π³ΡΠ°ΡΡΠΊΠΈ Π·Π°Π»Π΅ΠΆΠ½ΠΎΡΡΠ΅ΠΉ Π΄Π΅ΡΠΎΡΠΌΠ°ΡΡΠΉ Π²ΡΠ΄ ΠΊΡΡΠ° Π½Π°ΠΌΠΎΡΡΠ²Π°Π½Π½Ρ.ΠΠΎΡΠ»ΡΠ΄ΠΆΠ΅Π½ΠΎ ΠΊΠΎΠΌΠΏΠΎΠ·ΠΈΡΠ½Ρ ΡΡΡΠ±ΠΈ, Π²ΠΈΠ³ΠΎΡΠΎΠ²Π»Π΅Π½Ρ Π· Π²ΡΠ³Π»Π΅ΠΏΠ»Π°ΡΡΠΈΠΊΠ° KΠΠ£-4Π, Π° ΡΠ°ΠΊΠΎΠΆ ΡΠΊΠ»Π°Π΄ΠΎΠ²Ρ ΠΌΠ΅ΡΠ°Π»ΠΎ-ΠΊΠΎΠΌΠΏΠΎΠ·ΠΈΡΠ½Ρ ΡΡΡΠ±ΠΈ. Π Π΅Π·ΡΠ»ΡΡΠ°ΡΠΈ, ΠΎΡΡΠΈΠΌΠ°Π½Ρ Π΄Π»Ρ ΡΠ΅ΠΏΠ»ΠΎΠ²ΠΈΡ
Π½Π°Π²Π°Π½ΡΠ°ΠΆΠ΅Π½Ρ, Π΄ΠΎΠ±ΡΠ΅ ΡΠ·Π³ΠΎΠ΄ΠΆΡΡΡΡΡΡ Π· Π΄Π°Π½ΠΈΠΌΠΈ Π²ΡΠ΄ΠΎΠΌΠΎΠ³ΠΎ Π΅ΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΡ Ρ ΡΡΡΠ΅Π½Π½Ρ. ΠΠ°Π»Π΅ΠΆΠ½ΠΎ Π²ΡΠ΄ ΠΏΠ°ΡΠ°ΠΌΠ΅ΡΡΡΠ² Π½Π°Π²Π°Π½ΡΠ°ΠΆΠ΅Π½Ρ Π²ΠΈΠ·Π½Π°ΡΠ΅Π½Ρ ΠΊΠΎΠΌΠΏΠΎΠ·ΠΈΡΠ½Ρ ΡΠ° ΠΌΠ΅ΡΠ°Π»ΠΎ-ΠΊΠΎΠΌΠΏΠΎΠ·ΠΈΡΠ½Ρ ΡΡΡΡΠΊΡΡΡΠΈ Π· ΡΠΎΠ·ΠΌΡΡΠΎΡΡΠ°Π±ΡΠ»ΡΠ½ΠΈΠΌΠΈ Π²Π»Π°ΡΡΠΈΠ²ΠΎΡΡΡΠΌΠΈ.ΠΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΠΎ ΡΠΎΠ·ΠΌΡΡΠΎΡΡΠ°Π±ΡΠ»ΡΠ½Ρ ΡΡΡΡΠΊΡΡΡΠΈ ΠΌΠΎΠΆΡΡΡ Π±ΡΡΠΈ Π²ΠΈΠΊΠΎΡΠΈΡΡΠ°Π½Ρ Π΄Π»Ρ Π²ΠΈΡΡΡΠ΅Π½Π½Ρ ΠΏΡΠΎΠ±Π»Π΅ΠΌΠΈ ΠΊΠΎΠΌΠΏΠ΅Π½ΡΠ°ΡΡΡ ΠΏΡΡΠΆΠ½ΠΈΡ
Π΄Π΅ΡΠΎΡΠΌΠ°ΡΡΠΉ ΡΡΡΠ±ΠΎΠΏΡΠΎΠ²ΠΎΠ΄ΡΠ². Π ΡΡΡΡ ΠΌΠ΅ΡΠΎΡ Π·Π° Π΄ΠΎΠΏΠΎΠΌΠΎΠ³ΠΎΡ ΠΏΡΠΎΠ³ΡΠ°ΠΌΠ½ΠΎΠ³ΠΎ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΡ ASCP Π²ΠΈΠΊΠΎΠ½Π°Π½ΠΈΠΉ Π²Π°ΡΡΠ°Π½ΡΠ½ΠΈΠΉ Π°Π½Π°Π»ΡΠ· ΠΌΠΎΠ΄Π΅Π»ΡΠ½ΠΎΡ ΠΊΠΎΠ½ΡΡΡΡΠΊΡΡΡ. Π¨Π»ΡΡ
ΠΎΠΌ ΠΏΠΎΡΡΠ²Π½ΡΠ»ΡΠ½ΠΎΠ³ΠΎ Π°Π½Π°Π»ΡΠ·Ρ ΡΡΡΠΎΡ
Π²Π°ΡΡΠ°Π½ΡΡΠ² ΠΊΠΎΠ½ΡΡΡΡΠΊΡΡΡ ΠΎΡΡΠΈΠΌΠ°Π½Ρ ΡΡΡΡΠΊΡΡΡΠΈ ΠΏΠ°ΠΊΠ΅ΡΡΠ² ΡΠ°ΡΡΠ² Ρ ΡΡ
Π΅ΠΌΠΈ Π°ΡΠΌΡΠ²Π°Π½Π½Ρ, ΡΠΎΠ± Π·Π°Π±Π΅Π·ΠΏΠ΅ΡΠΈΡΠΈ Π·Π½Π°ΡΠ½Π΅ Π·Π½ΠΈΠΆΠ΅Π½Π½Ρ Π½Π°Π²Π°Π½ΡΠ°ΠΆΠ΅Π½Ρ Π½Π° ΠΎΠΏΠΎΡΠ½Ρ Π΅Π»Π΅ΠΌΠ΅Π½ΡΠΈ. ΠΠ° ΠΏΡΠΈΠΊΠ»Π°Π΄Ρ ΡΡΡΠ±ΠΎΠΏΡΠΎΠ²ΠΎΠ΄Ρ Π· ΠΏΡΠΎΡΡΠΊΠ°ΡΡΠΎΡ ΡΡΠ΄ΠΈΠ½ΠΎΡ ΠΏΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΠΎ Π·Π°ΡΡΠΎΡΡΠ²Π°Π½Π½Ρ ΡΠΎΠ·ΠΌΡΡΠΎΡΡΠ°Π±ΡΠ»ΡΠ½ΠΈΡ
Π±Π°Π³Π°ΡΠΎΡΠ°ΡΠΎΠ²ΠΈΡ
ΡΡΡΠ± Π΄ΠΎΠ·Π²ΠΎΠ»ΡΡ Π²ΠΈΠΊΠ»ΡΡΠΈΡΠΈ Π΄Π΅ΡΠΎΡΠΌΠ°ΡΡΡ Π²ΠΈΠ³ΠΈΠ½Ρ Ρ ΠΏΠΎΠΌΡΡΠ½ΠΎ Π·Π½ΠΈΠ·ΠΈΡΠΈ ΡΡΠ²Π΅Π½Ρ ΡΠΎΠ±ΠΎΡΠΈΡ
Π·ΡΡΠΈΠ»Ρ Ρ Π½Π°ΠΏΡΡΠΆΠ΅Π½Ρ.Π ΠΎΠ·ΠΌΡΡΠΎΡΡΠ°Π±ΡΠ»ΡΠ½Ρ Π±Π°Π³Π°ΡΠΎΡΠ°ΡΠΎΠ²Ρ ΡΡΡΠ±ΠΈ Π· ΠΊΠΎΠΌΠΏΠΎΠ·ΠΈΡΡΠ² Π²ΡΠ΄ΠΊΡΠΈΠ²Π°ΡΡΡ Π½ΠΎΠ²Ρ ΠΏΡΠ΄Ρ
ΠΎΠ΄ΠΈ Π΄ΠΎ ΠΏΡΠΎΠ΅ΠΊΡΡΠ²Π°Π½Π½Ρ ΡΡΡΠ±ΠΎΠΏΡΠΎΠ²ΠΎΠ΄ΡΠ² Ρ ΠΏΠΎΡΡΠ΄ΠΈΠ½ ΠΏΡΠ΄ ΡΠΈΡΠΊΠΎΠΌ. Π'ΡΠ²Π»ΡΡΡΡΡΡ ΠΌΠΎΠΆΠ»ΠΈΠ²ΠΎΡΡΡ ΡΡΠ²ΠΎΡΠ΅Π½Π½Ρ ΠΊΠΎΠ½ΡΡΡΡΠΊΡΡΠΉ Π· Π½Π°ΠΏΠ΅ΡΠ΅Π΄ Π·Π°Π΄Π°Π½ΠΈΠΌΠΈ (Π½Π΅ ΠΎΠ±ΠΎΠ²'ΡΠ·ΠΊΠΎΠ²ΠΎ Π½ΡΠ»ΡΠΎΠ²ΠΈΠΌΠΈ) ΠΏΠΎΠ»ΡΠΌΠΈ ΠΏΠ΅ΡΠ΅ΠΌΡΡΠ΅Π½Ρ, ΡΠ·Π³ΠΎΠ΄ΠΆΠ΅Π½ΠΈΠΌΠΈ Π· ΠΏΠΎΠ»ΡΠΌΠΈ ΠΏΠΎΡΠ°ΡΠΊΠΎΠ²ΠΈΡ
ΡΠ΅Ρ
Π½ΠΎΠ»ΠΎΠ³ΡΡΠ½ΠΈΡ
ΠΏΠ΅ΡΠ΅ΠΌΡΡΠ΅Π½Ρ, Π° ΡΠ°ΠΊΠΎΠΆ Π· ΠΏΠ΅ΡΠ΅ΠΌΡΡΠ΅Π½Π½ΡΠΌΠΈ ΡΠΏΠΎΠ»ΡΡΠ΅Π½ΠΈΡ
ΠΏΡΡΠΆΠ½ΠΈΡ
Π΅Π»Π΅ΠΌΠ΅Π½ΡΡΠ² Ρ ΡΡΡΠ°ΡΠΊΡΠ²Π°Π½Π½Ρ ΠΏΡΠΈ Π·ΠΌΡΠ½Ρ ΡΠ΅ΠΆΠΈΠΌΡ ΡΠΎΠ±ΠΎΡΠΈ. ΠΠ±Π»Π°ΡΡΡ Π·Π°ΡΡΠΎΡΡΠ²Π°Π½Π½Ρ ΠΏΠΎΠ΄ΡΠ±Π½ΠΈΡ
ΠΊΠΎΠ½ΡΡΡΡΠΊΡΡΠΉ Π½Π΅ ΠΎΠ±ΠΌΠ΅ΠΆΡΡΡΡΡΡ Β«Π³Π°ΡΡΡΠΈΠΌΠΈΒ» ΡΡΡΠ±Π°ΠΌΠΈ. ΠΡΡΠΈΠΌΠ°Π½Ρ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠΈ ΠΌΠΎΠΆΡΡΡ Π·Π½Π°ΠΉΡΠΈ Π·Π°ΡΡΠΎΡΡΠ²Π°Π½Π½Ρ Π² ΠΊΡΡΠΎΠ³Π΅Π½Π½ΡΠΉ ΡΠ΅Ρ
Π½ΡΡ
Add and Go: FRET Acceptor for Live-Cell Measurements Modulated by Externally Provided Ligand
A substantial number of genetically encoded fluorescent sensors rely on the changes in FRET efficiency between fluorescent cores, measured in ratiometric mode, with acceptor photobleaching or by changes in fluorescence lifetime. We report on a modulated FRET acceptor allowing for simplified one-channel FRET measurement based on a previously reported fluorogen-activating protein, DiB1. Upon the addition of the cell-permeable chromophore, the fluorescence of the donor-fluorescent protein mNeonGreen decreases, allowing for a simplified one-channel FRET measurement. The reported chemically modulated FRET acceptor is compatible with live-cell experiments and allows for prolonged time-lapse experiments with dynamic energy transfer evaluation