Search

Aim. One of the potential methods for treating cancer is photodynamic therapy (PDT), which involves the targeted destruction of cancer cells through the activation of photosensitizers (PS) using light irradiation. The aim of the study was to investigate the photodynamic properties of the squaraine dyes with hydrophilic and hydrophobic substituents. Methods. Fluorescence spectroscopy, UV-VIS spectroscopy, laser scanning confocal microscopy, cytotoxicity assay. Results. It was shown that all studied dyes have maximum absorption in the far-red region, which makes them suitable for PDT. It has been investigated that the squarains can form complexes with proteins, as indicated by changes in their fluorescence. The samples demonstrated minimal dark toxicity but exhibited cytotoxicity after irradiation. The cytotoxic effect of dyes with hydrophilic groups was reduced by binding to BSA. Meanwhile, a dye with hydrophobic substituents shows a photodynamic effect in the presence of albumin. All studied dyes can penetrate through the cellular membrane, stain the cell components in the cytoplasm, and do not accumulate in nuclei. Conclusions. All dyes showed low dark toxicity. However, cytotoxicity increased after irradiation with light of a wavelength of 670 nm. The binding of the dyes with hydrophilic substitutes to albumin negatively impacted their photosensitizing properties. At the same time, the dye with the highest tendency to aggregate exhibited the most significant cytotoxic effect. [ABSTRACT FROM AUTHOR]

We study the problem for homogeneous partial differential equations in two variables which are of second order with respect to time variable and of finite order with respect to another (spatial) variable, with homogeneous two-point conditions in time. We propose a method for the construction of nontrivial solutions of this problem when its characteristic determinant is nontrivial and the set of its zeroes is not empty. We apply this method to the construction of non-zero solutions of homogeneous two-point problems for some equations of mathematical physics. [ABSTRACT FROM AUTHOR]

Aim. Generation of monoclonal antibodies specific to Coenzyme A. Methods. Hybridoma technique. KLH carrier protein conjugated with CoA was used for immunization. Screening of positive clones was performed with BSA conjugated to CoA. Results. Monoclonal antibody that specifically recognizes CoA and CoA derivatives, but not its precursors ATP and cysteine has been generated. Conclusion. In this study, we describe for the first time the production and characterization of monoclonal antibodies against CoA. The monoclonal antibody 1F10 was shown to recognize specifically CoA in Western blotting, ELISA and immunoprecipitation. These properties make this antiboby a particularly valuable reagent for elucidating CoA function in health and disease. [ABSTRACT FROM AUTHOR]

Polymer composite materials with antibacterial properties based on poly(vinyl alcohol), which contain fine biocompatible mineral fillers and silver nanoparticles have been obtained. The method of preparation of polyacrylamide stabilized silver nanoparticles with shape of thin truncated triangular prisms has been developed. An approach to enhance adsorption capacity of the fillers to nanoparticulate silver was proposed. Studying bactericidal activity of obtained materials on model microorganisms (Escherichia coli) shown the high activity of the composites containing silver nanoparticles at the concentration of ≥ 0,018 %. [ABSTRACT FROM AUTHOR]

Aim. Since p70S6K1 plays a key role in protein synthesis and cell cycle regulation, it has been implicated in a number of human diseases, such as obesity, diabetes, and cancer. Increased activation of p70S6K1 has been reported in a variety of cancer types, so now it is being investigated as a potential therapeutic target. The purpose of this work was to create a novel more efficient approach of expression and purification of p70S6K1 constitutively active form by using Dual Bac-to-Bac protein expressing system. Methods. For the production of T-loop phosphorylated p70S6K1 protein we have used an insect Bac-to-Bac baculovirus expression system with pFastBac™ Dual vector, which possesses two multiple cloning sites controlled by polyhedrin promoter (PH) and p10 promoter, allowing the simultaneous expression of two heterologous genes. Therefore, we first cons tructed the pFastBac™ Dual plasmid loaded with 6Hisp70S6K1(ΔAID)T389D under the control of PH promotor together with GST PDPK1(ΔPH) controlled by p10. For the protein production Sf9 cells were infected with recombinant bacmid DNA.The purification of p70S6K1 was carried out using NiNTA affinity chromatography and subsequent dialysis. The quality and concentration of obtained protein was tested using SDS-PAGE under reducing conditions. To detect the activity of p70S6K1protein, in vitro kinase assay against kinase’s substrate — ribosomal protein S6, was used with following western blotting (WB). WBagainst phosphorylated by PDK1 sites of p70S6K1 was carried out to confirm the specificity and activity of the obtained recombinant kinase. Results. Here we demonstrate the novel approach of obtaining highly active recombinant p70S6K1 (His-actS6K1) using dual baculovirus vector coexpressing 6Hisp70S6K1(ΔAID)T389D together with GSTPDPK1(ΔPH), assuring the synthesis of both proteins by the infected cell. For this study the initial p70S6K1 was modified by addition of 6His tag at N terminal regulatory domain, deletion of C-terminal domain containing autoinhibitory motif (AID), and with mutation of T389 site to D389 to mimic the phosphorylation by mTORC1. This results in guaranteed and more efficient phosphorylation and following activation of p70S6K1, which was confirmed using in vitro kinase activity assay and WB. Conclusions. The current work was focused on the design of a new highly efficient system for the expression and purification of the recombinant His-actS6K1, using pFastBac™ Dual vector co expressing recombinant GST PDK1. Recent studies have shown that some cancer types demonstrate the enhanced expression and/or activation of p70S6K, hinting that it may serve as a biomarker for monitoring disease progression. With that in mind, our approach of large-scale expression of His-actS6K1 may offer a rapid, low-cost, and high yield method for producing constitutively active recombinant p70S6K1 and facilitate the development of its potential therapeutic regulators. [ABSTRACT FROM AUTHOR]

The retrospective analysis of invasive methods of prenatal diagnostics has been carried out in 346 women with hazard of fetal pathology. The following factors of hazard of inherent and heritable pathologies are established: complicated genetical and reproductive anamnesis (49.7%), extragynecological (37.0%) and gynecologial pathologies (13.6%), complicated course of pregnancy (48.6%). Using of invasive methods of prenatal diagnostics in the pregnant women of these groups made it possible to detect the innate malformations in 13.0% cases and heritable pathologies in 4.6% cases.

Aim: To identify novel tuberous sclerosis complex (TSC2) binding partners by yeast two-hybrid screening., Methods: The yeast two-hybrid system DupLEX-A developed by OriGene Technologies and Mouse embryo and HeLa cells cDNA libraries were used in this study. The "bait" constructs, containing full-length and truncated form of TSC2 were prepared. The expression of all constructs in yeast was confirmed by immunoblotting with specific anti-LexA antibodies. The suitability of generated constructs for screening was tested in autoactivation and nuclear translocation assays. Screening of mouse embryo and HeLa cDNA libraries with selected baits was carried out according to manufacturer's recommendations. Positive clones were selected using double selection procedure and further confirmed in mating assay. Isolated cDNA clones were identified by automated DNA sequencing and database searching., Results: Extensive screening of two cDNA libraries from mouse embryo and HeLa cells with TSC2 baits led to the isolation of 102 positives clones. The specificity of interaction between TSC2 and binding proteins of selected clones was confirmed by mating assay for 83 clones. Sequencing of these clones indicated that they encode already known and novel TSC2-binding partners., Conclusion: The isolation of several known TSC2-binding partners, such as several isoforms of 14-3-3, demonstrates the validity of generated bait constructs and screening conditions. In addition, we have found a number of novel interactors, which encode cytoskeletal proteins and signaling molecules, such as Ser/Thr phosphatases.