[Nd:YAG laser goniopuncture as a mandatory adjuvant procedure after non-penetrating deep sclerectomy (long-term observation results)]. / Nd:YAG-lazernaia goniopunktura kak obiazatel'naia ad''iuvantnaia protsedura posle nepronikaiushchei glubokoi skleréktomii (rezul'taty dolgosrochnogo nabliudeniia).

Today, Nd:YAG laser goniopuncture (LGP) is considered a mandatory non-penetrating deep sclerectomy adjuvant procedure. However, its indications and timing remain debatable. PURPOSE: To evaluate the effect of Nd:YAG laser goniopuncture on the long-term hypotensive effectiveness of non-penetrating deep sclerectomy. MATERIAL AND METHODS: The study included 114 patients after non-penetrating deep sclerectomy (NPDS). In the control group (n=58), Nd:YAG laser goniopuncture was performed within 3.4±1.9 (1.5-6.7) months, and in the main group (n=56) - within 1.12±0.08 (0.9-1.5) months after the surgery. Ultrasound biomicroscopy (UBM) was used to evaluate the semiotics of trabecular-Descemet's membrane (TDM), intrascleral canal (ISC) and filtering bleb. The follow-up period was 5 years. RESULTS: According to UBM data, the thickness (0.10±0.009 mm) and density (50±6%) of TDM (p=0.0001) increased before LGP in the control group, the height of ISC decreased to 0.49±0.19 (0.20-0.40) (p=0.03), the height of UBM scan - to 1.49±0.05 (1.41-2.9) (p=0.0001); IOP (P0) was 18.48±4.7 (11.2-22.9) mmHg (p=0.001). In the main group before LGP, TDM thickness was 0.08±0.006 mm, density was 40±5%, and IOP (P0) was 15.7±4.1 (9.1-18.5) mm Hg. Complete hypotensive success was achieved in 83.6% of cases in the control group and 96.2% in the main group in 6 months; 68.07% and 92.59% in 12 months; 41.3% and 75.8% in 24 months; 15.25% and 48.93% in 36; 15% and 34.8% in 60 months after the surgery, respectively (p=0.0001, 95% confidence interval). CONCLUSION: TDM is an additional level of retention of aqueous humor and plays key role in the formation of outflow pathways after NPDS. Performing LGP in the early postoperative period is an effective and safe adjuvant option, which excludes the influence of TDM on the formation of aqueous humor outflow pathways and significantly increases the long-term hypotensive efficacy of non-penetrating deep sclerectomy.

Design of Stable α-Helical Peptides and Thermostable Proteins in Biotechnology and Biomedicine.

α-Helices are the most frequently occurring elements of the secondary structure in water-soluble globular proteins. Their increased conformational stability is among the main reasons for the high thermal stability of proteins in thermophilic bacteria. In addition, α-helices are often involved in protein interactions with other proteins, nucleic acids, and the lipids of cell membranes. That is why the highly stable α-helical peptides used as highly active and specific inhibitors of protein-protein and other interactions have recently found more applications in medicine. Several different approaches have been developed in recent years to improve the conformational stability of α-helical peptides and thermostable proteins, which will be discussed in this review. We also discuss the methods for improving the permeability of peptides and proteins across cellular membranes and their resistance to intracellular protease activity. Special attention is given to the SEQOPT method (http://mml.spbstu.ru/services/seqopt/), which is used to design conformationally stable short α-helices.

[DYNAMICS AND MECHANISMS OF INTERACTION OF HETERO-HEXAMERIC TIP49a/b COMPLEXES WITH DS-DNA].

Evolutionary conserved TIP49a and TIP49b ATPases belong to the AAA+ superfamily of DNA-dependent ATPases that are involved in many cellular processes such as chromatin remodeling, regulation of transcription and cell division during mitosis, the maintenance of genome stability, snoRNP biogenesis, and participate in the formation of active form of telomerase. These proteins are involved in the complex networks of protein-protein interactions and, in spite of high structural similarity, in some cases, can perform opposite functions. Despite of the variety of their different activities, the exact mechanisms of action of TIP49a and TIP49b are still poorly understood. In this paper, by means of molecular docking approaches we first modeled the structures of hetero-hexameric TIP49 complexes with short ds-DNA fragments (20 base pairs with different GC content) within the central channel of hexameric ring. Using molecular dynamics simulations in the periodic water box (MD) we investigated conformational dynamics and mechanisms of DNA unwinding activity of these proteins. We shown that: a) the interaction between the positively charged protein loops and DNA within the central channel of protein ring leads to the partial unwinding of the DNA helix; b) DNA unwinding occurs only in the region within the protein ring, while the terminal parts of DNA outside the protein complex remain in its initial b-form conformation; c) the presence of ATP in the active sites of protein complex affects both the dynamics and the structure of DNA, leading to the breakage of some complementary bonds in AT-rich DNA sequences.

Possible Function of the ribT Gene of Bacillus subtilis: Theoretical Prediction, Cloning, and Expression.

The complete decipherment of the functions and interactions of the elements of the riboflavin biosynthesis operon (rib operon) of Bacillus subtilis are necessary for the development of superproducers of this important vitamin. The function of its terminal ribT gene has not been established to date. In this work, a search for homologs of the hypothetical amino acid sequence of the gene product through databases, as well as an analysis of the homolgs, was performed; the distribution of secondary structure elements was theoretically predicted; and the tertiary structure of the RibT protein was proposed. The ribT gene nucleotide sequence was amplified and cloned into the standard high-copy expression vector pET15b and then expressed after induction with IPTG in E. coli BL21 (DE3) strain cells containing the inducible phage T7 RNA polymerase gene. The ribT gene expression was confirmed by SDS-PAGE. The protein product of the expression was purified by affinity chromatography. Therefore, the real possibility of RibT protein production in quantities sufficient for further investigation of its structure and functional activity was demonstrated.