Development and pilot of equine facilitated physical therapy outcome measure tool for chronic low back pain patients.

Equine Facilitated Physical Therapy (EFPT) lacks consistent documentation due to being an unconventional physical therapy treatment to chronic low back pain patients (LBP) and lacking rehabilitation outcome measure tools for a stable (equestrian) environment. The objectives were to develop an online evaluation tool as well as to define inter- and intra-rater reliability to validate the outcome measurement tool "Evaluation of maintaining sitting position (on a horse) and walking (short distances)" designed for LBP patients in EFPT". A total of 48 movement related functions (n = 48), were derived from the International Classification of Functioning (ICF) and organized to an online evaluation tool. Depending on the state of validation two to six (2-6) raters scored randomized patient (n = 22) video material, recorded during a 12-week EFPT intervention, with the designed tool. Inter-rater agreement level between the experts reached good (α = 87) reliability for the scoring of the items and calculated per patient excellent (α = 100). Intra-rater reliability reached good (α = 87) and per patient good (α = 80) repeatability. For the healthy adults the reliability between raters reached acceptable (α = 72) levels and per rated excellent (α = 100). The developed assessment tool was found satisfactory to fulfil the requirement for the therapeutic practice. With the use of the tool physical therapist may detect postural changes for LBP patients as outcome report in EFPT. The tool may be used to identify treatment progress and to help design home exercises. The created tool will help to collect similar outcome measures from LBP patients in EFPT and to validate the treatment within industry.

[Realization of medical and medical psychological rehabilitation in sanatorium-resort organizations of Ministry of Defence of the Russian Federation]. / Organizatsiya meditsinskoi i mediko-psikhologicheskoi reabilitatsii v sanatorno-kurortnykh organizatsiyakh Minoborony Rossii.

Current conditions of military service demand higher standards of servicemen health status, whose professional activity in extreme combat conditions is accompanied by the impact of many adverse environmental factors on the body. One of the priority tasks of medical service of Armed Forces of the Russian Federation in 2023 yr. is to improve the system of curative and preventive measures in medical and medical psychological rehabilitation to recover health and increase working capacity of servicemen. The article includes an overview of measures given to servicemen for medical and medical psychological rehabilitation in sanatorium-resort organizations of Ministry of Defence in the Russian Federation for the period from 2013 to 2022 yrs.

Modular Nanotransporters Capable of Binding to SARS-CoV-2 Virus Nucleocapsid Protein in Target Cells.

On the basis of literature data, an antibody-like molecule, monobody, was selected that is capable of interacting with the nucleocapsid protein (N protein) of the SARS-CoV-2 virus with a high affinity (dissociation constant 6.7 nM). We have previously developed modular nanotransporters (MNTs) to deliver various molecules to a selected compartment of target cells. In this work, a monobody to the N protein of the SARS-CoV-2 virus was inserted in the MNT using genetic engineering methods. In this MNT, a site for the cleavage of the monobody from the MNT in endosomes was also inserted. It was shown by thermophoresis that the cleavage of this monobody from the MNT by the endosomal protease cathepsin B leads to a 12-fold increase in the affinity of the monobody for the N protein. Cellular thermal shift assay showed the ability of the obtained MNT to interact with the N protein in A431 cells transfected with the SARS-CoV-2 N protein fused to the mRuby3 fluorescent protein.

Intracellular Delivery of an Antibody-Like Molecule Capable of Inhibiting c-Myc.

A modular nanotransporter (MNT) carrying the sequence of an antibody-like molecule, anti-c-Myc nanobody, was synthesized and characterized. It was demonstrated that the created MNT is able to interact with the target protein, c-Myc oncogene, with a dissociation constant of 46 ± 14 nM, internalize into target cells, change Myc-dependent expression, and exert an antiproliferative effect.

Selection of an Amino Acid Site with One of the Fastest Cleavage Kinetics by the Endosomal Protease Cathepsin B for Potential Use in Drug Delivery Systems.

On the basis of known published data, six peptide sequences were selected that are potentially capable of being rapidly cleaved by the endosomal protease cathepsin B. For comparison, the cleavage of common linker sequences, polyglycine and polyglycine-serine, by cathepsin B was also studied. Different ends of these peptides were labeled with sulfoCyanine3 and sulfoCyanine5 fluorescent dyes, between which Förster resonant energy transfer (FRET) is possible. The kinetics of cleavage of peptides by cathepsin B was studied on a multimodal plate reader by FRET signal reduction. FKFL and FRRG cleavage sites have been shown to be the most suitable for potential use in various drug delivery systems. These sites are much more efficiently cleaved under slightly acidic conditions of endosomes than at neutral extracellular pH.

Quantitative Description of the N-Protein of the SARS-CoV-2 Virus Degradation in Cells Stably Expressing It under the Influence of New Modular Nanotransporters.

Two eukaryotic cell lines, A549 and A431, with stable expression of the nucleocapsid protein (N-protein) of the SARS-CoV-2 virus fused with the red fluorescent protein mRuby3 were obtained. Using microscopy, the volumes of the cytoplasm and nucleus were determined for these cells. Using quantitative immunoblotting techniques, the concentrations of the N-mRuby3 fusion protein in their cytoplasm were assessed. They were 19 and 9 µM for A549 and A431 cells, respectively. Using these concentrations, the initial rate of N-protein degradation in the studied cells was estimated from the decrease in cell fluorescence. In A549 and A431 cells, it was the same (84 nM per hour). The approach of quantitatively describing the degradation process can be applied to analyze the effectiveness of a wide class of antiviral drugs that cause degradation of viral proteins.

Modular Nanotransporters Capable of Causing Intracellular Degradation of the N-Protein of the SARS-CoV-2 Virus in A549 Cells with Temporary Expression of This Protein Fused with a Fluorescent Protein mRuby3.

Modular nanotransporters (MNTs) containing an antibody-like molecule, monobody, to the N­protein of the SARS-CoV-2 virus, as well as an amino acid sequence that recruits the Keap1 E3 ligase (E3BP) were created. This MNT also included a site for cleavage of the E3BP monobody from the MNT in acidic endocytic compartments. It was shown that this cleavage by the endosomal protease cathepsin B leads to a 2.7-fold increase in the affinity of the E3BP monobody for the N-protein. Using A549 cells with transient expression of the N-protein fused with the fluorescent protein mRuby3, it was shown that incubation with MNT leads to a significant decrease in mRuby3 fluorescence. It is assumed that the developed MNTs can serve as a basis for the creation of new antiviral drugs against the SARS-CoV-2 virus.

Delivery of Antibody-Like Molecules, Monobodies, Capable of Binding with SARS-CoV-2 Virus Nucleocapsid Protein, into Target Cells.

Based on previous studies, two antibody-like molecules, monobodies, capable of high-affinity interaction with the SARS-CoV-2 nucleocapsid protein (dissociation constant of tens of nM) were selected. For delivery to target cells, genetically engineered constructs containing monobody and TAT peptide, placed either at the N- or C-terminus of the resulting polypeptide, were produced and expressed in E. coli. The construct with the highest affinity to the SARS-CoV-2 nucleocapsid protein was revealed with the use of thermophoresis technique. Cellular thermal shift assay demonstrated the ability of this construct to interact with the nucleocapsid protein within HEK293T cells transfected with the SARS-CoV-2 nucleocapsid protein fused to the mRuby3 fluorescent protein. Replacement of TAT peptide to S10 shuttle peptide, containing endosomolytic peptide, significantly improved the penetration of the construct into the target cells.

Among Antibody-Like Molecules, Monobodies, Able to Interact with Nucleocapsid Protein of SARS-CoV Virus, There Are Monobodies with High Affinity to Nucleocapsid Protein of SARS-CoV-2 Virus.

Seven amino acid sequences of antibody mimetics molecules, monobodies, capable of interacting with the nucleocapsid protein of the SARS-CoV virus, were taken from the literature. Nucleotide sequences of monobody genes were obtained by gene synthesis, which were expressed in E. coli and isolated using Ni-NTA chromatography. It was shown by thermophoresis that three of the seven selected antibody-like molecules can interact with high affinity (dissociation constant of tens of nM) with the nucleocapsid protein of the SARS-CoV-2 virus. For the remaining four monobodies, only low affinity binding with a dissociation constant of several µM was found.

[Professor Veniamin Vasilievich Volkov (to the 100th anniversary of his birth)]. / Professor Veniamin Vasil'evich Volkov (k 100-letiyu so dnya rozhdeniya).

The article presents the main stages of the career of Professor V.V. Volkov - the Hero of Socialist Labor, USSR State Prize Laureate, Honored Scientist of the RSFSR, Honorary Doctor of the Military Medical Academy, and describes his scientific and practical contribution to the development of modern ophthalmology. The authors reviewed his life journey, clinical, scientific and educational activities.

Magnetic structures of Fe32+δGe33As2 and Fe32+δ'Ge35-xPx intermetallic compounds: a neutron diffraction and 57Fe Mössbauer spectroscopy study.

Fe32+δGe33As2 and Fe32+δ'Ge35-xPx are quasi-binary intermetallic compounds that possess a rare variant of intergrowth-type crystal structure, which is a combination of the column shaped Co2Al5 and MgFe6Ge6 structure type blocks. The compounds are antiferromagnets with the Néel temperatures around 125 K. Neutron powder diffraction experiments on the samples with δ≈ 0.1, δ'≈ 0.5 and x≈ 3 reveal commensurate magnetic ordering of low symmetry in both compounds and a non-monotonic change in the intensities of magnetic reflections. On the other hand, temperature dependence of the hyperfine fields obtained from 57Fe Mössbauer spectroscopy indicates a gradual, monotonic increase in local magnetic fields upon cooling. We interpret these results as a spin reorientation within the Co2Al5-type block of the crystal structure, with the possible formation of a non-collinear magnetic order at low temperatures. Between the compounds, the reorientation occurs at significantly different temperatures, however the resulting magnetic structures themselves are similar as well as the average values of the magnetic moments and the hyperfine fields.

New Recombinant Carriers Binding Specifically to the Epidermal Growth Factor Receptor.

New recombinant carriers-modular nanotransporters (MNTs)-with N-terminal ligand module to the epidermal growth factor receptor (EGFR) were developed and characterized. Human epidermal growth factor (hEGF) and antibody-like protein Z1907 were used as a ligand module. We demonstrated that MNTs are able to internalize in a receptor-specific manner into the target cancer cells and to accumulate in the target cell nuclei. Conjugation of MNTs with the Auger electron emitter 111In significantly enhanced the cytotoxic effect of 111In on the target cells. It was found that the transfer of EGF from the C-terminus to the N-terminus of the MNT enhanced the proliferation of target cells, whereas the use of Z1907 did not have a similar effect.

Stabilization of Modular Nanotransporters by Embedding Hemin in Them in a New Strain with Heme Receptor Expression.

It was found that the use of a new strain-producer Escherichia coli, expressing the heme receptor ChuA, enables obtaining a hemin-containing modular nanotransporter (MNT) for drug delivery into the nuclei of target cells. The hemin-containing MNT becomes stabilized, which leads to an increase in its thermal stability and prevents aggregation of this protein.

The Delivery of Biologically Active Agents into the Nuclei of Target Cells for the Purposes of Translational Medicine.

Development of vehicles for the subcellular targeted delivery of biologically active agents is very promising for the purposes of translational medicine. This review summarizes the results obtained by researchers from the Laboratory of Molecular Genetics of Intracellular Transport, Institute of Gene Biology RAS, which allowed them to design the core technology: modular nanotransporters. This approach ensures high efficacy and cell specificity for different anti-cancer agents, as they are delivered into the most vulnerable subcellular compartment within the cells of interest and makes it possible for antibody mimetics to penetrate into a compartment of interest within the target cells ("diving antibodies"). Furthermore, polyplexes, complexes of polycationic block copolymers of DNA, have been developed and characterized. These complexes are efficient both in vitro and in vivo and demonstrate predominant transfection of actively dividing cells.

[The role of the vascular surgeon in improvement of organization of dialysis help]. / Rol' angiokhirurga v uluchshenii organizatsii dializnoi pomoshchi.

Analysed herein are one-year results of formation of arteriovenous fistulas in 109 patients with end-stage chronic renal failure, as well as therapeutic decision-making after angiosurgical counselling of 144 patients presenting with 'problem' permanent vascular accesses. The counselling and formation of arteriovenous fistulas were carried out in conditions of interdepartmental collaboration between outpatient centres dealing with haemodialysis and vascular surgeons specialized in ultrasound mapping of peripheral vessels and performing different variants of arteriovenous fistulas. The angiosurgical care was as close to the patient as possible. Of the 109 operated patients, primary arteriovenous fistulas were made in 46 (42.2%) cases, secondary AVF - in 27 (24.8%) cases, and reconstruction of AVF - in 36 (33.0%) cases. Of the 144 patients with 'problem' permanent vascular assesses, correction of arteriovenous fistulas turned out impossible in 13 (9.1%). In the remaining 131 (90.9%) patients there was a possibility of different variants of open reconstruction of arteriovenous fistulas or performing angioplasty. Active policy of vascular surgeons in interdepartmental collaboration with nephrologists made it possible to bridge over the difficulties of patients routing which resulted in reduction of the terms of formation of arteriovenous fistulas by 2 months. Preventive arteriovenous fistulas were carried out in 17.4% of cases of primary permanent vascular assesses. During a year after formation of permanent vascular accesses, the number of patients with vascular catheters in ambulatory centres decreased from 22 to 17%. These positive changes in organization of the dialysis treatment made it possible to reduce the risks of infectious complications, to obtain adequate blood flow characteristics for haemodialysis procedures, as well as to decrease financial expenses and labour costs for AVF care.

Targeted Intracellular Delivery of Antibodies: The State of the Art.

A dominant area of antibody research is the extension of the use of this mighty experimental and therapeutic tool for the specific detection of molecules for diagnostics, visualization, and activity blocking. Despite the ability to raise antibodies against different proteins, numerous applications of antibodies in basic research fields, clinical practice, and biotechnology are restricted to permeabilized cells or extracellular antigens, such as membrane or secreted proteins. With the exception of small groups of autoantibodies, natural antibodies to intracellular targets cannot be used within living cells. This excludes the scope of a major class of intracellular targets, including some infamous cancer-associated molecules. Some of these targets are still not druggable via small molecules because of large flat contact areas and the absence of deep hydrophobic pockets in which small molecules can insert and perturb their activity. Thus, the development of technologies for the targeted intracellular delivery of antibodies, their fragments, or antibody-like molecules is extremely important. Various strategies for intracellular targeting of antibodies via protein-transduction domains or their mimics, liposomes, polymer vesicles, and viral envelopes, are reviewed in this article. The pitfalls, challenges, and perspectives of these technologies are discussed.

MNT Optimization for Intracellular Delivery of Antibody Fragments.

We studied the possibility of optimizing modular nanotransporters (MNTs) for the intracellular delivery of antibody fragments into the nuclei of cells of a specified type. Basic MNT with a reduced size retaining the desired functions was obtained, and the principal possibility of obtaining an MNT carrying an antibody fragment by microbiological synthesis was shown.

Modular Nanotransporter with P21 Fragment Inhibits DNA Repair after Bleomycin Treatment.

Modular nanotransporter (MNT) with C-terminal fragment of the p21 protein was synthesized and characterized, and its effect on DNA lesions was studied. This p21 fragment in MNT can significantly inhibit DNA repair in A431 human carcinoma cells after bleomycin treatment.

Study of Biodistribution of the Modular Nanotransporters after Systemic Administration in Murine Cloudman S91 Melanoma Model.

The distribution of modular nanotransporters (MNTs) that are used to deliver drugs into melanoma cell nuclei after their intravenous administration into mice with Cloudman S91 melanoma was studied. The modification of MNTs with polyethylene glycol (PEG) of different length and their administration during the treatment with docetaxel, nitroglycerin, and excess of nonspecific MNTs leads to an improved accumulation of MNTs in the tummor. Among the variants studied, the MNT with attached PEG with Mr 40 kDa exhibited the best properties.