Previously we suggested a new pharmaceutical derived from coordination complex of Co3+ with polyacrylic acid (PAA) exhibiting hemostatic and microbicidal activity, namely Hestatin. Differences in the physiological activity of Hestatin synthesized from PAA 10 kDa (Hestatin 10) and 200 kDa (Hestatin 200) were shown. We tested the acute toxicity of Hestatin and its effect on the healing rate of sterile wounds in rats. Free 10 kDa PAA, emulsion wax, emulsion wax carrying resveratrol, and dexpanthenol were tested for comparison. Hestatin 10 exhibited no acute toxicity when administered intragastrically at dosages of 5 g per kg. Hestatin 10 surpassed all tested drugs in its wound healing ability. Histological analysis of skin sections of rats in the area of healing defects showed an increased rate of synthesis of reticular fibers compared to the placebo. In the early stages of wound healing (inflammatory phase), Hestatin 10 stimulated taxis of mast cells (MCs) to the wound bottom but not to the wound perimeter. At the final stage of wound healing (remodeling phase), Hestatin 10 promoted MC evacuation from the skin defect area. This effect is the opposite of the well-known wound-healing agents (dexpanthenol and resveratrol), which enhance MC infiltration into the defect area in the remodeling phase.

The pW8-3C construct encoding the artificial tumor-targeting protein W8-3C with the addition of three residues of free Cys at the C terminus has been created and described for the first time. Using purified W8-3C protein, dispersions of nanoparticles 75.24 nm in diameter at a polydispersity index (Pdi) of 0.064 and Se content of 1.566 μg/mL were obtained and characterized for the first time. The dispersions remained stable during storage for 6 months at +4°C. For comparison, the maximum Se content in the nanoparticle dispersion obtained in the presence of W8-3C protein and Pluronic F-127 was 399 μg/mL. The cytotoxic activity of the obtained nanoparticles was studied on HeLa (cervical carcinoma), U87MG (glioma), MCF7 (breast carcinoma), and HCT116 (colon carcinoma) human tumor cell lines and compared with that on the diploid human fibroblast line WI-38 in vitro. It was shown that the IC50 of Se nanoparticles obtained using the W8-3C protein for tumor lines ranged from 5.25 to 8.37 μg/mL, while the IC50 for normal fibroblasts was 14.3 μg/mL (difference in values by a factor of 1.7–2.7 times).

For the first time, the pW8-3C construct encoding the artificial tumor-specific protein W8-3C with the addition of 3 residues of free Cys at the C end was created and described. Using purified W8-3C protein, dispersions of nanoparticles 75.24 nm in diameter at polydispersity index (Pdi) of 0.064 and Se content of 1566 µg/mL were obtained and characterized for the first time. The dispersions remained stable upon storage for 6 months at +4°C. For comparison, the maximum Se content in the nanoparticle dispersion obtained in the presence of W8-3C protein and Pluronic F-127 was 399 µg/ml. The cytotoxic activity of the obtained nanoparticles was studied on transplanted cells of human tumor lines: HeLa (cervical carcinoma), U-87 MG (glioblastoma), MCF-7 (breast carcinoma) and HCT-116 (colon carcinoma) and compared with that on the diploid human fibroblast line WI-38 in vitro. It was shown that the IC50 of Se nanoparticles obtained using the W8-3C protein for tumor lines ranged from 5.25 to 8.37 µg/ml, while the IC50 for normal fibroblasts was 14.3 µg/ml (difference in values by a factor of 1.7–2.7 times).

99mTc is a well-known radionuclide that is widely used and readily available for SPECT/CT (Single-Photon Emission Computed Tomography) diagnosis. However, commercial isotope carriers are not specific enough to tumours, rapidly clear from the bloodstream, and are not safe. To overcome these limitations, we suggest immunologically compatible recombinant proteins containing a combination of metal binding sites as 99mTc chelators and several different tumour-specific ligands for early detection of tumours. E1b protein containing metal-binding centres and tumour-specific ligands targeting integrin αvβ3 and nucleolin, as well as a short Cys-rich sequence, was artificially constructed. It was produced in E. coli, purified by metal-chelate chromatography, and used to obtain a complex with 99mTc. This was administered intravenously to healthy Balb/C mice at an activity dose of about 80 MBq per mouse, and the biodistribution was studied by SPECT/CT for 24 h. Free sodium 99mTc-pertechnetate at the same dose was used as a reference. The selectivity of 99mTc-E1b and the kinetics of isotope retention in tumours were then investigated in experiments in C57Bl/6 and Balb/C mice with subcutaneously transplanted lung carcinoma (LLC) or mammary adenocarcinoma (Ca755, EMT6, or 4T1). The radionuclide distribution ratio in tumour and adjacent normal tissue (T/N) steadily increased over 24 h, reaching 15.7 ± 4.2 for EMT6, 16.5 ± 3.8 for Ca755, 6.7 ± 4.2 for LLC, and 7.5 ± 3.1 for 4T1.

A cobalt polyacrylate tight complex was synthesized from polyacrylic acid (PAA) with average molecular masses of 9356 and 184,631 kDa. The source PAA with programmed molecular weight was synthesized under controlled conditions using isopropanol as a chain growth controlling agent. Tight complexes with Co2+ yielded 1:32 PAA units in the case of 10 kDa (Hestatin 10) and 1:19 in the case of 200 kDa (Hestatin 200). Purification of the tight complexes from the Co2+ in the weakly bound salt form was carried out by using sequential dialysis against deionized water. Testing of the haemostatic activity of the manufactured tight complexes on an animal model of capillary and venous bleeding was carried out; free PAA 10 and PAA 200 were used as the control samples. The experiment demonstrated high haemostatic efficacy of Hestatin 200, Hestatin 10, and PAA 10, whereas PAA 200 acted as an antithrombotic agent.

Probiotics can act as an alternative to antibiotics in animal feeding, but their use is minimal due to their expensive production. Dry grass is rich with bacteria beneficial for animal feeding and can be used as a probiotic. However, data about the quantitative dependence of the grass microbiome on environmental factors and seasons remain insufficient for preparing “grass-meal-based probiotics”. Four grass samples were collected in two geographically remote regions of Russia; their microbiome was characterized by metagenomic sequencing of 16S rDNA libraries and microbiological seeding, and biological testing of the grass meal was carried out on 6 groups of birds containing 20 Ross 308 cross broilers each for a period of 42 days. The positive control group (PC) obtained 16–25 mg/mL toltrazuril (coccidiostatic agent) and 0.5 mL/L liquid antibiotic enrostin (100 mg/mL ciprofloxacin and 106 MU/mL colistin sulfate in the commercial preparation) within the drinking water, while the negative control group (NC) obtained no medicines. Four experimental groups were fed the diet supplemented with 1% grass meal over the period of 7–42 days of life; no commercial medicines were used here. A spontaneous infection with Eimeria was registered in the NC control groups, which caused the loss of 7 chickens. No losses were registered in the PC group or the two experimental groups. In two other experimental groups, losses of coccidiosis amounted to 10% and 15%, respectively. All specimens of the grass meal demonstrated a significant effect on the average body weight gain compared to NC. Taken together, these observations support the hypothesis that the grass meal may substitute toltrazuril for protecting the chickens from parasitic invasion and increase average daily weight gain (ADG) as effectively as the antibiotic enrostin.

Feed phytases are purchased as a dry culture medium of secreting producers, mostly micellar fungi. These preparations are required to withstand heating up to 75–80 °C because they are intended for mixing with feed components with subsequent granulation by spray drying. For this reason, many phytases that have a high specific activity at 37 °C and correspond to the optimal pH of intestinal chyme are not used in practice. A novel expression system allowing accumulation of the phytase from Obesumbacterium proteus within yeast Yarrowia lipolytica was proposed. Encapsulation increases thermal stability of the enzyme from 55 °C up to 70 °C. The obtained preparation exhibited a high impact on the daily weight gain of a weaned mouse model fed a phosphorus-deficient diet at a dosage 165 phytase activity units (FYT)/kg, whereas a commercial phytase preparation—Ladozyme Proxi derived from Aspergillus ficuum—did not improve the daily weight gain even at the dosage of 15,000 FYT/kg.

The ability of gold polyacrylate (aurumacryl) to increase the sensitivity of murine tumor (В-16/F10 melanoma) to radiation treatment has been explored. Radiation therapy employed in combination with aurumacryl leads to a moderate but significant inhibition of tumor growth by 81–90

Three artificial proteins that bind the gadolinium ion (Gd3+) with tumour-specific ligands were de novo engineered and tested as candidate drugs for binary radiotherapy (BRT) and contrast agents for magnetic resonance imaging (MRI). Gd3+-binding modules were derived from calmodulin. They were joined with elastin-like polypeptide (ELP) repeats from human elastin to form the four-centre Gd3+-binding domain (4MBS-domain) that further was combined with F3 peptide (a ligand of nucleolin, a tumour marker) to form the F3-W4 block. The F3-W4 block was taken alone (E2-13W4 protein), as two repeats (E1-W8) and as three repeats (E1-W12). Each protein was supplemented with three copies of the RGD motif (a ligand of integrin αvβ3) and green fluorescent protein (GFP). In contrast to Magnevist (a Gd-containing contrast agent), the proteins exhibited three to four times higher accumulation in U87MG glioma and A375 melanoma cell lines than in normal fibroblasts. The proteins remained for >24 h in tumours induced by Ca755 adenocarcinoma in C57BL/6 mice. They exhibited stability towards blood proteases and only accumulated in the liver and kidney. The technological advantages of using the engineered proteins as a basis for developing efficient and non-toxic agents for early diagnosis of tumours by MRI as well as part of BRT were demonstrated.

Lake Vostok is the deepest lake of Antarctica but has poor accessibility for study due to a thick glacial cover, however, water samples of this lake have become available for study just recently. Previously, only the microbiome of the ice cover samples was characterized. Here we report results of bacteriological seeding with subsequent identification of the heterotrophic microorganisms (bacteria and micellar fungi) present by 16S rDNA sequencing as well as results of a direct molecular study of the water microbiome. Surprisingly, the data obtained gave evidence of a predominant occurrence of common chemoorganotrophs that were rather psychrotolerant than psychrophilic. We isolated and described strains belonging to eight heterotrophic microbial species able to grow in a rich medium: six bacterial strains belonging to the species Microbacterium testaceum and Microbacterium trichothecenolyticum, Brevundimonas diminuta, Sphingomonas oligophenolica, Sphingomonas sp. and Sphingobium limneticum; and two fungal strains belonging to Dendryphion sp. and Cladosporium fusiforme. Direct study of 16S rDNA purified water samples confirmed the predominance of the Brevundimonas, Microbacterium, Bradyrhizobium, and Bacillus (Bacillus cereus) genera.

In order to increase the bioavailability of water-insoluble pyropheophorbide-a (PPP-a) methyl ester, its liposomal form is prepared and the physicochemical and photochemical properties of this form are studied. The quantum yield of 1O2 is found to have a bell-shaped dependence on the concentration of PPP-a in the lipid phase of liposomes with the maximum at 31.6 µmol/g of lipids. The IR spectroscopy shows the photoinduced formation of aldehyde groups in the lipid phase of liposomes. The intracellular accumulation of PPP-a is confirmed by confocal microscopy.

An increase in the spread of antibiotic-resistant opportunistic microorganisms causes serious problems in the treatment of purulent infections, burns, and trophic ulcers. We tested the antimicrobial activity in vivo of three polyphenols, Resveratrol, Dihydroquercetin (Taxifolin), and Dihydromyricetin (Ampelopsin) from Norway spruce bark to promote the elimination of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans from wounds. Purulent infection was modelled on wounds in rats infected with suspensions containing 109 CFU (colony-forming unit)/mL of pathogens. The wound area was treated daily with solutions of the polyphenols or placebo for 14 days after the beginning of the treatment. The animals were examined daily, and each stage of the wound healing (inflammation, granulation, and maturation (marginal epithelialisation) was documented. The planimetric analysis of the wound recovery percentage was performed on the 3rd, 10th, and 14th day after the start of curing. Then, one echelon (three or four animals from each subgroup) was withdrawn from the experiment on days 3 (three animals), 10 (three animals), and 14 (four animals) for microscopy analysis of cytological composition of their wound defects by microscopy and microbiological analysis of their contamination with pathogens. Our results show that they are also able to suppress mast cell infiltration and stimulate lymphocyte and macrophage (monocyte) infiltration into the wound. Resveratrol stimulated the replacement of the scar with normal tissue (with a clear boundary between the dermis and epidermis) and the restoration of hair follicles. Resveratrol turned out to be significantly better than some commercial antimicrobial (Levomecol) and antifungal (Clotrimazole) ointments and can be proposed as a promising drug for topical use for the treatment of trophic ulcers and burns.

Previously, a method of synthesis of trimethyl-substituted furodihydroquinoline (FDHQ), a putative photosensitizer for the therapy of psoriasis, was disclosed in patent RU #2614248. FDHQ was proposed for substituting derivatives of the psoralen (8-methoxypsoralen, 8‑MOP; 5-methoxypsoralen, 5‑MOP; and trimethylpsoralen, TMP) actually applied for this purpose. FDHQ exhibited an advantage over psoralens in safety for the patient. However, practical use of FDHQ was found to be impossible due to its insufficient solubility in any clinically permitted solvents. To solve this problem, a synthesis of six earlier unknown derivatives of FDHQ with side substitutes in position 5 of the benzene nucleus (carbamate, acetamide, and sulfuric moieties) were synthesized: N-(6,8,8-trimethyl-8,9-dihydrofuro[3,2-h]quinoline-5-yl)acetamide (W); tert-butyl (6,8,8-trimethyl-8,9-dihydrofuro[3,2-h]quinoline-5-yl)carbamate (K); tret-butylacetyl (6,8,8-trimethyl-8,9-dihydrofuro[3,2-h]quinoline-5-yl)-carbamate (WK); N-methyl-N-(6,8,8-trimethyl-8,9-dihydrofuro[3,2-h]quinoline-5-yl)acetamide (WС1); N-octyl-N-(6,8,8-trimethyl-8,9-dihydrofuro[3,2-h]quinoline-5-yl)acetamide (WС8); sodium acetyl (6,8,8-trimethyl dihydrofuro[3,2-h]quinoline-5-yl)sulfamate (S). In vitro testing on T-cell lympholeukosis (Jurkat), В-cell lymphoma (Raji) cell lines and immortalized human fibroblasts demonstrated optimal photosensitizing properties in compound S. The ratio between dark toxicity and phototoxicity of this compound (irradiation with UV light with λ = 302 nm) on the chosen model cell lines suggests it as a putative efficient and safe medicine for phototherapy of psoriasis.

Staphylococcus aureus is an extremely infectious and malignant pathogen among many bacteria species. The aim of this work is to provide a robust classification model that would be able to identify S. aureus independent of the culture growth stage and the variations in bacteria concentration in suspension and also one that would be able to identify the pathogen among both taxonomically close species of the same genus and taxonomically distant species of different genera, using Fourier transform infrared spectroscopy (FTIR). In total, the spectra of 141 isolates of 17 bacteria have been used. Based on a combination of principal component analysis (PCA) and linear discriminant analysis (LDA), an identification model providing 100% sensitivity and 98% specificity was built. Inherent reliability and flexibility of the model have been shown. The proposed method of analysis allows us to get closer to the diagnostic requirements in the field of clinical microbiology, and it can be utilized for typing of other pathogenic bacteria species.

At present, there is a need for a simple, noninvasive, highly specific and sensitive diagnostic test for hepatobiliary system disorders. Compounds labeled with carbon isotopes are widely used in various diagnostic breath tests; they are safe and can reliably detect a metabolic disorder or enzyme deficiency. The aim of this study was to synthesize 13С- and 14С-labeled linoleic acids suitable for use in hepatobiliary breath tests in terms of purity. In the synthesis of 13С-labeled linoleic acid, the chemical yield for 1-bromo-8,11-heptadecadien was 86.4% and the chemical yield for barium carbonate-13С, 96.0%. In the synthesis of 14С-labeled linoleic acid, the chemical yield for 1-bromo-8,11-heptadecadien was 87.39%; for barium carbonate-14С it was 97.1%. The specific radioactivity of 14С-labeled linoleic acids was 45.36 ± 0.02 mCi/g. The radiochemical yield of the reaction was 96.0%. The proposed method is suitable for batch production.

В настоящее время для диагностики заболеваний печени и билиарной системы требуется разработка простого неинвазивного теста с высокой чувствительностью и специфичностью. Соединения, меченные изотопом углерода, уже имеют широкое применение в диагностике различных заболеваний методами дыхательных тестов, безопасны и способны достоверно выявлять метаболические нарушения или дефицит специфичных ферментов в органах. Целью работы было получить линолевую кислоту, меченную 13С и 14С, по степени очистки пригодную для проведения дыхательных тестов в целях диагностики заболеваний гепатобилиарной системы. В предложенном способе химический выход реакции синтеза 13С-линолевой кислоты по 1-бром-8,11-гептадекадиену составил 86,4%, по 13С-карбонату бария — 96,0%. Химический выход реакции синтеза 14С-линолевой кислоты по 1-бром-8,11-гептадекадиену составил 87,39%, по 14С-карбонату бария — 97,1%. Удельная радиоактивность 14С-линолевой кислоты составила 45,36 ± 0,02 мКи/г. Радиохимический выход реакции — 96,0%. Способ удобен для серийного выпуска готового продукта.

The poultry red mite Dermanyssus gallinae is commonly acknowledged as a serious biological threat to global poultry breeding. This is mostly because of the inefficiency of acaricidal treatments of poultry houses, which does not eliminate the mite population, only constrains it. However, the impact of D. gallinae on poultry welfare is poorly characterized. The biochemical and cytological characteristics of infection are limited mainly to anaemia, decreased levels of triglycerides in the blood and eosinophilia. For the first time, we report the adverse effects of D. gallinae infestation of industrial poultry houses on the functional state of poultry livers. Treatment of poultry houses with acaricidal powder preparation, Wooran-dust 0.7%, containing synthetic pyrethroids (permethrin, S-fenvalerate and tetramethrin), led to the elimination of the mite population and resulted in normalization of most of the haematological and biochemical blood parameters (haemoglobin, erythrocytes, neutrophils, basophils, lymphocytes and AST). Increased eosinophils and ALT were the only indicators of infection in the blood of laying hens, and they remained above normal range until the 22nd day after the beginning of treatment with the acaricidal agent.

Неинвазивные дыхательные тесты с применением изотопно-меченых соединений представляют собой новый высокоточный и безопасный метод функционального исследования печени и билиарной системы. Целью работы было провести биологические испытания острой и субхронической токсичности 13С-меченых линолевой и линоленовой кислот, синтезированных по оригинальной методике и предназначенных для проведения диагностических дыхательных тестов. При однократном внутрижелудочном введении изучаемых соединений лабораторным мышам линии BALB/c и крысам Wistar в дозах, превышающих диагностические в 500–2500 раз, образцы соединений не вызывали смертности экспериментальных животных. При проведении субхронического эксперимента на крысах при дозировках испытываемых соединений, в 5 и 25 раз превышающих терапевтическую дозу для человека, в течение 14 суток было выявлено отсутствие достоверных изменений у животных в экспериментальных группах по сравнению с контрольной по массе тела, гематологическим показателям (содержанию эритроцитов, лейкоцитов и тромбоцитов в крови) и биохимическим показателям сыворотки крови (уровню гемоглобина, общего белка, щелочной фосфатазы, аланинаминотрансферазы, аспартатаминотрансферазы, лактатдегидрогеназы, билирубина). Исследованные меченые кислоты безвредны в дозах, планируемых для перорального введения, и могут быть рекомендованы к доклиническим и клиническим испытаниям.

Noninvasive stable isotope breath tests allow highly accurate and safe estimation of liver and biliary tract function. The aim of this study was to test 13С-labeled linoleic and linolenic acids intended for diagnostic use for acute and subchronic toxicity. The acids were synthesized using the patented method. A single intragastric administration of the tested compounds to experimental BALB/c mice and Wistar rats in the amounts exceeding clinical doses 500 to 2500-fold did not cause animal death. In the subchronic toxicity test, the rats received 5 to 25 times higher doses than recommended for clinical use in humans. In a 14-day follow-up period, no significant differences were observed between the main and the control groups in terms of weight, blood count (red blood cells, white blood cells, platelets), and blood biochemistry (hemoglobin, total protein, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, bilirubin). The studied compounds are safe at doses intended for oral administration and are recommended for further preclinical and clinical trials.