A Novel C1q Domain-Containing Protein Isolated from the Mollusk Modiolus kurilensis Recognizing Glycans Enriched with Acidic Galactans and Mannans

C1q domain-containing (C1qDC) proteins are a group of biopolymers involved in immune response as pattern recognition receptors (PRRs) in a lectin-like manner. A new protein MkC1qDC from the hemolymph plasma of Modiolus kurilensis bivalve mollusk widespread in the Northwest Pacific was purified. The isolation procedure included ammonium sulfate precipitation followed by affinity chromatography on pectin-Sepharose. The full-length MkC1qDC sequence was assembled using de novo mass-spectrometry peptide sequencing complemented with N-terminal Edman’s degradation, and included 176 amino acid residues with molecular mass of 19 kDa displaying high homology to bivalve C1qDC proteins. MkC1qDC demonstrated antibacterial properties against Gram-negative and Gram-positive strains. MkC1qDC binds to a number of saccharides in Ca(2+)-dependent manner which characterized by structural meta-similarity in acidic group enrichment of galactose and mannose derivatives incorporated in diversified molecular species of glycans. Alginate, κ-carrageenan, fucoidan, and pectin were found to be highly effective inhibitors of MkC1qDC activity. Yeast mannan, lipopolysaccharide (LPS), peptidoglycan (PGN) and mucin showed an inhibitory effect at concentrations three orders of magnitude greater than for the most effective saccharides. MkC1qDC localized to the mussel hemal system and interstitial compartment. Intriguingly, MkC1qDC was found to suppress proliferation of human adenocarcinoma HeLa cells in a dose-dependent manner, indicating to the biomedical potential of MkC1qDC protein

Polyurethane–poly(2-hydroxyethyl methacrylate) semi- IPN–nanooxide composites

Two sets of hybrid polyurethane–poly(2-hydroxyethyl methacrylate) semi-interpenetrating polymer network–nanooxide composites with 0.25 or 3 wt% nanosilica or nanoalumina functionalised with OH, NH2 or CHLCH2 groups were prepared. A combination of atomic force microscopy, infrared spectroscopy, thermally stimulated depolarisation current measurement, differential scanning calorimetry and creep rate spectroscopy analysis of the nanostructure and properties of the composites was performed. The pronounced dynamic heterogeneity and the strong impact of oxide additives, basically suppression of the dynamics and temperature-dependent increasing modulus of elasticity, were observed. The effects correlated with either interfacial interactions (for silica) or the nanostructure (for alumina). A low oxide content strongly affected the matrix due to the formation of an unusual cross-linked, via double covalent hybridisation of three components, structure of the nanocomposites

CRISPR/Cas9-generated mouse model with humanizing single-base substitution in the Gnao1 for safety studies of RNA therapeutics

The development of personalized medicine for genetic diseases requires preclinical testing in the appropriate animal models. GNAO1 encephalopathy is a severe neurodevelopmental disorder caused by heterozygous de novo mutations in the GNAO1 gene. GNAO1 c.607 G>A is one of the most common pathogenic variants, and the mutant protein Gαo-G203R likely adversely affects neuronal signaling. As an innovative approach, sequence-specific RNA-based therapeutics such as antisense oligonucleotides or effectors of RNA interference are potentially applicable for selective suppression of the mutant GNAO1 transcript. While in vitro validation can be performed in patient-derived cells, a humanized mouse model to rule out the safety of RNA therapeutics is currently lacking. In the present work, we employed CRISPR/Cas9 technology to introduce a single-base substitution into exon 6 of the Gnao1 to replace the murine Gly203-coding triplet (GGG) with the codon used in the human gene (GGA). We verified that genome-editing did not interfere with the Gnao1 mRNA or Gαo protein synthesis and did not alter localization of the protein in the brain structures. The analysis of blastocysts revealed the off-target activity of the CRISPR/Cas9 complexes; however, no modifications of the predicted off-target sites were detected in the founder mouse. Histological staining confirmed the absence of abnormal changes in the brain of genome-edited mice. The created mouse model with the “humanized” fragment of the endogenous Gnao1 is suitable to rule out unintended targeting of the wild-type allele by RNA therapeutics directed at lowering GNAO1 c.607 G>A transcripts

Trends in the epidemiology of diabetic foot and lower limb amputations in Russian Federation according to the Federal Diabetes Register (2013–2016)

BACKGROUND: The epidemiological study of diabetic foot (DF) is very important because of high risk lower limbs amputations in patients with diabetes mellitus (DM). AIMS: The aim of the study was to evaluate the DF prevalence in adult patients with type 1 (T1) and 2 (T2) diabetes in Russian Federation for period 201316years. METHODS: We have used the database of the Russian Federal Diabetes register, 81st regions included in the online register. Indicators were estimated per 10,000 adult DM patients (18years). RESULTS: In 2016, the prevalence of DF in RF was T1 4,7%, T2 1,9%, with marked interregional differences: 0,1519,9%, 0,0710,3%, respectively. The DF prevalence in RF decreased: T1 506,3473,6, T2 214,60194,8. The incidence of new DF cases/per year was stable in adults with T1: 20,820,4/; increased in T2 13.214.2. The mean age of DF diagnosis increased by 2years for both DM types. The average DM duration of DF determine increased T1 15.419.0years, T2 7.410.1years. Proportion of DF forms: neuropathic with trophic ulcer 41.6%, neuropathic form (Charcot's foot) 17.9%, the neuroischemic 28.3%, ischemic 12.2%, in T2: 41.6%, 7,4%, 32,4%, 18,5%, respectively. The amount of new cases of amputations/per year in dynamics: T1 10,512,4, T2 9,610,9, with marked interregional differences 0.132.9% in T1, 0.04-6.0% in T2. The mean DM duration before amputation increased in T1 18.421.3years, in T2 9.19.9. The average amputation age: T1 51.7years, T2 66.2years. There was marked decrease in proportion of major amputations: T1 43,637,0%, T2 52.245.5 by redistribution in one toe amputations T14,010.0%, in T22,89.1%. CONCLUSIONS: The dynamic of new DF cases in adult patients in Russian Federation is stable at T1, in T2 tends to increase. The interregional differences in frequency of DF and amputations may be due to differences in the quality of specialized care, the lack or shortage of diabetic foot cabinets, treatment of patients with DF in general surgical practice in a number of regions, which is recognized as a less effective strategy. A positive fact that proportion of high amputations declines, DF develops in later age and longer diabetes duration, that may reflect the increasing effectiveness of preventive lower limbs in diabetes

Antimicrobial resistance in foodborne <i>Salmonella enterica</i> isolates in the Republic of Belarus

Introduction. Antimicrobial resistance is a global public health concern. Salmonella spp., which can be transmitted to humans through contaminated food, are among the most important foodborne pathogens worldwide. Materials and methods. The antimicrobial resistance of 358 bacterial isolates collected from food and water in the Republic of Belarus (Belarus) in 20182021 was studied by analyzing phenotypic and genotypic characteristics of antibiotic bacterial resistance. MALDI-TOF mass spectrometry was used to classify and identify bacteria. Phenotypic antimicrobial susceptibility of bacteria was measured by the minimum inhibitory concentration method using a Sensititre automated bacteriological analyzer and the disk diffusion test for 45 antimicrobial agents. Antimicrobial resistance genes in multidrug-resistant Salmonella isolates were identified by whole-genome sequencing. Results. The in vitro testing of phenotypic bacterial susceptibility showed high susceptibility to fluoroquinolones (97.2%), third-generation cephalosporins (93.9%), carbapenems (98.0%), ampicillin (81.8%), aminoglycosides (97.5%), tetracyclines (87.5%), chloramphenicol (93.8%), trimethoprim/sulfamethoxazole (co-trimoxazole) (95.3%) and colistin (85.2%). It was found that the antibiotic resistance mechanism in S. enterica was associated with the presence of genes blaTEM-1B (82%), blaTEM-1C (7.7%), blaSHV-12 (2.6%), blaDHA-1 (2.6%), blaCMY-2 (7.7%), qnrB2 (9.1%), qnrB4 (9.1%), qnrB5 (9.1%), qnrB19 (72.7%), aac(6)-Ib-cr (9.1%), aac(6)-Iaa (100%), aadA1 (13.2%), aadA2 (8.8%), tetB (74.3%), tetA (25.7%), tetM (2.9%), tetD (28.6%), mcr-9 (1.5%). Conclusion. All the bacterial isolates were phenotypically susceptible to first-line antibiotics used in treatment of salmonellosis: fluoroquinolones and third-generation cephalosporins. The whole-genome sequencing of multidrug-resistant Salmonella isolates (19.0%) detected resistance genes for 9 groups of antibiotics: aminoglycosides (100%), beta-lactams (57.4%), fluoroquinolones (16.2%), tetracyclines (51.5%), macrolides (1.5%), phenicols (30.4%), trimethoprim (13.0%), sulfonamides (47.8%) and colistin (1.4%). Thus, epidemiological surveillance of the Salmonella spread through the food chain is of critical importance for the monitoring of antimicrobial resistance among foodborne Salmonella

Селекция Cucumis sativus L. на устойчивость к фузариозу с применением фильтрата культуральной жидкости гриба Fusarium oxysporum Schlectend

Relevance Traditional breeding methods are based on crossing and selection of genotypes among hybrid offspring. In recent decades, along with traditional methods, more and more attention is paid to alternative methods of selection, based on biotechnological manipulations with plants. One of the most important methods of biotechnology is the method of cell selection, which is based on the replacement of the whole plant, as a unit of selection, on its cell. Applying biotechnology techniques from a single plant can produce millions of cells, which increases the chances of finding, eliminating the need for areas for the cultivation of tested plants. As well as accelerating the selection process due to the possibility to carry out the study in the offseason. Methods The studies used the linear material of C. sativus hybrids of All-Russian Scientific Research Institute of Vegetable Growing – Branch of the FSBSI Federal Scientific Vegetable Center and Agroholding "Poisk". Plants were cultivated in laboratory room conditions. As explants used hypocotyl 0.5-1 cm segments isolated from young plants. Results To obtain Cucumis sativus plants with increased resistance to Fusarium by cell selection method, it is recommended to alternate culturing of callus on a non – selective medium containing sucrose in a concentration of 30 g/l, agar – 7 g/l, 0.1 mg/l, NUC – 0.5 mg/l and the filter of the cultural fluid of the fungus in a concentration of 10% within 3 passages.Актуальность В последние десятилетия наряду с традиционными методами все больше внимания уделяется альтернативным методам селекции, в основе которых лежат биотехнологические манипуляции с растениями. Применяя методы биотехнологии из одного растения можно получить миллионы клеток, что увеличивает шансы поиска, исключая потребность в площадях для выращивания испытуемых растений, а также ускоряется селекционный процесс за счет возможности проводить исследования в межсезонье. Методика В исследованиях использовали линейный материал гибридов C. sativus селекции ВНИИО – филиала ФГБНУ ФНЦО и совместной селекции ВНИИО – филиала ФГБНУ ФНЦО с Агрохолдингом «Поиск». Материалом для исследования служили растения C. sativus, которые культивировали в вегетационных сосудах в условиях лабораторного помещения. В качестве эксплантов для получения пролиферирующей каллусной ткани, способной к морфогенезу, использовали гипокотильные сегменты размером 0,5-1 см, изолированные от молодых растений. Результаты Для получения растений Cucumis sativus L. с повышенной устойчивостью к фузариозу методом клеточной селекции рекомендуется чередование культивирования каллуса на неселективной и селективной средах, содержащих сахарозу в концентрации 30 г/л, агар – 7 г/л, БАП – 0,1мг/л, НУК – 0,5 мг/л и фильтрат культуральной жидкости гриба F. oxysporum в концентрации 10% в течение 3-х пассажей

Genetic and expression studies of SMN2 gene in Russian patients with spinal muscular atrophy type II and III

<p>Abstract</p> <p>Background</p> <p>Spinal muscular atrophy (SMA type I, II and III) is an autosomal recessive neuromuscular disorder caused by mutations in the survival motor neuron gene (<it>SMN1</it>). <it>SMN2 </it>is a centromeric copy gene that has been characterized as a major modifier of SMA severity. SMA type I patients have one or two <it>SMN2 </it>copies while most SMA type II patients carry three <it>SMN2 </it>copies and SMA III patients have three or four <it>SMN2 </it>copies. The <it>SMN1 </it>gene produces a full-length transcript (FL-SMN) while <it>SMN2 </it>is only able to produce a small portion of the FL-SMN because of a splice mutation which results in the production of abnormal SMNΔ7 mRNA.</p> <p>Methods</p> <p>In this study we performed quantification of the <it>SMN2 </it>gene copy number in Russian patients affected by SMA type II and III (42 and 19 patients, respectively) by means of real-time PCR. Moreover, we present two families consisting of asymptomatic carriers of a homozygous absence of the <it>SMN1 </it>gene. We also developed a novel RT-qPCR-based assay to determine the FL-SMN/SMNΔ7 mRNA ratio as SMA biomarker.</p> <p>Results</p> <p>Comparison of the <it>SMN2 </it>copy number and clinical features revealed a significant correlation between mild clinical phenotype (SMA type III) and presence of four copies of the <it>SMN2 </it>gene. In both asymptomatic cases we found an increased number of <it>SMN2 </it>copies in the healthy carriers and a biallelic <it>SMN1 </it>absence. Furthermore, the novel assay revealed a difference between SMA patients and healthy controls.</p> <p>Conclusions</p> <p>We suggest that the <it>SMN2 </it>gene copy quantification in SMA patients could be used as a prognostic tool for discrimination between the SMA type II and SMA type III diagnoses, whereas the FL-SMN/SMNΔ7 mRNA ratio could be a useful biomarker for detecting changes during SMA pharmacotherapy.</p

Profiles of Parental Burnout Around the Globe: Similarities and Differences Across 36 Countries

Parental burnout (PB) is a pervasive phenomenon. Parenting is embedded in cultural values, and previous research has shown the role of individualism in PB. In this paper, we reanalyze previously collected data to identify profiles based on the four dimensions of PB, and explore whether these profiles vary across countries’ levels of collectivistic-individualistic (COL-IND) values. Our sample comprised 16,885 individuals from 36 countries (73% women; 27% men), and we used a latent profile approach to uncover PB profiles. The findings showed five profiles: Fulfilled, Not in PB, Low risk of PB, High risk of PB and Burned out. The profiles pointed to climbing levels of PB in the total sample and in each of the three country groups (High COL/Low IND, Medium COL-IND, Low COL/High IND). Exploratory analyses revealed that distinct dimensions of PB had the most prominent roles in the climbing pattern, depending on the countries’ levels of COL/IND. In particular, we found contrast to be a hallmark dimension and an indicator of severe burnout for individualistic countries. Contrary to our predictions, emotional distance and saturation did not allow a clear differentiation across collectivistic countries. Our findings support several research avenues regarding PB measurement and intervention