Danilov, Lavrentiy Glebovich

Suchko P.A., Danilova A.A., Pinkovskaya E.V., Glotov O.S., Danilov L.G.

Uniparental disomy is a type of chromosomal variation leading to in which both homologous chromosomes or chromosomal regions are inherited from one parent. Such variations have been detected for all chromosomes. The frequency of uniparental disomies is estimated at 1 case per 2000 births. The causes of uniparental disomies include errors during meiosis, postzygotic errors, Robertsonian and reciprocal translocations. Clinical manifestations are associated with loss of heterozygosity for pathogenic genetic variants and defects in genomic imprinting.Currently, the diagnosis of uniparental disomy is performed using methods such as microsatellite analysis, chromosomal microarray analysis, methyl-sensitive PCR, methyl-specific amplification of a probe dependent on multiplex ligation and next-generation sequencing (NGS). The methods used nowadays separately do not allow for a definitive diagnosis of uniparental disomy. A combination of NGS method that simultaneously assesses the DNA methylation status and regions of loss of heterozygosity, in particular those based on fragmentation of genomic DNA by methyl-dependent restriction enzymes, with classical approaches such as methyl-sensitive PCR and microsatellite testing will enable rapid and accurate diagnosis of uniparental disomies.

Danilova A.A., Gusev K.A., Arkhipova N.O., Danilov L.G., Vishnyakov E.V., Maimistov D.N., Flisyuk E.V.

Introduction. Flavonoids, despite their pronounced therapeutic benefits, are limitedly used in medicine as active pharmaceutical ingredients (API) owing to a complex of unsatisfactory physicochemical properties. In order to study the mentioned problem, we decided to use quercetin as model compound with extremely low water solubility and dissolution in gastrointestinal (GI) tract media. Therefore, the main idea appears to study the possibility and prospects of hot melt extrusion (HME) application for enhancement the solubility properties of quercetin in the composition of solid dispersion system (SDS) based on hydrophilic polymeric carrier (polyvinylpyrrolidone vinyl acetate, PVP/VA). Consequently, there is a necessity to select effective drug-to-polymer ratio in solid dispersion for providing better solubility and dissolution properties.Aim. Assessment of PVP/VA effect on quercetin solubility properties in binary solid dispersion prepared by HME.Materials and methods. Quercetin substance with purity 98 % was purchased from Molekula Limited, United Kingdom. PVP/VA (copolymer of polyvinylpyrrolidone with vinyl acetate in the ratio of 60 : 40, VIVAPHARM® PVP/VA 64) as carrier was procured from JRS PHARMA (JRS PHARMA GmbH & Co. KG, Germany). Quercetin SDS were prepared using micro-conical twin screw compounder HAAKE™ MiniCTW (Thermo Fisher Scientific, Germany). The obtained samples were analyzed by phase-contrast microscopy, FTIR spectroscopy and differential scanning calorimetry (DSC). Quercetin quantitative content in SDS were determined by UV-spectrophotometry.Results and discussion. The improvement of water solubility and dissolution rates of quercetin SDS in comparison with pure substance was observed for all solid dispersion compositions irrespective to drug-to-polymer ratio. Notably, with reduction of quercetin content in SDS compositions the PVP/VA contribution was increased. We found partial or even complete amorphization of API in formulations with 1 % and 5 % quercetin content, resulting in the improvement of water solubility properties, stability of solutions and increased dissolution rates in GI tract media. Water solubility of 1 % SDS relative to pure substance was enhanced by 353-fold. At the same time, the complete release of quercetin from 1 % SDS was achieved in 40 minutes in the hydrochloric acid and citrate buffer, and also quercetin dissolution of 90 % in 60 minute was observed in phosphate buffer.Conclusion. 1 % quercetin SDS based on PVP/VA appears to be the most promising for solid dosage forms development.

Barinova V.V., Ivanov D.O., Bushtyreva I.O., Botasheva T.L., Dudurich V.V., Danilov L.G.

BACKGROUND: Numerous studies of the intestinal microbiome have shown its role in pathogenesis of diseases in children. However, the role of such a factor as the child’s gender is little taken into account in these studies. AIM: The purpose of this study to identify the features of the intestinal microbiome composition of children aged 1 month, born vaginally and breastfed, depending on the child’s gender. MATERIALS AND METHODS: the study included 103 children aged 4–6 weeks of life (group 1 — 46 girls, group 2 — 57 boys), examined at Professor Bushtyreva’s Clinic LLC from 2019 to 2020, each of whom underwent stool sampling for further sequencing of 16S rRNA. RESULTS: Results of 16s rRNA sequencing revealed that the proportion of Erysipelatoclostridium bacteria, that predispose to the development of allergic reactions and inflammatory bowel diseases, was significantly higher in boys than in girls (12.52 and 0.2% respectively, p = 0.020). The proportion of Lachnoclostridium bacteria, high amounts of which are associated with resistance to diseases of the nervous system, also differed significantly in the groups of boys and girls (0.01 and 5.78% respectively, p = 0.046). Analysis of correlation matrices revealed that the correlation adaptometry coefficient in the group of boys was almost 4 times higher than in girls (9.5 and 2.4 respectively). Analysis of morbidity in children under one year old revealed that allergies were almost 3 times more common in boys than in girls (33.3 and 13%). Episodes of acute intestinal infections in the first year of life were registered in 6 boys and only in 1 girl (10.5 and 2.2%). CONCLUSIONS: In boys at 1st month of life, born vaginally and breastfed, compared to girls, the proportion of bacteria of the genus Erysipelatoclostridium in the intestinal microbiome is higher, that is a risk factor for the development of allergic reactions and inflammatory bowel diseases. At the same time, the proportion of bacteria of the genus Lachnoclostridium, on the contrary, was 5 times higher in girls than in boys. The revealed differences can be used to select preventive probiotic therapy taking into account the child’s gender.

Novikova V.P., Polunina A.V., Bannova S.L., Balashov A.L., Dudurich V.V., Danilov L.G., Blinov A.E., Varlamova O.N., Seits A.V., Kukes E.A.

The new coronavirus infection (COVID-19) that emerged in 2019 is a infection caused by the SARS-CoV-2 virus. Although SARS-CoV-2 predominantly affects the respiratory system, numerous studies suggest significant gastrointestinal involvement in COVID-19, particularly in children. Aim. To evaluate the effect of the synbiotic Maxilac® in children with COVID-19 after antibacterial therapy (ABT). Materials and methods. A clinical post-approval open-label observational prospective single-center study with minimal intervention was conducted. The microbiome of children with COVID-19 was studied. Twelve children with COVID-19 complicated by a bacterial infection (pneumonia, sinusitis, otitis) were included, receiving ABT and then taking the synbiotic Maxilac® for 4 weeks. Patient data were collected: symptoms, medical history, clinical status, study of intestinal microbiota (IM) by 16S sequencing, stool test for zonulin, and review of drug therapy. The material was sampled in patients with COVID-19 at 3 time points: at the time of acute COVID-19 disease, at the time of recovery from COVID-19, and 4 weeks after starting the synbiotic therapy. Results. The administration of the synbiotic Maxilac® to patients who received ABT for bacterial complications of COVID-19 relieves clinical gastroenterological symptoms, prevents the development of dysbiosis, and normalizes intestinal permeability. The results indicated significant changes in IM after ABT in complicated COVID-19 and the normalizing effect of synbiotic Maxilac® on IM during administration for 30 days. The synbiotic Maxilac® in the group of children with COVID-19 who received ABT had no side effects and was safe. Conclusion. Children with COVID-19, complicated by a bacterial infection, treated with ABT are recommended to receive the synbiotic Maxilac® for at least 30 days after recovery.

Maksiutenko E.M., Barbitoff Y.A., Danilov L.G., Matveenko A.G., Zemlyanko O.M., Efremova E.P., Moskalenko S.E., Zhouravleva G.A.

In yeast Saccharomyces cerevisiae, there are two translation termination factors, eRF1 (Sup45) and eRF3 (Sup35), which are essential for viability. Previous studies have revealed that presence of nonsense mutations in these genes leads to amplification of mutant alleles (sup35-n and sup45-n), which appears to be necessary for the viability of such cells. However, the mechanism of this phenomenon remained unclear. In this study, we used RNA-Seq and proteome analysis to reveal the complete set of gene expression changes that occur during cellular adaptation to the introduction of the sup35-218 nonsense allele. Our analysis demonstrated significant changes in the transcription of genes that control the cell cycle: decreases in the expression of genes of the anaphase promoting complex APC/C (APC9, CDC23) and their activator CDC20, and increases in the expression of the transcription factor FKH1, the main cell cycle kinase CDC28, and cyclins that induce DNA biosynthesis. We propose a model according to which yeast adaptation to nonsense mutations in the translation termination factor genes occurs as a result of a delayed cell cycle progression beyond the G2-M stage, which leads to an extension of the S and G2 phases and an increase in the number of copies of the mutant sup35-n allele.

Kuznetsova X., Dodueva I., Afonin A., Gribchenko E., Danilov L., Gancheva M., Tvorogova V., Galynin N., Lutova L.

Spontaneous tumour formation in higher plants can occur in the absence of pathogen invasion, depending on the plant genotype. Spontaneous tumour formation on the taproots is consistently observed in certain inbred lines of radish (Raphanus sativus var. radicula Pers.). In this paper, using Oxford Nanopore and Illumina technologies, we have sequenced the genomes of two closely related radish inbred lines that differ in their ability to spontaneously form tumours. We identified a large number of single nucleotide variants (amino acid substitutions, insertions or deletions, SNVs) that are likely to be associated with the spontaneous tumour formation. Among the genes involved in the trait, we have identified those that regulate the cell cycle, meristem activity, gene expression, and metabolism and signalling of phytohormones. After identifying the SNVs, we performed Sanger sequencing of amplicons corresponding to SNV-containing regions to validate our results. We then checked for the presence of SNVs in other tumour lines of the radish genetic collection and found the ERF118 gene, which had the SNVs in the majority of tumour lines. Furthermore, we performed the identification of the CLAVATA3/ESR (CLE) and WUSCHEL (WOX) genes and, as a result, identified two unique radish CLE genes which probably encode proteins with multiple CLE domains. The results obtained provide a basis for investigating the mechanisms of plant tumour formation and also for future genetic and genomic studies of radish.

Kuznetsova X.A., Dodueva I.E., Afonin A., Gribchenko E., Danilov L.G., Gancheva M.S., Tvorogova V.E., Galynin N.A., Lutova L.A.

Spontaneous tumours in higher plants can develop depending on plant genotype without any pathogen invasion. Spontaneous tumour formation on the taproots is consistently observed in certain inbred lines of radish (Raphanus sativus var. radicula Pers.). In this work, we have sequenced the genomes of two closely related radish inbred lines that differ in their ability to spontaneously form tumours. We have identified numerous single nucleotide variants (amino acid substitutions, insertions or deletions) that are likely to be associated with the spontaneous tumour formation. Furthermore, we performed the identification of CLE and WOX genes the genomic sequences of the radish inbred lines and, as a result, identified two unique radish CLE genes which probably encode proteins with multiple CLE domains. The results obtained provide a basis for investigating the mechanisms of plant tumour formation and also for future genetic and genomic studies of radish.

Tsvetikova S.A., Kruglov E.E., Danilov L.G., Zilov D.S., Myakisheva Y.V., Makarova M.A., Kaftyreva L.A., Koshel E.I.

Objective: There is a large amount of data on fecal microbiome composition in inflammatory bowel disease and ulcerative colitis, but the mucosal microbiome is less described. Analysis of pathogenic determinants of microorganisms colonizing inflamed regions is promising for understanding their role in disease pathogenesis. The aim of this study is to compare the microbiome composition of ulcerated and non-inflamed areas and to evaluate the pathogenic potential of Escherichia coli isolates from damaged epithelial samples of ulcerative colitis patients. Methods: In this study, we investigated the mucosal microbiota of ulcers and non-inflamed areas of 25 patients with UC from the Southern European part of Russia. We described a composition of mucosal microbiome using 16S rRNA sequence analysis and characterized E. coli isolates from ulcers for antibiotic susceptibility, presence of virulence factor genes, and phylogenetic group distribution. Results and Discussion: Most of the E. coli isolates from ulcers were multidrug resistant, including 7 isolates with resistance to more than 7 antibiotics, and carried virulence factor genes (hly, 23%; pap, 38%; cnf, 42%). The combination of these markers and the determined phylogroup profile indicates a pathogenic potential of E. coli localized in patients’ ulcers and a possibility of development of intestinal and extraintestinal infections. Metagenomic analysis revealed a high similarity of microbial populations in non-inflamed areas and ulcers of different localization. Conclusions: The composition of the intestinal mucosal microbiota in ulcerative colitis is characterized by a low degree of variation between ulcerated and non-inflamed areas. At the same time, a pathogenic potential of E. coli isolates is observed. Our results support the importance of personalized antibiotic therapy prescription and patient monitoring to prevent opportunistic infections in the treatment of UC.

Volynets G.V., Nikitin A.V., Skvortsova T.A., Potapov A.S., Dudurich V.V., Danilov L.G., Kokiashvili V.S.

Relevance. The influence of the gut microbiota on the development of various diseases is of great interest to researchers. The conducted studies showed that in patients with chronic liver diseases, the dominant taxa of the gut microbiota were Bifidobacterium longum, Bifidobacterium adolescentis, Blautia massiliensis, and in healthy children - Neisseria flavescens. There is no comparative analysis of data on the taxonomic diversity of the intestinal microbiota in autoimmune and non-autoimmune liver diseases in children. Purpose of the study. To investigate differences in the taxonomic diversity of fecal microbiota in patients with autoimmune and non-autoimmune liver diseases, as well as to evaluate potential biomarkers of 16S rRNA gene amplicons in these diseases by comparing the taxonomic composition. Scope and methods of research. A metagenomic analysis of the intestinal microbiota of 24 children with chronic liver diseases (mean age 10.3±4.7 years) was carried out with the identification of the V3-V4 region of the 16S rRNA gene. The group included 18 children with autoimmune liver diseases and 6 children with non-autoimmune liver diseases. Research results. The conducted study revealed 684 types of microorganisms in the studied samples of patients’ faeces. The analysis of the conducted studies showed that no dominant taxa were found in the faecal samples of children with autoimmune liver diseases, while Veillonella dispar, Veillonella parvula, Cloacibacillus porcorum, Prevotella histicola and Bacteroides eggerthii were the dominant taxa in patients with non-autoimmune liver diseases. Conclusion. Studies have shown differences in the composition of the gut microbiota in children with autoimmune and non-autoimmune liver diseases.

Nikitin A.V., Volynets G.V., Potapov A.S., Dudurich V.V., Danilov L.G.

   The influence of the gut microbiota on the development of various diseases is of great interest to researchers. The conducted studies showed that in patients with chronic liver diseases, the dominant taxa of the gut microbiota were Bifidobacterium longum, Bifidobacterium adolescentis, Blautia massiliensis, and in healthy children — Neisseria flavescens. There is no comparative analysis of data on the taxonomic diversity of the intestinal microbiota in autoimmune and non-autoimmune liver diseases in children.   Purpose. To investigate differences in the taxonomic diversity of fecal microbiota in patients with autoimmune and non-autoimmune liver diseases, as well as to evaluate potential biomarkers of 16S rRNA gene amplicons in these diseases by comparing the taxonomic composition.   Material and methods. A metagenomic analysis of the intestinal microbiota of 24 children with chronic liver diseases (mean age 10,3 ± 4,7 years) was carried out with the isolation of the 16S rRNA gene region. The group included 18 children with autoimmune liver diseases and 6 children with non-autoimmune liver diseases.   Results. The conducted study revealed 684 types of microorganisms in the studied samples of patients’ feces. The analysis of the conducted studies showed that no dominant taxa were found in the fecal samples of children with autoimmune liver diseases, while Veillonella dispar, Veillonella parvula, Cloacibacillus porcorum, Prevotella histicola and Bacteroides eggerthii were the dominant taxa in patients with non-autoimmune liver diseases.   Conclusion. Studies have shown differences in the composition of the gut microbiota in children with autoimmune and non-autoimmune liver diseases.

Chirinskaite A.V., Rotov A.Y., Ermolaeva M.E., Tkachenko L.A., Vaganova A.N., Danilov L.G., Fedoseeva K.N., Kostin N.A., Sopova J.V., Firsov M.L., Leonova E.I.

Many retinal degenerative diseases result in vision impairment or permanent blindness due to photoreceptor loss or dysfunction. It has been observed that Pde6brd1 mice (rd1), which carry a spontaneous nonsense mutation in the pde6b gene, have a strong phenotypic similarity to patients suffering from autosomal recessive retinitis pigmentosa. In this study, we present a novel mouse model of retinitis pigmentosa generated through pde6b gene knockout using CRISPR/Cas9 technology. We compare this Pde6b-KO mouse model to the rd1 mouse model to gain insights into the progression of retinal degeneration. The functional assessment of the mouse retina and the tracking of degeneration dynamics were performed using electrophysiological methods, while retinal morphology was analyzed through histology techniques. Interestingly, the Pde6b-KO mouse model demonstrated a higher amplitude of photoresponse than the rd1 model of the same age. At postnatal day 12, the thickness of the photoreceptor layer in both mouse models did not significantly differ from that of control animals; however, by day 15, a substantial reduction was observed. Notably, the decline in the number of photoreceptors in the rd1 model occurred at a significantly faster rate. These findings suggest that the C3H background may play a significant role in the early stages of retinal degeneration.

Chen K., Zhurbenko P.M., Danilov L.G., Matveeva T.V., Otten L.

Horizontal gene transfer (HGT) plays an important role in plant evolution and plant development. Agrobacterium-mediated gene transfer leads to the formation of crown galls or hairy roots, due to expression of transferred T-DNA genes. Spontaneous regeneration of transformed cells can produce natural transformants carrying cellular T-DNA (cT-DNA) sequences of bacterial origin. HGT from Agrobacterium to dicots is remarkably widespread. The production of naturally genome modified plants could play a role in plant evolution and environment. Among these natural GMOs (nGMOs) there are the tea plants. Camellia sinensis var. sinensis cv. Shuchazao contains a single 5.5 kb cT-DNA fragment organized as imperfect inverted repeat with three inactive genes. 142 Camellia accessions, belonging to 10 of 11 species of the section Thea, were studied for the presence of cT-DNA alleles. All of them contain the cT-DNA insert, indicating that they are resulted from the single transformed event. Allele phasing showed that 82 accessions were heterozygous for T-DNA alleles, 60 others were homozygous. A phylogenetic analysis of all found alleles showed existence of two separate groups of them, further divided into subgroups. The alleles of the different Camellia species were distributed mosaically over groups, and different species showed very similar T-DNA alleles. This indicates that the taxonomy of Thea requires revision. The nucleotide divergence of the imperfect cT-DNA repeats indicates that the age of cT-DNA insertion is about 15 mya, which is earlier then emergence of section Thea [1]. We present a working model for the origin and evolution of nGMO plants derived from allogamous transformants.

Goleva O., Danilov L., Kusakin A., Eismont Y., Babachenko I., Tian N., Chukhlovin A., Krylov A., Glotov O.

The paper reports a familial case of HHV-6A chromosomal integration being an important and relevant issue of genetics and medicine. The study was aimed to test the hypothesis of HHV-6A chromosomal integration and vertical transmission in patient with persistent virus detection during recurrent respiratory diseases and the asymptomatic period when there were no health complaints. Sequencing of the patient’s father genome DNA was performed, and a phylogenetic tree was constructed by aligning 270 HHV-6A/B genome assemblies from the GenBank database. As a result, a familial case of ciHHV-6A transmission was identified. It was found that the detected ciHHV-6A observed on the phylogenetic tree was closely related to other two chromosomally integrated HHV-6A sequences reported by Moscow researchers. The study confirmed HHV-6A chromosomal integration. Further precise chromosome mapping of ciHHV-6A would be useful in terms of excluding probable somatic disorders associated with the chromosome structure alteration following HHV-6, particularly HHV-6A, integration, as well as for identification of insertion sites specific for various geographic locations.

Novikova V.P., Polunina A.V., Bannova S.L., Balashov A.L., Dudurich V.V., Danilov L.G., Blinov A.E., Varlamova O.N.

Background: studying the consequences of novel coronavirus infection is currently relevant. Many observations demonstrate that SARS- CoV-2 affects the gastrointestinal tract, gut microbiota composition, and intestinal permeability. Aim: to assess whether Maxilac® Baby synbiotic effects qualitative and quantitative composition of gut microbiota and intestinal permeability in children aged 3–14 with mild-to-moderate COVID-19. Patients and Methods: an open, observational, prospective, single-center study with minimal intervention was conducted. Group 1 included 16 children who received Maxilac® Baby 4 weeks after recovery. Group 2 children did not receive synbiotics after recovery. History and complaints were assessed, gut microbiome composition was determined, and fecal zonulin was measured at disease onset, time of recovery (day 14), and 4 weeks after recovery. Results: at disease onset, abdominal pain and diarrhea were reported in 16 children (50%), nausea in 8 children (25%), and vomiting in 1 child (3.1%). At the time of recovery, only abdominal pain was reported in both groups (4 patients each). Thirty days after recovery, re-appearance of intestinal symptoms was reported in group 2. No significant differences in the microbiome diversity profile at birth level were revealed between children with the COVID-19 at onset and healthy age-matched children. Taxa (species) whose predominance was typical for each group were identified. Conclusions: post-COVID-19 syndrome in children aged 3–14 diagnosed with mild-to-moderate disease who received Maxilac® Baby for 1 month after recovery is characterized by the lack of nausea, vomiting, and diarrhea, less abdominal pain, no changes in fecal zonulin levels, and biodiversity of gut microbiome, including those assessed by Shannon diversity index. KEYWORDS: SARS-CoV-2, COVID-19, intestinal permeability, gut microbiome, 16s rRNA sequencing, zonulin, novel coronavirus infection, post-COVID-19 syndrome. FOR CITATION: Novikova V.P., Polunina A.V., Bannova S.L., Balashov A.L., Dudurich V.V., Danilov L.G., Blinov A.E., Varlamova O.N. Gastrointestinal tract in children with novel coronavirus infection and post-COVID-19 syndrome. The role of synbiotics for improving clinical symptoms, gut microbiota, and intestinal permeability. Russian Journal of Woman and Child Health. 2023;6(3):283–289 (in Russ.). DOI: 10.32364/2618-8430-2023-6-3-10.

Volynets G.V., Potapov A.S., Nikitin A.V., Danilov L.G., Skvortsova T.A., Dudurich V.V.

   The value of the liver–gut axis is increasingly recognized as a major modulator of autoimmunity. There is no comparative analysis of data on the taxonomic diversity of the intestinal microbiota in chronic liver diseases in children.   Purpose. To investigate the taxonomic diversity of the intestinal microbiota in children with chronic liver diseases compared with healthy patients, to identify differences in bacterial diversity in autoimmune and non-autoimmune liver diseases, as well as the impact of immunosuppressive therapy on the intestinal microbiota.   Material and methods. A metagenomic analysis of the gut microbiota of 24 children with chronic liver diseases (mean age 10,3 ± 4,7 years) was carried out with the identification of the V3–V4 region of the 16S rRNA gene. The group included 18 children with autoimmune liver diseases and 6 children with non-autoimmune liver diseases. The control group consisted of fecal samples of 34 apparently healthy children.   Results. When comparing fecal samples of children with autoimmune liver diseases with samples of healthy children, the taxa of Bacteroides dorei, Collinsella aerofaciens, Ruminococcus caffidurs prevailed, and for children of the control group — Neisseria flavescens. When comparing samples of patients with non-autoimmune liver diseases and the control group, it was found that the taxa Bacteroides fragilis, Klebsiella pneumoniae, Bifidobacterium longum prevailed in healthy children. When comparing fecal samples from children with autoimmune and non-autoimmune liver diseases, it was found that Veillonella dispar, Cloacibacillus porcorum, Veillonella parvula, Prevotella histicola and Bacteroides eggerthii taxa dominate in patients with non-autoimmune diseases. No dominant taxa of the gut microbiota were found in children with autoimmune liver diseases. It has been established that the taxa Veillonella dispar, Faecalibacterium prausnitzii, Roseburia inulinivorans, Bacteroides xylanisolvens and Alistipes obesi prevail in patients receiving immunosuppressive therapy, and the taxa Phascolarctobacterium succinatutens, Bacteroides ovatus, Solobacterium mooreis and Holdemanella massilien prevail in patients not receiving immunosuppressive therapy.   Conclusion. A recent study of the gut microbiota in children with chronic liver disease shows differences in the imbalance of the gut microbiota compared to the results obtained in adults. The gut microbiota model is capable of distinguishing autoimmune liver diseases from non-autoimmune diseases. Immunosuppressive therapy is accompanied by the dominance of taxa that reduce the production of short-chain fatty acids.

Rush E.T., del Angel G., Dong J., Bates T., Steiner R.D., Cox A.

Sharma N., Heer K., Raychaudhuri S.

Katkowska M., Kosecka-Strojek M., Wolska-Gębarzewska M., Kwapisz E., Wierzbowska M., Międzobrodzki J., Garbacz K.

Objective: In the present study, we used phenotypic and molecular methods to determine susceptibility to oxacillin in coagulase-negative staphylococci (CoNS) and estimate the prevalence of strains with low-level resistance to oxacillin, mecA-positive oxacillin-susceptible methicillin-resistant (OS-MRCoNS), and borderline oxacillin-resistant (BORCoNS). Methods: One hundred one CoNS strains were screened for oxacillin and cefoxitin susceptibility using phenotypic (disk diffusion, agar dilution, latex agglutination, and chromagar) and molecular (detection of mecA, mecB, and mecC) methods. Staphylococcal cassette chromosome mec (SCCmec) typing was performed. Results: Sixteen (15.8%) CoNS strains were mecA-positive, and 85 (84.2%) were mec-negative. Seven (6.9%) were classified as OS-MRCoNS, accounting for 43.8% of all mecA-positive strains. Twelve (11.9%) mec-negative strains were classified as borderline oxacillin resistant (BORCoNS). Compared with MRCoNS and BORCoNS, OS-MRCoNS strains demonstrated lower resistance to non-beta-lactams. SCCmec type I cassette was predominant. The disc-diffusion method with oxacillin accurately predicted OS-MRCoNS strains but did not provide reliable results for BORCoNS strains. Meanwhile, the latex agglutination test and CHROMagar culture accurately identified BORCoNS but not OS-MRCoNS. Conclusions: Finally, our findings imply that the recognition of methicillin resistance in CoNS requires a meticulous approach and that further research is needed to develop unified laboratory diagnostic algorithms to prevent the misreporting of borderline CoNS.

Savenkova N.D., Leviashvili Z.G.

The article provides current information on the clinical forms of hypophosphatasia. The OMIM catalog lists 5 forms of hypophosphatasia: perinatal (lethal), infantile, childhood, adult, and odontohypophosphatasia. The ORPHA portal identifies 6 subtypes of the disorder, including adult, childhood, infantile, perinatal (lethal), and prenatal (benign) hypophosphatasia. M.E. Nunes (2023) identifies 7 forms of hypophosphatasia. International studies have established the pathogenesis, phenotypic variability, and severity of hypophosphatasia. A global consortium provides information on 446 mutation variants of the ALPL gene and 797 genotypes in pediatric and adult patients. The review presents updated diagnostic criteria for hypophosphatasia in children and adults with low alkaline phosphatase activity in the blood. Ten years of experience in 40 countries have proven the safety and efficacy of enzyme replacement therapy with Asfotase Alfa in children with perinatal, infantile, childhood hypophosphatasia, and odontohypophosphatasia. In the Russian Federation, enzyme replacement therapy with Asfotase Alfa for children with hypophosphatasia has been funded by the Circle of Kindness Foundation, established by the Ministry of Health, since 2021.

Veremeichik G.N., Solomatina T.O., Khopta A.A., Brodovskaya E.V., Gorpenchenko T.Y., Grigorchuk V.P., Bulgakov D.V., Bulgakov V.P.

Long-term cultured calli may experience a biosynthetic shift due to the IAA-dependent expression of the rolA gene, which also affects ROS metabolism. The “hairy root” syndrome is caused by the root-inducing Ri-plasmid of Rhizobium rhizogenes, also known as Agrobacterium rhizogenes. The Ri-plasmid contains genes known as rol genes or root oncogenic loci, which promote root development. The important implications of the rolA gene from the T-DNA include reduced plant size, resistance to infections, and the activation of specialised metabolism. Nevertheless, rolA does not belong to the plast gene group because its function is still uncertain. Recent investigations have shown two important effects of the rolA gene. First, the production of secondary metabolites has changed in long-term cultivated rolA-transgenic calli of Rubia cordifolia L. Second, the expression of both the rolA and rolB genes has a strong auxin-dependent antagonistic effect on reactive oxygen species (ROS) homeostasis. In this work, we attempted to elucidate two rolA gene phenomena: what caused the secondary metabolism of long-term cultured calli to change? How does the individual expression of the rolA gene affect ROS homeostasis? We analysed SNPs in the 5′ untranslated region and coding region of the rolA gene. These mutations do not affect the known essential amino acids of the RolA proteins. Notably, in the promoter of the rolA gene, an ACTTTA motif for auxin-mediated transcription factors was identified. Using two separate cell cultures, we demonstrated the strong auxin dependence of rolA gene expression. The expression of genes involved in ROS metabolism decreased in response to an auxin-mediated increase in rolA gene expression. Two assumptions can be made. The long-term cultivation of calli may cause changes in the hormonal state of the culture over time, which may modulate the action of the RolA protein. Moreover, auxin-dependent expression of the rolA gene led to a decrease in ROS metabolism. It can be assumed that the antagonistic interaction between rolA and rolB prevents strong rolB-induced auxin sensitivity and oxidative bursts to balance the cell state.

Bhutta N.K., Xu X., Jian C., Wang Y., Liu Y., Sun J., Han B., Wu S., Javeed A.

Gut microbiota regulates the immune system, the development and progression of autoimmune diseases (AIDs) and overall health. Recent studies have played a crucial part in understanding the specific role of different gut bacterial strains and their metabolites in different AIDs. Microbial signatures in AIDs are revealed by advanced sequencing and metabolomics studies. Microbes such as Faecalibacterium prausnitzii, Akkermansia muciniphila, Anaerostipes caccae, Bacteroides sp., Roseburia sp., Blautia sp., Blautia faecis, Clostridium lavalense, Christensenellaceae sp., Coprococcus sp., Firmicutes sp., Ruminococcaceae sp., Lachnospiraceae sp., Megamonas sp., Monoglobus sp., Streptococcus pneumoniae and Bifidobacterium sp. help maintain immune homeostasis; whereas, Prevotella copri, Ruminococcus gnavus, Lactobacillus salivarius, Enterococcus gallinarum, Elizabeth menigoseptica, Collinsella sp., Escherichia sp., Fusobacterium sp., Enterobacter ludwigii, Enterobacteriaceae sp., Proteobacteria, Porphyromonas gingivalis, Porphyromonas nigrescens, Dorea sp., and Clostridium sp. cause immuno-pathogenesis. A complex web of interactions is revealed by understanding the influence of gut microbiota on immune cells and various T cell subsets such as CD4+ T cells, CD8+ T cells, natural killer T cells, γδ T cells, etc. Certain AIDs, including rheumatoid arthritis, diabetes mellitus, atopic asthma, inflammatory bowel disease and non-alcoholic fatty liver disease exhibit a state of dysbiosis, characterized by alterations in microbial diversity and relative abundance of specific taxa. This review summarizes recent developments in understanding the role of certain microbiota composition in specific AIDs, and the factors affecting specific regulatory T cells through certain microbial metabolites and also focuses the potential application and therapeutic significance of gut microbiota-based interventions as novel adjunctive therapies for AIDs. Further research to determine the precise association of each gut bacterial strain in specific diseases is required.

Brzezinski M., Argudo P.G., Scheidt T., Yu M., Hosseini E., Kaltbeitzel A., Lemke E.A., Michels J.J., Parekh S.H.

Filatova E.N., Utkin O.V., Khrulev A.E., Zaitseva N.N.

Human herpesvirus 6A (HHV6A) and human herpesvirus 6B (HHV6) are ubiquitous viruses that infect more than 95% of the population. Clinical manifestations of HHV6 infection and associated diseases are diverse, which may depend on virus molecular genetic characteristics (genovariants). Estimating the significance of the molecular genetic diversity is complicated due to the lack of proper classification. The aim of the study was to develop an intraspecies HHV6A and HHV6B classification. Using 50 and 207 HHV6A and HHV6B full-genome sequences retrieved from the NCBI Nucleotide database, various fragments of virus genome were analyzed. Multiple sequence alignment was performed using MAFFT L-INS-i algorithm; F81 nucleotide substitution model and maximum likelihood method were used to construct dendrograms. Nucleotide substitutions were determined relative to reference sequences X83413 (HHV6A) and AF157706 (HHV6B). Genovariants were defined based on the nucleotide substitutions in variable positions of the genomic fragment. The results were confirmed by constructing dendrograms. An opportunity of using fragments of HHV6A and HHV6B genomes to construct an intraspecies classification was assessed. Fragments U90 (part206) and U90B(part431) were selected as optimal. Based on the nucleotide sequences of the fragments, the intraspecies classification for HHV6A and HHV6B was constructed, including seven genovariants of each virus. The genovariants were characterized by unique nucleotide composition in the signature positions. A minimum (0.001 or less for both viruses) nucleotide diversity within the isolated genovariants was established. The classification reflects the phylogenetic relationships of circulating and inherited chromosomally integrated forms of HHV6A and HHV6B: divergence of HHV6A genovariants depending on its persistence form and integration site and coevolution of two HHV6B forms within several genovariants. Further studies on virus molecular genetic diversity in different regions of Russia and abroad may supplement the classification. The method of HHV6A and HHV6B classification is characterized by simplicity, technological accessibility and can be implemented in laboratories of different levels of technical equipment. The classification can be used to analyze an effect of virus molecular genetic diversity on the clinical characteristics of associated diseases, optimize the epidemiological surveillance system and develop new approaches for diagnostics, prevention, and treatment of HHV6 infection.

Gupta P., Pasmanik-Chor M., Zemach H., Barash I., Teper D., Sessa G.

AbstractPantoea agglomeranspv.betae(Pab) induces tumor-like galls in beet and gypsophila, a process mediated by the secretion of effector proteins viaPab’s type III secretion system (T3SS). The molecular mechanisms underlyingPab-induced gall formation remain largely unexplored. This study delves into the cellular architecture and transcriptional profile ofPab-mediated galls, comparing host responses to wild-typePaband a T3SS-inactive mutant,hrcC−. Morphological analysis using scanning electron microscopy and cross-sectional visualization of infected beet leaf tissues revealed thatPab-induced gall-like structures are linked to cell hyperplasia and tissue ruptures, contingent on T3SS activity. Comparative transcriptome analysis of wild-typePabandhrcC−Pab-infected beet leaves at 12 and 48 hours unveiled significant transcriptional reprogramming, with nearly 2,000 differentially expressed genes at 48 hours post inoculation. Enrichment analyses identified the upregulation of pathways related to signal transduction, defense, carbohydrate metabolism, and cell wall modulation in wild-typePab-infected leaves compared to controls. Particularly notable was the significant upregulation of numerous genes associated with cell wall loosening by wild-typePab, suggesting an initial rearrangement of cell wall architecture facilitates gall formation. Furthermore, transcriptome analysis demonstrated that wild-typePabsuppresses the expression of the betalain biosynthetic geneDOPA 4,5-DIOXYGENASE, leading to reduced betalain accumulation in infected tissues compared to the mutant strain. These findings offer fresh insights into the transcriptional and physiological manipulation of host tissue during the early stages ofPab-induced gall formation.

Дейнеко Е.В.

Развитие и совершенствование методов молекулярной и клеточной биологии существенно расширило возможности исследователей по модификации геномов растительных клеток и послужило основой для развития новых технологий получения рекомбинантных белков, используемых в фармацевтике и других отраслях народного хозяйства, а также стимулировало создание новых высокоурожайных сортов важных сельскохозяйственных культур, устойчивых к неблагоприятным абиотическим и биотическим факторам среды. Перенос генов в растительный геном из других гетерологичных систем поставил перед исследователями ряд вопросов, связанных с функционированием трансгенов в новом окружении генома-реципиента, а также с их влиянием на функционирование собственных генов растения. За последние сорок лет с момента получения первого трансгенного растения возможности этих технологий были существенно углублены и расширены за счет разработки методов геномного редактирования, основанных на системе CRISPR/Cas. Это позволило не только изменять функционирование целевых генов путем нокаутов или исправлять нежелательные мутации, но и вносить гены интереса в заданные исследователем районы-мишени растительного генома. В предлагаемом обзоре рассматриваются основные этапы исследований по модификации геномов растений за последние сорок лет, с акцентом не только на практическую значимость созданных агробиотехнологий, но и на важность для фундаментальных исследований функционирования генов и выявления структурных особенностей организации генома растений.

Huang S., Li F., Quan C., Jin D.

Roca Paixao J.F., Déléris A.

Abstract Mobile elements known as T-DNAs are transferred from pathogenic Agrobacterium to plants and reprogram the host cell to form hairy roots or tumors. Disarmed non-oncogenic T-DNAs are extensively used to deliver transgenes in plant genetic engineering. Such T-DNAs were the first known targets of RNA silencing mechanisms, which detect foreign RNA in plant cells and produce small RNAs that induce transcript degradation. These T-DNAs can also be transcriptionally silenced by the deposition of epigenetic marks such as DNA methylation and the dimethylation of lysine 9 (H3K9me2) in plants. Here, we review the targeting and the roles of RNA silencing and DNA methylation on T-DNAs in transgenic plants as well as during pathogenesis. In addition, we discuss the crosstalk between T-DNAs and genome-wide changes in DNA methylation during pathogenesis. We also cover recently discovered regulatory phenomena, such as T-DNA suppression and RNA silencing-independent and epigenetic-independent mechanisms that can silence T-DNAs. Finally, we discuss the implications of findings on T-DNA silencing for the improvement of plant genetic engineering.

Chung G., Piano F., Gunsalus K.C.

ABSTRACTTelomeres are eukaryotic chromosome end structures that guard against sequence loss and aberrant chromosome fusions. Telomeric repeat motifs (TRMs), the minimal repeating unit of a telomere, vary from species to species, with some evolutionary clades experiencing a rapid sequence divergence. To explore the full scope of this evolutionary divergence, many bioinformatic tools have been developed to infer novel TRMs using repetitive sequence search on short sequencing reads. However, novel telomeric motifs remain unidentified in up to half of the sequencing libraries assayed with these tools. A possible reason may be that short reads, derived from extensively sheared DNA, preserve little to no positional context of the repetitive sequences assayed. On the other hand, if a sequencing read is sufficiently long, telomeric sequences must appear at either end rather than in the middle. The TeloSearchLR algorithm relies on this to help identify novel TRMs on long reads, in many cases where short-read search tools have failed. In addition, we demonstrate that TeloSearchLR can reveal unusually long telomeric motifs not maintained by telomerase, and it can also be used to anchor terminal scaffolds in new genome assemblies.

Glotov O.S., Zhuchenko N.A., Balashova M.S., Raspopova A.N., Tsai V.V., Chernov A.N., Chuiko I.V., Danilov L.G., Morozova L.D., Glotov A.S.

Hypophosphatasia (HPP) is a rare inherited disorder characterized by the decreased activity of tissue-nonspecific alkaline phosphatase (TNSALP), caused by mutations in the ALPL gene. The aim of this study was to conduct differential diagnostics in HPP patients using whole-exome sequencing (WES). The medical records of HPP patients and the genetic testing of the ALPL gene were reviewed. Seven patients were recruited and underwent WES using the Illumina or MGI sequencing platforms. All of the exome samples were matched onto a GRCh38.p13 reference genome assembly by using the Genome Analysis ToolKit (GATK) and the BWA MEM read aligner. We present the clinical and molecular findings of the seven patients referred for genetic analyses due to a clinical and biochemical suspicion of HPP. In two patients out of three (with identified heterozygous variants in the ALPL gene), we also identified c.682T>A in exon 3 of the WNT10A gene and c.3470del in exon 23 of the SMC1A gene variants for the first time. In four patients, variants in the ALPL gene were not detected, but WES allowed us to identify for the first time rare variants (c.5651A>C in exon 36 of the TRIO gene, c.880T>G in exon 6 of the TRPV4 gene, c.32078-1G>T in intron 159 of the TTN gene, c.47720_47721del in exon 235 of the TTN gene, and c.1946G>A in exon 15 of the SLC5A1 gene) and to conduct differential diagnostics with HPP. Using WES, for the first time, we demonstrate the possibility of early differential diagnostics in HPP patients with other rare genetic diseases.

Xie S., Xiao H., Zhang F., Lan Y., Luo M.

Wdowiak K., Tajber L., Miklaszewski A., Cielecka-Piontek J.

The co-administration of curcumin and hesperetin might be beneficial in terms of neuroprotective activity; therefore, in this study, we attempted to develop a fixed-dose formulation comprising these two compounds in an amorphous state. The aim of obtaining an amorphous state was to overcome the limitations of the low solubility of the active compounds. First, we assessed the possibility of using popular sweeteners (erythritol, xylitol, and sorbitol) as plasticizers to reduce the glass transition temperature of PVP K30 to prepare the polymer–excipient blends, which allowed the preparation of amorphous solid dispersions via hot-melt extrusion at a temperature below the original glass transition of PVP K30. Erythritol proved to be the superior plasticizer. Then, we focused on the development of fixed-dose amorphous solid dispersions of curcumin and hesperetin. Powder X-ray diffraction and thermal analysis confirmed the amorphous character of dispersions, whereas infrared spectroscopy helped to assess the presence of intermolecular interactions. The amorphous state of the produced dispersions was maintained for 6 months, as shown in a stability study. Pharmaceutical parameters such as dissolution rate, solubility, and in vitro permeability through artificial membranes were evaluated. The best improvement in these features was noted for the dispersion, which contained 15% of the total content of the active compounds with erythritol used as the plasticizer.

Danilova A.A., Gusev K.A., Maimistov D.N., Flisyuk E.V.

The pharmacological activity of flavonoids as potential candidates for the development of drugs based on biologically active substances of plant origin is confirmed by research data. The low bioavailability of flavonoids attributed to poor solubility and absorption in the gastrointestinal tract is a limiting factor in their use. Microand nano-milling technologies; complexation with surfactants, cyclodextrins, and chelating agents; and production of solid disperse systems are currently used to improve the physicochemical properties of polyphenolic compounds. Development of solid dispersion systems using hot melt extrusion is the most promising approach from the viewpoint of industrial implementation. The present review is devoted to consideration of current approaches to the use of hot extrusion of polyphenolic compounds to improve their pharmacokinetic and technological properties.

Xing X., Hu T., Wang Y., Li Y., Wang W., Hu H., Wei Q., Yan Y., Gan D., Bao C., Wang J.

Radish (Raphanus sativus L.) is a vegetable crop with economic value and ecological significance in the genus Radish, family Brassicaceae. In recent years, developed countries have attached great importance to the collection and conservation of radish germplasm resources and their research and utilization, but the lack of population genetic information and molecular markers has hindered the development of the genetic breeding of radish. In this study, we integrated the radish genomic data published in databases for the development of single-nucleotide polymorphism (SNP) markers, and obtained a dataset of 308 high-quality SNPs under strict selection criteria. With the support of Kompetitive Allele-Specific PCR (KASP) technology, we screened a set of 32 candidate core SNP marker sets to analyse the genetic diversity of the collected 356 radish varieties. The results showed that the mean values of polymorphism information content (PIC), minor allele frequency (MAF), gene diversity and heterozygosity of the 32 candidate core SNP markers were 0.32, 0.30, 0.40 and 0.25, respectively. Population structural analysis, principal component analysis and genetic evolutionary tree analysis indicated that the 356 radish materials were best classified into two taxa, and that the two taxa of the material were closely genetically exchanged. Finally, on the basis of 32 candidate core SNP markers we calculated 15 core markers using a computer algorithm to construct a fingerprint map of 356 radish varieties. Furthermore, we constructed a core germplasm population consisting of 71 radish materials using 32 candidate core markers. In this study, we developed SNP markers for radish cultivar identification and genetic diversity analysis, and constructed DNA fingerprints, providing a basis for the identification of radish germplasm resources and molecular marker-assisted breeding as well as genetic research.

Xu C., Li M., Gu T., Xie F., Zhang Y., Wang D., Peng J.

Abstract Background Chromosomal microarray analysis (CMA) is a valuable tool in prenatal diagnosis for the detection of chromosome uniparental disomy (UPD). This retrospective study examines fetuses undergoing invasive prenatal diagnosis through Affymetrix CytoScan 750 K array analysis. We evaluated both chromosome G-banding karyotyping data and CMA results from 2007 cases subjected to amniocentesis. Results The detection rate of regions of homozygosity (ROH) ≥ 10 Mb was 1.8% (33/2007), with chromosome 11 being the most frequently implicated (17.1%, 6/33). There were three cases where UPD predicted an abnormal phenotype based on imprinted gene expression. Conclusion The integration of UPD detection by CMA offers a more precise approach to prenatal genetic diagnosis. CMA proves effective in identifying ROH and preventing the birth of children affected by imprinting diseases.

Park H.Y., Lim Y., Jung M., Sathiyamoorthy S., Heo S.H., Park B., Shin Y.

Chavli E.A., Klaasen S.J., Van Opstal D., Laven J.S., Kops G.J., Baart E.B.

Zhang B., Zhang X., Luo Z., Ren J., Yu X., Zhao H., Wang Y., Zhang W., Tian W., Wei X., Ding Q., Yang H., Jin Z., Tong X., Wang J., et. al.

Gut microbiota and circulating metabolite dysbiosis predate important pathological changes in glucose metabolic disorders; however, comprehensive studies on impaired glucose tolerance (IGT), a diabetes mellitus (DM) precursor, are lacking. Here, we perform metagenomic sequencing and metabolomics on 47 pairs of individuals with IGT and newly diagnosed DM and 46 controls with normal glucose tolerance (NGT); patients with IGT are followed up after 4 years for progression to DM. Analysis of baseline data reveals significant differences in gut microbiota and serum metabolites among the IGT, DM, and NGT groups. In addition, 13 types of gut microbiota and 17 types of circulating metabolites showed significant differences at baseline before IGT progressed to DM, including higher levels of Eggerthella unclassified, Coprobacillus unclassified, Clostridium ramosum, L-valine, L-norleucine, and L-isoleucine, and lower levels of Eubacterium eligens, Bacteroides faecis, Lachnospiraceae bacterium 3_1_46FAA, Alistipes senegalensis, Megaspaera elsdenii, Clostridium perfringens, α-linolenic acid, 10E,12Z-octadecadienoic acid, and dodecanoic acid. A random forest model based on differential intestinal microbiota and circulating metabolites can predict the progression from IGT to DM (AUC = 0.87). These results suggest that microbiome and metabolome dysbiosis occur in individuals with IGT and have important predictive values and potential for intervention in preventing IGT from progressing to DM.

Manchia M., Paribello P., Pisanu C., Congiu D., Antoniades A., Vogazianos P., Tozzi F., Pinna F., Aristodimou A., Caria P., Dettori T., Frau D.V., Cocco C., Noli B., Panebianco C., et. al.

Excessive predominance of pathological species in the gut microbiota could increase the production of inflammatory mediators at the gut level and, via modification of the gut–blood barrier, at the systemic level. This pro-inflammatory state could, in turn, increase biological aging that is generally proxied by telomere shortening. In this study, we present findings from a secondary interaction analysis of gut microbiota, aging, and inflammatory marker data from a cohort of patients with different diagnoses of severe mental disorders. We analyzed 15 controls, 35 patients with schizophrenia (SCZ), and 31 patients with major depressive disorder (MDD) recruited among those attending a community mental health center (50 males and 31 females, mean and median age 46.8 and 46.3 years, respectively). We performed 16S rRNA sequencing as well as measurement of telomere length via quantitative fluorescence in situ hybridization and high-sensitivity C-reactive protein. We applied statistical modeling with logistic regression to test for interaction between gut microbiota and these markers. Our results showed statistically significant interactions between telomere length and gut microbiota pointing to the genus Lachnostridium, which remained significantly associated with a reduced likelihood of MDD even after adjustment for a series of covariates. Although exploratory, these findings show that specific gut microbiota signatures overexpressing Lachnoclostridium and interacting with biological aging could modulate the liability for MDD.

Shao D., Liu L., Tong H., Shi S.

Pyrroloquinoline quinone disodium (PQQ·Na2) has been considered a human food supplement for human health promotion with its antioxidant properties. To determine whether PQQ·Na2 had similar functions to improve the antioxidant ability of layers and eggs, 180 laying hens were fed with 0 or 0.4 mg/kg PQQ·Na2 diets. Supplementation with PQQ·Na2 increased the albumen height, Haugh unit of the eggs. PQQ·Na2 also led to a higher glutathione peroxidase (GSH-Px) concentration in plasma and a lower malondialdehyde (MDA) content in the liver and egg yolk. Similarly, liver gene and protein expression of nuclear factor erythroid 2-related 2 (Nrf2) and heme oxygenase 1 (HO-1) were up-regulated by PQQ·Na2. Moreover, PQQ·Na2 increased the abundance of Firmicutes, Microbacterium, Erysipelatoclostridium, Mailhella, Lachnospiraceae_UCG-010, and Herbaspirillum in gut. Overall, these results suggested PQQ·Na2 increased the antioxidant ability of layers and eggs which might be in connection with the activation of the Nrf2/HO-1 pathway and optimized gut microflora.

Кашеварова А.А., Толмачева Е.Н., Маркова Ж.Г., Миньженкова М.Е., Твеленева А.А., Малышева О.В., Юров И.Ю., Шилова Н.В., Лебедев И.Н.

Хромосомный микроматричный анализ представляет наиболее динамично развиваемую область современной клинической цитогенетики, обеспечивая эффективную и высокоразрешающую диагностику несбалансированных микроструктурных хромосомных перестроек. Настоящие рекомендации определяют показания к назначению хромосомного микроматричного анализа для постнатальной и пренатальной диагностики конститутивных хромосомных аномалий, а также рассматривают вопросы интерпретации клинической значимости выявляемых хромосомных вариантов, и медико-генетического консультирования семей пациентов с хромосомными болезнями. Делаются акценты на приоритетности экспертного мнения и необходимости диалога молекулярного цитогенетика и врача-генетика в оценке патогенетической значимости хромосомных вариантов и тактики дальнейшего лабораторного обследования пациента. Chromosomal microarray analysis is the state-of-the-art of clinical cytogenetics, providing efficient and high-resolution diagnostics of unbalanced chromosomal rearrangements. Current recommendations define the indications for chromosomal microarray analysis in postnatal and prenatal diagnostics of constitutive chromosomal anomalies. Approaches to interpretation of the clinical significance of detected chromosomal variants, as well as genetic counseling for families of patients with chromosomal diseases are discussed. Emphasis is given to the priority of expert opinion and collaboration between molecular cytogeneticist and clinical geneticist in assessing the pathogenetic significance of chromosomal variants and the strategy for subsequent laboratory examination of the patient.

Zhu X., Cai J., Wang Y., Liu X., Chen X., Wang H., Wu Z., Bao W., Fan H., Wu S.

The prevalence of non-alcoholic fatty liver disease (NAFLD) is increasing annually, and emerging evidence suggests that the gut microbiota plays a causative role in the development of NAFLD. However, the role of gut microbiota in the development of NAFLD remains unclear and warrants further investigation. Thus, C57BL/6J mice were fed a high-fat diet (HFD), and we found that the HFD significantly induced obesity and increased the accumulation of intrahepatic lipids, along with alterations in serum biochemical parameters. Moreover, it was observed that the HFD also impaired gut barrier integrity. It was revealed via 16S rRNA gene sequencing that the HFD increased gut microbial diversity, which enriched Colidextribacter, Lachnospiraceae-NK4A136-group, Acetatifactor, and Erysipelatoclostridium. Meanwhile, it reduced the abundance of Faecalibaculum, Muribaculaceae, and Coriobacteriaceae-UCG-002. The predicted metabolic pathways suggest that HFD enhances the chemotaxis and functional activity of gut microbiota pathways associated with flagellar assembly, while also increasing the risk of intestinal pathogen colonization and inflammation. And the phosphotransferase system, streptomycin biosynthesis, and starch/sucrose metabolism exhibited decreases. These findings reveal the composition and predictive functions of the intestinal microbiome in NAFLD, further corroborating the association between gut microbiota and NAFLD while providing novel insights into its potential application in gut microbiome research for NAFLD patients.

Garbiec E., Rosiak N., Zalewski P., Tajber L., Cielecka-Piontek J.

Genistein, an isoflavone known for its antioxidant and antidiabetic effects, suffers from the drawback of low solubility. To overcome this limitation, co-amorphous systems were synthesized by incorporating amino acids that were chosen through computational methods. The confirmation of the amorphous state of lysine and arginine-containing systems was ascertained by X-ray powder diffraction. Subsequently, the characterization of these systems was extended by employing thermo-gravimetry, differential scanning calorimetry, Fourier-transform infrared spectroscopy, and scanning electron microscopy. The investigation also included an assessment of the physical stability of the samples during storage. The apparent solubility of the systems was studied in an aqueous medium. To evaluate the in vitro permeability through the gastrointestinal tract, the parallel artificial membrane permeability assay was employed. The biological properties of the systems were assessed with regard to their antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl and cupric ion-reducing antioxidant capacity assays, as well as their ability to inhibit α-glucosidase. The systems’ glass transition temperatures were determined, and their homogeneity confirmed via differential scanning calorimetry analysis, while Fourier-transform infrared spectroscopy analysis provided data on molecular interactions. Stability was maintained for the entire 6-month storage duration. The co-amorphous system containing lysine displayed the most pronounced apparent solubility improvement, as well as a significant enhancement in antioxidant activity. Notably, both systems demonstrated superior α-glucosidase inhibition relative to acarbose, a standard drug for managing type 2 diabetes. The results indicate that co-amorphous systems with lysine and arginine have the potential to significantly enhance the solubility and biological activity of genistein.

Gusev K.A., Aliev A.R., Generalova Y.E., Aksenova N.А., Rechkalov G.V., Maimistov D.N., Alekseeva G.M., Flisyuk E.V.

Introduction. Ebastine is a second-generation antihistamine drug available in the form of orally disintegrating tablets and film-coated tablets. Ebastine substance exhibits high bioavailability, but low solubility in water and gastrointestinal tract media. The technology of solid dispersions based on polymer carriers by hot melt extrusion is proposed to solve the problem of ebastine low solubility.Aim. Composition development of extrudate and its production technology to create an amorphous solid dispersion of ebastine in oder to increase the recovery rate and bioavailability.Materials and methods. Ebastin micronized (JSC "Active Component", Russia); ebastin crystalline (Arevipharma GmbH, Germany); VIVAPHARM® PVP/VA 64 (JRS Pharma GMbH & Co. KG, Germany). Extrudates were obtained on a HAAKE™ miniCTW co-rotating twin-screw laboratory extruder (Thermo Fisher Scientific, Germany). Extrudates were studied by differential scanning calorimetry, synchronous thermal analysis, powder X-ray diffraction and FTIR-spectroscopy. The quantitative content of the active ingredient was determined by spectrophotometry. The content of related impurities in the amorphous solid dispersion of ebastine was determined by HPLC.Results and discussion. The technology of amorphous solid dispersion of ebastine by hot melt extrusion was developed. The pharmacokinetic properties of ebastine were significantly improved. The process of obtaining solid dispersion with 20 % of ebastine was optimized in order to reduce the content of impurities in the extrudate.Conclusion. The maximum concentration of ebastine for proper quality amorphous solid dispersion based on PVP/VA64 amounted to 20 %. Obtaining a solid dispersion by hot melt extrusion with ebastine content in PVP/VA64 higher than 30 % is impossible because the melt does not possess the glass transition property.

Kuznetsova K., Efremova E., Dodueva I., Lebedeva M., Lutova L.

Background: A feature of higher plants is the modular principle of body organisation. One of these conservative morphological modules that regulate plant growth, histogenesis and organogenesis is meristems—structures that contain pools of stem cells and are generally organised according to a common principle. Basic content: The development of meristems is under the regulation of molecular modules that contain conservative interacting components and modulate the expression of target genes depending on the developmental context. In this review, we focus on two molecular modules that act in different types of meristems. The WOX-CLAVATA module, which includes the peptide ligand, its receptor and the target transcription factor, is responsible for the formation and control of the activity of all meristem types studied, but it has its own peculiarities in different meristems. Another regulatory module is the so-called florigen-activated complex, which is responsible for the phase transition in the shoot vegetative meristem (e.g., from the vegetative shoot apical meristem to the inflorescence meristem). Conclusions: The review considers the composition and functions of these two functional modules in different developmental programmes, as well as their appearance, evolution and use in plant breeding.

Novikova V.P., Polunina A.V., Bannova S.L., Balashov A.L., Dudurich V.V., Danilov L.G., Blinov A.E., Varlamova O.N.

Background: studying the consequences of novel coronavirus infection is currently relevant. Many observations demonstrate that SARS- CoV-2 affects the gastrointestinal tract, gut microbiota composition, and intestinal permeability. Aim: to assess whether Maxilac® Baby synbiotic effects qualitative and quantitative composition of gut microbiota and intestinal permeability in children aged 3–14 with mild-to-moderate COVID-19. Patients and Methods: an open, observational, prospective, single-center study with minimal intervention was conducted. Group 1 included 16 children who received Maxilac® Baby 4 weeks after recovery. Group 2 children did not receive synbiotics after recovery. History and complaints were assessed, gut microbiome composition was determined, and fecal zonulin was measured at disease onset, time of recovery (day 14), and 4 weeks after recovery. Results: at disease onset, abdominal pain and diarrhea were reported in 16 children (50%), nausea in 8 children (25%), and vomiting in 1 child (3.1%). At the time of recovery, only abdominal pain was reported in both groups (4 patients each). Thirty days after recovery, re-appearance of intestinal symptoms was reported in group 2. No significant differences in the microbiome diversity profile at birth level were revealed between children with the COVID-19 at onset and healthy age-matched children. Taxa (species) whose predominance was typical for each group were identified. Conclusions: post-COVID-19 syndrome in children aged 3–14 diagnosed with mild-to-moderate disease who received Maxilac® Baby for 1 month after recovery is characterized by the lack of nausea, vomiting, and diarrhea, less abdominal pain, no changes in fecal zonulin levels, and biodiversity of gut microbiome, including those assessed by Shannon diversity index. KEYWORDS: SARS-CoV-2, COVID-19, intestinal permeability, gut microbiome, 16s rRNA sequencing, zonulin, novel coronavirus infection, post-COVID-19 syndrome. FOR CITATION: Novikova V.P., Polunina A.V., Bannova S.L., Balashov A.L., Dudurich V.V., Danilov L.G., Blinov A.E., Varlamova O.N. Gastrointestinal tract in children with novel coronavirus infection and post-COVID-19 syndrome. The role of synbiotics for improving clinical symptoms, gut microbiota, and intestinal permeability. Russian Journal of Woman and Child Health. 2023;6(3):283–289 (in Russ.). DOI: 10.32364/2618-8430-2023-6-3-10.