Haemophilus influenzae is one of the most common causative agents of community-acquired pneumonia (CAP). Gene recombinations and high polymorphism make it difficult to diagnose the pathogen even by methods of molecular genetics. The development of DNA microarrays for H. influenzae detection in clinical samples of patients with CAP is promising. The aim of this work was to evaluate the possibility of detection of H. influenzae in clinical samples of patients with CAP using DNA microarray. Oligonucleotide probes were selected using disprose program and sequences from NCBI Nucleotide. Probes capable of cross-linking with H. haemolyticus and H. parainfluenzae DNA were removed in order to exclude nonspecific interaction. Probes were tested as part of DNA microarray design using samples of nucleic acid from tracheal aspirates of children with CAP. H. influenzae detection frequency in clinical samples was determined using sputum samples, tracheal aspirates and pharyngeal swabs of children and adults with CAP. The prevalence of the hybridization signal of the specific probes over 3 standard deviations of the hybridization signal of the negative control probes was interpreted as positive. The results were validated by PCR. 22 probes for H. influenzae detection with DNA microarray were selected. The hybridization signal of probes exceeded the threshold while testing samples containing H. influenzae DNA and did not exceed the threshold value while testing negative samples. H. influenzae detection frequency among patients with CAP was assessed. The results can be used for development of diagnostic tools for establishing the etiological factor of CAP.
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 013_Филатова_148-158.pdf | 808.63 KB |
The aim of this study was to evaluate the effects of two variants of the Daedaleopsis confragosa fungus extract, F-1368 strain, differing in polysaccharide concentration, on five human cancer cell lines (U-87 MG, C-33 A, SK-Mel-28, MDA-MB-231, SW620) in vitro. Standard cytotoxicity assessment methods, including MTT-assay and clonogenic assay, were employed. Additionally, an experiment was conducted on laboratory SCID mice with heterotopic xenografts of human glioblastoma U-87 MG, where the test preparations were administered subcutaneously into the tumor area. Tumor sizes were measured using a caliper, and upon euthanasia, xenografts were histologically examined with hematoxylin-eosin staining under a light microscope. Results from MTT and clonogenic assays demonstrated that F-1368 extracts reduced the viability, mitochondrial function, and proliferative activity of tumor cells in vitro. However, a threefold increase in polysaccharide concentration in one of the extracts did not significantly enhance its cytotoxicity against tumor cells in vitro. Furthermore, one extract was tested on U-87 MG cell xenografts, revealing a reduction in tumor growth in SCID mice. The maximum tumor growth inhibition index for U-87 MG cells reached 50.7% at 21 days post-commencement of extract injections. These findings suggest that the D. confragosa F-1368 strain holds promise for investigating both in vitro and in vivo models of antitumor activity and identifying potential bioactive molecules or compounds.
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| 012_Соловьева_139-147.pdf | 641.57 KB |
The aim was to study macrolide resistance of erm B -positive H.pylori in patients with newly diagnosed and recurrent helicobacteriosis in Nizhny Novgorod. Materials and methods: PCR detection of H.pylori DNA in gastric juice and biotopes of the mucous membrane of the gastric antrum was performed in 3450 patients with gastroenterological diseases. To identify the ermB gene associated with macrolide resistance, domestic commercial PCR test systems were used. Results: from 2005 to 2023 there was a progressive reduction in visits for gastric and duodenal ulcers. The dynamics of primary and secondary genetic H.pylori macrolide resistance increased continuously from 2011 to 2014 and stabilized thereafter. From 2014 to 2023, the proportion of patients with ermB gene positive H.pylori infection increased approximately 2-fold (to 17.0% in 2018). In patients with a history of eradication therapy, the minimum detection rate of the ermB gene was established in 2011. Since 2012, there has been an increasing detection rate of secondary H.pylori macrolide resistance to 35.4% in 2022. Conclusions: during the observation period, a progressive reduction in the proportion of patients with gastric and duodenal ulcers among patients with gastroenterological diseases was revealed. The study of primary and secondary H.pylori macrolide resistance showed an increase in the detection rate of resistant isolates in Nizhny Novgorod. The study of genetic H.pylori macrolide resistance is necessary when re-identifying in patients with a history of eradication therapy and in patients who took clarithromycin for other reasons for the optimal selection of drugs included in the re-treatment regimen of infection.
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| 011_Перфилова_129-138.pdf | 771.68 KB |
Gastric cancer (GC) is one of the most common cancer types in the world with a high mortality rate. It is assumed that polymorphisms of the NFKB1 gene that disrupt its expression predispose to the development of epithelial cancer, including GC. The aim of this study is to explore the association of polymorphism rs28362491 of NFKB1 gene with the risk of GC development for individuals from the Volga-Ural region of Russia. The samples for the study were the DNA of 374 patients with GC and 365 healthy donors of various ethnicities (Russians, Tatars, Bashkirs). It was shown that in all studied groups the most common heterozygous genotype ID of the polymorphic locus rs28362491 of the NFKB1 gene, alleles I and D occur with similar frequencies. Also, it has been established that for Tatars, allele D of the rs28362491 polymorphic locus of the NFKB1 gene is a marker of an increased risk of developing GC, and allele I and genotype II are markers of a reduced risk of developing GC. Meta-analysis showed statistically significant differences in the distribution of allele frequencies of the polymorphic locus rs28362491 of the NFKB1 gene between patients with cancer and controls when combining samples of Tatars and Bashkirs. We hypothesize that polymorphism rs28362491 of the NFKB1 gene may contribute to the genetic structure of susceptibility to GC.
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| 010_Нургалиева_120-128.pdf | 624.21 KB |
Epigenetic regulation of spatiotemporal gene expression in ontogenesis is determined by programmed species-specific activations of retroelements in successive cell divisions. Evolutionary selection of this genome control mechanism is aimed at achieving a mature state, after which unprogrammed activation of retroelements occurs, which expression products stimulate interferon response, aseptic inflammation and aging-associated diseases development, such as atherosclerosis. Interferon in atherosclerosis stimulates pro-inflammatory macrophage phenotype, which contributes to pathological immune response, foam cell formation and atherosclerosis progression. Activation of retroelements occurs under the influence of viral infections, which role in atherosclerosis development has been proven, which confirms my hypothesis. Dysfunctional foam macrophages produce HERV-K102, which stimulates innate immunity, HERV-K HML2 expression correlates with macrophage immune activation and interferon response. Data were obtained on association with atherosclerosis of microRNAs derived from retroelements, which are involved in the disease pathogenesis due to their influence on cholesterol metabolism (miR-498, -520d), immune processes (miR-1257, -28, -2909), activation of DNMT1 (miR-1264) and EZH2 (miR-630), gene expression in endothelial cells (10 specific miRNAs), vascular smooth muscle cells (14 specific miRNAs) and macrophages (miR-320b, -326, -378, -384), contributing to pathological phenotype of these cells. In atherosclerosis microRNAs derived from retroelements interact with circular RNAs (miR-495, -576, -579, -630, -633, -637, -942) and long non-coding RNAs (miR-326, -4731, -495, - 616, -641, -664a) the key sources of which are retroelements. Role of ANRIL, NEAT1, PAPIA, MAARS, VINAS, H19, AK136714, MIAT, and interaction of Alu elements with ANRIL and NEAT1, identified in atherosclerosis development. The data obtained can become the basis for targeted effect on retroelements activation in atherosclerosis using microRNAs.
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| 09_Мустафин_108-119.pdf | 449.97 KB |
The focus of this review is on the study of aberrant metabolism in brain cells in Parkinson's disease. Parkinson's disease is the second most prevalent neurodegenerative disorder, characterized by the aggregation of the pathological protein α-synuclein, loss of dopaminergic neurons in the compact part of the substantia nigra, leading to a combination of motor and non-motor symptoms. While the hallmark motor symptoms of PD are well-documented, emerging research sheds light on intricate metabolic changes occurring at the cellular level, providing new insights into the pathophysiology of the disease. Studying the role of endogenous small molecules in protein-metabolite intermolecular interactions, conformational rearrangements of protein molecules, especially membrane receptors and transporters in regulating blood-brain barrier permeability, modulation of signaling transduction processes in neuroinflammation and neurodegeneration, remains pertinent.
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| 08_Колотьева_90-107.pdf | 1.89 MB |
Injury to the proximal part of the equine suspensory ligament (SL), called proximal suspensory desmitis (PSD), commonly causes lameness in horses. PSD is extremely difficult to manage and treat, with present methods often unable to achieve full recovery, especially in chronic cases. The present study was the first to use gene therapy to restore moderate and severe injuries of the proximal suspensory ligament in horses. Plasmid DNA encoding species specific bone morphogenetic protein 2 (bmp2) and vascular endothelial growth factor (vegf164) was injected into the site of proximal suspensory ligament injury, followed by box rest and a controlled exercise program. Clinical observations and ultrasound imaging were used to evaluate effectiveness over a period of 12 months. No negative side effects were observed. Clinical improvements were observed, especially in the forelimb affected horses, by day 30. In horses with chronic hindlimb PSD, few clinical improvements were reported. Echogenicity and the fiber alignment scoring improved but no concomitant changes to cross section area, dorsopalmar thickness or lateromedial width of the proximal suspensory ligament were observed. The transfer of bmp2 and vegf164 genes into the equine PSL exhibited beneficial effects in horses with acute or subacute forms of lesions, primarily in the forelimb.
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| 07_Закирова_74-89.pdf | 1.15 MB |
Heavy metals, due to their ability to bioaccumulate and are highly toxic even in low concentrations, are the most dangerous environmental pollutants, especially in industrialized countries. The purpose of the study was to evaluate the degree of cadmium accumulation, as well as the expression of the Mt1a and Mt2a genes, in the kidneys of rats using two experimental models of subchronic intoxication with cadmium chloride. A total of 80 adult white outbred rats of both sexes were equally distributed into four groups: a control group (negative control), group 1 (0.001 mg/kg/day CdCl2), group 2 (0.01 mg/kg/day CdCl2), 3rd group (0.1 mg/kg/day CdCl2). After three months of exposure, 10 animals (5 males and 5 females) were randomly selected from each group and euthanized, followed by kidney samples for cadmium analysis and gene expression assessment. The remaining animals (n = 40) were left for an additional 30 days without treatment, before being sacrificed to collect tissue. The results showed that 1 month after cadmium withdrawal, the processes of redistribution of the metal in the body are still ongoing, which is expressed in a greater accumulation of cadmium in the kidneys. We also recorded an increase in the Mt1a gene expression and a decrease in the Mt2a gene expression in the kidneys of animals that went through the remission stage compared to animals without it. These data suggest that even after Cd withdrawal, there may be long-term negative effects on the kidneys.
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| 06_Валова_66-73.pdf | 719.38 KB |
Obesity is recognized as a multifactorial health condition characterized by excess body fat accumulation. Orlistat is a well-known effective anti-obesity therapeutic drug, however, like many other medications on the market, it has certain unpleasant side effects. Medical herbs have recently acquired popularity in the treatment of obesity. The current project's intention was to evaluate the effect of treatment with Artemisia annua extract (AAE) to ameliorate hepato-renal dysfunction in obese rats. 40 male Sprague Dawley (SD) rats were divided into 4 groups (n = 10). The 1st group (Gp1) served as a negative control, and Gp2 was used as a positive control and given a high-fat diet (HFD) for a period of 12 weeks. For a period of 8 weeks, Gp3 and Gp4 received HFD and daily treatments of orlistat (30 mg/kg) or AAE (150 mg/kg), respectively. Hematological, biochemical, and histopathological parameters were determined. The results demonstrated that obese rats, Gp2, had hepato-renal impairment. Moreover, hepato-renal dysfunctions were exacerbated when orlistat was administered to obese rats of Gp3. In contrast, AAE-treated obese rats, Gp4, have shown alleviated hematological changes and resulted in considerable improvements in hepato-renal function. Taken together, AAE administration demonstrated potential ameliorative effects against hepato-renal dysfunctions in obese rats when compared to treatment with orlistat.
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| 05_Hasan_47-65.pdf | 3.26 MB |
Herniated (prolapsed) discs HD is a multifactor disease that afflicts around 9% of the global population and occurs when the nucleus pulposus NP bulges and pushes on the ruptured annulus fibrosus AF, releasing substances that may irritate the surrounding nerves and cause inflammation that leads to extensive histopathological and immunological changes. The aim of this study is to demonstrate the etiological role of IL-10, IL-17 levels, CD4+, and CD68 in the pathogenesis of herniated disc in Iraqi patients. The immunohistochemical analysis was performed on 30 excised HD specimens taken during endoscopic discectomy, and stored in the fixative solution (formalin 10%). Cytokines (IL-10 and IL-17) were measured by using their Enzyme-Linked Immunosorbent Assay ELISA kits, in sera of 40 patients who were diagnosed with HDs as a case group, in addition to 20 healthy as a control group. The results revealed that the CD4+ expression was negative in 70% of the HD samples, while only 8 HD samples were negative for CD68 expression. On the other hand, results of ELISA analysis recorded high levels of IL-10 with low levels of IL-17 in HD patients sera compared with control sera. According to the above it can concluded that immunohistochemical analysis of HD tissues showed infiltration of CD4+ and CD68. Also compared to controls high levels of IL-10 in HD samples combined with low levels of IL-17.
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| 04_Dhilal_40-46.pdf | 748.85 KB |
