Background: the nucleus pulposus NP pulls on the ruptured annulus fibrosus AF, bulging the intervertebral disc IVD and releasing chemicals that may irritate nerves and cause inflammation and pain. This produces histological changes to the IVD, including less gelatinous NP, cracks and fissures, decreased matrix water content, and proteoglycan composition changes. Aim of study: this study aims to investigate the histopathological changes in the Herniated Disc HD tissue, as well as cartilage histopathology grade and stage assessment. Materials and methods: fourty tissue samples of lumbar HD obtained after the operations were kept in 10% formalin. Then all HD sections were processed, and embedded, after that, 5 μm thick glass mounted sections were stained with Hematoxylin and Eosin and Alcian blue (pH 0.02) and examined microscopically for histopathological changes and grading scoring was also conducted based on cell density, structural alterations of collagen fibers, and proteoglycan degeneration. Results: hemorrhage with fibrin deposition, mucous degeneration around chondrocyte clones and degeneration, increased chondrocyte density, expanded lacunae with degenerated chondrocytes, fiber disorientation, cleft formation, mucoid matrix changes, and inflammatory cell infiltration with fibrocyte prefiltration were the most histopathological changes in HD samples. Along with necrotic chondrocyte increase and tissue fiber alterations. Histological cell density grade indicated different-sized clones. All HD tissues revealed collagen fiber structural alterations, with gradients (52.5%) being most common. All HD samples showed mucous (proteoglycans) degradation, especially in gradients abundantly present and intermediate between 1 and 3 (45% and 42.5%). Conclusions: histopathological changes in intervertebral disc associated with HD infection.
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 02_Al-Muathen_14-23.pdf | 3.84 MB |
A varicocele is the most common rectification cause of male infertility. It is an abnormal dilation of the veins of the pampiniform plexus in the spermatic cord. It is estimated to affect 10-15% of men and adolescents. The aim of the current study was to investigate the immunological parameters, gene expression profiles, and their correlations in varicocele and non-varicocele oligospermic patients compared to healthy controls, providing insights into the molecular mechanisms underlying male infertility. A total of 120 males were involved in this study over the period from October 2023 to January 2024. Blood and semen samples were collected from patients who were diagnosed with varicocele confirmed oligospermia (40 samples) and non-varicocele oligospermic patients (40 samples), in addition to 40 apparently healthy males as control. The level of IL-18 and IL-37 was estimated in the serum of tested groups using Enzyme Linked Immunosorbent Assay (ELISA) technique. The serum levels of the pro-inflammatory cytokine IL-18 were significantly elevated in all patient groups compared with control. Interestingly, the anti-inflammatory cytokine IL-37 was also significantly increased. The gene expression of heat shock protein A2 (HSPA2) was detected using reverse transcriptase polymerase chain reaction (RT-PCR). The fold of gene expression results by RT-PCR technique revealed a significant downregulation of HSPA2 (0.3453 ± 0.311-fold) gene expression in varicocele patients and a 0.154 ± 0.13-fold decrease in non-varicocele oligospermia compared with control. However, no significant correlation was found between the expression of HSPA2 and the levels of IL-18 and IL-37.
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| 01_Abed_5-13.pdf | 862.76 KB |
MicroRNAs play a crucial role in the regulation of biological processes variety associated with neoplasm development and progression, such as cell proliferation and differentiation, apoptosis, angiogenesis, inflammation, migration, invasion and metastasis, epithelial-mesenchymal transition and others. The purpose of this work was to investigate the DNA methylation level of miR-152 in 25 paired tissue samples from patients with an established diagnosis of ovarian cancer and various histological and clinical characteristics by the MS-HRM method. Our results indicate a higher frequency of the miR-152 methylation in ovarian tumor tissues (51.5% ± 5.4) compared to normal tissues (43.9% ± 7.2), however, the differences did not reach the statistical level significance, p = 0.5. There was no relationship between the metastatic process in the tumor depending on the level of methylation (46.5% ± 11.9 in patients with metastases vs 45.2% ± 7.8 in cases without metastases). One patient with the highest methylation level of all samples researched – 89.91%, despite a good response to primary therapy, had a relapse of the disease after 7 years. In addition, there is a tendency for a lower level of miR-152 methylation in patients with a complete response to therapy, in contrast to women with a partial response or the tumor process stabilization. Thus, our research provides evidence in favor of the suppressor function of the miR-152 in tumor, and its possible role in sensitivity to polychemotherapy, however, the results did not reach a statistical level of significance and additional studies on larger material are required.
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| 014_Прокофьева_159-167.pdf | 892.81 KB |
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 |
