Current issue

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.

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.

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.

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.

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 1^st 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.

Breast cancer (BC) is one of the most common causes of death among women in the world. This study investigated the role of global DNA methylation (5mC) and hypoxia-inducible factor 1 alpha (HIF-1α) levels in BC disease progression. Blood samples were collected from 40 patients with benign breast tumors, 40 patients with malignant breast tumors, and 40 healthy subjects. Patients with malignant breast tumors were divided into two groups: women in stage II (low-level), and patients in stages III and IV (high-level). Genomic DNA was isolated from whole blood samples from the subjects and used for global DNA methylation. Furthermore, the levels of HIF-1α expression were measured. The results showed that the levels of 5mC in patients with BC and benign breast tumors were considerably lower (0.538 ± 0.03 and 0.432 ± 0.04%, respectively), compared to the control (0.619 ± 0.05%). Furthermore, there was a significant (p ≤ 0.05) decrease in levels of 5mC in BC patients at stages III and IV compared to control. However, there were no significant differences between low-level and high-level stages. The HIF-1α levels of patients in both the benign breast tumors and BC were insignificant (923.35 ± 72.42 and 1386.03 ± 102.01pg/ml, respectively), compared to the control (825.5 ± 62.36 pg/ml). Although, BC patients at low levels showed no significant difference in HIF-1α levels compared with patients at high levels. The findings indicated that variations in 5mC levels across different stages and types of breast tumors may serve as a prognostic indicator for the development of BC and also implicated HIF-1α in the development of BC.