A chronic periprosthetic infection after knee replacement typically requires two-stage treatment. However, the inter-stage rehabilitation protocol for patients with an articulating spacer has not been adequately developed. This study aimed to determine whether adding interactive biofeedback walking training on the Walker View treadmill enhances the effectiveness of a standard recovery program following the first stage of two-stage revision treatment. The prospective randomized controlled trial included 87 patients who had undergone removal of their endoprostheses and placement of articulating spacers. The treatment group (n = 43) had the standard 21-day rehabilitation program combined with Walker View sessions, while the control group (n = 44) only followed the program. We assessed knee joint movement volume, quadriceps EMG amplitude, stride length, walking speed, postural stability, and SF-36, WOMAC, and KSS scores. By the end of the rehabilitation course, the results registered in the treatment group were better than in the control group: flexion — 78 ± 6° versus 71 ± 7° (p = 0.01); EMG amplitude — 179 ± 16 versus 165 ± 16 μV (p = 0.01); step length — 54.2 ± 5.0 versus 49.5 ± 5.0 cm (p = 0.01); walking speed — 0.70 ± 0.05 versus 0.65 ± 0.05 m/s (p = 0.02); overall stability — 80 ± 8% versus 72 ± 7% (p = 0.01); physical component SF-36 — 51 ± 8 versus 47 ± 7 points (p = 0.01). The differences in WOMAC and KSS scores were insignificant (p = 0.06 and p = 0.07). The inclusion of Walker View sessions in the inter-stage rehabilitation program yields more pronounced improvements in mobility, neuromuscular function, walking, and balance restoration.
VIEWS 509
Systemic osteoporosis is one of the most significant systemic factors capable of worsening bone tissue quality and affect osseointegration of dental implants. The increase in the number of patients of older age groups in need of implant treatment determines high clinical significance of preoperative assessment of the implant bed condition in this category of patients. The study aimed to assess clinical and morphological features of bone tissue in the dental implantation zone in patients with osteoporosis and determine the correlation of those with the primary stability of implants and early marginal bone remodeling. A total of 84 patients aged 55–75 were included in a prospective pilot study, who were planned for implantation in the areas of maxillar and mandibular premolars and molars: 42 with the confirmed systemic osteoporosis and 42 having no signs of osteoporosis. All the patients underwent CBCT with the bone quality and type determination in accordance with the Lekholm and Zarb classification; histological and histo-morphometric analysis of bone biopsy specimens was performed in a subgroup of 32 patients. Primary stability was assessed by the resonance frequency analysis; clinical monitoring was conducted after 2, 6, and 12 months. The D3–D4 type bones were more often found in patients with osteoporosis, along with lower bone density and implant stability quotient (ISQ) values, reduced bone volume fraction (BV/TV), trabecular thinning, and increased RANKL/OPG ratio. There was a positive correlation between the BV/TV, radiological bone density, and primary stability of implants. Systemic osteoporosis degrades the implant bed quality, therefore, the implantation protocol personalization and comprehensive preoperative assessment are required.
VIEWS 501
Drug-induced xerostomia is common among elderly patients taking multiple medications. The condition significantly affects dental health and quality of life. This study aimed to evaluate the clinical and laboratory characteristics of oral fluid (OF) in xerostomia patients taking xerogenic medications, and to assess associations between total xerogenic load, salivary flow rates, and OF composition. The study included 60 people aged 45–75 years. The treatment group consisted of 40 patients with at least 3 months of dry mouth history and routine intake of two or more medications with known xerogenic potential. The control group included 20 healthy individuals exhibiting no signs of xerostomia and not taking medications routinely. We used the Xerostomia Inventory questionnaire to collect data from the participants; they also underwent clinical dental examination and sialometry for unstimulated and stimulated oral fluid (OF). The fluid samples were examined in the laboratory to determine pH, buffer capacity, total protein content, alpha-amylase activity, glucose and lactate levels. Compared to the control group, patients in the treatment group showed marked hyposalivation, decreased OF pH and buffer capacity, increased total protein content and alpha-amylase activity, and tended more often to have multiple caries lesions, candidal stomatitis, and atrophic changes in the oral mucosa. Thus, drug-induced xerostomia is accompanied by pronounced quantitative and qualitative changes in OF as well dental health and quality of life deterioration. A comprehensive clinical and laboratory assessment of OF provides an objective measure of xerostomia severity and enables compilation of tailored prevention and treatment programs.
VIEWS 587
Studying molecular mechanisms of carcinogenesis, including abnormalities of the homologous recombination (HR) system, is an important objective when studying malignization. Dysfunction of HR genes, such as BRCA1/2, contributes to genomic instability and the development of more aggressive tumor clones. The use of chemical carcinogens, such as dimethylbenz(a)anthracene (DMBA), allows one to simulate tumorigenesis processes and assess changes in expression of  repair genes. It is important to study such changes to understand the mechanisms underlying adaptation of tumor cells to genotoxic stress and develop personalized approaches to cancer treatment. The study aimed to assess the expression of major HR genes in chemotherapy-induced carcinogenesis in mice. The study involved female outbred ICR laboratory mice (CD-1; n = 20). Two groups of animals were formed: the control group and the treatment group that was administered DMBA. Histological analysis of autopsy specimens was conducted to identify tumors. Gene expression levels were assessed using RT-PCR, and testing for chromosomal aberrations was performed using digital PCR. Tumors were found in four animals. Zero expression of the genes Brca1, Brca2, Cdk12, Chek2, Palb2, Bard1, Brip1 and Rad paralogues was observed in three tumor samples. One sample showed high expression of the genes Cdk12 (14.3), Chek1 (27.6), Rad51d (38.5). Predominance of deletions in the test genes was reported in the majority of cases. Thus, tumorigenesis is associated with the decrease in expression of major repair genes, chromosomal aberration formation, which can contribute to the emergence of more aggressive clones and increase sensitivity to chemotherapy drugs.
VIEWS 523
The menopausal transition is accompanied by the decrease in estrogen levels and changes in the estrogen to androgen ratio, resulting in dysregulation of multipotent mesenchymal stem cell (MSCs) differentiation in the subcutaneous adipose tissue, reduction of their adipogenic potential, adipocyte hypertrophy, and metabolic disorder progression. Menopausal hormone therapy (MHT) is used to manage menopausal symptoms. However, the effects of exogenous hormones on MSCs are still poorly understood. The study aimed to assess adipogenic differentiation of the subcutaneous adipose tissue MSCs and their sensitivity to testosterone, 17β-estradiol, and parathyroid hormone (PTH) in postmenopause. A total of six patients with benign gynecological disorders were included in the study, among them two were of reproductive age, one was perimenopausal, and three were postmenopausal. The MSCs adipogenic differentiation was performed throughout 14 days with the addition of testosterone, 17β-estradiol, or PTH, 10 nM each, then the proportion of cells containing lipid droplets was assessed. The adipogenesis level in control samples was 26–30% in patients of childbearing age and 12–42% in postmenopausal ones, with the pronounced interindividual variability. Hormonal stimulation considerably suppressed MSCs adipogenesis in postmenopause: testosterone reduced adipogenesis to 46–56% of control levels, estradiol to 51–84%, PTH to 53–66%, while patients of childbearing age showed a less pronounced effect (65–85%). The obtained data demonstrate a shift in hormonal sensitivity of MSCs from subcutaneous adipose tissue in postmenopause and suggest that MHT may exert an additional inhibiting effect on adipogenesis through suppression of MSCs differentiation.
VIEWS 702
For several decades, Wilson’s disease (WD) has remained the focus of attention for a wide range of specialists, including hepatologists, general practitioners, neurologists, geneticists, etc. However, despite significant advances in understanding its molecular basis, establishing clear correlations between the genotype and clinical phenotype of the disease remains a key unresolved issue. The study aimed to identify patterns between genetic variants in the ATP7B gene and the WD clinical manifestations using next-generation sequencing. The data from 81 WD patients, who were followed up between 2015 and 2019, were used in the study. Molecular genetic testing of biomaterial (blood) samples was performed by NGS. The analysis of the molecular genetic testing results using targeted NGS revealed 31 pathogenic variants. The following variants were the most frequent: c.3207C>A (p.His1069Gln) — 51.85% alleles, c.3190G>A (p.Glu1064Lys) — 8.64% alleles, and c.3402delC (p.Ala1135fs) — 6.17% alleles. A moderate correlation between genotype and phenotype was established: pathogenic variants (nonsense, frameshift, splicing) in the homo- or compound heterozygous state are associated with severe liver damage, severe degree of cirrhosis, and lower cholinesterase levels. The data obtained emphasize the importance of molecular genetic diagnosis for clarifying the diagnosis of WD and predicting the disease severity.
VIEWS 727
The PI3K/AKT/mTOR signaling pathway is a key regulator of cell growth, and its dysregulation is involved in oncogenesis. Existing methods for assessing mTOR activity have design flaws. The aim of this work was to develop and validate a novel multiplex RT-qPCR assay for relative quantification of mTOR gene expression normalized to RPLP0 and TBP. Primers and probes were designed in silico. Validation was performed using the human SCP-1 cell line. Specificity was assessed in 10 separate and 10 multiplex runs. Analytical sensitivity and efficiency were determined from 27 technical replicates using a protocol without an elongation step. Specificity of amplification was assessed by agarose gel electrophoresis, and quantitative analysis was performed in real-time PCR using FAM (mTOR), HEX (RPLP0), and ROX (TBP) fluorescence channels. The assay showed 100% specificity. Stable detection was achieved at 125,000 cells/mL. Amplification efficiencies were 73–81%. The variation of mTOR expression normalized to RPLP0 ranged from –21.5% to 26.4%, and normalized to TBP from –14.3% to 19.2%. Normalization to the geometric mean of both reference genes provided the best reproducibility, with an interquartile range from –9% to 23.4%. The developed assay demonstrates high specificity, sensitivity, and reproducibility, making it a reliable tool for subsequent clinical research.
VIEWS 847