Pregnant women are susceptible to SARS-CoV-2 infection due to physiological features of the immune system. The aim of our study was to evaluate the levels of interleukin-1β (IL-1β) in peripheral blood of pregnant women during SARS-CoV-2 infection with mild to moderate forms of the disease. We examined 60 pregnant women with COVID-19, including 14 women in the 1st trimester, 17 in the 2nd trimester, and 29 in the 3rd trimester. The comparison group consisted of 20 pregnant women without clinical and laboratory signs of acute respiratory infections. IL-1β concentrations were determined in serum using ELISA technique. Statistica 10.0 (StatSoft, USA) was used for statistical data processing. The significant error level was considered to be p < 0.05. The study results showed higher levels of IL-1β in blood of SARS-CoV-2-infected pregnant women compared to pregnant women without clinical and laboratory evidence of acute respiratory infections (p = 0.005). COVID-19 is characterized by a decreased IL-1β level in blood during the 3rd trimester of gestation as compared to the 1st trimester (p = 0.01). Pregnant women with COVID-19, compared with pregnant women without detectable signs of acute respiratory infections, had higher levels of red blood cells (p = 0.043), total protein (p = 0.006), direct bilirubin (p = 0.02), high-sensitivity C-reactive protein (p < 0.001), ferritin (p = 0.016), prothrombin time (p = 0.018) and blood clotting INR values (p = 0.013). COVID-19 in pregnant women is associated with lower levels of lymphocytes (p = 0.004), serum iron (p < 0.001) and transferrin (p = 0.003), transferrin saturation coefficient (p = 0.026) and total iron-binding capacity of blood serum (p = 0.021). IL-1β in SARS-CoV-2-infected pregnant women had an inverse correlation with biochemical and coagulation blood parameters: transferrin (p = 0.006), total iron-binding capacity of serum (p = 0.03), latent iron-binding capacity of serum (p = 0.009), fibrinogen (p = 0.002). IL-1β in the blood of SARS-CoV-2-infected pregnant women is associated with the level of direct bilirubin (p = 0.045) in moderate COVID-19, C-reactive protein (p = 0.047) and alanine aminotransferase (p = 0.024) in mild forms of the disease. Moderate COVID-19 in pregnant women is associated with a decreased leukocyte counts (p = 0.038), and an increase in prothrombin time (p = 0.046). Thus, a relationship was established between the concentration of IL-1β in blood, blood coagulation and biochemical parameters in women with COVID-19 at different stages of pregnancy, as well as dependence on the clinical severity of COVID-19.
This study aimed to evaluate the levels of tumor necrosis factor-α (TNFα), receptor activator of nuclear factor-κB ligand (RANKL), and calprotectin in gingival crevicular fluid (GCF), as well as serum C-reactive protein (CRP), in periodontitis patients with and without T1DM compared with periodontally healthy controls. A total of 90 participants were enrolled and divided into three groups (n = 30 each): healthy controls, periodontitis patients without diabetes, and periodontitis patients with T1DM. Clinical periodontal parameters, including probing depth (PD), clinical attachment loss (CAL), plaque index (PI), and bleeding on probing (BOP), were recorded. GCF samples were analyzed for TNFα, RANKL, and calprotectin using ELISA technique, while serum CRP and HbA1c levels were measured using automated analyzers. Statistical comparisons were performed using ANOVA and post-hoc tests.Clinical periodontal parameters were significantly worse in both periodontitis groups compared with controls (PD: 3.00±0.00 vs 4.53±0.51, and 4.63±0.49 mm; CAL: 2.00±0.00 vs 3.53±0.51 and 3.63±0.49 mm; p < 0.0001). TNFα levels were significantly elevated in periodontitis patients with T1DM (15.11±2.98 pg/mL) and without diabetes (13.90±2.91 pg/mL) compared with controls (8.72±2.35 pg/mL; p = 0.000). RANKL levels were highest in the diabetic periodontitis group (314.98±74.65 pg/mL) compared with controls (270.46±35.89 pg/mL; p = 0.021). Calprotectin levels showed a significant increase in periodontitis patients with T1DM (0.285±0.149 ng/mL) compared with controls (0.223±0.018 ng/mL; p = 0.017). A strong positive correlation was observed between TNFα and RANKL in both periodontitis groups (r = 0.812, p < 0.001). Periodontitis is associated with significant elevations in local inflammatory and bone-resorptive biomarkers, with T1DM further amplifying neutrophil-mediated inflammation and osteoclastogenic activity. GCF biomarkers, particularly TNFα, RANKL, and calprotectin, may serve as valuable adjunctive tools for periodontal disease assessment in diabetic patients.
Despite its economic importance and health benefits, strawberries (Fragaria × ananassa) may cause IgE-mediated allergic reactions, primarily manifesting as oral allergy syndrome. The primary sensitizing factor is the Fra a 1allergen. This protein is a member of the PR-10 (pathogenesis-related proteins) family, being a structural homologue of the major birch pollen allergen Bet v 1. This study assessed the allergenic potential of the Fra a 1 strawberry allergen and analyzed the effect of heat treatment on its structural and functional properties. We have obtained a recombinant Fra a 1 allergen. Fluorescence spectroscopy has demonstrated the ability of this protein to bind a wide range of fatty acids, thus confirming functional activity of the recombinant analogue. Circular dichroism demonstrated that heating to 98 °C caused irreversible changes in the protein's secondary structure. ELISA testing demonstrated that pre-boiling (99 °C, 15 minutes) leads to a significant partial reduction (up to 30%) of the Fra a 1 ability to bind cross-reactive polyclonal anti-Bet v 1 IgG, whereas binding to specific IgE from sera of patients with pollen and food allergy was decreased in only half the cases. A study of proteolysis kinetics revealed rapid in vitro degradation of the allergen by pepsin, which was not accelerated by its preheating. Furthermore, Fra a 1 exhibited relative resistance to in vitro digestion by lysosomal enzymes of human macrophages compared to birch Bet v 1 and soybean Gly m 4. These data indicate partial thermal stability of the Fra a 1 allergen, its ability to retain IgE-binding activity in some sensitized individuals, and its susceptibility to proteolytic degradation that may modulate its immunogenicity. The results of our study are important for assessing the risks associated with consumption of heat-treated strawberries and developing strategies to reduce allergenicity.
The multiple functions of neutrophilic granulocytes (NG) are associated with various subsets and phenotypes of these cells. Previously, in children with severe purulent inflammatory diseases, a correlation was found between the severity of disease and contents of CD66b+CD16+CD33+HLA-DR+NG (APC-NG) subset. Thus, the determination of the APC-NG subset may serve as a valuable diagnostic tool and an efficiency index during complex therapy in severe purulent-inflammatory diseases. Our goal was to assess the dynamic changes in APC-NG levels in children with AHO in comparison with the dynamics of indicators of clinical effectiveness during immunomodulatory therapy with a synthetic thymic hexapeptide (HP). Materials and methods. The study group (SG) included 15 children aged 10-15 years with acute hematogenic osteomyelitis (AHO), the comparison group (CG) consisted of 13 conditionally healthy children. The contents of NG expressing CD66b, CD16, CD33, HLA-DR (FC500, Beckman Coulter), and expression density of these molecules was evaluated in peripheral blood (PB) of patients and controls. The PB samples were collected before surgery, on the 10th day of therapy, and before discharge. The immunological parameters of control samples were studied once. The amounts of APC-NG were found to be increased by 18 times in study group before treatment, being decreased by 1.9 times on the 10th day after the beginning of treatment, and by 3.8 times at the time of discharge (20-25th day). However, the content of APC-NG after treatment did not reach the CG levels. Moreover, a decreased number of APC-NG correlated with positive clinical dynamics: there was an earlier regression of general and local symptoms. A delay in positive changes of APC-NG was observed in the absence of clinical efficiency, thus requiring a revision of therapeutic strategy, and changes of antibacterial therapy. Conclusion: The study demonstrated high clinical efficacy of thymic HP in the complex treatment of children with AHO and a diagnostic potential of the APC-NG contents as a diagnostic marker for monitoring efficacy of postsurgical treatment of pediatric AHO patients. This marker may also serve as a tool that allows for timely correction of AHO treatment at the in- and outpatient stages.
Malignant tumors of the central nervous system (CNS) are poorly immunogenic, thus limiting the effectiveness of therapies and contributes to a dismal prognosis for patients with these malignancies. Therefore, the development of new approaches to treatment of these diseases is becoming increasingly important. Atypical cyclin-dependent kinase 5 (CDK5) is involved in tumor progression and metastasis, as well as in developing immune escape by modifying the expression of MHC-I molecules and immune checkpoints, being an important candidate for regulating the molecular mechanisms of tumor growth. However, the influence of CDK5 on the functional state of T cell subsets and expression of key cytotoxic effectors in the microenvironment of primary and metastatic CNS cancers remains poorly understood. The aim of this study was to evaluate the effect of Cdk5 gene knockdown in tumor cells of murine model upon functional status of cytotoxic T lymphocytes in the microenvironment of CNS cancers in order to compare the experimental results with clinical data of patients with CNS tumors. The objectives of our study were as follows: to analyze the expression profile of genes associated with the T cell response using scRNA-seq in a murine model of the disease with CDK5 knockdown; to validate changes in infiltration and effector molecule expression using immunohistochemistry (IHC) technique; bioinformatic evaluation of correlations between CDK5 expression levels and cytotoxic genes in cohorts of patients with brain metastases and primary glioblastoma using bulk- and scRNA-seq; to assess the impact of effector molecule expression on overall patient survival. Our results indicate that CDK5 expression levels influence the transcriptional activity of genes encoding key T cell effector molecules such as perforin or granzymes. The results of this analysis may provide a more accurate prognosis of patient survival based on the level of expression of CDK5 and effector molecules. Further studies are required to elucidate the conditions and characteristics that mediate the immunomodulatory effect of CDK5.
