Solid-phase Enzyme Immunoassay Research Articles

The role of interleukin-6 in the pathogenesis of protein-energy malnutrition in patients treated with haemodialysis

BACKGROUND. Protein-energy malnutrition (PEM) often develops in patients receiving long-term treatment with programmed haemodialysis (HD). Its main causes are decreased intake of basic nutrients, increased losses, disorders inherent to the terminal renal failure itself (including chronic inflammation), as well as the influence of factors associated with the HD procedure. THE AIM: to clarify the role of interleukin-6 (IL-6 ) in the pathogenesis of BEN in patients treated with programmed haemodialysis. PATIENTS AND METHODS. We examined 645 patients receiving HD treatment, including 300 men and 345 women aged 56.8±12.8 years, the duration of renal replacement therapy was 8.4±5.3 years. Nutritional status was assessed according to International Society of Renal Nutrition and Metabolism (ISRNM) recommendations. Serum IL-6 levels were determined by a three-step "sandwich" version of a solid phase enzyme immunoassay using mono- and polyclonal antibodies to IL-6 using a commercial kit "Interleukin-6-IFA-BEST" from Vector-Best, Russia, under the manufacturer's instructions. The reference values for IL-6 are 0-7 pg/ml. RESULTS. The prevalence of BEN was 24.9 % (160 patients). Mean IL-6 concentration was 6.47±2.64 pg/ml in patients without evidence of BEN, and 23.20±10.40 pg/ml in patients with BEN, p<0.001. Elevated IL-6 levels revealed statistically significantly lower levels of total protein, albumin, prealbumin, total cholesterol, transferrin and blood lymphocyte counts. Patients with elevated IL-6 levels were also characterized by statistically significantly lower values of body mass index, skeletal muscle mass index and skeletal muscle mass index. CONCLUSION. The results of this study suggest that the high prevalence of PEM in patients treated with HD is closely related to an imbalance of pro- and anti-inflammatory cytokines. An increase in the duration of renal replacement therapy is accompanied by an increase in serum IL-6 levels. Therefore, this cytokine can be considered as a therapeutic target for prevention and treatment of sarcopenia in dialysis patients.

Efficacy of narrow-band medium-wave phototherapy in correcting the cytokine status of eczema patients

Background: Eczema is one of the most common skin diseases. Despite the success of systemic and local pharmacotherapy, effective treatment of patients with eczema is hampered by the lack of clear ideas about the etiology and the multifactorial nature of the disease pathogenesis. The research results available to date indicate that it is possible to increase the effectiveness of treatment of eczema patients using phototherapy methods. Aim: To evaluate the possibilities of narrow-band phototherapy with a wavelength of 311 nm (UVB-311 nm) in increasing the effectiveness of drug therapy in patients with eczema. Methods: The dermatological index of the symptom scale (DISS) was used to assess the clinical manifestations of eczema. Serum levels of cytokines – interleukins (IL) -1β, -2, -6, -10, interferon gamma (INF-γ), tumor necrosis factor alpha (TNF-α) were determined by solid-phase enzyme immunoassay. UVB-311 nm therapy procedures (25 sessions) were performed with the Dermalight®1000 device (Germany). The results of the treatment were evaluated after 2 and 4 months from the beginning of treatment. Results: The study included 66 patients with eczema (mean age 58.6±3.4 years, 44 males, 22 females), randomized into 2 groups. Group 1 included 33 patients who received only basic drug therapy (BDT). In the 2nd group, there were 33 patients who, in addition to BDT, received narrow-band phototherapy UVB-311 nm (РT+BDT). In the BDT group, the indicators of DISS after 2 months decreased by 1.33 times (p=0.01), while in the РT+BDT group - by 1.95 times (p<0.001). The level of cytokines in the BDT group did not change significantly (p>0.05) after 2 months. In the РT+BDT group, after 2 months, the level of IL-1ß decreased by 1.25 times (p=0.001), IL-2 - by 1.44 times (p<0.001), IL-6 - by 1.69 times (p=0.001), INF-γ – by 1.28 times (p=0.001), TNF-α – by 1.29 times (p=0.002);the level of IL-10 increased by 1.22 times (p=0.002). Conclusions: Basic drug therapy contributes to a moderate decrease in the DISS by 1.33 times, but does not provide a significant decline in the pro-inflammatory cytokine potential of the blood. The use of UVB-311 nm phototherapy contributes to a more significant decrease in the levels of the complex of proinflammatory cytokines (IL-1β, -2, -6, TNF-α, INF-γ) and an increase in anti-inflammatory IL-10, which favorably affects the clinical course of dermatosis, manifesting by a decrease in the DISS by 1.95 times.

Цитокиновые реакции при дислипопротеинемиях у взрослых жителей Европейского Севера России в зависимости от индекса массы тела

An increase in proinflammatory cytokine content in the blood is a stereotypical inflammatory response, regardless of the etiology or type of inflammation. Since cytokines exhibit pleiotropy, they serve as signalling molecules in intercellular interactions and stimulate the functional activity of cells, their proliferation and differentiation. The purpose of this study was to identify the frequency of elevated cytokine levels in dyslipoproteinemia in adults with different body weight living in the European North of Russia. Materials and methods. The study involved 313 residents of Arkhangelsk aged 45–60 years: with normal body weight (211 subjects; body mass index up to 30 kg/m2) and obesity (102 subjects; body mass index 31–40 kg/m2). The enzymatic and turbidimetric methods were applied to evaluate lipid profile parameters using an automatic biochemical analyser; cytokine levels were determined using solid-phase enzyme immunoassay. Results. In subjects with normal body weight, elevated blood concentrations of the cytokines under study coinciding with dyslipoproteinemia were detected in 2–11 % of cases. In obesity, elevated cytokine levels were 2–7 times more frequent; the sharpest differences were observed regarding IL-6, whose elevated concentrations reached 44 % of cases. A significant increase in IL-1ß concentrations in the blood of the subjects is associated with an increase in the levels of low-density lipoproteins (LDL) and very-low-density lipoproteins (VLDL) as well as triglycerides (TG). The frequency of increased TNF-α levels was the most significant (36.28 %) at abnormally high TG concentrations. Elevated IL-6 levels were most often associated with an increase in the blood concentration of several transport forms: LDL, VLDL, oxidized LDL, and TG. In over 40 % of cases, increased IL-10 was observed in the blood of individuals with abnormally high levels of LDL and VLDL. Thus, obese residents of the European North of Russia are characterized by a more pronounced cytokine response. At the same time, cytokine response (irrespective of the body mass) is primarily associated with an increase in the blood levels of LDL, VLDL and oxidized LDL.

Investigation of anti-PF4 versus anti-PF4/heparin reactivity using fluid-phase enzyme immunoassay for 4 anti-PF4 disorders: classic heparin-induced thrombocytopenia (HIT), autoimmune HIT, vaccine-induced immune thrombotic thrombocytopenia, and spontaneous HIT

BackgroundFour platelet-activating anti-platelet factor 4 (PF4) disorders have been recognized: classic heparin-induced thrombocytopenia (cHIT), autoimmune heparin-induced thrombocytopenia (aHIT), spontaneous heparin-induced thrombocytopenia (SpHIT), and vaccine-induced immune thrombotic thrombocytopenia (VITT). All test immunoglobulin G (IgG) positive using solid-phase enzyme immunoassay (solid-EIA) against PF4/heparin (PF4/H) and/or PF4 alone. Fluid-phase EIA (fluid-EIA) should better discriminate between anti-PF4 and anti-PF4/H antibodies since conformationally altered PF4 bound to solid phase is avoided. ObjectivesTo compare anti-PF4 vs anti-PF4/H antibody profiles for anti-PF4 disorders using solid- and fluid-EIA. MethodsWe developed a novel fluid-EIA to measure anti-PF4 vs anti-PF4/H antibodies. ResultsUsing fluid-EIA, 27 of 27 (100%) cHIT sera tested IgG positive with PF4/H, but only 4 of 27 (14.8%) tested positive against PF4 alone; all 27 exhibited heparin-enhanced binding. In contrast, 17 of 17 (100%) VITT sera tested IgG positive against PF4 alone, with markedly reduced binding against PF4/H; this distinct VITT antibody profile was not evident using solid-EIA. All 15 aHIT sera and all 11 SpHIT sera tested IgG positive against PF4 alone, with variable reactivity in PF4/H-EIA (heparin-enhanced binding in 14 of 15 and 10 of 11 aHIT and SpHIT sera, respectively). Remarkably, 1 SpHIT patient with a VITT-mimicking fluid-EIA profile (PF4 >> PF4/H) also clinically resembled patients with VITT (postviral cerebral vein/sinus thrombosis), with anti-PF4 reactivity correlating inversely with platelet count recovery; moreover, the single aHIT patient with a VITT-mimicking fluid-EIA profile also developed postviral cerebral vein/sinus thrombosis. ConclusioncHIT and VITT sera showed opposite fluid-EIA profiles (cHIT: PF4/H >> PF4, with most testing negative against PF4 alone; VITT: PF4 >> PF4/H, with most testing negative against PF4/H). In contrast, all aHIT and SpHIT sera reacted against PF4 alone but with variable (usually enhanced) reactivity against PF4/H. VITT-mimicking clinical/serologic profiles occurred in only a minority of patients with SpHIT and aHIT.

Some features of the immune system response in the new coronavirus infection COVID-19

AIM: to study the cytokine link of immunity in patients with the new coronavirus infection COVID-19 and the possibility of using these data to predict the risk of lung tissue damage.
 MATERIALS AND METHODS: Patients with diagnosed new coronavirus infection COVID-19 were divided into 4 groups depending on the severity of the disease: 1 asymptomatic patients; 2 patients with lung tissue damage in the amount of 25%; 3 patients with lung tissue damage 50%; 4 patients with lung tissue damage 75% (according to the results computed tomography). Immunological studies were performed upon admission to the clinic, then after 7 days and before discharge. The concentration of cytokines in blood serum was studied by the method of solid-phase enzyme immunoassay using kits manufactured by Vector Best.
 RESULTS: In all groups, a low content of proinflammatory cytokines was found: tumor necrosis factor and interferon-, interleukin-1, interleukin-2. The median level of interleukin-6 in all groups during the entire follow-up period was within the normal range, however, in some patients there was a multiple excess of its concentration. In groups with 50% and 75% lung tissue damage, an increase in the concentration of interleukin-8 was shown. Statistically significant differences were noted only for the level of interleukin-8 between the group with an asymptomatic form of infection and the CT-2 and CT-3 groups. It was found that the concentration of interleukin-1 below 3.58 pg/ml is a significant risk factor for severe disease. In all groups of subjects, low levels of interleukin-4 and interleukin-10 were noted, while there was no significant difference between the groups. Positive correlations were revealed between the degree of lung tissue damage and the level of interleukins 6, 2 and 8. Negative correlations were established between the degree of lung tissue damage and the content of interleukin-1.
 CONCLUSION: There was no significant activation of the cytokine link of adaptive antiviral immunity in the examined contingent. Serum cytokine levels were within normal values and had no significant intergroup differences. An increase in IL-8 in groups with clinically pronounced signs of infection indicates the activation of indicators characterizing innate immunity.

Оцінка ефективності комбінованої терапії препаратами рекомбінантного гормона росту та вітаміну D дітей препубертатного віку із соматотропною недостатністю

The treatment of somatotropic insufficiency (SI) remains one of the most important problems in paediatric endocrinology. Since 1985, recombinant growth hormone (rGH) has been widely used to correct the growth of children with SI. Sufficient clinical experience has been gained in the use of rGH preparations, and the efficacy and safety of rGH therapy in children and adolescents with various forms of short stature has been proven. However, there are a number of factors that can have a significant impact on the achievement of final height in adulthood, including a wide range of sensitivity to exogenous growth hormone (GH), the degree of endogenous GH deficiency, patient compliance, and other uncertain factors. Patients with GH deficiency require long-term treatment, the effectiveness of which decreases significantly over time, prompting the search for optimal treatment options. Purpose - to evaluate the effectiveness of combined therapy of pre-pubertal children with SI with rGH and vitamin D drugs. Materials and methods. 23 pre-pubertal children with isolated SI were examined, who took monotherapy with rGH for 3 years. When the effectiveness of monotherapy decreases, vitamin D are added to the treatment complex. The level of 25(OH)D in blood serum is determined by the immunochemiluminescence method on microparticles (Abbott, USA). The levels of GH and insulin-like growth factor-1 were studied by solid-phase enzyme immunoassay using Immulite 2000 XPI kits. Results. At the end of the first year of rGH monotherapy, all patients showed a significant (p<0.05) increase in height. However, already after the second and third years of treatment, height velocity (HV) decreased in all patients. The increase in HV in the third year of treatment with rGH was significantly lower (p<0.05) than in the first year. Thanks to the addition of vitamin D, the further decrease in HV was stopped after the third year of monotherapy with rGH. The increase in HV during the first year of combined therapy was significantly (p<0.05) greater than during the third year of treatment with rGH monotherapy. The same trend was observed after the second year of combined therapy. Conclusions. In the presence of a decrease in the effectiveness of therapy with rGH after the first years of treatment, it is considered appropriate to use combined therapy of children with SI with rGH and vitamin D. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of the participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

CLINICAL, FUNCTIONAL AND IMMUNOLOGICAL CHARACTERISTICS OF SEVERE BRONCHIAL ASTHMA

Bronchial asthma is the most common and socially significant disease in humans. Among patients who cannot achieve control, a special group is patients with severe asthma. Objective: comprehensive assessment of clinical, functional, immunological features and pharmacotherapy of severe bronchial asthma in real clinical practice to optimize basic pathogenetic therapy. Materials and methods: 83 patients diagnosed with severe asthma were examined. Patients with severe asthma are divided into 2 groups: patients with and without fixed airway obstruction. Plasma concentrations of cytokines IL-4, IL-5, IL-9, IL-13, periostin, cathepsin S, TGF- were determined by solid-phase enzyme immunoassay. Immune status was investigated on the NAVIOS Flow Cytometer. Results: In both groups of severe bronchial asthma, we found a decrease in T helper and immunoregulatory index levels with simultaneous increases in cytotoxic T lymphocytes, natural T killers, naive T lymphocytes, activated T and B lymphocytes, and phagocytic index compared to controls. There were no differences in immune status between the groups and the resulting changes were independent of the presence or absence of fixed obstruction. In both study groups, we found an increase in cathepsin S and TGF- in plasma compared to control. We identified the most significant risk factors for the formation of fixed obstruction: taking SABA more than 4 inhalations per day (OR = 4.2) and FeNO concentration more than 20 ppb (OR = 6.0). There was a significant improvement in the clinical condition of patients with severe asthma with fixed obstruction while taking genetically engineered biological therapy for a year. Conclusion: To date, there is no unambiguous idea of the mechanisms of implementation of pathobiochemical reactions in the bronchial wall in severe asthma, which lead to the development of fixed airway obstruction. Severe asthma is variable and depends on the correct choice of management tactics.

Blood biomarkers and Ki-67 proliferation marker in breast cancer

Metastasis is the leading cause of death in patients with breast cancer (BC). It is known that the lesion of regional lymph nodes by tumor cells is more common in tumors with higher proliferative activity. Moreover, there is literature evidence on effects of cytokines and proteins upon the migration potential of the tumor. The aim of our work was to study the correlation between the concentrations of cytokines, proteins, and expression of Ki-67 proliferation marker in breast cancer with histology of non-specific invasive carcinoma.On the basis of pathological findings, 16 patients had metastases in regional lymph nodes (group I), and 18 patients had no detectable metastases (group II). Solid-phase enzyme immunoassay was used to determine concentrations of 14 cytokines in the supernatants of immunocompetent blood cells, i.e., IL-2, IL-6, IL-8, IL-10, IL-17, IL-18, IL-1β, IL-1ra, TNFα, IFNγ, G-CSF, GM-CSF, VEGF and MCP-1, and concentrations of 6 proteins were determined in blood serum: estrogen and progesterone receptors, cadherin-E (CDH1), plasminogen activator type 1 (PAI-1), mucin 1 (MUC1), heat shock protein 90αA1 (HSP90αA1). Immunohistochemical study of Ki-67 expression was performed in paraffin sections of tumors using monoclonal antibodies.The study showed that Ki-67 expression in tumor tissues and blood concentrations of IL-6, IL-8, IL-1β and TNFα were higher in group I patients. On the contrary, blood concentrations of CDH1 and PAI-1 were higher in group II patients. It was found that Ki-67 showed both inverse correlations with CDH1 and PAI1, and direct correlations with IL-8 and TNFα. CDH1 had a direct correlation with PAI1, and inverse correlations with IL-6, IL-1β and TNFα. The studied cytokines showed direct correlations with each other. The analysis of ROC curves showed good quality and optimal values of the cut-off points for Ki-67 expression, cytokine and protein concentrations, thus allowing best prediction for detectable lymphatic metastasis.On the basis of these results, a quotient was proposed, which represents a ratio of CDH1 contents to the sum of IL-1β and TNFα concentrations in blood samples, which can help identification of the patients with breast cancer at risk for lymphatic metastasis.

Взаимосвязь амилоидогенеза и таупатии с клиническими исходами ишемии головного мозга. Экспериментальное исследование

Currently, it has been established that increased formation of β-amyloid and tau protein play a significant role in the pathogenesis of cerebral ischemia. At the same time, these molecules can act as prognostic markers of the severity of cerebral ischemia. The aim of the study. To evaluate the peculiarities of changes in the concentration of β-amyloid and tau protein in blood serum and cerebrospinal fluid in rats with experimental subtotal cerebral ischemia, as well as to establish their relationship with the severity of post-ischemic clinical changes. Materials and methods. Subtotal cerebral ischemia was modeled by simultaneous occlusion of common carotid arteries in Wistar rats. The study assessed changes in neurological, sensorimotor and cognitive deficits. The content of β-amyloid and tau protein was assessed in blood serum and cerebrospinal fluid on the 1st, 3rd, 5th and 7th days of ischemia. During the analysis, species-specific kits for solid-phase enzyme immunoassay were used. Results. It was found that in animals with subtotal cerebral ischemia, as the duration of the ischemic period increases from day 1 to day 7, the concentration of β-amyloid and tau protein in both blood serum and cerebrospinal fluid progressively increases. It is worth noting that these changes strongly correlated with changes in the clinical manifestations of cerebral ischemia — neurological, sensorimotor and cognitive deficits.

Changes in Neuronal Antibody and S100β Protein Content as a Risk Factor for Occupational Neurointoxication

The production of the vinyl chloride (VC) still extremely popular in many countries of the world today. The aim of this research was to study changes in the content of S100β protein and antibodies to nerve tissue proteins as regulators of the nervous system in workers in the production of vinyl chloride, depending on the severity of the pathological process. Men working in the production of VC were examined. There are 53 healthy trained workers without signs of disorders of neuro-mental status and 26 patients with initial manifestations of neurointoxication with vinyl chloride among them. The concentration of S100β protein and neurotrophic antibodies (NAB) in blood serum was determined by solid-phase enzyme immunoassay. The more pronounced increase in S100β protein was found in persons with initial manifestations of neurointoxication, compared with trained healthy workers. The involvement of autoimmune reactions in the processes of regulation of the nervous system through the production of antibodies (AB) to certain structures of the nervous tissue has been proved. It is shown that the intensity of autoimmune reactions and the spectrum of produced AB working in the production of VC depend on the duration of exposure and the stage of development of the neurointoxication. It was also established that the change in the synthesis of NAB to specific proteins of nervous tissue is associated with the deterioration of neurological status in trained healthy workers. The obtained results allow us to assess the degree of interest of certain structures of the nervous tissue – bioregulators of the nervous system in the development of neurological disorders. Revealed an increase in serum concentrations of NAB to MAG, and in persons with clinical manifestations of asthenic disorder with autonomic dysfunction – NAB to DNA, NAB to S100β in trained healthy workers. These indicators can be regarded as a risk factor for occupational neurointoxication. The results obtained will improve the quality of diagnosis, especially in the early stages of the disease and, as a result, the formation of the most optimal treatment tactics.

Assessment of the Myocardial Stress Biomarker NT-proBNP in Real Clinical Practice

Subjects and Methods. The study involved 35 vascular surgery patients of varying degrees of cardiological risk. Blood specimens were collected from each patient at 3 time-points: 1. prior to surgery (NT-proBNP1), 2 — after the procedure (NT-proBNP2), 3 — before the discharge from the hospital (NT-proBNP3). Each specimen was split into equal aliquots for biomarker quantification using two different techniques (ELISA using domestic reagents — for the 1st series of analyses, and ICLA using an imported kit — for the 2nd series). Perioperative cardiovascular complications were recorded. The consistency of the measurement results obtained by two different methods was evaluated using the Bland–Altman technique. A discrimination ability of independent variables in relation to a binary dependent variable was studied using ROC analysis.Results. In the 1st series, ranges of the biomarker were as follows: NT-proBNP1 — 24–774 pg/ml, NT-proBNP2 — 41.2–889.1 pg/ml, NT-proBNP3 — 39.3–1013.3 pg/ml. In the 2nd series, NT-proBNP1 was 31.2–2087.0 pg/ml, NT-proBNP2 — 32.5–3754.0 pg/ml, NT-proBNP3 — 34.1–2728.0 pg/ml. In the Bland–Altman analysis, 97.03% of the values fell within the lower and upper limits of consistency (±1.96 SD of the average difference), which indicated comparability of the results in the series, but the values of NT-proBNP in the 1st series were lower than in the 2nd ones. Cardiovascular complications were registered in 3 (8.5%) patients. In the 1st series, NT-proBNP1 > 218 pg/ml predicted cardiovascular complications with a sensitivity of 66.7% and a specificity of 81.3% (AUC 0.844, 95% CI 0.681–0.944, P = 0.0003). In the 2nd series, NT-proBNP1 > 315 pg/ml predicted cardiovascular complications with a sensitivity of 66.7% and a specificity of 75.0% (AUC 0.828, 95% CI 0.663–0.934, P = 0.001).Conclusion. The domestic ELISA kit for solid-phase enzyme immunoassay proved its clinical informativeness for quantitation of NT-proBNP demonstrating its value for diagnostic and prognostic purposes, or scientific studies. The novel domestic technique provides consistently reproducible results, although with lower reference values as compared to the standard immunochemiluminescence assay.

Platelet and coagulation hemostasis status in patients with atrial fibrillation depending on warfarin dosing method

The aim. To evaluate indicators of induced platelet aggregation and D-dimer level in patients with atrial fibrillation (AF) depending on the method of warfarin (WF) dosing. Materials and methods. The study involved 110 patients with AF (mean age 68.72 ± 0.79; men – 57, women – 53). Patients with AF were divided into two groups: the main group – 50 patients with AF and genotype-guided dosing method, the control group – 60 patients with AF and traditional dosing method. CYP2C9, CYP4F2, VKORC1 genetic polymorphisms were determined using multiplex real time polymerase chain reaction, D-dimer concentration – by the method of solid-phase enzyme immunoassay; ADP- and epinephrine-induced aggregation indicators – by the method of G. Born. Results. The percentage (by 24 %), time (by 3 min 6 s) and rate in 30 s (by 19.5 %/min) of ADP-induced platelet aggregation were significantly lower in patients with AF and genotype-guided WF dosing method. D-dimer concentration in patients of the main and control groups did not differ significantly. However, significantly fewer patients with elevated D-dimer values were registered in the group with genotype-guided WF dosing method compared to the group with traditional dosing method: 0 (0.00 %) vs. 7 (11.67 %) at 500 ng FEU/ml cut-off (χ2 = 4.43, P < 0.05), 1 (2.00 %) vs. 9 (15.00 %) at 390 ng FEU/ml cut-off (χ2 = 4.16, P < 0.05), 0 (0.00 %) vs. 7 (11.67 %) at age-adjusted cut-off (χ2 = 4.43, P < 0.05). Conclusions. The obtained results may indicate a potentially lower risk of thrombotic events in patients with AF and genotype-guided WF dosing method compared to the group with traditional dosing method, which confirms the benefit of the widespread use of the pharmacogenetic testing in clinical practice.

Associations of bone mass and polygenic risk of osteoporosis with indicators of arterial wall condition

Background: The identification of genetic factors that are simultaneously responsible for the predisposition to the development of cardiovascular diseases (CVD) and osteoporosis (OP) is important for the prevention of both conditions.Aim: The aim of this study is to evaluate three genetic risk scales (GRS) that previously showed an association with bone mineral density (BMD) and fracture risk, as well as to study the associations of these GRS with vascular wall pathology.Materials and methods: 250 female outpatients (aged 45 to 69) were enrolled into a cross-sectional study. The intima-media thickness (IMT), the presence and number of atherosclerotic plaques (AP) were studied using duplex scanning. Pulse wave velocity (PWV), augmentation index (AI) were measured by applanation tonometry. Coronary vessels calcium deposits were registered by multispiral computed tomography (MSCT) using the Agatston calcium index (CI). The BMD of the spine, hip neck (HN) and proximal hip (PH) was measured using double energy x-ray absorptiometry. Bone resorption marker type-1 collagen C-terminal telopeptide (CTx) was assessed solid-phase enzyme immunoassay. The genetic study included DNA extraction from whole blood samples. Targeted sequencing was performed on the Nextseq550 sequencer (Illumina, USA). Statistical analysis was carried out using the SAS software package for Windows, version 9.0 (SAS Institute Inc., USA).Results: The chance of detecting low bone mass increased more than 4 times at values of IMT ≥0.9 mm (OR=4.17; 95%CI [1.2–14.4], p<0.02), 2.4 times in the presence of AP in the carotid arteries (OR=2.45; 95%CI [1.12–4.88], p><0.05), by 6.7 times with an Agatstone CI ≥ 100 units (OR=6.68; 95%CI [1.56–28.7], p><0.001), 1.4 times (OR=1.43; 95%CI [0.56–3.68], p><0.438) with a PWV ≥10 m/s, 1.2 times (OR=1.2; 95%CI [0.601–2.43], p><0.60) with increased AI ≥ 27%. According to multivariate linear regression analysis (adjusted for age, duration of postmenopause, marker of bone resorption CTx), a significant association of all GRS with BMD in all parts of the skeleton was revealed. Both univariate and multivariate regression models adjusted for several covariants (age, total cholesterol, systolic blood pressure) showed a reliable association of GRS62 with the presence of plaques and GRS63 — with coronary artery CI. Conclusion: The results of the study demonstrated the association of polygenic genetic risk of GRS-based OP with BMD and vascular wall status indicators in women in the peri and postmenopausal periods.>< 0.02), 2.4 times in the presence of AP in the carotid arteries (OR=2.45; 95%CI [1.12–4.88], p< 0.05), by 6.7 times with an Agatstone CI ≥ 100 units (OR=6.68; 95%CI [1.56–28.7], p< 0.001), 1.4 times (OR=1.43; 95%CI [0.56–3.68], p< 0.438) with a PWV ≥10 m/s, 1.2 times (OR=1.2; 95%CI [0.601–2.43], p<0.60) with increased AI ≥ 27%. According to multivariate linear regression analysis (adjusted for age, duration of postmenopause, marker of bone resorption CTx), a significant association of all GRS with BMD in all parts of the skeleton was revealed. Both univariate and multivariate regression models adjusted for several covariants (age, total cholesterol, systolic blood pressure) showed a reliable association of GRS62 with the presence of plaques and GRS63 — with coronary artery CI.>< 0.60) with increased AI ≥ 27%. According to multivariate linear regression analysis (adjusted for age, duration of postmenopause, marker of bone resorption CTx), a significant association of all GRS with BMD in all parts of the skeleton was revealed. Both univariate and multivariate regression models adjusted for several covariants (age, total cholesterol, systolic blood pressure) showed a reliable association of GRS62 with the presence of plaques and GRS63 — with coronary artery CI.Conclusion: The results of the study demonstrated the association of polygenic genetic risk of GRS-based OP with BMD and vascular wall status indicators in women in the peri and postmenopausal periods.

Relationships between the factors of innate immune system and results of neurocognitive function parameters in the persons living with HIV infection: interim results of the study

HIV infection is closely associated with damage to the nervous system, even despite usage of antiretroviral therapy. Thus, the patients are at a high risk for neurological complications. Due to limited permeability of the blood-brain barrier, the opportunity of its damage, both directly by the proteins of HIV-infected cells and indirectly, by immune system, e.g., inducing the production of inflammatory mediators, the involvement of the central nervous system in pathogenesis of HIV infection seems to be only a matter of time. HIV-associated neurocognitive disorders (HAND) comprise a poly-etiological pathological condition associated with direct and indirect damage to neurons in HIV infection. The role of interaction between the innate immune system and nervous system mediators in viral infections suggests that the specific changes in content and ratios of pro- and anti-inflammatory cytokines that are among the most polypotent mediators of immune regulation can be detectable in the persons prone to the development of HAND in HIV infection. The purpose of the present study was to characterize the features of serum content and the relationship between the indexes of innate immunity factors with the results of neurocognitive testing in the persons living with HIV.
 123 persons were included in the study, of which 93 subjects live with HIV infection. They were divided into 3 groups according to the stages of HIV infection, according to the Russian Classification of HIV infection. 30 persons represented a group of healthy volunteers, comparable in age and gender with the members of main groups, and served as a control group. Determination of IL-1 and IL-10 contents in blood serum was carried out by solid-phase enzyme immunoassay. The level of CD4+T lymphocytes was determined by flow cytometry. Neurocognitive testing using Schulte tables, the Munsterberg test was conducted by a medical psychologist, according to generally accepted methods.
 A significant increase in the serum IL-10 content in subjects from the main groups was revealed as compared with the control group, as well as in group III compared with the values groups I and II. The levels of IL- 1, CD4+T lymphocytes were significantly lower in group III, compared with all other groups. The personal performance index determined by testing with Schulte tables was reduced in all groups, relative to the controls, with a significant predominance of the results in groups I and II over the results of the participants from group III. The scores in Munsterberg test were equally diminished in all the main groups compared to the control values. When assessing the Spearman coefficient, the presence of various correlation profiles was established, depending on the stage of the disease.

Hormonal status and cardiorespiratory adaptation of late premature newborns in the early neonatal period

The growth of late preterm labor in recent years and the increased frequency of neonatal complications in 34–36-week gestation newborns explain the interest of researchers in the problem of late prematurity. The loss of the last weeks of pregnancy by late prematurity interruptsthe formation of hierarchical connections between regulatory links and target organs, but the questionsremain open, how significant is the effect of late prematurity on the hormonal status of the newborn and what are the immediate and long-term consequences of hormonal disorders that occurred in the early neonatal period.Purpose. This study analyzes the effect of hormonal status on cardio-respiratory adaptation of late premature newborns in the early neonatal period.Materials and methods. The study included 105 newborns with a gestation period of 34–36 weeks. A comprehensive assessment of the early adaptation period was carried out, the levels of TSH, T3, T4 total, T4 free, cortisol were determined. Blood sampling was carried out immediately after birth from the umbilical cord vein and on the 4th day after birth from the subcutaneous veins. The concentration of hormones was determined by solid-phase enzyme immunoassay using standard sets (Alkor Bio, Russia).Results. The results of the work established a relationship between the thyroostatic state, cortisol concentration and the course of cardio-respiratory adaptation in late premature newborns. Newborns in need of respiratory and cardiotonic therapy had reduced concentrations of T3, T4 free and cord blood cortisol. On the 4th day of life, newborns in need of respiratory therapy showed a decrease in free T4 and an increase in cortisol, children receiving cardiotonic support had a reduced concentration of free T4.Conclusion. Hormonal maladaptation can be considered as an additional factor in the pathogenesis of respiratory disorders in late premature newborns along with a reduced gas exchange area in the lungs, immaturity of sodium pumpsin alveolocytes and surfactant deficiency.

Role of neuroproteins in retinal neurodegeneration in diabetic retinopathy

Aim. To evaluate the serum levels of S100b protein, brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF) in patients with type 2 diabetes mellitus, as well as to establish a correlation between the studied parameters and retinal neurodegeneration.Methods. In total, 80 patients with confirmed type 2 diabetes (experimental group) and 30 healthy volunteers (control group) were included in the study. All patients were examined using an RTVue-100 optic coherence tomography scanner (Optovue, USA) and a MAIA microperimeter (CenterVue, Italy). The serum levels of S100b, BDNF, and NGF were determined by a sandwich-type solid-phase enzyme immunoassay using reagents produced by R&D Diagnostics Inc. (USA).Results. The serum level of S100b protein was found to be significantly (p = 0.001) higher in the experimental group, comprising 56.86 (31,12–104,02) pg/ml, compared to that in the control group of 45.19 (37.27–51.79) pg/ml. Conversely, the brain-derived neurotrophic factor (BDNF) showed a significant (p = 0.02) decrease to the level of 27.38 (25.15–29.12) pg/ml in the experimental group compared to the level of 30.19 (27.38–32.14) pg/ml in the control group. The serum level of nerve growth factor (NGF) in the experimental group was found to be 7.53 (5.63–10.54) pg/ml, thereby exceeding significantly (p = 0.02) the level of 5.96 (4.77–8.13) pg/ml in the control group.Conclusions. The results obtained indicate an imbalance in the serum levels of the studied neuroproteins in patients with type 2 diabetes mellitus and retinal neurodegeneration signs. The identified variations require further research in order to determine the feasibility of their use as additional criteria for predicting the development of retinal neurodegeneration in such patients.

Anti-interferon alpha autoantibodies and their significance in COVID-19

During the last two years, treatment of patients with novel coronavirus infection COVID-19 remains an urgent health problem. Interferon proteins are known to play a significant role in antiviral immunity. Some pathological conditions are accompanied by production of neutralizing autologous immunoglobulins against own host interferons (auto-IFN-Abs). There is evidence that autoantibodies against interferons alpha and omega are detected in patients with life-threatening course of COVID-19 pneumonia. The aim of our study was to analyze prevalence of autoantibodies against interferon alpha in patients with COVID-19 coronavirus infection and assess their impact on clinical course of the disease. We examined 70 patients with severe COVID-19, who received inpatient treatment at the intensive care units. Serum autoantibodies against interferon alpha were determined on day 850 after disease onset by using solid-phase enzyme immunoassay (ELISA). Patients were divided into 2 groups: those with and without (group 2) autoantibodies against interferon alpha (group 1). Anti-COVID serum from 57 donors was used a control. Among patients, autoantibodies against interferon alpha were detected in 13 (18%) subjects, which level ranged from 26.8 to 1000 ng/ml. Among donors, auto-IFN-Abs were detected in 5 (8.8%) subjects at trace concentrations (from 1.65 to 12.0 ng/ml). Respiratory failure developed significantly more often in patients with auto-IFN-Abs. While analyzing laboratory parameters, it was noted that the concentration of C-reactive protein was significantly higher in the group of patients with auto-IFN-Abs. Mortality rate of patients with high auto-IFN-Abs levels was 60%. In conclusion, it was found that serum autoantibodies against IFN alpha in COVID-19 patients caused lung damage that significantly more often required hardware respiratory support, so comparable by duration with it for patients without auto-IFN-Abs. High concentrations of auto-IFN-Abs (more than 100 ng/ml) in patients with COVID-19 can be considered as a predictor of unfavorable disease outcome.

Диагностическое значение иммунорегуляторных медиаторов в зависимости от степени распространенности опухолевого роста при почечно-клеточном раке

Introduction. In recent years, it has been established that the study of the content of molecular genetic biomarkers in blood serum, tissues characterizing the activity of tumor-infiltrating immune cells or tumor microenvironment makes it possible to evaluate the effectiveness of immunogenic effects of antitumor therapy, as well as to predict the development of its complications. The diagnostic value of immunoregulatory mediators in peripheral blood as prognostic biomarkers of immunotherapy in patients with renal cell carcinoma (RCC) has been confirmed by a small number of studies. This limits the further development of effective strategies for the treatment of patients with renal cell carcinoma. Additional research is needed to find molecular markers. The purpose of the study – to establish the diagnostic value of changes in the serum content of tumor microenvironment mediators from the stage of the disease in patients with RCC. Material and methods. The study included 66 patients with RCC and 30 practically healthy individuals who made up the control group. Additionally, by the method of solid-phase enzyme immunoassay, the serum content of seven mediators produced by tumor microenvironment cells was determined: proinflammatory cytokines – TNFa, INFy; chemokine – MCP1; colony-stimulating factors – G- CSF GM- CSF; growth factors – VEGF, TGF-β1. Results. It was found that at the 1st and 2nd stages of the disease, the level of cytokines/chemokines and colony-stimulating factors in the blood serum increases in comparison with the group of conditionally healthy individuals, with the exception of the content of INFy. With the progression of tumor growth in patients with stages 3 and 4 of the disease, a statistically significant high content of cytokines/chemokines and colony-stimulating factors remains in the blood serum, the level of INFy decreases and the concentration of growth factors (VEGF, TGF-β1) increases. The calculated AUC data showed a good quality of the prediction model, which gives reason to consider the increase in the serum content of mediators of the tumor microenvironment of RCC as an indicator of the stage of the disease and the prevalence of tumor growth, as well as a promising laboratory marker for predicting the effectiveness of immunotherapy treatment technologies. Conclusions. The growth of tumor cells in the initial stages of RCC is accompanied by a significant increase in the content of pro-inflammatory cytokines/chemokines and colony-stimulating factors in the blood serum. During the progression of tumor growth in patients with RCC, there is an activation of the release of growth factors such as TGF-β1 and VEGF into the bloodstream. Mediators of the tumor microenvironment entering the peripheral bloodstream (cytokines/chemokines, colony-stimulating and growth factors) are prognostic indicators for evaluating the effectiveness of immunotherapy technologies for the treatment of tumor diseases.

On the Issue of Evaluating the Effectiveness of Vaccination of Employees of Medical Organizations against COVID-19

Relevance . Active mass vaccination of the population against a new COVID-19 is being carried out on the territory of the Russian Federation, which is recognized as a priority strategy for the country's healthcare for the near and long-term periods. One of the main risk groups that are subject to priority vaccination is medical workers. Aims. To evaluate the effectiveness of vaccination of medical organizations ' employees against COVID-19 based on the results of a 6-month prospective follow-up. Materials and methods. The observation group consisted of 356 employees of a medical organization who were vaccinated against COVID-19 with the drug «Gam-Covid-Vac» from December 2020 to April 2021. The effectiveness of vaccination of employees was evaluated by the coefficient of IgG positivity to SARS-CoV-2 by solid-phase enzyme immunoassay 3 weeks after the first administration and 3–4 weeks after the second administration of the vaccine and then 1 time per month. Employees who were revaccinated after 5–6 months. After the initial vaccination, they were examined 10–14 days after the introduction of the first component «Gam-Covid-Vac». A total of 1921 serum samples were studied. A specific T-cell immune response was determined in two study participants without seroprotection after administration of two components of the «Gam-Covid-Vac» vaccine and eight employees with the elimination of IgG antibodies 4–5 months after vaccination using ELISPOT technology. In addition, 92 blood serum samples of 32 employees from the observation group were examined for specific antibodies to adenovirus by indirect enzyme immunoassay. From December 2020 to June 2021, the study participants were subjected to dynamic clinical observation, once a week they were examined by PCR to detect SARS-CoV-2 RNA in smears from the pharynx and nose (a total of 5696 samples). Results . After the completed course of immunization, the formation of both a humoral (in 99.4% of cases) and cellular immune response (100% among the studied samples) was confirmed. In the next 6 months after vaccination, cases of coronavirus infection were registered in 4.8% of those vaccinated, including 1 person – in the first month after vaccination and 16 – 3–5 months after vaccination. In all cases, the disease occurred in the form of an acute respiratory infection of mild or moderate severity and was characterized by a shorter period of virus isolation compared to similar data on the persistence of the virus in unvaccinated patients (15 days in vaccinated compared to 22 days in unvaccinated). It was found that the presence of immunity to adenovirus infection during vaccination with the drug «Gam-Covid-Vac» did not affect the possibility of forming an immune response to COVID-19. In the group of persons re-vaccinated with the first component of «Gam-Covid-Vac» after 5–6 months. after the initial vaccination, an immune response was received during the follow-up period. Conclusion. Thus, according to the results of the study, a high immunological and epidemiological effectiveness of vaccination against COVID-19 with the drug «Gam-Covid-Vac» was established in a group of medical workers, and the effectiveness and safety of the administration of a booster dose of the vaccine after primary vaccination was also shown.

On the use of laser therapy in dentistry

The aim of the research: to optimize the prevention of dental complications by using electromagnetic low-intensity laser radiation (LILR) techniques on dental and periodontal tissues. Research methods. In 138 dental patients, a clinical and functional assessment of the dental apparatus was carried out, dental indices were studied, the results of vascular endothelial parameters (VEGF-A, sVEGF-R1, sVEGF-R2) and immune parameters (FNF, TNF, IFN-γ, neutrophils) of blood serum (solid-phase enzyme immunoassay with a standard set), as well as VAS pain syndrome parameters were analyzed before/after laser therapy of different wavelength, mode and range (red, infrared). Results. The study made it possible to assess the effectiveness of treatment with LILR with one wavelength and in combination of different lengths of laser radiation as independent factors of preventive exposure in dentistry patients. After applying the techniques of transcanal treatment, dental periapical tissues were treated with laser radiation, including root canals (difficult teeth roots), using the C11 attachment from the «Lazmik-01» device. The study used a modulated red spectrum of LILR in continuous and pulsed mode (l = 635 nm) and an infrared pulsed spectrum (l = 904 nm) in a continuous mode with a sequential exposure to red (l = 635 nm) pulsed and infrared continuous laser radiation (l = 904 nm). Emphasis was placed on pain syndrome, vascular endothelial and immune shifts in dental and periodontal tissues before/after using laser therapy in dental patients. Conclusion. The combination of the red spectrum of LILR in pulsed mode (l = 635 nm) with a sequential exposure to infrared laser radiation (l = 904 nm) in continuous mode provides high vascular endothelial responses in the dental and periodontal tissues of the dental apparatus, and the use of red-spectrum laser radiation in continuous mode (l = 635 nm) in dental patients provides antimicrobial and sanitizing effects, including at the level of the root canals, which is more significant than after the use of infrared laser radiation in pulsed mode, when analgesic results are higher.