Enzyme Immunoassay Method Research Articles

THE ROLE OF ENDOTOXICOSIS AND INFLAMMATION IN DEEPENING THE PANCREATIC FUNCTIONAL INSUFFICIENCY IN CHRONIC PANCREATITIS IN COMBINATION WITH TYPE 2 DIABETES.

Aim: To analyze the state of parameters of inflammation, endotoxicosis, and their influence on the functional capacity of the pancreas in the comorbid course of chronic pancreatitis and type 2 diabetes mellitus (DM2). Materials and methods: 115 patients with CP in the phase of mild therapeutic exacerbation in combination with DM2 in the stage of subcompensation were examined. To assess the impact of comorbid DM2 on the clinical condition of patients with CP, a comparison group of 25 patients with CP in the exacerbation phase was included in the study. The assessment of the presence and depth of pancreatic exocrine insufficiency (PEI) was carried out according to the "gold standard" - determination of the content of fecal α-elastase-1, which was determined by the method of enzyme immunoassay using standard kits. As the main criterion for diagnosis and monitoring of DM, the measurement of HbA1c was used, which was determined by the method of ion exchange chromatography. C-reactive protein (CRP) was determined by the immunoturbidometry method by photometric measurement of the antigen-antibody reaction to human CRP antibodies; reference values of CRP in blood serum are up to 3 mg/l. Endogenous intoxication (EI) was assessed based on the levels of medium-mass molecules (MMM) - MMM1 and MMM2 at wavelengths 254 and 280 nm. The level of circulating immune complexes (CIC) was determined by the method of selective precipitation in 3.75% ethylene glycol followed by photometry. Results: Moderate and moderate inverse correlations were established between CRP and fecal α-elastase in CP and CP-DM2 comorbidity (r=-0.423 and r=-0.565, p<0.05). This proved a reliable influence of the depth of inflammation according to the content of CRP on the increase in PEI according to the level of fecal α-elastase, which was higher in the CP-DM2 comorbidity compared to CP. A deeper level of secretory insufficiency of the pancreas was established in CP with concomitant DM2, which deepened when the CRP level increased, compared to that in isolated CP: an increase in the strength of reliable direct moderate HbA1c-CRP correlations in patients with CP in combination with DM2 was proved in relation to such cases isolated CP (respectively r=0.313 and r=0.410, p<0.05). Conclusions: We proved a reliable influence of the index of endogenous intoxication on the level of PEI according to the level of fecal α-elastase, which was higher in the CP-DM2 comorbidity compared to isolated CP: moderate and medium-strength inverse correlations were established IEI-fecal α-elastase in patients with CP and CP-DM2 comorbidity (r=-0.471 and r=-0.517, p<0.05). An increase in the strength of reliable direct, moderate, and moderate correlations between the levels of HbA1c and the index of endogenous intoxication in patients with isolated CP and CP-DM2 comorbidity (r=0.337 and r=0.552, p<0.05), which proved a deeper level of secretory pancreas insufficiency with concomitant DM2, which worsened with increasing endotoxicosis according to the value of the index of endogenous intoxication.

THE STRUCTURAL AND FUNCTIONAL STATE OF THE PANCREAS AND LIVER IN CHRONIC PANCREATITIS IN COMBINATION WITH CHRONIC VIRAL HEPATITIS C DEPENDING ON THE INDEX OF THE SURVEY ACCORDING TO THE INTERNATIONAL CAGE QUESTIONNAIRE.

The aim: To conduct a comparative analysis of parameters of the structural and functional state of the liver and pancreas in patients with chronic pancreatitis in comorbidity with treated etiologically chronic viral hepatitis C, depending on the results of testing according to the international CAGE questionnaire. Materials and methods: 100 ambulatory patients with CP with concomitant HCV, treated etiotropically, were examined. All patients were examined ac-cording to generally accepted algorithms. To establish the role of alcohol on the formation of CP and the condition of patients with treated HCV, latent craving for alcohol was verified using the international CAGE questionnaire. The study of the density of the liver parenchyma and the liver of the patients was carried out not only according to the ultrasound data in the B-mode, but also with the simultaneous measurement of the shear wave elastography (SWE) method on the Ultima PA scanning ultrasound device with the further determination of the median of the parameters, which characterizes the stiffness in kilopascals (kPa). Determination of the presence and depth of pancreatic exocrine insufficiency (PEI) was carried out by the content of fecal elastase-1 (FE-1), which was determined by the enzyme immunoassay method. Results: Screening-testing of patients with CP on the background of etiotropically treated HCV using the CAGE scale made it possible to state that 65.0% of such patients had a hidden craving for alcohol, and 21.0% of this cohort were women, which needs to be taken into account in the management of such patients. It has been proven that in the group of patients with CAGE≥2.0, the level of functional and structural changes in the liver and liver was significantly more severe (according to the deepening of the PEI, a decrease in fecal α-elastase by 13.01%, according to an increase in the total index of the coprogram by 15.11% and the total US-indicator of the pancreas structure by 28.06%, and the total US-indicator of the liver structure - by 40.68% (p<0.05) and corresponded to the average degree of severity of the process in panceas according to the criteria of the Marseille-Cambridge classification, and in the group with CAGE<2.0 - only a mild degree. Conclusions: The negative effect of the factor of increased alcohol use according to CAGE was proven by increasing the density of the echostructure of the liver by 5.73% (p<0.05), and the liver by 5.16% (p<0.05). According to the results of the correlation analysis of the dependence of the structural state of the liver and PW of the studied patients on the value of the CAGE scale, which was R=0.713, p<0.05, and R=0.686, p<0.05, respectively, it was established that there is a strong direct dependence of the structural state of the liver and PW from the value of the CAGE questionnaire, which proved an independent, reliably significant role of alcohol consumption for patients with a comorbid course of CP and HCV.

Protective Antibody Titer against Hepatitis B in Healthcare Workers: A Cross-sectional Study from Tertiary Care Hospital of Kashmir

Introduction: Hepatitis B is not only the most transmissible blood borne viral infection, but also the only one that is preventable by vaccination. In developing countries, Hepatitis B vaccination coverage among Healthcare Workers (HCWs) is very low for various reasons. Aim: To evaluate Hepatitis B Virus (HBV) immunisation status and Hepatitis B surface antibody (anti-HBs) titer among HCWs in a tertiary care hospital in Kashmir. Materials and Methods: This cross-sectional study was conducted in the Department of Microbiology, Government Medical College, Srinagar, India from April 2019 to June 2019. Serum samples were collected from 196 HCWs and their vaccination history was collected. Those who had taken all three doses of hepatitis B were considered to be fully vaccinated those that had taken two doses as partially vaccinated. Triple serology was done for all which included testing for HBV, Hepatitis C Virus (HCV) and Human Immunodeficiency Virus (HIV) infection, one HCW turned out to be Hepatitis B Surface Antigen (HBsAg) positive and was dropped out of study. Those negative (195/196) were further tested for anti-HBs titer by enzyme immunoassay method. Parametric data are expressed as mean±Standard Deviation (SD) and categorical values as percentages. Results: Out of 195 HCWs selected for the study, 112 (57.4%) were males and 84 (43%) were females with mean age 38.4±6.9 years. Among them fully vaccinated were 46 (23.5%), partially vaccinated were 84 (43.07%) and not vaccinated were 65 (33.33%). Anti-HBs titers were protective in 54 (27.7%) which belonged 81.5% (44/54) to vaccinated category and 18.5% (10/54) to partially vaccinated category. Conclusion: Fully vaccinated HCWs (95.6%) had a protective antibody titer but unfortunately there are low vaccination rates among HCWs. There is a need for well-planned and clear policies for HBV screening and vaccination in HCWs, especially those who are at a greater risk of exposure to blood or other potentially infectious material.

CHANGES IN THE SERUM LEVEL OF LEPTIN AND TRANSFORMING GROWTH FACTOR-Β1 IN PATIENTS WITH ARTERIAL HYPERTENSION ON A BACKGROUND OF ABDOMINAL OBESITY.

The aim: Study of the levels of leptin and the growth modulator TGF-β1 in the blood serum of patients with hypertension, which occurs on the background of AO and without it. Materials and methods: Carbohydrate metabolism was studied by the enzymatic method, the level of insulin in the blood (by the enzyme immunoassay method), the oral glucose tolerance test and the calculation of the NOMA index. Results: The data obtained in the work indicate a significant role of leptin in the formation of hypertension itself and the development of obesity, carbohydrate and lipid metabolism disorders. The increased level of transforming growth factor-β1 in the blood of such patients can be used as a fairly informative marker of the unfavorable prognosis of these diseases. Conclusions: 1. In the control group, there was a significant increase in the initial values of heart rate, average levels of SBP and DBP, the frequency of hy-percholesterolemia and insulin resistance was established. 2. Significant disorders of lipid and carbohydrate metabolism and leptin synthesis were found in patients with hypertension, which occurs against the background of AO. 3. When analyzing the level of leptin depending on gender, a statistically significant increase in the level of blood leptin was found in the group of women with AH with AO compared to women with AH without AO and the control group. 4. A significant increase in the level of transforming growth factor-β1 in blood serum of patients with hypertension was established.

STUDY OF IMMUNE-INFLAMMATORY RESPONSE CHANGES IN ORAL FLUID IN PATIENTS WITH DISEASES OF PERIODONTAL TISSUES IN COMBINATION WITH GENERAL SOMATIC PATHOLOGY.

The aim: To determine the concentration of markers of the immune-inflammatory response (IL-1β, IL-10, IL-1β / IL-10, hsCRP) in oral fluid in patients with diseases of periodontal tissues in combination with general somatic pathology. Materials and methods: The study was conducted at Danylo Halytsky Lviv National Medical University, Department of therapeutic dentistry FPGE, Lviv, Ukraine. The patients were divided into two groups: the main group - 144 patients (with periodontal tissue diseases on the background of general somatic pathology) and the control group - 30 somatically and dentally healthy persons, in whose oral fluid was determined the concentration of IL-1β, IL-10, hsCRP by the enzyme immunoassay method. Results: As a result of our research, it was found that in people with periodontal tissue diseases, against the background of general somatic pathology, there is an activation of the immune-inflammatory response, which aggravates the course of general somatic and dental diseases in this contingent of patients. Conclusions: Therefore, in patients with periodontal tissue diseases on the background of somatic diseases, a significant increase in the level of the pro-in-flammatory cytokine IL-1β and hsCRP was determined against the background of a decrease in the anti-inflammatory cytokine IL-10 in the oral fluid compared to the values in the control group.

РАПСОВЫЙ ЖМЫХ И ШРОТ: ПЕРВЫЕ РЕЗУЛЬТАТЫ МИКОТОКСИКОЛОГИЧЕСКОГО ОБСЛЕДОВАНИЯ

The article presents the results of a survey of 26 production batches of rapeseed cake and meal for microscopic fungi and mycotoxin contamination. The indicator of the total number of fungi in most samples corresponded to the regulated norm of 5000 CFU/g, Penicillium spp., Aspergillus spp. prevailed in the composition of the mycobiota, Cladosporium spp., Scopulariopsis spp. were detected less often, and flour fungi and yeasts were found in small amounts. The presence of the anthraquinone toxin emodin in all samples was established by the method of indirect competitive enzyme immunoassay, alternariol, mycophenolic acid and ergoalkaloids were detected somewhat less frequently, in isolated cases – aflatoxin B1, cyclopiazonic acid and T-2 toxin. The prospects for further research and the need to assess the potential of toxin formation by sanitary-significant micromycetes are discussed.

CLINICAL EFFICACY OF TREATMENT OF WOMEN WITH CHRONIC RECURRENT UROGENITAL CHLAMYDIA INFECTION.

Aim: To to increase the efficacy of the treatment of women diagnosed with complicated urogenital chlamydia infection based on the study of the immune status, clinical and pathogenetic features of the course, development and implementation of pathogenetically substantiated therapy methods. Materials and methods: Laboratory diagnostics of chlamydia infection was conducted with the use of direct immunofluorescence, enzyme immunoassay and cytological method. The dynamics of clinical symptoms was studied in the patients of Group I (64 women) who received Wobenzym in combination with Doxycycline and Group II (64 women) who were treated with conventional therapy (CT). Results: Clinical effectiveness of the treatment of women in Groups I and II was analyzed. The treatment according to our developed method was well tolerated by all the patients. No manifestations of the disease exacerbation which could be associated with the inclusion of these drugs in the comprehensive treatment were noted. The insignificant effectiveness of our treatment in the examined patients with complicated urogenital chlamydia infection in Groups I and II was distributed as follows: only 1 (1.56%) woman (Group I) had an insignificant improvement in clinical manifestations after the treatment according to our method. An insignificant improvement in the clinical manifestations after the treatment with conventional therapy (Group II) was observed in 4 women constituting 6.25%. Unfortunately, a small percentage of women in Groups I and II with no changes in clinical symptoms after the treatment was observed. In particular, the absence of treatment effectiveness was noted in 3 (4.69%) women after the application of our method and in 6 women (9.38%) who were treated by the conventional therapy. Therefore, these women were prescribed an additional course of treatment. Conclusions: The dynamics of clinical symptoms was found to be significantly more pronounced and fast in the patients of Group I compared to the patients in Group II who were treated with conventional therapy.

Association of Proliferative Diabetic Retinopathy in the Elderly with the Blood Complement System

A common complication of diabetes mellitus, mainly type 2, is diabetic retinopathy, among which the most unfavorable form for complete loss of vision is considered to be proliferative diabetic retinopathy. The participation of the complement system in the development of proliferative diabetic retinopathy is mainly devoted to experimental work on a limited number of blood complement components and without assessing their risk in the pathogenesis of the disease. The purpose of the study was to determine the association of components of the blood serum complement system with the development of proliferative diabetic retinopathy in the elderly. In clinical conditions, 115 patients 60–74 years old suffering from proliferative diabetic retinopathy and 48 patients of the same age with the absence of this ophthalmopathology were examined. The components of the blood complement system were studied by enzyme immunoassay and hemolytic method. The relative risk of the influence of the complement components was calculated according to the generally accepted method. There was a statistically significant increase in the blood serum of patients with proliferative diabetic retinopathy of most components of the complement system, with the exception of the C1 ing. and C5 components. The content of the C3a component increased especially to 127.6 ± 4.7 ng / ml versus 30.4 ± 3.5 ng/ml in the control, the C5a component to 5.6 ± 0.5 ng/ml versus 2.4 ± 0.3 ng/ml, and factor H to 228.7 ± 4.9 versus 106.3 ± 3.8 mng / ml, respectively. The highest value of the relative risk among the studied components is inherent in the C3a component of the blood complement with a reliable confidence interval of 4,451–5,103. The development of proliferative diabetic retinopathy in the elderly is associated with an increased content of C3a, C5a components and factor H in the blood serum, which can be used to develop targeted therapy for this disease.

JS-HR-9: A NEW ERA OF PROGRESS IN PRIMARY ALDOSTERONISM TREATMENT, A NEW ALDOSTERONE ASSAY, AND A CLINICAL PRACTICE GUIDELINE

Primary aldosteronism (PA), one of the mineralocorticoid receptor (MR)-associated hypertension, is a highly prevalent secondary hypertension that accounts for 5–15% of all hypertension and is a severe hypertension that is prone to cerebrovascular and cardiovascular disease. Therefore, it is expected that the pathophysiology and treatment strategies for PA will be elucidated. In recent years, the major changes in PA have been in treatment and diagnosis in Japan. Bilateral or non-surgical unilateral PA is treated with mineralocorticoid receptor antagonist (MRA). Esaxerenone, a non-steroidal MRA, is now widely used in Japan for hypertension including PA. It has been reported that add-on treatment using esaxerenone with maximal tolerable doses of a renin-angiotensin system inhibitor reduced the urinary albumin creatinine ratio in patients with type 2 diabetes mellitus (ESAX-DN), suggesting a renoprotective effect against diabetic nephropathy. Several clinical studies of another non-steroidal MRA, finerenone(FIDELIO-DKD, FIGARO-DKD, and FIDELITY), have shown its renoprotective effect against diabetic nephropathy as well as cardiovascular events, particularly hospitalization for heart failure in patients with type 2 diabetes and chronic kidney disease. Although there are no reports of clinical studies on finerenone for PA, finerenone may be used in the future for type 2 diabetes patients with PA, as obesity, glucose intolerance, and sleep apnea are common complications in patients with PA. Secondly, radiofrequency ablation of macroscopic adrenal tumors has been reported as an alternative treatment for PA to lower blood pressure and plasma aldosterone levels. This treatment has been covered by health insurance providers in Japan since April 2022, but long-term outcomes need to be validated. In diagnosis, serum and urine aldosterone has been measured in Japan since early 2021 by the two-step sandwich chemiluminescent enzyme immunoassay (CLEIA) method. This new CLEIA method correlates well with LC-MS/MS values and is expected to be more accurate. As CLEIA serum aldosterone levels are lower than those measured with a previous radioimmunoassay, the Japan Endocrine Society issued the 2021 Primary Aldosteronism Clinical Guidelines, which included a provisional positive value for the diagnosis of PA. More than ever, the diagnosis of PA needs to be carefully considered. This CLEIA method is currently available only in Japan. This presentation will cover the latest treatments, diagnostic procedures, and other hot topics in PA as described above.

Sustained high HIV incidence among men who have sex with men in Jiangsu province, China: based on the limiting-antigen avidity EIA method and six consecutive surveys, 2016-2021.

The epidemic of HIV infection among men who have sex with men (MSM) is a major public health concern in some parts of China, but data on trends in HIV incidence are limited. This study aimed to examine the trends in HIV incidence and factors associated with recent HIV infection among MSM in Jiangsu province, China, based on the limiting-antigen avidity enzyme immunoassay (LAg-Avidity-EIA) method. Six consecutive surveys were implemented among MSM throughout Jiangsu province from 2016 to 2021. Participants were recruited in three ways. Socio-demographic and behavioral information were collected through face-to-face interviews. Venous blood samples were taken to test for HIV and syphilis. HIV incidence was estimated using the LAg-Avidity-EIA method. Chi-square trend tests were used to observe trends over the years. Multivariate regression analyses were used to identify factors associated with recent HIV infection. A total of 15,401 participants were enrolled in the study. The prevalence of HIV infection ranged from 8.0 to 9.8%, with no consistent rise or fall over the years (P = 0.189). HIV incidence ranged from 5.0 to 9.0%, and no uptrend or downtrend was shown (P = 0.418). MSM who lived locally for more than 2 years (aOR = 1.366, P = 0.019), had a lack of comprehensive HIV knowledge (aOR = 1.643, P = 0.031), had engaged in unprotected anal intercourse (UAI) in the past 6 months (aOR = 7.373, P < 0.001), had been tested for HIV within 12 months (aOR = 1.292, P = 0.035), and tested positive for syphilis (aOR = 2.840, P < 0.001) were likely to be recently infected with HIV. HIV incidence among MSM has remained at a high level in Jiangsu province. In China, health education, condom use, and HIV/syphilis testing should continue to be top priorities for HIV prevention among MSM to reduce HIV transmission.

Validation of dried blood spots samples for the screening of hepatitis B virus surface by enzyme immunoassay method

Dried Blood Spots (DBS) are blood collection carriers that facilitate the storage and transport of samples. Used for quality control during sero-epidemiological investigations, DBS eluate are not the conventional specimen indicated by manufacturers for enzyme immunoassay method (EIA) for hepatitis B virus surface (HBs antigen).The aim of our study was to evaluate DBS eluates used as a matrix for EIA of HBs antigen in a reference laboratory.This study took place from August 2016 to November 2017 at the Centre for Diagnosis and Research on AIDS and other infectious diseases (CeDReS) in Abidjan, Côte d'Ivoire. We used a panel of 149 whole blood samples from blood donors. The DBS performed with these samples were analyzed after elution with the HBsAg (version ULTRA) ELISA, Dia.Pro Diagnostic Bioprobes S.R.L., Sesto San Giovanni, Italy. The technical performance (sensitivity and specificity and kappa coefficient) of the test performed on DBS was determined for different ratios (optical density/threshold value) compared to the results obtained on the plasma used as reference.We obtained a sensitivity of 100% with DBS for all ratios.The specificity increased according to the ratio of optical density of the individual EIA reaction to the threshold value, with 6.09%, 47.0%, and 83.0%, respectively, for ratios of 1.0, 3.0, and 5.0. Best performance was observed at ratio of 10.0 with a sensitivity and specificity of 100%.In conclusion, DBS eluate can be used for the diagnosis of viral hepatitis B and would be useful for conducting sero-epidemiological investigations. However, ratio giving best performance must be determined for each enzyme immunoassay method kits.

Serum oxytocin correlated with later logical memory in older Japanese women: A 7-year follow-up study

ObjectivesThis study aimed to investigate the longitudinal relationship between serum oxytocin and logical memory among older adults in rural Japan and clarify sex differences in this relationship. MeasurementsThe first survey was conducted from October 2009 to March 2011 (Time 1) and the second from November 2016 to September 2017 (Time 2). The final analysis for Time 1 included 385 participants (median age 75 years, interquartile range [IQR] 70–81 years) and that for Time 2 included 76 participants (median age 80 years, IQR 76–83 years). We assessed cognition, logical memory, and living conditions, and measured serum oxytocin levels. Logical memory was evaluated using the Wechsler Memory Scale-Revised Logical Memory II delayed recall part A (LM II-DR). Serum oxytocin was measured using the enzyme immunoassay method. ResultsThe median (IQR) oxytocin level among men (n = 20) was 34 (16–78) pg/mL at Time 1 and 53 (28–140) pg/mL at Time 2. The median (IQR) oxytocin level among women (n = 56) was 117 (35–412) pg/mL at Time 1 and 76 (32–145) pg/mL at Time 2. The median oxytocin level among women at Time 2 was significantly lower than that at Time 1 (p = 0.004). The multivariate analysis showed that for women, LM II-DR score at Time 2 was positively associated with oxytocin level at Time 1 (p = 0.042) and negatively associated with age (p = 0.02). ConclusionsOur study suggests that maintaining high oxytocin levels in older women may prevent age-related decline in logical memory.

Performance evaluation of the bio-rad BioPlex 2200 multiplex system in the detection of measles, mumps, rubella, and varicella-zoster antibodies

BackgroundMeasles, mumps, rubella, and varicella-zoster (MMRV) immunity testing is important in occupational screening of healthcare workers and at-risk populations. ObjectivesThe goal of this study was to compare the performance of the Bio-Rad BioPlex 2200 MMRV multiplex fluorescence immunoassay (MFI) against two enzyme immunoassay (EIA) methods: the Bio-Rad Evolis Twin Plus measles, mumps, and varicella-zoster (MMV) IgG assay and the Abbott Architect Rubella IgG assay. Study DesignClinically uncharacterized serum specimens were obtained and analyzed using the Bio-Rad BioPlex assay and compared against the EIA methods. ResultsThe Bio-Rad BioPlex demonstrated total agreement of 85.5% (95% confidence interval (CI), 78.0 to 90.7%), 92.7% (95% confidence interval (CI), 86.7 to 96.1%), 92.4% (95% confidence interval (CI), 86.9 to 95.7%), and 98.8% (95% confidence interval (CI), 93.7 to 99.8%) for measles, mumps, varicella-zoster, and rubella, respectively, against the current EIA methods. Furthermore, precision testing agreed with the manufacturer's package insert in 10 of 13 pooled samples. ConclusionThese data indicate that the Bio-Rad BioPlex has comparable performance to the EIA methods.

Plasma levels of galectin-3 in residents of the Podillya region of Ukraine without signs of cardiovascular pathology carriers of different variants of the coding gene (LGALS-3, rs 2274273)

Annotation. The purpose of the study is to investigate the plasma level of galectin-3 in postmenopausal women without signs of cardiovascular pathology and its possible association with the structural and functional parameters of the myocardium in carriers of different polymorphic variants of the coding gene (LGALS-3, rs2274273). 67 women of postmenopausal age (average age 56.43±0.64 years) without signs of cardiovascular pathology residents of the Podillya region of Ukraine were examined. In addition to the general clinical examination, the research used the enzyme immunoassay method to determine the level of galectin-3 in blood plasma, genotyping of the LGALS3 gene at the rs2274273 locus by means of polymerase chain reaction, and ultrasound of the heart. Statistical processing of the obtained results was carried out using the package of statistical programs SPSS, STATISTICA v. 10.0. By means of correlation analysis with the calculation of Spearman's rank correlation coefficients (r), an assessment of the possible association of the plasma level of the biomarker and the structural and functional indicators of the heart was carried out. The critical value of the significance level (p) was ≤0.05. In the examined population of women, there was no significant difference between the frequency of occurrence of the GA and GG genotypes carriers of the galectin-3 gene locus rs2274273: 49.25% (n=33) and 40.30% (n=27), respectively (p&gt;0.05). The AA genotype variant was found in 10.45% (n=7) of individuals, which is significantly less frequent than both the GA and GG genotypes (p&lt;0.01). The frequency of distribution of genotypes corresponded to the Hardy-Weinberg equilibrium. The average level of galletin-3 in blood plasma in women was 6.68±0.30 ng/ml, it was within the generally accepted norms and did not reliably differ from the corresponding indicators in men of the same region. No significant difference in plasma concentrations of galectin-3 was found in carriers of different polymorphic variants of the galectin-3 gene. Correlation between the level of galectin-3 in the blood plasma and most indicators of intracardiac and systemic hemodynamics in women without cardiovascular pathology was not found. In the future, it is planned to study the formation of prerequisites for chronic heart failure (CHF) in women with EH based on indicators of the plasma level of galectin-3 in carriers of polymorphic variants of the coding gene (LGALS-3, rs2274273).

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 &lt; 0.05), 1 (2.00 %) vs. 9 (15.00 %) at 390 ng FEU/ml cut-off (χ2 = 4.16, P &lt; 0.05), 0 (0.00 %) vs. 7 (11.67 %) at age-adjusted cut-off (χ2 = 4.43, P &lt; 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.

The relationship of melatonin concentration in preterm infants and adverse outcomes in the late neonatal period.

The aim of research was to assess the melatonin concentrations in the early neonatal period as a predictor of adverse outcomes of late neonatal period in preterm infants and to estimate its optimal predictive cut-off values. A total of 115 preterm infants admitted to the neonatal intensive care unit were screened for eligibility, five did not meet the criteria, six parents declined the participation. So, a total of 104 preterm infants with gestational age 25-34 weeks were included in research. The concentration of melatonin in urine was determined by the Enzyme Immunoassay method (Human Melatonin Sulfate ELISA kit, Elabscience, China). The Mann-Whitney U-test and analysis of the receiver operating characteristic (ROC) curve were used in statistical analysis. Analysis of the ROC curves has revealed optimal cut-off values for urinary melatonin concentration to predict late outcomes. Melatonin concentration below 3.58 ng/ml with sensitivity of 72% can predict development of retinopathy of prematurity (ROP) (AUC = 0.73; 95% confidence intervals (CI) 0.61-0.86). Good diagnostic accuracy (AUC = 0.80; 95% CI 0.67-0.93) has been shown for bronchopulmonary dysplasia (BPD). The optimal cut-off value for melatonin concentration in BPD prediction is 3.71 ng/ml (sensitivity 80%, specificity 64%). Urinary melatonin concentration below 3.79 ng/ml can be associated with late-onset sepsis (AUC = 0.76; 95% CI 0.64-0.87; sensitivity 72%; specificity 62%). There were no significant associations between melatonin concentration and necrotizing enterocolitis (P = 0.912). Urinary melatonin concentration below the certain cut-off values in the early neonatal period may serve as one of the predictors of adverse outcomes such as BPD, ROP, and late-onset sepsis in the late neonatal period in preterm infants.

An Overview of ELISA-Based Initial Velocity Methods to Measure the Immunoreactive Fraction, Association Rate, And Equilibrium Constants of Monoclonal Antibodies

Three biochemical parameters—the on-rate (k+1), the dissociation rate (k-1), and the equilibrium (Ka) constants—describe the antibody interaction with a ligand. In addition, the antibody immune reactive fraction (IRF) is important because it affects k+1 and Ka constant values. The on-rate and dissociation constants are kinetic, but the equilibrium constant is a thermodynamic parameter, and the methods for their determination are different. Among the available methods to determine these parameters, only surface plasmon resonance (SPR)-based techniques allow calculation of the three antibody constants in a single experiment. However, conventional immunoassays are also suitable to determine antibody constants. Here, we describe some enzyme immunoassays (ELISA) methods developed in our laboratory to, i) quantitate mAb in culture supernatants from the initial velocity of the progress curve, ii) determine mAb IRF by a two-step ELISA method, iii) calculate mAb on-rate constant by kinetic ELISA based on time-course data of ligand binding to capture mAb, and, iv) describe a competitive-inhibition ELISA to measure the inhibition constant (Ki) of mAb binding to plate-bound ligand, in the presence of competing concentrations of mAb and soluble ligand.

Detection of immunoglobulins specific to SARS-CoV-2 virus S-protein in mixed saliva

BACKGROUND: Since the beginning of the COVID-19 pandemic, scientists have been concerned about finding an effective alternative to blood-based diagnostics of antibodies to SARS-CoV-2. According to the results of numerous studies by domestic and foreign authors, saliva is a valuable diagnostic material. Mixed saliva implies the possibility of painless and non-invasive sampling and opens up great opportunities for diagnosing COVID-19 in vulnerable populations, such as children.&#x0D; AIM: Development of a method for detecting immunoglobulins specific to the SARS-CoV-2 virus in saliva after infection with COVID-19 and vaccination.&#x0D; MATERIALS AND METHODS: The study involved 43 people aged 20 to 67 years, divided into 3 groups: recovered from COVID-19; those who believe they have not had COVID-19; vaccinated with various drugs against COVID-19. Samples of unstimulated mixed saliva and capillary blood from a finger were obtained from each subject, which were subsequently examined by methods of qualitative and quantitative enzyme immunoassay.&#x0D; RESULTS: A high level of correlation between the content of local IgG and IgA in saliva was demonstrated, while the relationship between the content of IgA in blood and saliva was shown to be of medium strength. A higher number of seropositive patients was demonstrated in the group vaccinated with vaccines against SARS-CoV-2, compared with those who recovered from COVID-19, which is consistent with the anamnesis data.&#x0D; CONCLUSIONS: This work demonstrates and confirms that saliva in the oral fluid is a valuable diagnostic material for the study of local antibodies. The data obtained will allow the development of unique test systems for studying humoral immunity to the SARS-CoV-2 virus, which can become an alternative replacement for the usual determination of virus-specific immunoglobulins in the blood.

Features of the epidemiological history of patients with Lyme borreliosis

In the treatment of Lyme borreliosis (LB), early diagnosis is a key component. The epidemiological history of the patient plays one of the leading roles in suspected borreliosis and at the same time is an important criterion for clinical diagnosis.Purpose. Analysis of epidemiological history data in outpatient cases of LB in Moscow.Material and methods. We conducted a retrospective, non-randomized, single-center cohort study, based on the analysis of data from 660 outpatient records of patients, referred by local physician with tick bite or suspicion of LB, and consulted by an infectious disease specialist on the basis of the Infectious clinical hospital No.1 in Moscow. For further analysis, patients were divided by age and the presence or absence of LB. Confirmation of LB was based on clinical and/or laboratory criteria using methods of enzyme immunoassay and immune blot.Results. We have updated the epidemiological features of the LB for Moscow, such as: the relative number of cases in the administrative districts of Moscow, places and seasonality of tick bites. Among children, compared with adults, cases of a tick bite without the development of LB was predominant, which must be taken into account in order to avoid overdiagnosis of LB. In patients with LB, an indication of the fact of tick bite in the anamnesis was much less common than going to its habitats.Conclusion. Our data clarify the epidemiological features of LB for residents of Moscow. In the absence of a history of indications of tick bite, an informative sign for clinical diagnosis is a visit to its habitats, including not only trips to wooded areas, but also to suburban areas and parks.

Diagnosing myeloma in general practice: how might earlier diagnosis be achieved?

<h3>Abstract</h3> <h3>Background</h3> Netrin-1 is a laminin class protein that guides the axonal during the first embryonic development, has pushing and pulling properties, and has axonal chemoattractant activity. Netrin-1 has been shown to increase the effect of fibrosis in mouse lung and human SSc lung cell cultures. In this study, we aimed to investigate the relationship between Netrin-1 and Systemic sclerosis (SSc) and to emphasize the role of Netrin-1 in the pathophysiology of SSc by increasing the known VEGF and M2 macrophage expression, which supports the fibrotic process. <h3>Methods</h3> The study included 56 SSc patients with a mean age of 48·.08±13.59 years and 58 healthy volunteers with a mean age of 48.01±11.59 years. SSc organ involvements were scanned retrospectively from patient files, and patients were grouped according to SSc complications. Calculation of Netrin-1 levels was performed using a quantitative sandwich enzyme immunoassay method with an ELISA kit (Elabscience, Texas, USA; catalog number: E-EL-H2328; lot number: GZWTKZ5SWK). The modified Rodnan skin score (mRSS) was used for skin thickness scoring in SSc patients. <h3>Results</h3> The median value of Netrin-1 was found to be significantly higher in SSc (268.8 [82.75-1006.64]) than in controls (108.63 [21.02-351.49]) (p&lt;0.0001). In ROC analysis, a cut-off value of 354.24 for Netrin in SSc was found to provide a sensitive confidence interval with 32.8% sensitivity and 98.3% specificity (AUC[95% CI]: 0.746-0.895, p&lt;0.0001). There was no significant correlation between Netrin-1 level, organ involvement in SSc, and mRSS (p&gt;0.05). <h3>Conclusion</h3> We found a significant relationship between Netrin-1 levels and SSc disease in this study. Our study is the first clinical study in which Netrin-1 elevation was demonstrated in SSc patients.