Enzyme Immunoassay Method Research Articles

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.

Enzyme immunoassays for the detection of mycotoxins in plant-based milk alternatives: pitfalls and limitations

Plant-based milk alternatives (PBMAs) are a potential source of mycotoxin uptake. To ensure food safety, simple and rapid testing methods of PBMAs for mycotoxins are therefore required. This study investigated the applicability of enzyme immunoassay (EIA) methods for direct testing of PBMAs without sample extraction. Mycotoxin analyses included aflatoxin B1 (AFB1), sterigmatocystin (STC), ochratoxin A (OTA), deoxynivalenol (DON), and T-2/HT-2-toxin (T-2/HT-2). It was found that the PBMA matrix negatively affected the EIA to varying degrees, thus affecting the reliability of the results. A dilution of PBMAs of at least 1:8 was necessary to overcome matrix interference. This resulted in calculated detection limits of 0.4 µg/L (AFB1), 2 µg/L (STC), 0.08 µg/L (OTA), 16 µg/L (DON), and 0.4 µg/L (T-2/HT-2). After analysis of 54 PBMA products from German retail stores, positive results in at least one test system were obtained for 23 samples. However, most positive results were near the calculated detection limit. Control analyses of selected samples by LC–MS/MS for AFB1, STC, and OTA qualitatively confirmed the presence of trace amounts of STC in some samples, but quantitative agreement was poor. It was concluded that the high diversity of ingredients used in PBMAs led to a highly variable degree of sample matrix interference even in a 1:8 dilution. Since the use of higher dilutions conflicts with the need to achieve low detection limits, the application of EIA for routine mycotoxin analysis in PBMA for mycotoxins requires further study on the development of a feasible sample preparation method.

Phenytoin concentration in people with epilepsy: a comparative study in serum and saliva

In clinical practice, therapeutic drug monitoring (TDM) makes it possible to measure the concentration of drugs in serum or saliva, the purpose of which is to reduce adverse effects and optimize pharmacological therapy. The objective was to determine the concentrations of Phenytoin in saliva and serum of people with epilepsy. Cross-sectional, descriptive study with dynamic recruitment of 30 people with epilepsy (n = 30; 17 men, 56.7% and 13 women, 43.3%; mean age 33.9 ± 11.83 years). Serum and saliva samples were collected at trough levels from patients, who were under phenytoin treatment for at least three months. Drug levels were assessed by the Cloned Donor Enzyme Immunoassay method. Phenytoin levels were found in saliva between 0.01 to 3.56 mg/L and in serum between 0.09 to 36.60 mg/L. Pearson’s analysis showed an association between the estimated serum and saliva phenytoin concentrations (R2 0.7026; 95% CI 0.685-0.921), with a significant statistical correlation (p &amp;lt; 0.05). The Bland-Altman test broke concordance, the difference between the two saliva/serum methods is within 95% confidence. It is concluded that there is an association and concordance between the concentrations of phenytoin in serum and saliva, therefore, this technique can be useful in the clinical monitoring of phenytoin.

Myocardial infarction and cannabis consumption: Two years of data collection in CHU Amiens-Picardie

Cannabis is the most illicit drug consumed in France, with tetrahydrocannabinol (THC) levels increasing over past decades. The lack of epidemiological data concerning myocardial infarction (MI) occurring under cannabis use raises the question of imputability of THC as a cause of these serious cardiovascular events. In this context, it is essential to understand patient's characteristics in the North French region, where cardiovascular morbidity and mortality (20%) are greater than entire France (ARS, 2017). We performed a retrospective clinical study including patients in the Amiens University Hospital cardiac intensive care unit (CICU) for MI in a context of cannabis consumption between January 2020 and March 2022. Cannabis status was first determined using enzyme immunoassay method in urines (Atellica™ CH (Siemens®). Then blood quantifications of THC, 11-OH-THC and THC-COOH by LC-MS/MS (8060 Shimadzu®) was operated in case of positivity of urine samples. One-hundred-and-five patients were included. Median delay between hospitalisation and sample collection was 5 h 50. Twenty-nine patients (27.6%) were positive for cannabis urine testing. Their mean age was 34.5 years and 82.8% were men. Blood concentrations for THC, 11-OH-THC and THC-COOH were performed for 22 patients (with 7 uncollected blood samples). Eighteen were blood positive with respective mean (min; max) concentrations at 1.29 ng/mL (0.2; 3.7), 1.07 ng/mL (0.3; 6.7) and 21.7 ng/mL (1.3; 88.3), meaning patients were under cannabis influence during blood sampling. Similar results in literature showed that MI-cannabis induced risk population is mostly young men (Yang, Am J Med, 2020, 5:605–612). Knowing that chest pain occurs only few hours after smoking (Chetty, CJC Open, 2020, 1:12–21) and THC is rapidly relocated in organs (Goullé, Ann Pharm Fr, 2008, 4:232–244), it can be demonstrated that the patient's THC positivity is due to a short delay between smoking, chest pain and blood test. Ten blood samples were collected less than one hour after hospitalisation, revealed a well-known patients management of MI in young adult population by emergency and CICU services. Cannabis consumption declarative habits were collected for 14 patients: 7 patients smoked more than one joint per day (1 to 10) and the others smoked more than 1 joint per week. However, a very few data about cannabis kinetic were available, except time-lapse between chest pain and blood dosage for 9 patients, with a mean time of 16 hours (2h10; 48 h), with however a great variability. No patient was positive for others illicit drugs. Regarding clinical and biological data, mean body mass index was 26.41 kg/m2 and mean blood pressure was normal (121/81 mmHg). Nevertheless, cardiac biomarkers were impaired with troponin peak at 48,550 ng/L (3; 376890) and myoglobin at 723 μg/L (21; 7890). Tobacco use was the most frequently risk factor found in our patients: 93% were current smokers, as seen in others clinical studies (Aggarwal, JRSM Cardiovasc Dis, 2012, 1:8). However, it is difficult to determine whether tobacco or cannabis is the cause of MI due to the consumption mode (Chetty, CJC Open, 2020, 1:12–21). Other cardiovascular risk factors were identified: 7 obese patients, 5 high blood pressure patients and 4 diabetic patients. Left ventricular ejection fraction (LVEF) was preserved (LVEF > 50%) for 25 patients. Stent placement was the most common medical treatment. Unfortunately, one death has occurred. One of the limits of the study is the low number of patients. This study shows that characteristics of patients with MI induced by cannabis in North French region is similar to other global clinical studies. However, if cannabis use is lower than the total French average, risk factor prevention is essential in the French region with high cardiovascular morbidity.

An autopsy case of fatal insulin preparation overdose

Insulin preparations, a group of drugs used to treat diabetes, are of great forensic interest because they are difficult to detect in blood while they are sometimes intentionally administered in large doses for suicide or murder. In this study, we validated a quantification method for insulin glargine and insulin lispro using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the blood sample from a case of suicidal overdose with insulin. Case: A woman in her 30s had a history of mental illness and diabetes. A day before her death, she informed her partner that she had self-administered large doses of insulin preparations and prescription drugs. Bovine insulin was used as the internal standard (IS). The blood sample (250 μL) was added to the diluent and IS solution, followed by 250 μL each of hexane and N,N-dimethylformamide, and the mixture was centrifuged (10000 g × 10 min). The supernatant was collected and 900 μL of 5% ammonium solution was added. The extract was analyzed using Nexera 30A, LCMS8060 (Shimadzu), L-column3 ODS C18 metal-free column (150 × 2.1 mm, 3 μm), ionization method: electrospray ionization+, multiple reaction monitoring conditions: insulin glargine [M + 6H]6+ 1011.5 > 1179.4, insulin lispro[M + 6H]6+ 968.9 > 217.0, bovine insulin (IS) [M + 6H]6+ 956.3 > 1120.9. The autopsy revealed no evidence of fatal injuries. Coronary artery stenosis was observed as a finding of endogenous disease, but there were no significant histopathological changes in the myocardium. Biochemical tests revealed the following: blood glucose level, 6 mg/dL; glucose in the vitreous region, 6 mg/dL; glycated hemoglobin (HbA1c), 11.2%; blood insulin level estimated using the chemiluminescent enzyme immunoassay method, 1.03 μU/mL (6.27 pmol/mL); and C-peptide concentration, 0.6 ng/mL (16.8 pmol/L). A drug screening test and simple quantification using LC-MS/MS detected many prescription drugs, but none of them were at intoxicating concentrations. The insulin analysis showed insulin glargine concentrations of 429 μU/mL in femoral blood and 1362 μU/mL in cardiac blood, and insulin lispro was detected in femoral and cardiac blood below the lower limit of quantification (<50 μU/mL). The cardiac/peripheral blood ratio, a measure of postmortem redistribution, was 3.2. Although the therapeutic ranges of insulin glargine and insulin lispro have not been defined, the femoral and cardiac blood concentrations of insulin lispro in this case was lower than the serum concentration (155 μU/mL) listed in the package insert. However, while the femoral and cardiac blood insulin glargine concentrations were considerably higher than the serum concentration (15 μU/mL) listed in the package insert. Considering that insulin is easily degraded in whole blood due to hemolysis, it is possible that insulin concentrations were even higher at the time of administration and death than those observed in autopsy specimens on the third postmortem day. However, since insulin lispro is a fast-acting analog of insulin (1–3 hours), it may have contributed to hypoglycemia but was not the direct cause of death, and hypoglycemia due to insulin glargine overdose was the most likely cause of death in this case. We quantified insulin lispro and insulin glargine in a case of suicide to determine insulin glargine's involvement in causing death. In future, it is necessary to examine postmortem degradation and redistribution of insulin and consider the appropriate concentration in postmortem samples of insulin poisoning.

Laboratory diagnostics and pathogenetic therapy of pneumococcal respiratory tract infections

The article presents the results of diagnostic measures and pathogenetic therapy of pneumococcal infections of the respiratory tract. All patients included in the examination underwent X-ray examination or computed magnetic resonance imaging of the paranasal sinuses. 100 % of the subjects were examined for the presence of antibodies to Human gamma herpesvirus 4 (enzyme immunoassay method), the study of blood serum for IgM and IgG and the test for determining the avidity of Ig G antibodies to γ-herpesviruses. The data obtained indicate the effectiveness of pathogenetic therapy in the treatment of pneumococcal infections of the upper respiratory tract.

Identification of the risk of IHD development in young people, formation of a diagnostic model

Background and Aims : To study the level of biomolecules characterizing the main pathogenetic links of coronary atherosclerosis. To develop a model for diagnosing the risk of coronary heart disease in young people based on significant biochemical parameters.Methods: The study included 49 people with IHD and 127 people without IHD. All participants signed an Informed Consent. A survey was conducted using validated questionnaires. Anamnestic data, anthropometry, functional examinations were collected from the all participants. Total antioxidant activity, MDA-oxLDL, SOD, MCP-1, TNF-alpha, IL-1, IL-6, IL-8, total cholesterol, TG, HDL and LDL were determined in blood serum using enzyme immunoassay, multiplex assay and enzymatic method.Results: The analysis of the data obtained revealed a difference in concentrations between the studied subgroups for the total antioxidant activity, SOD, MCP-1 and IL-6. In patients with IHD, the level of the total antioxidant activity was significantly lower by 2.9 times and MCP-1 by 1.4 times, the concentration of SOD and IL-6 was higher by 16 and 30%, respectively. For interpretation, we used a scatter diagram of the density of values - SHAP, reflecting the degree of influence of each parameter on the resulting variable. The greatest contributions to the model project were made by: the total antioxidant activity, SOD, MCP-1, IL-6 and HDL.Conclusions: The data obtained by us can be recommended for the development of a coronary heart disease risk meter in young people, which includes the most significant laboratory parameters. This study was conducted within the framework of the project RSF 21-15-00022. Background and Aims : To study the level of biomolecules characterizing the main pathogenetic links of coronary atherosclerosis. To develop a model for diagnosing the risk of coronary heart disease in young people based on significant biochemical parameters. Methods: The study included 49 people with IHD and 127 people without IHD. All participants signed an Informed Consent. A survey was conducted using validated questionnaires. Anamnestic data, anthropometry, functional examinations were collected from the all participants. Total antioxidant activity, MDA-oxLDL, SOD, MCP-1, TNF-alpha, IL-1, IL-6, IL-8, total cholesterol, TG, HDL and LDL were determined in blood serum using enzyme immunoassay, multiplex assay and enzymatic method. Results: The analysis of the data obtained revealed a difference in concentrations between the studied subgroups for the total antioxidant activity, SOD, MCP-1 and IL-6. In patients with IHD, the level of the total antioxidant activity was significantly lower by 2.9 times and MCP-1 by 1.4 times, the concentration of SOD and IL-6 was higher by 16 and 30%, respectively. For interpretation, we used a scatter diagram of the density of values - SHAP, reflecting the degree of influence of each parameter on the resulting variable. The greatest contributions to the model project were made by: the total antioxidant activity, SOD, MCP-1, IL-6 and HDL. Conclusions: The data obtained by us can be recommended for the development of a coronary heart disease risk meter in young people, which includes the most significant laboratory parameters. This study was conducted within the framework of the project RSF 21-15-00022.

Clinical significance of dynamics between oxidative stress levels during mechanical lung ventilation in premature infants with persistent pulmonary hypertension

The aim of this study was to determine the clinical significance of the levels and dynamics of the urinary 8-hydroxy-2-deoxyguanosine, degree of pulmonary hypertension on choosing the duration and form of respiratory support in premature infants with respiratory distress syndrome in combination with perinatal asphyxia.&#x0D; Materials and methods. The levels of the urinary 8-hydroxy-2-deoxyguanosine (8-OHdG), ng/ml were determined by enzyme immunoassay (ELISA) method and the degree of pulmonary hypertension (PH) - by echocardiography in 60 premature new-borns at 26-32 weeks of gestation on the 1st and the 3rd–5th days of life. A comparative analysis of indicators was carried out in 2 groups: I – 32 children with respiratory distress syndrome (RDS); II – 28 children with RDS combined with perinatal asphyxia. All infants received a respiratory support, depending on the clinical condition, blood gas analyses, and the chest X-ray.&#x0D; Results. The level and dynamics of the urinary 8-OHdG correlated to degree of PH, form, and duration of respiratory support. Children of the group II on the 1st day of life had echocardiogram indicators of mild PH and indicators of the urinary 8-OHdG 2.27±0.39 ng/ml; by 3rd–5th days of life – level of the urinary 8-OHdG increased to 4.10±0.42 ng/ml and the level of PH – to moderate, required 76.2 % longer respiratory support compared to children of the group I.&#x0D; Conclusions. Prematurely born infants with RDS in combination with perinatal asphyxia have a higher oxidative stress (OS) levels and more significant pulmonary hypertension, which requires a longer respiratory support by 76.2 % and more often use of high frequency oscillatory ventilation (HFOV) by 9.3 %

Herpes Simplex Virus Seroprevalence among Pregnant Finnish Women and Their Spouses-A Six-Year Follow-Up Cohort Study.

The aim was to evaluate the herpes simplex virus (HSV) seroprevalence and seroconversion among 285 pregnant women and their 120 male spouses in Finland during a six-year follow-up (FU) between 1998–2008. We also studied the effect of sexual habits, pregnancy, and other demographic factors on the acquisition of HSV infection. Combined HSV-1 and HSV-2-IgG antibodies were assessed in the first baseline serum samples with an indirect enzyme immunoassay method. The individuals with seronegative or borderline HSV serology at baseline were additionally tested using their latest FU serum sample available. The overall HSV seroprevalence during the FU was 58.9% (168/285) among the women and 53.3% (64/120) among their spouses. The seroconversion rate was 11.4% (15/132) and 12.5% (8/64) among women and their spouses, respectively. Both spouses were HSV seropositive in 39.2% (47/120). To determine the HSV-2 seroprevalence, we also tested all HSV-seropositive participants using HSV-2-specific antigen. HSV-2 seropositivity was detected in 10.9% (44/405) of the participants. The age (p = 0.006) and history of genital warts (p = 0.006) of the women were associated with combined HSV-1 and/or HSV-2 seropositivity, while a younger age was related to HSV seroconversion (p = 0.023). Among the male spouses, HSV seropositivity was associated with the practice of oral sex (p = 0.033). To conclude, women of childbearing age acquire primary HSV infections and the presence of HSV in oral epithelium is common among HSV-seropositive individuals.

The efficiency of using maternal markers in early detection and management of congenital anomalies in Majmaah city, KSA

Nowadays the serum analytic investigations have become the first step, noninvasive, detecting method of choice of foetal abnormalities specially for at-risk patients.This study volenteers were Saudi pregnant women in King Khalid General Hospital (KKGH) at Majmaah city, Riyadh, Saudi Arabia.The study intended to judge and evaluate using the concentrations of Beta Human Chorionic Gonadotropin (βHCG) in serum as efficient maternal markers for early detection as well as management of foetal abnormalities and pregnancy complications.A total of 201 serum samples were studied. 101 sera of pregnant women were taken in the first trimester, while the other 100 non-pregnant females were acted as control volenteers. The sensitive Enzyme immunoassay (ELISA) method was applied for quantitative scoring of beta HCG levels. Pregnant women were followed up by a consultant obstetrician till delivery. Trisomy 21 and blighted ovum disorders were selected as cases of interest.Serum βHCG concentrations multiple of median (MoM) for the mothers of affected babies compared with the healthy ones. Defrences were significant as p: value showed 0.001 for each appraisal.Non-pregnant serum beta HCG (2.1 IU/ml) compared with only three weeks of gestation (576 IU/ml) for early detection of pregnancy. Upon comparison a significant high level of β HCG was recorded for the three weeks gestation with (p,value:0.001) which clearly indicates the existence of pregnancy.The use of β HCG could be the method of choice for early detection of pregnancy. This will save the embryo life during the first trimester of gestation especially in the radiology departments.

Antibody Response to SARS-CoV-2 Vaccination in Patients With Lymphoproliferative Disorders and Plasma Cell Dyscrasias: Anti-Lymphoma Therapy as a Predictive Biomarker of Response to Vaccination.

We retrospectively analyzed SARS-CoV-2 vaccination antibody responses in a cohort of 273 patients with lymphoproliferative disorders or plasma cell dyscrasias who were seen at a single tertiary cancer center. Semi-quantitative anti-spike protein serologic testing was performed with enzyme immunoassay method. We found that the antibody response rate to SARS-CoV-2 vaccination was 74.7% in our patient cohort with no difference based on gender, age or race. The highest response rate was found in patients with Multiple Myeloma (MM) (95.5%). The response rates found in Diffuse Large B-Cell Lymphoma (DLBCL), Chronic Lymphocytic Leukemia (CLL), and Low-Grade Non-Hodgkin Lymphoma (LG-NHL) were 73.2%, 61.5% and 53% respectively. We also evaluated the effects of receiving active chemo-immunotherapy on SARS-CoV-2 vaccination antibody response. We found that the patients on treatment had lower response than the patients off treatment (62.1% versus 84.4% p<0.001). Thirty-four of 58 LG-NHL patients were receiving anti-lymphoma treatment with a lower SARS-CoV-2 vaccination response as compared to the patients who were not on treatment (29.4% v 87.5% p<0.001). We observed a similar pattern in CLL patients receiving treatment (48.1 v 76.0 p:0.049). We found that only disease type and treatment status (on-treatment vs. off- treatment), but not gender, age or race were significant predictors of non-response in the multivariable logistic regression model. The interaction between disease type and treatment status was not statistically significant by multivariate analysis. In conclusion, receiving anti-cancer treatment was found to play a significant role in decreasing the response to COVID-19 vaccination.

Heart rate variability and cortisol levels in school-age children with different cognitive tests

Background. An urgent task of age-related physiology is to study the functional state of the body of school-age children in cognitive activity due to the large academic load and the use of information and computer technologies in the educational process to identify the characteristics of the reactivity of the body of students when performing cognitive load of various types, including on electronic devices, is necessary for the organization of the school educational environment.The aim. To assess the nature of vegetative, cardiovascular and hormonal reactivity in cognitive load of various types in school-age children.Materials and methods. By methods of heart rate variability analysis, electrocardiography, tonometry and enzyme immunoassay of cortisol in saliva, 117 school-age children were examined while performing cognitive load of various types.Results. There is a change in heart rate variability indicators while performing cognitive load. Oral counting causes an increase in sympathetic influences on the heart rate with a decrease in parasympathetic activity, as well as a shift in the vagosympathetic balance. Operation on the electronic devices causes a decrease in the total power density of the spectrum and an increase in the index of low-frequency and highfrequency vibrations ratio due to a decrease in parasympathetic activity. Two types of reaction were revealed: type I – an increase in the concentration of cortisol in saliva, an increase in sympathetic effects on Heart rate with a simultaneous decrease in parasympathetic activity (counting), a decrease in the total power of the spectrum density (laptop), a decrease in parasympathetic activity (tablet). Type II – a decrease in hormone levels and a decrease in very low-frequency vibrations and parasympathetic activity, regardless of the type of load presentedConclusion. The results obtained indicate that the nature of the reactivity of heart rate indicators and the stress hormone cortisol in students depends not so much on which electronic device it is performed on, but on the type of cognitive load.

Taking caffeinated beverages at rest affects hormone metabolism

It is generally accepted, that drinking tea and coffee causes an increase in cortisol levels in majority of people, due to the presence of alkaloids in these drinks, main of which is caffeine. The literature data about the ability of alkaloids to increase the cortisol secretion, and the high consumption of these beverages by men in our population, as well as the relatively scarce information on their effects on the body, forced us to focus in this study on examination the role of tea and coffee on the change in cortisol and testosterone levels in young men after taking strong tea or coffee at rest. İn this study we examined influence of coffee and tea on 21 healthy young men on testosterone and cortisol levels in the blood. For this, Hema kit (Russia) enzyme immunoassay method was used for testosterone definition, and Steroid ELISA-cortisol kit (Russia) – for serum cortisol levels in blood serum. We found out that cortisol may be lowered or raised depending on individuals, while testosterone does not change significantly either under action of coffee or tea.

Dynamics of Oxidative Stress in Helicobacter pylori-Positive Patients with Atrophic Body Gastritis and Various Stages of Gastric Cancer.

Gastric cancer is a global health problem. The pathogenesis of this disease remains unclear. This study included 198 H. pylori (+) men aged 45 to 60 years old. Group A included 63 practically healthy men, group B included 45 men with severe atrophic body gastritis, group C included 37 men with epithelial gastric cancer stages I–II according to TNM, and group D included 54 men with epithelial gastric cancer stages III–IV according to the TNM scale. The content of malondialdehyde (MDA), diene conjugates (DCs), superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and glutathione peroxidase (GPO) was detected using an enzyme immunoassay (ELISA) or spectrophotometric methods in the blood plasma. The concentrations of MDA and DC were increased in the patients of group B compared with group A, and in patients of groups C and D compared with groups A and B. The ratio of MDA/SOD and MDA/CAT was decreased in the patients in group D compared with the patients in group C, and was significantly higher compared with group A. The ratios of MDA/GPO and MDA/GST increased linearly and were at a maximum in groups C and D. Our work determined that indicators of oxidative stress may be the biochemical substrate, which brings together the various stages of the Correa cascade, and may explain disease progression. The dynamics of changes in the content of SOD and CAT in the plasma in patients with gastric cancer may be a target of future investigations.

Antibody Sustainability in SARS-CoV-2 Healthcare Professionals' Patient Cohort

In this study, it was aimed to evaluate one-year follow-up of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) specific antibodies formed against the virus binding site, in a coronavirus disease-2019 (COVID-19) positive case cohort (n= 413) between the period March 2020 to December 2020 in Manisa Celal Bayar University Hospital, until July 2021. SARS-CoV-2 antibodies were determined by the chemiluminescent enzyme immunoassay (CLIA) method. Values of 1.0 and above were considered positive. Chi-square tests and Joinpoint regression analysis (version 4.7.0) were used in the statistical analyses. The mean age of the participants was 34.9 ± 9.3 and 60.2% of them were women. Between 21-30 days after the diagnosis of COVID-19, total antibody level was above the threshold value in 72.2% (n= 126) of the participants, while this rate increased to 79.1% (n= 240) in 31-60 day interval. In the following period, this rate decreased to 38.8% (n= 108) in days 211st to 240th. Antibody response could not be detected in 76 (20.7%) of 367 employees who have initially been followed up. The percentage of total antibody positivity prevalence ranged from 98.9% to 96.1% in the 31-210th day after diagnosis, in the follow-up of 291 employees whose total antibody positivity was detected after diagnosis. According to the results of the Joinpoint regression analysis, after the diagnosis of COVID-19, the curve showing the percentage of antibody positivity was broken at two points: The first breaking point was observed in 181-210th days (6-7 months) (p= 0.069), and the second breaking point was in 271-300th days (9-10 months) (p< 0.001). As a result, the highest antibody positivity rates were detected after the 30th day of the disease onset and antibody positivity was maintained in the first seven months after diagnosis; the antibody positivity rate decreased to 25% at the end of the first year.

Association between the clinical-functional parameters and lipid markers of systemic inflammation in mild asthma complicated with obesity

Aim. To identify the features of the association between the clinical-functional conditions and lipid markers of systemic inflammation (fatty acids, eicosanoids, plasmalogens) of patients with mild asthma and obesity. Materials and methods. Obese (n=80) and nonobese (n = 55) adults with mild, partially controlled asthma aged 28 to 65 years, and nonobese (n = 30) healthy controls had venous blood samples analysed. The ACQ-5 test, spirometry were evaluated. Serum thromboxane B2 (ТХВ2) and leukotriene B4 (LTB4) were analyzed by the enzyme immunoassay method. Fatty acidsas methyl esters (FAMEs) and plasmalogens in the form of dimethyl acetals (DMA) were estimated in blood plasma by gas chromatography-mass spectrometry. Results. In obese patients with asthma, the ACQ-5 test was lower in comparison to nonobese patients. FEV1 decreased by 8 %, FVC by 12% (p &lt; 0.05). A decrease in the level of DMA16: 0 by 17.5 % and DMA18: 0 by 25 % was established in obese asthma patients. The ratio of DMA16:0/FAME 16: 0 declined by 21 %, DMA18:0/FAME18: 0 − by 28 %. A fall in eicosapentaenoic acid (20: 5n3) and a change in the 20:4n6/20:5n3 ratio were found. The level of ТХВ2 and LTB4 was increased regardless of body weight. There was a positive correlation between body mass index and levels of LTB4 (r = 0.46), ТХВ2 (r = 0.49); and a negative correlation between ACQ and ТХВ2 (r = - 0.58). Indicators of lung function (FEV1, FVC, VC) had correlative links with DMA18:0, DMA20:0, eicosapentaenoic acid. Conclusion. We showed the change in the inflammatory fatty acids and eicosanoids synthesis and plasmalogen deficiency in obese-asthma. Established relationships between clinical and functional parameters and lipid markers of systemic inflammation, plasmalogen deficiency in patients with asthma and obesity reveal new pathogenetic mechanisms of the systemic inflammatory process in chronic respiratory diseases.

Clinical Evaluation of a Novel Stool Antigen Test Using Bioluminescent Enzyme Immunoassay for Detecting Helicobacter pylori.

Background BLEIA ™ “EIKEN” Helicobacter pylori antigen (B[EIA]) is based on the bioluminescent enzyme immunoassay (BLEIA) method that was newly developed with high sensitivity in detecting Helicobacter pylori (H. pylori) antigen in feces. Methods In the project for H. pylori screening and treatment in Saga Prefecture in 2019, 141 students received the stool H. pylori antigen test as a secondary test. For 141 students, a comparative test was conducted between B (EIA) and extracorporeal diagnostic agents that were marketed in Japan as of 2019. The detection performance of H. pylori ATCC43504 standard strain and H. pylori antigen in commercial human fecal specimens were conducted. Results The comparison of B (EIA) with Quick Chaser TMH. pylori (Q [IC]) revealed positive and negative concordance ratios of B (EIA) to Q (IC) of 100.0% (110/110) and 71.0% (22/31), respectively. A comparative test was conducted between B (EIA) and extracorporeal diagnostic agents that were marketed in Japan as of 2019, and B (EIA) was most sensitive on “detecting H. pylori antigen of ATCC43504 standard strain” and “detecting H. pylori antigen in commercial human fecal specimens,” compared with other kits. Nine dissociated specimens that were negative for Q (IC) and positive for B (EIA) were confirmed. The measured value of B (EIA) in the dissociation samples were 1.3–87.4 cutoff index in the range that can be evaluated as negative by other fecal H. pylori antigen test kits, all the dissociation samples were H. pylori antigen-positive cases, and finally the cause of result divergence was presumed as false negative due to insufficient sensitivity of Q (IC). Conclusion B (EIA) that is based on the BLEIA method, which applies firefly luciferase luminescence, is more sensitive than stool antigen test kits that are currently marketed in Japan and is very useful in diagnosing H. pylori infection, especially in situations where noninvasive tests are preferred, such as in children.

Türkiye’de bir üçüncü basamak hastanede sağlık çalışanlarında SARS-CoV-2 seroprevalansı

Objective: The aim of this study is to investigate the previous four months (March-July 2020) SARS-CoV-2 infection rate, seroprevalence and the variables affecting these in HCWs in a university hospital. Methods: The present study is a SARS-CoV-2 seroprevalence study on HCWs working in a tertiary hospital during the first stage (March-July 2020) of the outbreak in Turkey. The presence of IgM and IgG antibodies against the spike structure of the virus was investigated by the chemiluminescent enzyme immunoassay (CLIA) method using the commercial antibody kit (COV2T, Siemens®, Tarrytown, NY, US). Participants’ socio-demographic characteristics, health status, lifestyle, risky occupational and social and personal protective equipment (PPE) usage were independent variables of the study. Chi-square test and Fisher’s exact test were used in univariate analyzes, and accepted type 1 error value was 0.05. The analyzes were made using the SPSS 23.0 package program. Results: 1177 out of a total of 1702 health workers participated in the study. Participation rate was 69.1% . The mean age of the study group was 35.3 ± 9.8 and 62.7% were females. SARS-CoV-2 infection rate detected by nucleic acid amplification test (NAAT-PCR) or antibody test (Elisa) was (18/1177) 1.5%; The seroprevalence of SARS-CoV-2 was 1.01%. 17% of the entire SARS-CoV-2 cases were asymptomatic. The highest infection prevalence was significantly higher in auxiliary health workers (3.7%) compared to other groups. The presence of symptoms HCW’s and their family members that did not exist before in the last 15 days, being overweight or obese and consulting as contacted person in survelliance unit were significantly related to having SARS-CoV-2 infection (p

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.

Markers of oxidative damage lipids and DNA in men with type 1 diabetes mellitus and different levels of albuminuria

BACKGROUND : Diabetic nephropathy is a specific kidney damage that affects up to 40% of type 1 diabetes mellitus patients. There is still insufficient knowledge about oxidative stress at the different levels of albuminuria. AIM : To assess the indicators of oxidative damage to lipids, DNA and antioxidant defense in men with type 1 diabetes mellitus and albuminuria different levels. MATERIALS AND METHODS: The study was conducted during 2018–2019. The main group included 56 men of reproductive age with type 1 diabetes mellitus (T1DM) divided into 2 groups: 24 patients with albuminuria level A1 (group A1) and 32 patients with albuminuria level A2 (group A2). The control group consisted of 28 healthy men. The oxidative stress indicators content, as well as the activity of antioxidant defense system various links was evaluated. Spectrophotometric, fluorimetric and enzyme immunoassay methods were used. RESULTS : In patients of the A1 group there were higher values of the median of primary — conjugated dienes, secondary — ketodienes and conjugated trienes, final — Schiff bases products and 8-hydroxy-2 ’ -deoxyguanosine in comparison with the control. Similar changes in patients of the A2 group were found. Intergroup differences related to increased median values of TBARs and 8-hydroxy-2 ’ -deoxyguanosine in patients of the A2 group compared with A1 group. The activity of the antioxidant defense system in A1 group relative to the control by increased values of the glutathione peroxidase, reduced glutathione, oxidized glutathione median, and retinol was characterized. A2 group had higher values of glutathione peroxidase, reduced glutathione and oxidized glutathione medians in comparison with controls. Correlation analysis in A1 group showed the relationships between the duration of the disease and the products of lipid peroxidation, glycated hemoglobin with 8-hydroxy-2 ’ -deoxyguanosine. In A2 group, there was a relationship between the albumin/creatinine ratio and the duration of the disease, glomerular filtration rate and creatinine level. CONCLUSION : In men with T1DM, regardless of the albuminuria level, there are higher values of the oxidative damage lipids and DNA parameters, as well as the presence of close relationships between these parameters and the duration of the disease, which can be used to develop potential strategies for the prevention and early therapy of diabetic nephropathy.