UCHL1 and Proteasome in Blood Serum in Relation to Dietary Habits, Concentration of Selected Antioxidant Minerals and Total Antioxidant Status among Patients with Alzheimer’s Disease

Alzheimer’s disease (AD) is an incurable neurodegenerative disease. It is the most common form of dementia among the elderly population. So far, no effective methods of its treatment have been found. Research to better understand the mechanism of pathology may provide new methods for early diagnosis. This, in turn, could enable early intervention that could slow or halt disease progression and improve patients’ quality of life. Therefore, minimally invasive markers, including serum-based markers, are being sought to improve the diagnosis of AD. One of the important markers may be the concentration of UCHL1 and the proteasome in the blood serum. Their concentration can be affected by many factors, including eating habits. This study was conducted in 110 patients with early or moderate AD, with a mean age of 78.0 ± 8.1 years. The patients were under the care of the Podlasie Center of Psychogeriatrics and the Department of Neurology (Medical University of Białystok, Poland). The control group consisted of 60 healthy volunteers, matched for gender and age. The concentration of UCHL1 and the 20S proteasome subunit were measured by surface plasmon resonance imaging (SPRI). In addition, a nutritional interview was conducted with patients with AD, which assessed the frequency of consumption of 36 groups of products. In the group of patients with AD, compared to the control group, we showed a significantly higher concentration of UCHL1 (56.05 vs. 7.98 ng/mL) and the proteasome (13.02 vs. 5.72 µg/mL). Moreover, we found a low negative correlation between UCHL1 and the proteasome in the control group, and positive in the AD group. The analysis of eating habits showed that the consumption of selected groups of products may affect the concentration of the tested components, and therefore may have a protective effect on AD.

Influence of the Zinc and Fibre Addition in the Diet on Biomechanical Bone Properties in Weaned Piglets

The effects of the zinc and fibre source in piglets’ diet on the bone mineral content, density, and strength parameters of the femur were investigated using 24 piglets fed a diet supplemented with either lignocellulose (LC) or potato fibre (PF). Half of each group of piglets consumed a diet with ZnSO4 monohydrate or with zinc glycinate (ZnGly). The diets contained similar amounts of lysine, energy, and fibre. Bone mineral content and density were over 9% higher in pigs receiving diets with ZnGly than in animals fed diets with ZnSO4. Moreover, ZnGly strongly improved maximum and elastic strength (by 25.7 and 20.0%, respectively, p < 0.0001) and bone stiffness (by 29.4%, p < 0.0001). Only the mass of the femur was affected by the type of fibre in the diet, as the femurs of piglets fed diets with LC were over 7% (p < 0.0001) heavier than in piglets fed diets with PF. The intake of digestible zinc and the zinc content in the blood serum were positively correlated with the measured bone parameters and, depending on the parameter, “r” ranged from 0.749 to 0.866 and from 0.400 to 0.479, respectively. It can be concluded that bone parameters are affected more strongly by the organic than inorganic source of zinc.

Study of the resistance of cows with mastitis depending on the age of lactation

Relevance. The breeding stock and young cattle in the conditions of industrial technology of animal husbandry experience an increased antigenic load, which leads to disturbances in the immune status and the development of inflammatory processes manifested by endometritis and mastitis, diarrheal and respiratory syndromes. It was relevant to conduct a comprehensive assessment of the immunological reactivity of the animal’s body according to the state of resistance in cattle with clinical and subclinical mastitis, depending on the age of lactation. As a result of complex studies, the specific and nonspecific reactivity of the body of cattle in different periods of lactation was studied.Methods. The object of the study were cows of a black-and-white Holstein breed (n = 450). Groups of animals were formed according to the age of lactation:1st lactation, 2–3rd lactation, 3–4th lactation and for health reasons: clinically healthy animals, with subclinical mastitis and clinical mastitis. Milk samples were taken from cows to count the number of somatic cells, and blood to determine the quantitative content of IgG immunoglobulin and indicators of nonspecific resistance.Results. It was found that with mastitis in cows of different lactation ages, almost identical changes in the immune status were observed. Quantitative determination of IgG levels in animal blood serum is the basis for assessing the immunity of cattle. The immune status in clinically healthy animals was characterized by a higher level of the total amount of IgG immunoglobulin in the blood serum (amounted to 22.25 ± 0.70 mg/ml), whereas in cows with a subclinical form of mastitis, the level of IgG immunoglobulin decreased by 12.3%, with a clinical form of mastitis — by 17.2%. The results also showed that in cows of different lactations with subclinical and clinical mastitis, the bactericidal activity (by 10.8%) and the lysozyme activity of the blood increased significantly (by 8.22%). In cows with clinical mastitis — by 8.6% and 9.7% respectively, compared with the indicators in groups of healthy animals. Thus, mastitis, occurring in acute and subacute form, leads to a decrease in the resistance of the cows’ organism.

Analysis of the Increase of Vascular Cell Adhesion Molecule-1 (VCAM-1) Expression and the Effect of Exposure in a Hyperbaric Chamber on VCAM-1 in Human Blood Serum: A Cross-Sectional Study

Background and Objectives: Vascular cell adhesion molecule-1 (VCAM-1) was identified as a cell adhesion molecule that helps to regulate inflammation-associated vascular adhesion and the transendothelial migration of leukocytes, such as macrophages and T cells. VCAM-1 is expressed by the vascular system and can be induced by reactive oxygen species, interleukin 1 beta (IL-1β) or tumor necrosis factor alpha (TNFα), which are produced by many cell types. The newest data suggest that VCAM-1 is associated with the progression of numerous immunological disorders, such as rheumatoid arthritis, asthma, transplant rejection and cancer. The aim of this study was to analyze the increase in VCAM-1 expression and the impact of exposure in a hyperbaric chamber to VCAM-1 levels in human blood serum. Materials and Methods: The study included 92 volunteers. Blood for the tests was taken in the morning, from the basilic vein of fasting individuals, in accordance with the applicable procedure for blood collection for morphological tests. In both groups of volunteers, blood was collected before and after exposure, in heparinized tubes to obtain plasma and hemolysate, and in clot tubes to obtain serum. The level of VCAM-1 was determined using the immunoenzymatic ELISA method. Results: The study showed that the difference between the distribution of VCAM-1 before and after exposure corresponding to diving at a depth of 30 m was at the limit of statistical significance in the divers group and that, in most people, VCAM-1 was higher after exposure. Diving to a greater depth had a much more pronounced impact on changes in VCAM-1 values, as the changes observed in the VCAM-1 level as a result of diving to a depth of 60 m were statistically highly significant (p = 0.0002). The study showed an increase in VCAM-1 in relation to the baseline value, which reached as much as 80%, i.e., VCAM-1 after diving was almost twice as high in some people. There were statistically significant differences between the results obtained after exposure to diving conditions at a depth of 60 m and the values measured for the non-divers group. The leukocyte level increased statistically after exposure to 60 m. In contrast, hemoglobin levels decreased in most divers after exposure to diving at a depth of 30 m (p = 0.0098). Conclusions: Exposure in the hyperbaric chamber had an effect on serum VCAM-1 in the divers group and non-divers group. There is a correlation between the tested morphological parameters and the VCAM-1 level before and after exposure in the divers group and the non-divers group. Exposure may result in activation of the endothelium.

Investigation of the Potential of Heterophil/Lymphocyte Ratio as a Biomarker to Predict Colonization Resistance and Inflammatory Response to Salmonella enteritidis Infection in Chicken

Salmonella causes significant economic loss to the poultry industry and represents a real threat to human health. The region of difference 21 (ROD21) pathogenicity island removal is a genetic mechanism by which Salmonellaenteritidis (SE) invades the intestinal epithelium and induces systemic infection in mice. The heterophil/lymphocyte (H/L) ratio reflects the chicken’s robustness and immune system status. The H/L ratio is considered a disease resistance trait, and it could be used as a marker for selecting Salmonella resistance in live chickens. However, the association of the H/L ratio with Salmonella resistance and the inflammatory response remains to be elucidated. Moreover, the kinetics of ROD21 excision in the intestine and immune organs of chickens is unknown. Therefore, this study aimed to investigate the bacterial load, the ROD21 excision, the IL-1β, IL-8, and INF-γ blood serum concentration kinetics, and the association with the H/L ratio in chicken at 1, 3, 7, and 21 days post-SE infection. The results showed a significant correlation between the H/L ratio and the bacterial load in the ileum and caecum at 7 dpi. The ROD21 pathogenicity island absolute and relative excision in the caecum were positively correlated at 1 dpi but negatively correlated at 7 dpi with the H/L ratio. However, in the liver, we found the opposite tendency. The association of the H/L ratio with IL-1β, IL-8, and INF-γ blood serum concentrations showed that a low H/L ratio is correlated with increased IL-1β and INF-γ at 21 dpi. This study confirmed that the H/L ratio is associated with robustness and Salmonella-resistance in chicken. The methodology used in this study can separate individuals into susceptible and resistant and can help in the selection and breeding of Salmonella-resistant chickens.

Development of the ELISA Test System for Quantitative Determination of IgG to the Varizella zoster virus in Human Serum and Assessment of its Diagnostic Efficiency

Relevance. The introduction of Varicella vaccine prophylaxis explains the need to develop a methodology for monitoring the vaccination effectiveness and the intensity of population immunity. This problem can be solved using quantitative immunoassay methods. Aim. Development of an enzyme-linked immunosorbent assay for the concentration of class G immunoglobulins (AB) to Varicella zoster virus (VZV) determining and assessing its functional characteristics and diagnostic efficiency. Materials and methods. Recombinant antigen GE VZV. WHO International Standard for Antibodies to VZV W1044. Blood serum samples from healthy people and patients with Chickenpox and Herpes zoster, blood serum samples containing IgG antibodies to herpes simplex viruses of the first and second types, cytomegalovirus, Epstein-Barr virus. Anti-VZV ELISA (IgG) reagent kit (Euroimmun, Germany). Indirect enzyme-linked immunosorbent assay. Immunization of animals with recombinant antigen GE, isolation, and purification of specific antibodies. Conjugation of monoclonal antibodies to human IgG with antibodies to antigen GE and with horseradish peroxidase. Results. An enzyme-linked immunosorbent assay in «an indirect» format has been developed to determine the specific antibodies to VZV concentration (IU/ml) in human serum/plasma. An artificial calibrator for determining the concentration of AB-VZV had been synthesized and standardized according to the International WHO-standard W1044. The main functional characteristics of the developed enzyme-linked immunosorbent assay are determined in accordance with GOST 51352-2013. The diagnostic kit was tested on blood serum samples from children with chickenpox (n = 43), adults with Herpes zoster (n = 158), healthy individuals (n = 781). The diagnostic sensitivity of the test system was 85%, the diagnostic specificity was 87% according to the ROC analysis. The absence of cross-reactivity of the test system was shown on samples with serological markers of other herpesvirus infections (n = 94). Comparative trials of the developed test system and its commercial analog, the Anti-VZV ELISA (IgG) reagent kit, did not reveal statistically significant differences between their functional characteristics. Conclusions. The developed test system for determining of the AB-VZV concentration in human serum/plasma in terms of its functional characteristics meets the GOST requirements, is characterized by high diagnostic efficiency, can be used to monitor the effectiveness of vaccine prophylaxis and strength of population immunity, as well as to assess the immune response in chickenpox and Herpes zoster.

Protein linking fatty acids and its genetic regulation in children having food allergy

Objective: Study the effect of the Ala54Thr FABP gene polymorphism on the produce of the intestinal FABP fraction in blood serum. urine and coprofiltrate in children having food allergies.Methods: The content of the FABP intestinal fraction in urine, feces, and blood serum was determined using ELISA method. The study of FABP genes polymorphism (G163A, Ala54Thr) was carried out using PCR method.Results: Statistically significant increase of the FABP level in blood serum, urine and feces in children with FA was detected in various biological fluids. The distribution of FABP2 alleles and genotypes obeyed the Hardy-Weinberg law (χ 2 = 0; p = 1,000) and did not significantly differ from the distribution of genotypes in children having FA andin the control sample (p = 0.638).Conclusions: The study did not reveal an association of the pathological genotype FABP G163A, (Ala54Thr) with the hyperproduction of the FABP intestinal fraction in children having FA, confirming the diagnostic significance of this marker increase during exacerbation of the disease.

Electroanalysis of Fentanyl and Its New Analogs: A Review

The review describes fentanyl and its analogs as new synthetic opioids and the possibilities of their identification and determination using electrochemical methods (e.g., voltammetry, potentiometry, electrochemiluminescence) and electrochemical methods combined with various separation methods. The review also covers the analysis of new synthetic opioids, their parent compounds, and corresponding metabolites in body fluids, such as urine, blood, serum, and plasma, necessary for a fast and accurate diagnosis of intoxication. Identifying and quantifying these addictive and illicit substances and their metabolites is necessary for clinical, toxicological, and forensic purposes. As a reaction to the growing number of new synthetic opioid intoxications and increasing fatalities observed over the past ten years, we provide thorough background for developing new biosensors, screen-printed electrodes, or other point-of-care devices.

Mitochondrial DNA polymorphism and myocardial ischemia: Association of haplogroup H with heart failure

The pathogenesis of atherosclerosis and ischemic heart disease is associated with oxidative stress and mitochondrial dysfunction. Mitochondrial DNA encodes subunits of mitochondrial respiratory chain and is highly polymorphic in human populations. Mitochondrial DNA can be considered a candidate genetic locus for predisposition to cardiovascular diseases.Aim. To analyze the associations of the mitochondrial genome polymorphism and chronic heart failure in ischemic heart disease.Material and Methods. The study included two groups of individuals: patients with a combination of ischemic heart disease and chronic heart failure (n = 175) and a population sample of residents of Tomsk (n = 424). Percentages of patients with chronic heart failure of NYHA classes II, III, and IV were 37%, 50%, and 13%, respectively. All patients underwent echocardiographic examination; body mass index and the lipid fractions in blood serum were determined. The average was 55.4 years in patients and 47.6 years in the population sample. Polymorphism of mtDNA was studied by sequencing the hypervariable segment of D-loop of mtDNA and subsequent classification of mtDNA haplotypes into the known haplogroups. The mtDNA haplogroup frequencies were compared between the samples using the Chi-square test. The associations of genotype with quantitative trait variability were analyzed by variance analysis.Results. Male patients showed a higher frequency of haplogroup H compared to the population (45.86% in patients and 35.4% in population) and a higher total frequency of haplogroup H subgroups except the most frequent subgroup H1 (36.94% and 25.22%, respectively). The values of significance level (p-value) and odds ratio (OR) were determined as follows: p = 0.04; odds ratio OR = 1.55 (95% confidence interval (CI) 1.02–2.34) for haplogroup H as a whole; p = 0.02; OR = 1.74 (95% CI 1.12–2.70) for haplogroup H without subgroup H1. Analysis of quantitative traits revealed the associations of the same genetic marker (mtDNA haplogroup H) with the levels of high-density lipoproteins (p = 0.03) and triglycerides (p = 0.02) in blood serum of men in the population sample.Conclusion. The obtained results suggested that the most frequent European mtDNA haplogroup H may be a risk factor for the complications of ischemic heart disease in men.