Immunological Parameters Research Articles

Analysis of Natural Antibodies during the Development of Phantom Pain Syndrome.

We determined natural antibodies (n-Abs) to the regulators of the main systems of biochemical homeostasis: β-endorphin, serotonin, dopamine, histamine, orphanin, angiotensin, GABA, glutamate, bradykinin, vasopressin, thrombin, and α-2-macroglobulin in individuals with phantom pain syndrome (PPS), resulting from amputation after injury. It was established that each patient has an individual immunoprofile, but for all of them there was a significant increase in the level of antibodies to serotonin, histamine, and angiotensin, which reflect the chronicity of the pain syndrome and do not depend on the self-assessment of the severity of PPS. Determination of the role of regulators of biochemical homeostasis in the development of phantom pain showed that, at high, moderate, and weak severity of PPS, the biogenic amine and angiotensinergic systems are activated. A decrease in PPS intensity normalizes deviations in all immunological parameters. The levels of n-Abs for the pain (β-endorphin) and analgesic (orphanin) systems are significant only at low PPS. Monitoring the individual profile of n-Abs to endogenous regulators allows us to obtain an objective picture of the pain status of the patient's body.

Diagnostic value of anti-hexokinase 1 and anti-kelch-like 12 antibodies in primary biliary cholangitis patients

Abstract Objectives Anti-mitochondrial antibody (AMA) is not always present in patients with primary biliary cholangitis (PBC). We aimed to determine the additional value of anti-hexokinase 1 (anti-HK1) and anti-kelch-like 12 (anti-KLHL12) antibody in PBC and analyzed the biochemical and immunological parameters of 212 subjects, including PBC patients and healthy controls. Methods Serum anti-gp210 and sp100 antibodies were determined by an immunoblotting test (IBT). Enzyme-linked immunosorbent assay (ELISA) was employed to evaluate anti-HK1 and anti-KLHL12. The diagnostic value of anti-HK1 and anti-KLHL12 to PBC was analyzed by constructing a receiver operating characteristic (ROC) curve. Results ROC analyses didn’t show a very good performance of serum anti-HK1 for PBC diagnosis; the AUC was 0.664 with a sensitivity of 53.3 % and a specificity of 79.2 %. Regarding anti-KLHL12, ROC analysis yielded an AUC of 0.626, with a sensitivity of 45.7 % and a specificity of 93.8 %. For AMA-negative PBC patients, the AUC increased to 0.790 for KLHL12, and 0.708 for HK1. AMA combined with anti-HK1 or anti-KLHL12 antibody significantly improved the diagnostic sensitivity of PBC from 82 to about 95 %, respectively. In AMA-negative PBC patients, the sensitivities for anti-HK1 (62.50 %) and anti-KLHL12 (75 %) antibodies were higher than for anti-gp210 (37.5 %) and anti-sp100 antibody (43.75 %). When these four antibodies were combined, the overall sensitivity increased to 87.5 %. Conclusions The determination of anti-HK1 and anti-KLHL12 facilitates the diagnosis of PBC, particularly in AMA-negative patients. Adding anti-HK1 and anti-KLHL12 antibodies to clinical detection enables early diagnosis and timely treatment, potentially improving patient prognosis.

Effect of dietary seaweed Caulerpa racemosa on growth, biochemical, non-specific immunity, and disease resistance to Pseudomonas aeruginosa in Cirrhinus mrigala

BackgroundThe green seaweed Caulerpa racemosa possesses highly potential elements in animal forages and human use since ancient times. The current study was designed to investigate the antioxidants, phytochemical properties of C. racemosa, as well as their effects on growth parameters, biochemical components, non-specific immunological parameters, and disease resistance to P. aeruginosa in Cirrhinus mrigala. The experimental group, divided into five groups as T1–T5. T2–T5, was given C. racemosa extract at concentrations of 0.5%, 2.5%, 4.5%, & 6.5% supplemented with basal diets. T1's group food is devoid of extract and acts as a control, and the trial lasted forty-five days.ResultsC. racemosa exhibited dose-dependent antioxidant activity. The highest levels of DPPH (61.21%), ABTS (70.55%), and H2O2 (66.55%) scavenging activities were obtained at 100 µg/ml 100 μg/ml. GC–MS analysis revealed phytoconstituents in the crude extract, such as palmitolinoleic acid, butanoic acid, arachidonate, linoleic acid, vaccenic acid, octadecenoic acid, trans-phytol, and eicosanoids. Among these different concentrations, 4.5% C. racemosa supplemented diet produced a significantly higher growth metrics of C. mrigala’s (WG, SGR, FCR), biochemical constituents, when compared to other concentrations. In a post-challenge trial, 4.5% C. racemosa extract meals increased C. mrigala SOD, CAT, non-specific immune response (lysozyme, NBT, phagocytic activity), and haematological (RBC, WBC, HCT & Hb) parameters when compared to other experiments.ConclusionsThe findings revealed that 4.5% of C. racemosa may be supplied in the diet of C. mrigala to regulate better survival growth performance and haematological parameters.

Impact of Some Immunological Parameters (antioxidant – cytokines) in Cutaneous leishmaniasis in a Sample of Patients in the Al-Ramadi City.

Impact of Some Immunological Parameters (antioxidant – cytokines) in Cutaneous leishmaniasis in a Sample of Patients in the Al-Ramadi City.

Utilizing outer membrane proteins (OMPs) as a vaccine to manage motile Aeromonas septicemia (MAS) in Nile tilapia (Oreochromis niloticus) with evaluating the vaccine immunoprotective effectiveness

AbstractMotile aeromonads are an opportunistic and heterogenic bacteria that cause motile Aeromonas septicemia (MAS) which is accompanied with variety of clinical signs and lesions in fish. It has been found that their outer membrane proteins (OMPs) are highly immunogenic and may be used as a vaccine candidate. The present study was undertaken to evaluate OMP protection against Aeromonas hydrophila (A. hydrophila) strain ATCC 7966 (A5 and A8) and A. hydrophila strain DSM 30,187 (A6). Pathogenicity tests revealed that out of 12 isolated strains, seven were highly pathogenic, three were moderate, and two were avirulent. SDS-PAGE analysis of OMPs showed several protein bands (7–22) of different molecular weights ranging from 14 to 346 kDa. The common protein band was 39 kDa. It seems that pathogenicity and the number of protein bands are related in the highly pathogenic strains. Challenged A. hydrophila for Nile tilapia (O. niloticus) previously actively immunized with OMPs (A4, A5, A6, A8, and A11) showed a significant increase in relative percent survival (RPS) ranging from 50 to 100%. Immunological parameters such as immunoglobulin M (IgM), antioxidants (SOD, GSH, and CAT), and nitric oxide (NO) and expression of pro-inflammatory cytokines, interleukin 1β (IL-1β), and tumor necrosis factor-α (TNFα) showed a significant increase in OMP-immunized fish. Challenged A. hydrophila for O. niloticus previously passively immunized with OMPs (A4, A5, A6, A8, and A11) in fish-raised sera showed a significant increase in RPS (20 to 100%) in the vaccinated groups. It is concluded that OMPs from Aeromonas spp. have been identified as suitable target for protection development, especially in the case of heterogenic bacteria.

Retinal Vascular Abnormalities and Clinical Parameters in Systemic Sclerosis

Background: Systemic sclerosis is a complex autoimmune disease characterized by vasculopathy, fibrosis, and immune dysregulation. Ocular manifestations in these patients are increasingly recognized, suggesting potential correlations between systemic vascular abnormalities and ocular microvascular changes. Advancements in molecular immunology and imaging technology using ocular coherence tomography (OCT) have unveiled intricate pathways underlying possible disease pathogenesis. Understanding the interplay between retinal vascular abnormalities and molecular immunology parameters could provide insights into disease mechanisms and potential biomarkers. Purpose: The aim of this study was to investigate vascular abnormalities, detected with optical coherence tomography angiography (OCT-A), in systemic sclerosis patients and to find correlations between the severity of the disease detected with molecular immunology findings and OCT-A parameters. Methods: A group of 32 systemic sclerosis patients were compared with 9 healthy controls. Ganglion cell complex thickness (GCC), retina thickness of the fovea and parafovea, nerve fiber layer thickness (RNFL) and cup/disc area ratio were investigated using OCT. Vessel density (VD) of the superficial (SCP) and deep capillary plexus (DCP) of the whole macular area and ETDRS grid, size of the foveal avascular zone (FAZ) and vessel density of the radial peripapillary capillary plexus (RPCP) were evaluated using OCT-A. Modified Rodnan skin score (mRSS), capillaroscopy and disease duration were used to stage disease severity. Results: There was a statistically significant reduction in retina thickness of the fovea and parafovea, VD of the whole DCP, VD of the SCP and DCP in ETDRS grid in the patient group compared to controls (p < 0.001). The patients presented a significant enlargement of the FAZ (p 0.005). No significant correlation between OCT and OCT-A parameters and disease severity scores was found. Conclusions: OCT-A could represent a non-invasive tool to detect retinal microvascular damage in systemic sclerosis.

Effect of Toxicity of Chromium (VI) Stressors Alone and Combined to High Temperature on the Histopathological, Antioxidation, Immunity, and Energy Metabolism in Fish Phoxinus lagowskii

Fish in aquatic ecosystems are often impacted by environmental stressors like temperature fluctuations and exposure to heavy metals. Chromium (Cr6+) is a known environmental pollutant that poses a threat to aquatic life. Various environmental factors, such as water temperature, have been found to affect the toxicity of dissolved chemicals in aquatic ecosystems. We investigated the toxicity of combinations of different concentrations of hexavalent chromium (Cr6+) with high temperatures in fish. Hematological indices demonstrated changes in white blood cells (WBCs), hematocrit (HCT), red blood cells (RBCs), and hemoglobin (Hb) levels during the exposure. The qualitative and semi-quantitative analyses of different tissues confirmed that higher concentrations of Cr6+ caused more significant damage than lower concentrations, with evident alterations observed in circulatory and regressive aspects. Furthermore, brain acetylcholinesterase levels decreased in both single heavy metal exposure and combined exposure at a high temperature. The activity of antioxidant oxidase and immunological parameters increased in all treatment groups compared with the control group following long-term exposure. A significant and increased effect of Cr6+ in the high-temperature groups was observed on the evaluated biomarkers, suggesting a possible synergistic effect between Cr6+ and increased temperature. The integrated biomarker response (IBR) reported the highest level of stress at 10 mg/L Cr6+ combined with high temperature. The IBR analysis revealed that the highest activity of response enzymes, such as acid phosphatase (ACP), superoxide dismutase (SOD), and glutathione S-transferases (GST), was observed in the liver, whereas the gills displayed alkaline phosphatase (ALP), GST, and SOD activity, and the kidneys demonstrated SOD, ACP, and aspartate aminotransferase (AST) to be most active. Through histopathology, antioxidant enzymes, and metabolism- and immunity-related enzymes, we determined that high temperatures enhance the potential toxicity of Cr6+ in fish. We recommend conducting a thorough assessment of the impact of climate change, particularly temperature fluctuations, when studying the toxic effects of metal pollution, like chromium, in aquatic ecosystems.

Bioaccumulation of arsenic in fish (Labeo rohita) in presence of periphyton: ameliorative effect on oxidative stress, physiological condition, immune response and risk assessment.

The present study explores the use of periphyton to ameliorate toxic properties of arsenic (As) to Labeo rohita and also assesses the human food safety aspects. Fish were introduced to arsenite [As(III)] contaminated water (0.3 and 3mg/L) along with periphyton. Biochemical, physiological and immunological parameters, including gene expression, were assessed after 30days of exposure. Periphyton incorporation significantly improved (p < 0.05) the adverse effects of As on respiration, NH3 excretion and brain AChE activity by reducing oxidative stress and As bioaccumulation. The presence of periphyton in As(III) exposed fish (3mg/L) increased the immune response (Immunoglobulin M and Complement C3) in the serum and the regulation of the respective immune genes in the anterior kidney was found to be similar to the control. A speciation study using LC-ICP-MS confirmed the high accumulation of As by periphyton (5.0-31.9μg/g) as arsenate [As (V)], resulting in a lower amount of As in fish muscle. The calculated human health risk indices, Target Hazard Quotient (THQ) and Target Cancer risk (TCR) indicate that fish grown in periphyton-treated water may lower the human health risks associated with As. The study signifies the importance of periphyton-based aquaculture systems in As contaminated regions for safe fish production with enhanced yield.

Concentrations of selected immunological parameters in the serum and processing fluid of suckling piglets and the serum and colostrum of their mothers

BackgroundBlood sampling from neonatal piglets is related to multiple disadvantages. Therefore, a new, alternative matrix is required to assess piglets’ early immune status efficiently. The present study aimed to assess the usefulness of processing fluid for determining selected piglets’ immune parameters. 264 pigs − 31 sows, 146 male piglets, and 87 female piglets from commercial indoor farrow-to-finish pig herd were included in this study. 264 serum, 31 colostrum, and 146 processing fluid samples were collected. Serum was collected from all animals, colostrum was collected from sows, and processing fluid was collected from male piglets only. Using commercial ELISA tests, the concentration of various immunoglobulins, cytokines, and acute phase proteins was assessed in each matrix. Statistical analyses were employed to determine differences in the concentration of measured indices between piglets’ serum and processing fluid and correlations in the concentration of tested indices between particular sets of matrices.ResultsStatistical analyses did not reveal significant differences in the IgG, IgA, IL-1β, IL-4, IL-6, and IFN-γ concentration between piglets’ serum and processing fluid (p > 0.05). A positive correlation (p < 0.05) regarding the concentration of some indices between processing fluid and samples collected from sows was also observed.ConclusionsProcessing fluid can be considered a promising alternative to blood for assessing some immunological indices in piglets, such as IgG, IgA, IL-1β, IL-4, IL-6, and IFN-γ, and, possibly, in the indirect assessment of some indices in lactating sows, including IgA, IL-1β, IL-4, IL-6, IL-8, IFN-γ, or Pig-MAP.

Abstract RF02-07: Precocious modulation of metabolic and immunological parameters predicts tumor response to fasting-mimicking diet plus chemotherapy in patients with early stage TNBC

Abstract Background: Preclinical studies showed that severely calorie restricted fasting-mimicking diets (FMDs) enhance the antitumor efficacy of chemotherapy (CT) or immunotherapy (IO) in murine triple negative breast cancer (TNBC) models. These effects are mediated by a combination of blood glucose reduction and positive immunomodulatory effects. Moreover, combining fasting and metformin produced synergistic anticancer effects in a broad range of tumor models. The BREAKFAST trial was designed to investigate if FMD, plus/minus metformin, could increase the antitumor activity of neoadjuvant CT in patients with stage I-III TNBC. Methods: BREAKFAST (NCT04248998) is a randomized, non-comparative, phase II, pilot trial that enrolled stage I-III (cT&amp;gt;1 cm) TNBC patients (pts) candidate to receive 4 cycles of neoadjuvant CT with doxorubicin-cyclophosphamide every 3 weeks, followed by 12 cycles of weekly paclitaxel. Pts were randomized 1:1 to receive: CT plus 5-day FMD every 3 weeks, up to 8 cycles (arm A); CT plus FMD plus daily metformin (1700 mg) (arm B). The primary study endpoint was the rate of pCR in either experimental arm. Secondary/exploratory endpoints included safety, compliance, and biomarker analyses based on blood metabolomics and tumor transcriptomics analyses at different timepoints. Results: Between June 2020 and February 2022 we enrolled 30 pts. Then, the study was interrupted after the introduction of chemo-immunotherapy (CT-IO) as a standard neoadjuvant therapy for early stage TNBC pts. Among 30 pts, 13 were treated with CT plus FMD, while 17 pts received CT plus FMD plus metformin. Overall, pCR rate was 56.6%, i.e., significantly higher than pCR rates reported with anthracycline-taxane CT alone in previous phase II/III trials (26-39%), with no significant differences among treatment arms (p=0.49). The FMD acutely reduced blood glucose, insulin and LDH levels, which reflects a reduction in systemic and/or tumor glucose metabolism. Of note, precocious LDH reduction was more pronounced in patients undergoing pCR. RNA-seq analysis of tumor samples revealed a significant downmodulation of glycolysis and TCA cycle pathways after one treatment cycle, paralleled by an increase of intratumor activated T cells, memory T cells and NK cells, as estimated by deconvolution analyses of tumor transcriptomic data. Of note, these changes were observed only in patients achieving pCR. While intratumor metabolic changes were similar in the two treatment arms, the modulation of intratumor immunity was more pronounced in patients not receiving metformin. Conclusion: Preoperative CT plus cyclic FMD (plus/minus metformin) results in excellent pCR rates in localized TNBC patients. Early on-treatment downregulation of systemic and intratumor metabolic parameters related to glucose metabolism predicts pCR, and this is independent of metformin use. Based on results of this study, we recently initiated a large, multicentric trial, namely the BREAKFAST-2 (NCT05763992) study, which will investigate if adding cyclic FMD to neoadjuvant CT-IO increases pCR rates in ~ 145 pts with stage II-III TNBC. Citation Format: Francesca Ligorio, Giovanni Fucà, Andrea Vingiani, Fabio Iannelli, Riccardo Lobefaro, Leonardo Provenzano, Lucrezia Zanenga, Cristina Ferraris, Antonino Belfiore, Silvia Brich, Alessia Bertolotti, Gianfranco Scaperrotta, Catherine Depretto, Antonia Martinetti, Elisa Sottotetti, Paola Antonia Corsetto, Giulia Valeria Bianchi, Giuseppe Capri, Secondo Folli, Saverio Minucci, Marco Foiani, Massimiliano Pagani, Giancarlo Pruneri, Filippo De Braud, Claudio Vernieri. Precocious modulation of metabolic and immunological parameters predicts tumor response to fasting-mimicking diet plus chemotherapy in patients with early stage TNBC [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr RF02-07.

The Influence of Probiotic Lactobacilli on COVID-19 and the Microbiota.

This comprehensive review explores the potential of using lactobacilli as a probiotic in the management of COVID-19. Our findings suggest that lactobacilli show promise in reducing the risk of death, gastrointestinal and overall symptoms, and respiratory failure, as well as in lowering cytokines and inflammatory markers associated with the disease. The molecular mechanisms by which lactobacilli protect against COVID-19 and other viral infections may be related to the reduction in inflammation, modulation of the immune response, and direct interaction with viruses to produce antiviral substances. However, the selected studies demonstrate the presence of mixed findings for various clinical, biochemical, hematological, and immunological parameters, which may be attributed to methodological differences among studies. We highlight the importance of clearly describing randomization processes to minimize bias and caution against small sample sizes and inappropriate statistical tests that could lead to errors. This review offers valuable insights into the therapeutic potential of lactobacilli in the context of COVID-19 and identifies avenues for further research and applications. These findings hold promise for the development of novel approaches to managing COVID-19 and warrant further investigation into the potential benefits of lactobacilli in combating the disease.

ВЛИЯНИЕ РЕВМАТОИДНОГО АРТРИТА НА ТЕЧЕНИЕ ЗАБОЛЕВАНИЙ ПАРОДОНТА

The most common chronic systemic autoimmune disease of connective tissue is rheumatoid arthritis, the incidence of which is up to 50% (in Russia). The etiology of rheumatoid arthritis is still not known for certain. Scientists pay special attention to searching and studying the factors influencing the occurrence of this disease. Researchers are interested in periodontitis as a likely factor in the development of rheumatoid arthritis. The literature presents limited data on the study of the characteristics of the course of dental diseases in patients with rheumatoid arthritis, which indicates the need for further study of this issue and clarification of the relationships between the intensity of caries, the condition of periodontal tissues and the temporomandibular joint in this category of patients. The study of these issues is of scientific and practical interest, as it allows us to improve the methods of diagnosis and monitoring of patients with rheumatoid arthritis by a dentist. The purpose of this study is to determine the effect of rheumatoid arthritis on the course of periodontal diseases. Materials and methods. The methodology of the scientific research is based on the analysis of domestic and foreign literature in the eLIBRARY databases; basic and additional methods of examining the patient were carried out. During the clinical examination, we assessed the general condition of the patient with rheumatoid arthritis, his dental and periodontal status. Results and discussion. A clinical examination of a patient with somatic pathology was carried out. Microorganisms of periodontal pockets were determined, the data obtained were compared with a patient who does not have a systemic disease. Conclusions. Patients with rheumatoid arthritis generally have a thin gingival biotype. Immunological parameters of oral fluid (IgA) in rheumatoid arthritis decrease, which indicates a lack of local and general immunity and leads to damage to periodontal tissue. In the future, this scientific research will help us develop optimal methods for diagnosing and treating patients with autoimmune disease.

Specific Immune Marker Associated With Coronavirus Disease 2019

Background: The infection that caused the pneumonia, the virus known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the source of coronavirus disease 2019 (COVID-19) is the source of the global pandemic. The pneumonia cases began in Wuhan, China in December 2019 and had no apparent reason. Essential inflammatory mediators Chemokines are necessary for an immune response to eliminate pathogens. However, the fundamental cause of hyperinflammation is their excessive release. Chemokines may be directly responsible for the acute respiratory illness syndrome in the current COVID-19 outbreak. Objective: The current study aimed to estimate hematological changes with immunogenic markers CD177, GNLY, and CXCR4 gene expression in COVID-19 patients, and compare these parameters among patients and healthy individuals. Materials and Methods: The study was conducted at Al-Yarmouk Teaching Hospital and Medical Teaching Laboratories. Finally, to detect the immune markers level in samples of patients, by RT-PCR. Results: The results revealed the gene expression results in patients with COVID-19 means Ct difference of CD177 (24.70) fold more than (10.5) elevated when compared with the mean of Ct control healthy group is (28.21) fold of gene expression is 1.00 of CD177 when our study showed increased gene expression the mean of GNLY Ct receptor was reached to (28.70 ng/L) with fold more than 15.0 in COVID-19 patients, while mean Ct in healthy control was (32.61 ng/L) with fold 1.00, the current study found mean CT of CXCR4 gene expression ( 31.2) with fold (1.1) compared with healthy control CT(31.3) fold of gene expression was (1.0). Conclusion: It was concluded that COVID-19 caused significant changes in many immunological parameters and found the titer of immune markers was higher in infection compared with control.

Pirarucu requires taurine to maximize growth and antioxidant status when fed diets high in plant-based ingredients

Taurine (Tau) is a free conditionally essential amino acid for most fish species that plays a significant nutritional role, mainly in carnivorous fish species. However, limited studies are still available regarding the role of Tau on pirarucu nutrition, an important Amazonian fish species. Therefore, this study used diets high in plant-based ingredients to investigate the effect of taurine (Tau) supplementation on juvenile pirarucu's growth, metabolism, and health (n = 115, Weight = 504.12 ± 19 g). Fish were fed five diets containing increasing levels of Tau: 0.95, 5.1, 9.6, 12.9, and 17.2 g/kg in a completely randomized design with three replicates per treatment. After a 60-day feeding trial, growth, hematology, blood chemistry, Tau content in muscle, carcass, and liver, antioxidant status, and immunological parameters were evaluated. The control group (0.95 g/kg Tau) showed the lowest growth. According to the segmented regression model, the recommended Tau level for maximizing the development of juvenile pirarucu was 4.49 g/kg of feed. Additionally, fish fed the lowest dietary Tau level showed the highest ROS (reactive oxygen species) concentration. Dietary Tau levels did not affect hematological parameters and blood chemistry (p > 0.05). Fish fed 9.6 g/kg of Tau showed increased catalase activity, while the control group had the worst result in NBT (nitroblue tetrazolium reduction test). Fish fed with the highest dietary Tau levels (12.9 and 17.2 g/kg) showed increased total serum immunoglobulin concentration. These results suggest that the optimal inclusion level of Tau in diets for juvenile pirarucu is 4.49 g/kg of feed and that this amino acid acts as a mitigator of oxidative stress.

Immune system parameters in patients with chronic opistorchiasis based on genetic polymorphism associated with carbohydrate and lipid metabolism disorders

A study assessing differences in the immunopathogenesis of chronic opisthorchiasis with or without genetic polymorphisms in patients associated with predisposition to type II diabetes mellitus and lipid metabolism disorders was carried out. Venous blood samples from 89 patients were collected to analyze immunological parameters and genetic polymorphisms by using pyrosequencing. In the presence of mutations affecting regulation of carbohydrate metabolism, patients with opisthorchiasis had less pronouncedly increased phagocytic activity, a lower monocyte count, higher total T-lymphocyte count, lower T-cytotoxic cell and B-lymphocyte counts, lower IgA, IgG but higher IgM concentration. This indicates a moderately compromised nonspecific resistance, imbalanced effector T- and B-immunity, but in most cases (PPARG, TCF7L2 rs12255372, CDKAL1, CDKN2A/2B, SLC30A8) aggravated humoral immune reaction to invasion particularly revealed by lower B-lymphocyte count. The presence of polymorphisms that alter lipid metabolism regulation bidirectionally affects the parameters of immune response. An increased B-lymphocyte count and other indicators of humoral immune activation to invasion, which are detected in groups with mutations in the APOE (rs429358), PCSK9, ABCA1, APOC3 rs2854117 genes, can contribute to a more effective response to sustained antigenic stimulation. Changes in the T-lymphocytes subpopulations, characteristic of opisthorchiasis invasion, are aggravated in the presence of mutations in the PCSK9, ABCA1, and APOC3 rs2854117 genes. Mutations in the genes APOC3 rs5128, LPL rs268 activate patients’ nonspecific resistance, although this effect may also be associated with exhaustion of neutrophil bactericidal reserve. In general, minor alleles of the APOE (rs429358), APOC3 rs2854117, LPL rs268, LPL rs328, PON1 rs662 genes can be considered “protective” for the immunopathogenesis of chronic opisthorchiasis. Thus, patients with chronic opisthorchiasis with different genotypes predisposing to carbohydrate and lipid metabolism disorders had significant differences in immune system parameters, which can also affect the disease course. Mutations in different loci of the PCSK9, ABCA1, APOE, APOC3, and PON1 genes have opposite effects on the analyzed immune parameters. In the presence of mutations in other genes (PPARG, TCF7L2 rs12255372, CDKAL1, CDKN2A/2B, SLC30A8, LPL), opisthorchiasis invasion leads to more pronounced alterations in immune response; mutations in the lipoprotein lipase gene may have some “protective” immune-related effect in opisthorchiasis. The effect of all studied genetic polymorphisms associated with a predisposition to developing type II diabetes mellitus was predominantly negative.

Impact of a varied set of stimuli on a suite of immunological parameters within peripheral blood mononuclear cells: toward a non-animal approach for assessing immune modulation by materials intended for human use.

Introduction: In toxicology, steps are being taken towards more mechanism-focused and human relevant approaches to risk assessment, requiring new approaches and methods. Additionally, there is increasing emphasis by regulators on risk assessment of immunotoxicity. Methods: Here we present data from a peripheral blood mononuclear cell (PBMC) system whereby a varied set of stimuli, including those against the TCR and Toll-like receptors, enable readouts of cytokine and prostaglandin E2 (PGE2) production with monocyte, T cell and B cell viability, proliferation, and associated activation markers. In addition to results on the impact of the stimuli used, initial profiling data for a case study chemical, curcumin, is presented, illustrating how the system can be used to generate information on the impact of exogenous materials on three major constituent immune cell subsets for use in risk assessment and to direct follow-on studies. Results: The different stimuli drove distinct responses, not only in relation to the "quantity" of the response but also the "quality". Curcumin had a limited impact on the B cell parameters measured, with the stimuli used, and it was noted that in contrast to T cells where there was either no impact or a reduction in viability and proliferation with increasing concentration, for B cells there was a small but significant increase in both measurements at curcumin concentrations below 20µM. Similarly, whilst expression of activation markers by T cells was reduced by the highest concentration of curcumin, they were increased in B cells. Curcumin only impacted the viability of stimulated monocytes at the highest concentration and had differential impact on different activation markers. Levels of all cytokines and PGE2 were reduced at higher concentrations. Discussion: Although the platform has certain limitations, it nevertheless enables assessment of healthy baseline monocyte, T-, and B-cell responses, and scrutiny of the impact of different stimuli to detect potential immune suppression or enhancement from exogenous materials. In the case of curcumin, a pattern of responses indicative of immune suppressive / anti-inflammatory effects was detected. It is an accessible, highly modifiable system that can be used to screen materials and guide further studies, providing a holistic, integrated picture of effects.

Bighorn Sheep (Ovis canadensis) with Higher Whole Blood Selenium Levels Have Improved Survival and Altered Immune Responses.

Bighorn sheep (Ovis canadensis) are herbivorous ungulates that live in forage-poor areas of the American west. The trace minerals that herbivores derive from forage are important for immune function. Therefore, identifying trace minerals that affect immune function in bighorn sheep could provide important insights into disease susceptibility and population health in threatened populations. We sought to determine whether trace mineral composition in blood or plasma correlates to survival and determine whether immunologic parameters correlate with any trace minerals that affect survival. We used data collected from 2016 to 2018 as part of a large study on bighorn sheep in southeastern Oregon and northern Nevada, US. We measured the survival of 135 bighorn sheep during the 8-mo monitoring period, including general metrics of immune function and trace mineral levels. We found that animals with higher selenium had improved survival over the monitoring period, with higher peripheral blood mononuclear cell activity (lymphocytes and monocytes) and lower bacterial killing ability in an in vitro assay. This suggests that bighorn sheep may have altered immune function when selenium levels are low, making them more likely to die during the 8-mo monitoring period. Future work should consider whether habitat management strategies that increase selenium intake might improve disease resistance and survival in bighorn sheep in selenium-poor areas.

Synergistic Effect of Dietary Supplementation with Sodium Butyrate, β-Glucan and Vitamins on Growth Performance, Cortisol Level, Intestinal Microbiome and Expression of Immune-Related Genes in Juvenile African Catfish (Clarias gariepinus).

The effect of dietary supplementation with sodium butyrate, β-glucan and vitamins (A, D3, E, K, C) on breeding indicators and immune parameters of juvenile African catfish was examined. The fish were fed with unenriched (group C) and enriched feed with a variable proportion of sodium butyrate/β-glucan, and constant content of vitamins (W1-W3). After the experiment, blood and the middle gut were collected. The microbiome of the gut was determined using Next Generation Sequencing (NGS). Liver tissue was collected for determination of expression of immune-related genes (HSP70, IL-1β, TNFα). W2 and W3 were characterized by the most favorable values of breeding indicators (p < 0.05). The highest blood cortisol concentration was in group C (71.25 ± 10.45 ng/mL), and significantly the lowest in W1 (46.03 ± 7.01 ng/ mL) (p < 0.05). The dominance of Cetobacterium was observed in all study groups, with the largest share in W3 (65.25%) and W1 (61.44%). Gene expression showed an increased number of HSP70 genes in W1. IL-1β and TNFα genes peaked at W3. The W3 variant turns out to be the most beneficial supplementation, due to the improvement of breeding and immunological parameters. The data obtained can be used to create a preparation for commercial use in the breeding of this species.

Prebiotic, probiotic and marine algae suplementation in juvenile tilapia diet

This study evaluated the probiotic mannan oligosaccharide (MOS), and marine algae for Nile tilapia (Oreochromis niloticus) feeding. The parameters analyzed were fish-growth rate, hematological and immunological parameters, and intestinal microbiota. Also, fish were submitted to experimental infection challenge with Aeromonas hydrophila to evaluate the immune response. Nile tilapia juveniles (weight 8.86 ± 3.22 g) were used in the six treatments with four replicates, for 63 days. The treatments were: control basal diet; PAS-TR: basal diet plus 0.04 g·kg-1 of the probiotic (Bacillus cereus and Bacillus subtilis); MOS: basal diet plus 4 g·kg-1 of the prebiotic mannooligosaccharide, 4 g·kg-1 of the prebiotic Kappaphycus alvarezii. Two more diets were formulated by a combination of PAS-TR + MOS (4 +4 g·kg-1), and PAS-TR + KAP (4 +4 g·kg-1). For the challenge experiment, the fish were fed for 21 days, infected via intraperitoneal injection with A. hydrophila (1 x 106 UFC·mL-1), and the mortality rate was registered for 15 days post-infection. Results indicated the capacity of probiotic to remain in the gut for 63 days, and it was inhibited neither by autochthonous microbiota nor by prebiotics used. The feed additives tested for Nile tilapia did not cause a beneficial or adverse effect on growth or hematological variables. However, these treatments protected the fish from A. hydrophyla infection, proved by higher survival rate, and relative protection levels. We concluded that probiotic PAS-TR and prebiotics MOS and KAP, combined or not as symbiotics, may promote immune protection and reduce the mortality rate of A. hydrophyla infection.

Chronic cypermethrin induced toxicity and molecular fate assessment within common carp (Cyprinus carpio) using multiple biomarkers approach and its novel therapeutic detoxification

Cypermethrin (CYP) is a chemical of emerging concern which has persistent and bioaccumulating impacts as it can be found extensively in freshwater ecosystem and agricultural products. It has exposure risk and toxic effects over human edible fish, as common carp. Four groups were designed for toxicity assessment and detoxification approach: control group (CL), CYP exposure group (CYP), CYP + 10% M. oleifera leaves and 10% M. oleifera seeds (CMO group), 10% M. oleifera leaves and 10% M. oleifera seeds (MO group). Trial period was forty days during which cohort of 240 fish in CYP and CMO group was exposed to 1/5 of 96h LC50 of CYP (0.1612 μg/L). CYP-exposed carp exhibited lower growth parameters, but carp fed with 10% M. oleifera seeds and leaves showed significant improvement in growth rate (SGR, RGR) and weight gain (WG) as compared to the control group. CYP exposure negatively affected haemato-biochemical parameters. Moreover, CYP exposure also led to oxidative stress, damaged immunological parameters, genotoxicity and histopathological damage in liver and intestinal cells. Whereas, M. oleifera supplementation has ameliorated these conditions. Thereby, supplementation with M. oleifera is potential and novel therapeutic detoxication approach for common carp and human health against persistent and bioaccumulating emerging chemicals.