Commercial Enzyme-linked Immunosorbent Assay Research Articles

Longitudinal humoral immune response and maternal immunity in horses after a single live-attenuated vaccination against African horse sickness during the disease outbreak in Thailand.

African horse sickness (AHS) has become a newly emerging disease after an outbreak in northeastern Thailand in March 2020. Mass vaccination in horses with live-attenuated AHS virus (AHSV) vaccine is essential for AHS control and prevention. This study aimed to monitor the longitudinal humoral immune response before and after a single vaccination using a live-attenuated vaccine against AHS in stallions, mares, and pregnant mares, including maternal immunity in foals born from pregnant mares during the outbreak in Thailand. A total of 13 stallions and 23 non-pregnant and 21 pregnant mares were vaccinated with live-attenuated AHSV vaccines. Serum samples from selected horses were collected on the day of vaccination and 1, 6, 8, 9, 10, and 12-months post-vaccination. Furthermore, seven serum samples of foals born from vaccinated pregnant mares were collected on parturition date and 1, 3, and 6-months old. The antibody titer against AHS in all collected serum samples was evaluated using a commercial enzyme-linked immunosorbent assay kit. All data were analyzed for mean and standard deviation for each group of samples using a spreadsheet program. Antibody titers between times were analyzed using a one-way analysis of variance as repeated measurement, and antibody titers between horse groups were analyzed using a general linear model for statistically significant differences when p < 0.05. In stallion and non-pregnant mare groups, there were no statistically significant differences in antibody titers in all 6 time periods after vaccination. The antibody titer in the pregnant mare group showed a non-statistically significant difference between each gestation stage, except at 8 months post-vaccination. Furthermore, increasing antibody titers on days 1 and 3 after receiving colostrum in foals indicate the major role of transcolostral antibody transfer for AHS. This study demonstrated that a single AHS vaccination using a live-attenuated vaccine could stimulate high antibody titers sufficient for AHS control and prevention during the outbreak in Thailand. Similarly, the antibody response of vaccinated horses of both genders, including various stages of pregnant mares, was statistically not different.

Influence of two anti-tumor drugs, pazopanib, and axitinib, on the development and thyroid-axis of zebrafish (Danio rerio) embryos/larvae.

In recent years, the potential toxicities of different pharmaceuticals toward the thyroid system have received increasing attention. In this study, we aim to evaluate the toxic effects of pazopanib and axitinib, two anti-tumor drugs with widespread clinical use, on thyroid function in the zebrafish model. We measured levels of thyroid-related hormones using the commercial Enzyme-Linked Immunosorbent Assay (ELISA) kit. Whole-mount in situ hybridization (WISH) analysis was employed to detect target gene expression changes. Morphology of the thyroid were evaluated by using transgenic Tg (tg: EGFP) fish line under a confocal microscope. The relative mRNA expression of key genes was verified through quantitative real-time polymerase chain reaction (RT‒qPCR). The size and number of the follicles was quantified whereby Hematoxylin-Eosin (H & E) staining under a light microscope. The results revealed that fertilized zebrafish embryos were incubated in pazopanib or axitinib for 96 hours, development and survival were significantly affected, which was accompanied by significant disturbances in thyroid endocrine system (e.g., increased thyroid-stimulating hormone (TSH) content and decreased triiodothyronine (T3) and thyroxine (T4) content, as well as transcription changes of genes associated with the hypothalamus-pituitary-thyroid (HPT) axis. Moreover, based on whole-mount in situ hybridization staining of tg and histopathological examination of zebrafish embryos treated with pazopanib and axitinib, we observed a significantly abnormal development of thyroid follicles in the Tg (tg: EGFP) zebrafish transgenic line. Collectively, these findings indicate that pazopanib and axitinib may have toxic effects on thyroid development and function, at least partially, by influencing the regulation of the HPT axis. Thus, we believe that the potential thyroid toxicities of pazopanib and axitinib in their clinical applications should receive greater attention.

Formulation of Recombinant H1N1 Hemagglutinin in MF59 and Alum Adjuvants: A Comparison of the Vaccines Potency and Efficacy in BALB/C Mice.

In this study, we reported the expression and potency of the recombinant H1N1 hemagglutinin (HA) vaccine as our in-house vaccine in a BALB/c mouse model. Recombinant H1N1 HA was produced in SF9 cell line, purified and formulated in MF59 adjuvant. Experimental mice were injected on days 0 and 14 with MF59-formulated vaccine, alum-based vaccine, and phosphate-buffered saline (PBS). Interleukin (IL)-2, IL-4, and interferon (IFN)-γ were assessed with commercial enzyme-linked immunosorbent assay (ELISA). Antibody responses and cytotoxic T lymphocyte (CTL) activity were assessed by hemagglutination inhibition and granzyme B ELISA, respectively. Moreover, the mice were challenged to show the vaccine efficacy. A considerable rise in IFN-γ and IL-4, as well as IFN-γ/IL-4 ratio, was observed in comparison with the alum-based vaccine and PBS group. Furthermore, our candidate vaccine showed superiority in humoral immune responses and CTL activity versus the alum-based vaccine and PBS group. The challenge showed that the survival rate in the vaccinated groups revealed a significant increase as compared with that in the PBS group. In conclusion, our candidate vaccine showed a robust Th1 response and CTL activity the alum-based vaccine. Moreover, a significant humoral immune response and a higher survival rate were detected in our vaccine as compared with the alum-based vaccine. It seems that the superiority of the MF59-based vaccine is due to the type of vaccine formulation in the candidate vaccine.

Distribution and clinical significance of anti-carbamylation protein antibodies in rheumatological diseases among the Chinese Han population.

Several studies have demonstrated that anti-carbamylation protein antibodies (Anti-CarPA) are persistent in patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), systemic sclerosis (SSC), primary Sjögren's syndrome (pSS), and interstitial lung disease associated with RA (RA-ILD). However, the relationship between anti-CarPA and other rheumatic diseases (RDs) and non-RA-ILD is not known till now. This study sought to examine the presence of anti-CarPA in Chinese Han patients with RDs and its clinical significance. The study included 90 healthy controls (HCs) and 300 patients with RDs, including RA, SLE, polymyositis/dermatomyositis (PM/DM), pSS, SSC, spondyloarthritis (SpA), anti-neutrophil cytoplasmic autoantibodies associated with vasculitis (AAV), undifferentiated connective tissue disease (UCTD), and Behcet's disease (BD). Antibodies against carbamylated human serum albumin were detected using commercial enzyme-linked immunosorbent assay kits. Correlations between clinical and laboratory parameters were analyzed. Serum levels of anti-CarPA in RA (34.43 ± 33.34 ng/ml), SLE (21.12 ± 22.23 ng/ml), pSS (16.32 ± 13.54 ng/ml), PM/DM (30.85 ± 17.34 ng/ml), SSC (23.53 ± 10.70 ng/ml), and UCTD (28.35 ± 21.91 ng/ml) were higher than those of anti-CarPA in the HCs (7.30 ± 5.05 ng/ml). The concentration of serum anti-CarPA was higher in patients with rheumatic disease-related interstitial lung disease (RD-ILD), especially RA-ILD, PM/DM-ILD, and pSS-ILD. Patients with RD-ILD who tested positive for anti-CarPA were more likely to have a more severe radiographic classification (grades II, p = 0.045; grades III, p = 0.003). Binary logistic regression analysis suggested that anti-CarPA had an association with ILD in RA (p = 0.033), PM/DM (p = 0.039), and pSS (p = 0.048). Based on receiver operating characteristics (ROC) analysis, anti-CarPA cutoffs best discriminated ILD in RA (>32.59 ng/ml, p = 0.050), PM/DM (>23.46 ng/ml, p = 0.038), and pSS (>37.08 ng/ml, p = 0.040). Moreover, serum levels of anti-CarPA were correlated with antibodies against transcription intermediary factor 1 complex (anti-TIF1) (R = -0.28, p = 0.044), antibodies against glycyl-transfer ribonucleic acid synthetase (anti-EJ) (R = 0.30, p = 0.031), and antibodies against melanoma differentiation-associated gene 5 (anti-MDA5) (R = 0.35, p = 0.011). Serum anti-CarPA could be detected in patients with RA, PM/DM, pSS, SSC, and UCTD among the Chinese Han population. And it may also assist in identifying ILD in patients with RA, PM/DM, and pSS, which emphasized attention to the lung involvement in anti-CarPA-positive patients.

Dengue seroprevalence study in Bali.

Dengue infection poses significant public health problems in tropical and subtropical regions worldwide. The clinical manifestations of dengue vary from asymptomatic to severe dengue manifestations. This serological survey highlighted the high incidence of asymptomatic cases. This study aimed to determine the prevalence of dengue in healthy and ill adults in Bali. Cross-sectional seroprevalence surveys were performed between July 2020 and June 2021 among healthy and ill adults in Denpasar Bali. Blood samples were collected from 539 randomly selected urban sites in Denpasar. Immunoglobulin G antibodies against the dengue virus were detected in serum using a commercial enzyme-linked immunosorbent assay kit. Overall, the dengue seroprevalence rate among the 539 clinically healthy and ill adults was high (85.5%). The median age was 34.1 (18-86.1). Most of the participants in the study were younger than 40 years (61.2%). Men were the dominant sex (54.5%). The study found a significant association between dengue seropositivity among people aged > 40 years and healthy status (p = 0.005; odds ratio [OR] = 0.459 and p < 0.001; OR = 0.336, respectively). The study reported that as many as 60% of the subjects had a history of previously suspected dengue infection. This study reflected the proportion of asymptomatic dengue patients requiring better assessment with a serological test. The current study highlighted that real cases of dengue infection may be higher than reported, with a high prevalence of dengue seropositivity and a relatively dominant proportion of asymptomatic cases. The study guides physicians to be aware of every dengue infection in tropical countries and prevent the spread of the disease.

One-pot synthesis of AuPt@FexOy nanoparticles with excellent peroxidase-like activity for development of ultrasensitive colorimetric lateral flow immunoassay of cardiac troponin I

Detection of cardiac troponin I (cTnI) plays a critical role in diagnosing acute myocardial infarction (AMI). In this report, a new kind of spherical AuPt@FexOy core@shell nanoparticles (termed as AuPt@FexOy NPs) were one-pot synthesized by a redox interaction-engaged strategy (RIES) without the addition of any surfactants or reducing agents. The as-synthesized AuPt@FexOy NPs not only retain the plasmonic activity of gold nanoparticles (AuNPs), but also possess excellent catalytic activities of platinum nanoparticles (PtNPs) and FexOy nanoclusters. The features of AuPt@FexOy NPs enable greatly enhance the colorimetric detection sensitivity of lateral flow immunoassay (LFIA) through integrating AuPt@FexOy NPs labeling procedure and catalyzing oxidation of chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) signal amplification strategy. The as-developed colorimetric LFIA (termed as AuPt@FexOy-LFIA) exhibits the limit of detection (LOD) as 26.0 pg mL−1 cTnI under the TMB signal amplification mode. In particular, the detection results of cTnI in 40 clinical seral samples by AuPt@FexOy-LFIA are correlated well with those of cTnI in the same samples by commercial enzyme-linked immunosorbent assay (ELISA) detection kit (R2 = 0.97, slope = 1), demonstrating the highly reliable analytical performance and good application prospect of AuPt@FexOy-LFIA.

Poster 151: Femoral Cartilage T2* Relaxation Times Correlate With Inflammatory Biomarker MCP-1 Levels In Serum, But Not Synovial Fluid, After ACL Injury In A Skeletally Immature Porcine Model

Poster 151: Femoral Cartilage T2* Relaxation Times Correlate With Inflammatory Biomarker MCP-1 Levels In Serum, But Not Synovial Fluid, After ACL Injury In A Skeletally Immature Porcine Model

Paper 09: Serum Biochemical Biomarker Profiles Linked to Poor Knee Joint Health Are Not Associated with OA-Related Symptoms at 12 Months Post-ACLR

Objectives: Biochemical joint tissue changes are associated with the development of posttraumatic osteoarthritis (PTOA). Serial assessments of serum biomarkers related to cartilage degradation are predictive of symptomatic progression of idiopathic knee OA but have not been explored in patients with symptomatic PTOA following anterior cruciate ligament (ACL) injury. Both serum and synovial biomarker profiles of inflammation and cartilage degradation post-ACL injury and ACL reconstruction (ACLR) have been associated with deleterious changes of magnetic resonance imaging markers of tibiofemoral cartilage composition (i.e., lower proteoglycan density and poor collagen organization) which precede radiographic PTOA development. However, it is unclear if biochemical changes post-ACL injury and ACLR are also associated with clinically relevant knee OA symptoms. The purpose of this study was to determine if changes in biochemical biomarker profiles preoperatively to 6-months post-ACLR were associated with clinically relevant knee symptoms related to PTOA at 12-months post-ACLR. Methods: Participants with a primary history of ACL injury who were planning on undergoing ACLR with a bone-patellar tendon-bone autograft were included in the prospective, longitudinal cohort study. Serum was collected from participants at three separate study visits: i) preoperatively within 15 days of ACL injury; ii) 6 months post-ACLR; and iii) 12 months post-ACLR. Blood samples from the antecubital fossa were collected in 5 mL serum separation tube vacutainers, centrifuged at 3000 rpms and -4°C, and stored in aliquots at -80°C both preoperatively and 6 months post-ACLR. At 12 months post-ACLR, participants completed the Knee Injury and Osteoarthritis Outcome survey. Upon completion of the study, serum samples were batch processed using commercial enzyme-linked immunosorbent assays (ELISA) to determine biomarker concentrations of monocyte chemoattractant protein 1 (sMCP-1), cartilage oligomeric matrix protein (sCOMP), matrix metalloproteinase 3 (sMMP-3), and type-II collagen degradation to synthesis ratio (sC2C:sCPII) which assess inflammation, cartilage breakdown, cartilage degradation, and cartilage turnover, respectively. KOOS subscale scores were calculated on a scale of 0-100. Participants were classified as Symptomatic or Asymptomatic based on Englund et al. cut off scores for each KOOS subscale which were previously validated in patients with radiographic and symptomatic knee OA after a traumatic knee injury. A K-means cluster analysis was used to determine serum biochemical biomarker profiles based on z-score changes in sMCP-1, sCOMP, sMMP-3, sC2C:sCPII preoperatively to 6 months post-ACLR. One-way ANOVAs were used to determine statistically significant differences between biochemical biomarker changes and characterize the profiles of each group. Independent t-tests were used to assess differences in KOOS subscale scores between each group profile and P-values were Bonferroni corrected for multiple comparisons ( p&lt;0.0125). Cohen’s d effect sizes were calculated to determine the magnitude of differences between groups. Odds ratios (OR) and 95% confidence intervals (CIs) were also calculated to determine the odds of being classified as Symptomatic and reporting clinically relevant, OA-related symptoms. Results: Thirty-six participants completed all study visits and were dichotomized into groups based on changes in biochemical biomarker profiles over 6 months post-ACLR (Table 1). Group 1 (n=15) was characterized by increases in sMCP-1 and sCOMP preoperatively to 6-months post-ACLR (sMCP-1: p&lt;0.001, d=1.75, 95% CI=[0.96-2.52] and sCOMP: p&lt;0.001, d=1.45, 95% CI=[0.70-2.20]) and Group 2 (n=21) was characterized by decreases in sMCP-1 and sCOMP preoperatively to 6-months post-ACLR (Figure 1). No statistically significant differences were found for changes in sMMP-3 ( p=0.18, d=0.46, 95% CI=[-0.22-1.13]) or sC2C:sCPII ( p=0.10, d=-0.57, 95% CI=[-1.25-0.11]) preoperatively to 6-months post-ACLR between groups. Group 1 characterized by serum biochemical biomarker profiles with increasing sMCP-1 and sCOMP preoperatively to 6-months post-ACLR did not demonstrate statistically significant differences in self-reported knee pain ( p=0.72, d=-0.12, 95% CI=[-0.79-0.54]), symptoms ( p=0.96, d=0.02, 95% CI=[-0.64-0.68]), activities of daily living ( p=0.89, d=-0.05, 95% CI=[-0.71-0.62]), sport ( p=0.99, d=0.01, 95% CI=[-0.66-0.67]), or quality of life ( p=0.99, d=0.00, 95% CI=[-0.66-0.67) at 12 months post-ACLR (Figure 2) compared to Group 2 characterized by serum biochemical biomarker profiles with decreasing sMCP-1 and sCOMP preoperatively to 6-months post-ACLR. There were no statistically significant associations between serum biochemical biomarker groups and symptomatic OA status (OR=2.13, 95% CI=0.51-9.01). Conclusions: Pre-operative and post-ACLR changes in biochemical biomarker sampling revealed a group of ACLR patients with increased biomarker concentrations associated with inflammation (sMCP-1) and cartilage breakdown (sCOMP). Serum biomarker changes from the pre-operative to the 6-month post-ACLR time points had no predictive value with respect to patient reporting of clinically relevant knee OA symptoms 12 months post-ACLR. Therefore, serum biochemical changes post-ACLR may play a role in worsening knee joint health as reported in prior studies, but not poor symptom development related to PTOA. Future research is needed to assess the mechanistic role of biochemical changes in symptomatic PTOA development. [Table: see text]

Diabetic retinopathy and its relation to serum brain-derived neurotrophic factor level

Diabetes mellitus (DM) is a metabolic disorder of the proteins, lipids, and carbohydrates, results in hyperglycemia. Abnormalities in the function of insulin on target cells, its release from beta cells, or both may contribute to DM. The purpose of this research was to assess the progression of diabetic retinopathy (DR) to the levels of serum brain-derived neurotrophic factor (BDNF), glycated hemoglobin (HbA1c), and biochemical parameters. The study included 44 normal control subjects, 44 diabetic participants, who were separated into four groups based on their diabetes status and the results of fundoscopic examination. A commercial enzyme-linked immunosorbent assay kit was used to measure the levels of BDNF in the serum. The analysis revealed that diabetics had significantly lower serum BDNF levels than non-diabetics (p&lt; 0.001). Also, there was a significant reduction in BDNF levels with the development of proliferative diabetic retinopathy in comparison with diabetics without DR (p &lt; 0.001). In conclusion, serum BDNF levels decreased significantly in diabetics with and without DR compared to apparently healthy individuals, as well as with the progression of DR.

Brucellosis detection and the role of Brucella spp. cell wall proteins.

Brucellosis remains an endemic zoonotic disease in many developing countries, causing great harm to public health and devastating losses to livestock. One of the main reasons for the low effectiveness of anti-brucellosis measures is the lack of reliable methods for diagnosing infected animals throughout their lifespan. Classical serological tests, such as the tube agglutination test, rose Bengal plate test, and complement fixation test, as well as commercial enzyme-linked immunosorbent assay kits, are based on the detection of antibodies to the cell wall polysaccharide antigens of Brucella spp. smooth strains. As a result, they do not exclude cross-reactions with related bacteria and fail to differentiate between infected and vaccinated animals. Over the past decades, many attempts have been made to identify immunoreactive and pathogen-specific protein antigens. To date, several studies have investigated Brucella spp. recombinant proteins, including cell wall proteins, as the best antigens for diagnosing brucellosis in animals and humans. However, the available results on the specificity and sensitivity of serological tests based on cell wall proteins are ambiguous and sometimes contradictory. This review aims to provide an overview of the current state of knowledge of the diagnostic value of outer membrane and/or periplasmic proteins of Brucella spp. The goal is to identify future developments that may lead to reliable antigens for serological tests.

Seroprevalence of Human Cystic Echinococcosis and Risk Factors in Nahavand, Hamadan, Western Iran

Background: Hydatid cyst is a global infectious disease caused by the larval stage of Echinococcus granulosus. The annual incidence of human cystic echinococcosis (CE) in Iran varies from 0 to 61 in 100,000 cases, depending on geographical conditions, epidemiological factors, and diagnostic methods. Studies showed that E. granulosus infection is one of the most important health problems in most parts of the country. Methods: This descriptive cross-sectional study was conducted on 400 cases referring to Shahid Ghodoosi and Shahid Alimoradi hospital laboratories in Nahavand, Hamadan, Western Iran. IgG antibodies against hydatid cysts were evaluated by the commercial enzyme-linked immunosorbent assay (ELISA) kit, and socio-demographic data were collected using a questionnaire. Then, data were statistically analyzed by SPSS software. Results: Out of 400 subjects in this study, five (1.25%) were positive for CE IgG antibodies by the ELISA method. Two (40%) females and three (60%) males were reported positive for CE IgG via the ELISA Hydatid test. Rural residents consisted of the majority of positive cases (80%). Nearly all of the hydatid-infected subjects were categorized in the age group above 50. Moreover, housewives and self-employed businesses were identified as the two occupations with the highest antibody titers against E. granulosus. In addition, no significant statistical difference was observed in variables. Conclusion: Although the frequency of hydatid cyst infection in Nahavand is lower than the average proportion confirmed by previous studies in the country, due to the numerous records of hydatid cyst disease in the region, the establishment of health programs is regarded essential to control this parasitic infection.

Serum Zonulin and Claudin-5 but not Interferon-Gamma and Interleukin-17A Levels Increased in Children with Specific Learning Disorder: A Case-Control Study.

Gut-blood and blood-brain barrier permeabilty (gut-brain axis) has been attracting increased attention in the etiology of neurodevelopmental disorders. In this study, we aimed to investigate serum levels of zonulin (a biomarker of intestinal permeability), claudin-5 (a biomarker of blood-brain barrier permeability), and interferon-gamma and interleukin-17A in children with specific learning disorder. Forty-three children with DSM-5 diagnosis of specific learning disorder and 43 healthy children were included in this study. Serum levels of zonulin, claudin-5, interferon-gamma, and interleukin-17A were measured using commercial enzyme-linked immunosorbent assay kits. Serum zonulin and claudin-5 levels of the study group were significantly higher than the control group according to the multivariate analysis of covariance test while controlling for age, gender, and body mass index. However, serum interferon-gamma and interleukin-17A levels were not significantly different between the two groups. There was no correlation either between zonulin and interferon-gamma and interleukin-17A or claudin-5 and interferon-gamma and interleukin-17A. Gut-blood and blood-brain barrier permeability may be disrupted in subjects with special learning disorder. Further research is needed to determine whether zonulin and claudin-5 may be biomarkers, and some dietary interventions or specific agents such as zonulin or claudin-5 inhibitors could be used in the management of neurodevelopmental disorders including special learning disorder.

Nanozyme-Catalyzed Metasurface Plasmon Sensor-Based Portable Ultrasensitive Optical Quantification Platform for Cancer Biomarker Screening.

Developing plasmonic biosensors that are low-cost, portable, and relatively simple to operate remains challenging. Herein, a novel metasurface plasmon-etch immunosensor is described, namely a nanozyme-linked immunosorbent surface plasmon resonance biosensor, for the ultrasensitive and specific detection of cancer biomarkers. Gold-silver composite nano cup array metasurface plasmon resonance chip and artificial nanozyme-labeled antibody are used in two-way sandwich analyte detection. Changes in the biosensor's absorption spectrum are measured before and after chip surface etching, which can be applied to immunoassays without requiring separation or amplification. The device achieved a limit of alpha-fetoprotein (AFP) detection < 21.74 fM, three orders of magnitude lower than that of commercial enzyme-linked immunosorbent assay kits. Additionally, carcinoembryonic antigen (CEA) and carbohydrate antigen 125 (CA125) are used for quantitative detection to verify the universality of the platform. More importantly, the accuracy of the platform is verified using 60 clinical samples; compared with the hospital results, the three biomarkers achieve high sensitivity (CEA: 95.7%; CA125: 90.9%; AFP: 86.7%) and specificity (CEA: 97.3%; CA125: 93.9%; AFP: 97.8%). Due to its rapidity, ease of use, and high throughput, the platform has the potential for high-throughput rapid detection to facilitate cancer screening or early diagnostic testing in biosensing.

A survey for antibodies against Fasciola hepatica in cattle and sheep in Finland indicates a low level of exposure

BackgroundFasciolosis is a parasitic infection caused by the liver fluke Fasciola hepatica that can have a major economic impact on livestock industry. The prevalence of the disease has recently been increasing in many North European countries. The objective of this study was to determine the prevalence of antibody against F. hepatica in Finnish cattle herds and sheep flocks during 2019 by using a commercial enzyme-linked immunosorbent assay (ELISA). Randomly selected bulk tank milk samples were obtained from 660 dairy herds. Blood samples were collected at slaughterhouses from 1944 suckler cows from 309 herds and from 1120 sheep from 95 flocks.ResultsAntibodies against F. hepatica were found in 0.45% (95% confidence interval (CI): 0.15–1.33) of the dairy herds and 0.97% (95% CI: 0.33–2.82) of the suckler cow herds. The seropositive herds were located in eastern and central Finland. None of the sampled sheep flocks tested had antibodies against F. hepatica (95% CI: 0–3.89). The results of the assays were compared with meat inspection data received from the slaughterhouses. All positive herds also had liver condemnations due to F. hepatica based on the meat inspection reports.ConclusionsCompared to other North European countries, the prevalence of fasciolosis in Finland can be considered low, and according to meat inspection reports, there are no indications of the prevalence increasing in Finland.

1492-P: IL-8/CXCL8 May Identify a New Type 1 Diabetes Endotype

Recent research focused on identifying mechanisms explaining the observed heterogeneity of type 1 diabetes (T1D) in age at onset, residual β-cell function and rate of disease progression. Mechanisms include changes in both the immune system and β-cells, leading to the loss of tissue-specific immune tolerance and disease onset. Interleukin-8 (IL-8; CXCL8) is a pro-inflammatory chemokine released by macrophages, endothelial, epithelial and airway smooth muscle cells playing a major role in the innate immune response. Macrophages release IL-8 at the site of an injury where they recruit and activate neutrophils by interacting with the IL-8 receptors CXCR1 and CXCR2. An increase in circulating IL-8 has been shown in several autoimmune disorders. IL-8 levels, but not IL-6 or TNF-α, are elevated in adolescents with T1D, and also associated with insulin resistance, suggesting a role for IL-8 beyond acute inflammation in this setting. In vitro experiments revealed that IL-8 transcription is induced by hyperglycaemia. In the present study newly diagnosed T1D patients within the first year of disease onset (mean age 16.2 ± 7.6 yrs and % HbA1c of 10.7 ± 2.7) (N=42) were studied. Clinical and laboratory features included age, gender, body mass index (BMI), disease duration, HbA1c, and C-peptide. Serum IL-8 and myeloperoxidase (MPO) were measured with commercial enzyme-linked Immunosorbent assays. When compared to normal subjects, T1D patients showed a significantly higher IL-8 values (median [IQR] 112.0 [64.74-311.8] vs 43.05 [30.94-49.70], p-value= 0.002, respectively). Furthermore, T1D patients showed a significantly higher MPO levels than controls (median [IQR], 95327 [55718 - 168974] vs 42729 [24147 - 95327], p-value= 0.0349, respectively). This study shows for the first time that IL-8 and MPO are elevated in patients with recent onset T1D compared to normal subjects. There is also a trend for a positive correlation between IL-8 and HbA1c levels suggesting that elevated IL-8 values are associated with poor metabolic control. Disclosure G.Alhamar: None. S.Fallucca: Consultant; Dompé. S.Pieralice: None. L.Valente: None. P.Pozzilli: None.

Cissampelos pareira ethanolic extract modulates hormonal indices, lipid profile, and oxidative parameters in transient infertility-induced female albino rats

Cissampelos pareira is a medicinal plant with conflicting local claims regarding its usage in the management of fertility disorder. This study investigates the effect of the ethanolic extract of C. pareira on fertility indices in female albino rats. Ethanolic extract of C. pareira was administered orally for 14 days to female rats placed in different groups. Levonorgestrel (0.14mg/g) was orally administered to certain rat groups for 7 days to induce transient infertility before other treatments. The rats were sacrificed at the completion of their respective oral administration route. Blood samples were collected through cardiac puncture for hormonal and lipid profiling, which were analysed using commercial standard enzyme-linked immunosorbent assay (ELISA) kit. Organs were harvested for the assessment of oxidative parameters. Phytochemical screening reveals the presence of alkaloid, saponin, glycoside, tannin, steroid and flavonoid in the extract. The extract has no significant effect on progesterone, significantly reduced testosterone (p&lt;0.05), but increased prolactin and estradiol concentrations (p&lt;0.05). No significant hormonal difference in groups pre-administered levonorgestrel. The extract significantly elevated serum triglyceride, but reduced cholesterol level (p&lt;0.05), even in groups pre-administered with levonorgestrel. The extract significantly reduced H2O2 and SOD activities (p&lt;0.05) in the ovary and kidney respectively. Following levonorgestrel pre- administration, the extract significantly reduced H2O2 and GSH in the liver and kidney, elevated SOD activities in the liver, ovary and brain (p&lt;0.05). This study demonstrated the antifertility properties of the ethanolic extract of Cissampelos pareira, shown by its effects of the hormonal and lipid profile, with moderate antioxidant effect.

Comparative Analysis of Different ELISA Methods for the Serodiagnosis of Przhevalskiana silenus Infestation in Goats

Przhevalskiana silenus (warble fly) grubs cause myiasis in goats, in mountainous and semi-mountainous areas and different regions in Pakistan, and cause substantial losses to livestock. The palpation method for detecting warble flies generally neglects the infestation intensity; therefore, the development of a reliable and efficient diagnostic technique is extremely necessary. This study compared three indirect enzyme-linked immunosorbent assay (ELISA) methods for detecting anti-P. silenus antibodies using the hypodermin C (HyC) purified from Hypoderma spp. Larvae collected in cattle (local isolate, Microbiology Laboratory, PMAS-Arid Agriculture University, Rawalpindi), the crude antigen from the first instar stage of P. silenus, and a commercial Bovine Hypodermosis Antibody ELISA kit (IDEXX Laboratory), for accurately estimating the seroprevalence of goat warble fly infestation (GWFI) in the Pothwar plateau, Punjab, Pakistan. The ELISA with the crude antigen of P. silenus proved very sensitive and specific, 91% and 93%, respectively. The optical density exhibited a monthly variation, and the antibody titer began increasing from June, continually increased from July to December, and gradually decreased thereafter until March. The study confirmed the endemic status of GWFI in the Pothwar region and identified that ELISA based on the crude antigen of P. silenus was a more sensitive and specific immunodiagnostic method for determining seroprevalence, and could be employed for initiating nationwide eradication campaigns.

Effects of Polydextrose Supplementation on Intestinal Function in Hemodialysis Patients: A Double-Blind, Randomized, Placebo-Controlled Trial

Intestinal constipation is a frequent complication in hemodialysis (HD) patients. Polydextrose (PDX), a nondigestible oligosaccharide, has been reported as a fermentable fiber with potential benefits. This study aimed to investigate the possible influence of PDX supplementation on intestinal function in HD patients. This randomized, double-blind, placebo-controlled trial included 28 patients who received daily oral supplementation with 12g of PDX or placebo (corn starch) for 2months. ROME IV criteria were used to define constipation and questionnaires were applied to patient assessment of constipation symptoms (PAC-SYM) and their impact on the patient assessment of constipation quality of life. The Bristol scale was used to assess stool consistency. Commercial Enzyme-Linked Immuno Sorbent Assay kits were used to evaluate the interleukin-6 and tumor necrosis factor-α plasma levels. 25 patients completed the study; 16 in the PDX group [7 females, 48.5years (IQR=15.5)] and 9 in the control group [3 females, 44.0years (IQR=6.0)]. According to ROME IV criteria, 55% of patients were diagnosed with constipation. PAC-SYM faecal symptoms domain was reduced after 2months of PDX supplementation (P=.004). We also observed a significant reduction in the PAC-QoL-concerns domain (P=.02). The average values for PAC-SYM and patient assessment of constipation quality of lifewere reduced significantly after intervention with PDX. There were no significant changes after the intervention period concerning biochemical variables, food intake, and inflammation markers. No adverse effects were observed during the supplementation period. The results of the present study suggest that short-term PDX supplementation may have favourable results on intestinal function and the quality of life of chronic kidney disease patients in HD.

Increasing Levels of Macrophage Migration Inhibitory Factor in COVID-19 Infection and Its Pathophysiological Role; Cut-off Value Determination Might Be Clinically Misleading

Background: The serious outbreak of coronavirus disease 2019 (COVID-19) has provoked deep concern throughout the world. The pathophysiologic network leading to severe conditions has still unsolved gaps. Considered a pleiotropic, multifaceted cytokine, macrophage migration inhibitory factor (MIF) has distinct functions, which seem to stand at the edge of distinct known mechanisms involving in COVID-19 pathogenesis. Additionally, MIF is a key mediator of acute respiratory distress syndrome and lung injury. Objectives: The current study aimed to evaluate the serum levels of MIF in COVID-19 patients, particularly in severe cases. Methods: This case control study was performed on the sera of 60 randomly selected COVID-19 patients as case group and 30 randomly selected healthy individuals as control group during November 2020 till April 2021 at Mashhad University of Medical Sciences. The case group included 30 outpatients with mild disease and 30 hospitalized severe subjects. A commercial enzyme-linked immunosorbent assay was utilized to measure serum MIF. Data were analyzed using SPSS version 16 with student t-test and chi-squared test considering a P &lt; 0.05 as statistical significance level. Results: There was no statistical difference between two groups regarding demographic variables. According to the obtained data, significantly higher MIF levels were observed in the affected subjects than the healthy individuals, particularly in severe COVID-19 subjects (severe: 65.31 ± 6.2 ng/mL, mild: 40.45 ± 6.6 ng/mL, healthy: 20.63 ± 6.1 ng/mL P &lt; 0.0001). The receiver operating characteristic (ROC curve) drawn for the present study illustrates that MIF amounts differentiate COVID-19 severe and mild cases with high accuracy (90.8%) (sensitivity:86.6%, specificity:96.6%). Conclusions: There might be an association between MIF concentration with respiratory failure and disease exacerbation due to COVID-19 infection. Therefore, MIF can act as a marker of clinical severity for COVID 19 infection. However, due to variations in MIF amounts a definite cut-off value might be specific to the study and should be considered with caution. MIF is supposed to be one of the most important cytokines in COVID-19 pathogenesis and might be a target for therapeutic approaches with MIF inhibitors in possible upcoming disease peaks.

Canine Toxoplasmosis: Cytokine Profile in Seropositive Stray Dogs

Toxoplasma gondii is a ubiquitous protozoan parasite that may cause devastating conditions in animals, including humans. Dogs are the main host of T. gondii; however, little information is available on the cytokine expression profile in T. gondii–seropositive dogs. The present study aims to evaluate the levels of certain cytokines in serum collected from T. gondii–seropositive stray dogs. With a commercial enzyme-linked immunosorbent assay kit, we investigated the presence of antibodies for T. gondii in stray dogs in Hefei, Anhui, P. R. China, and found 26 of those tested were seropositive. For proinflammatory cytokines, higher levels of interleukin (IL)-6 were detected in the sera of T. gondii–seropositive dogs compared with seronegative dogs. Additionally, the concentration of anti-inflammatory cytokine IL-10 in seropositive dogs was slightly lower.