PSX-A-8 Late-Breaking: Development of multiplex assays for equine cytokines IL-1β, IL-4, IL-6, IL-8, IL-10, and TNFα

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 371-371
Author(s):  
Sarah H White-Springer
Keyword(s):  
Polyclonal Antibodies ◽  
Positive Control ◽  
Viable Option ◽  
Meso Scale Discovery ◽  
Multiplex Assays ◽  
Tnf Α ◽  
Before And After ◽  
Bronchoalveolar Fluid ◽  
Cytokine Assay ◽  
Necrosis Factor

Abstract Equine-specific assays to quantify cytokine concentrations are limited and often have a restricted range such that physiological concentrations of many cytokines are below detectable limits of the assay. We aimed to develop custom multiplex assays for equine interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-10, and tumor necrosis factor (TNF) α using the Meso Scale Discovery U-PLEX platform. Equine-specific ELISA kits containing unlabeled and biotinylated polyclonal antibodies and the specific recombinant equine cytokine were purchased for each cytokine. Each biotinylated antibody was coupled to a linker specific for a unique spot within each well of the U-PLEX plates. The unlabeled antibodies were conjugated with electrochemiluminescent labels to serve as detection antibodies. Each cytokine assay was optimized individually prior to optimization in multiplex. Two preliminary experiments were performed: 1) multiplexed conjugation of equine IL-10 and TNFα to the U-PLEX plates; and 2) multiplexed conjugation of equine IL-1β, IL-4, IL-6, and IL-8 to the U-PLEX plates. Standard curves were run at concentrations ranging from 0 to 5,000 pg/mL for TNFα and IL-8, to 12,500 pg/mL for IL-10, to 25,000 pg/mL for IL-1β and IL-6, and to 50,000 pg/mL for IL-4. The minimum average concentrations measured by the standard curves were 0.065, 0.006, 0.017, 0.00013, 0.196, and 0.050 pg/mL for IL-1β, IL-4, IL-6, IL-8, IL-10, and TNFα, respectively. Test samples of equine serum (n = 5) and bronchoalveolar fluid (n = 3) before and after exercise and of ConA-stimulated equine peripheral mononuclear cell supernatants (positive control) were analyzed for each multiplexed assay. With the exception of serum from one horse, all samples ran within detectable limits of each assay. This preliminary work indicates the U-PLEX platform is a viable option to simultaneously quantify concentrations of multiple equine cytokines, allowing for expansion of research efforts focused on understanding immune responses in the horse.

scholarly journals EKSPRESI Tumor Necrosis Factor alpha (TNF-α) DAN Transforming growth factors beta (TGF-β) PADA PERIODONTITIS APIKALIS KRONIS AKIBAT INDUKSI Enterococcus faecalis PADA TIKUS WISTAR

2019 ◽  
Vol 7 (2) ◽  
pp. 66
Author(s):  
Richard Fritzgerald ◽  
Cecilia Lunardhi ◽  
Ruslan Effendy ◽  
Tamara Yuanita
Keyword(s):  
Tissue Damage ◽  
Treated Group ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Transforming Growth Factors ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Factor Alpha ◽  
Necrosis Factor

Background. Root canal treatment is a main role in decreasing infection from root canal and pulp. The main cause of periapical damage mostly are bacteries. E.faecalis is a bactery that is found as an etiology of endodontic treatment failure. Cell wall of this bacteria is containing Lipoteichoic acid (LTA). LTA can penetrate into the periradicular tissue, act as endotoxin in host and cause periradicular inflammation then lead to bone destruction. LTA stimulates immunology reaction that produce Tumor Necrosis Factor alpha (TNF-α) and Transforming growth factors beta (TGF-ß). TNF-α is a main mediator and also have an important role in inflamation response otherwise TGF-ß is working as a multifunction  regulator of cell growth and differentiation during reforming and remodelling.  Purpose. The aim of this study is to know about the expression of TNF-α and TGF-ß during the periapical tissue damage due to induction of E.faecalis. Method. This study used laboratory experimental with the post test only control group design. A total of 30 male rats were randomly divided into 3 main groups, Group A (control negative) : normal tooth. Group B (control positive) : every tooth was induced only by sterile BHI-b. Group C (treated group) : every tooth  was induced by 10 μl BHI-b E.faecalis ATCC212(106 CFU). The animals were sacrificed 21 days later and prepared for histological examination of tissue damage, then we did the immunohistochemistry  followed by calculation on the light microscope. Result. The analysis revealed that the expression of TNF-α at treated group are higher than negative control and positive control but the expression of  TGF-ß at treated group are higher than the negative control group but lower than positive control. Conclusion. From this study we know that the expression of TNF-α and TGF-ß are changing during the periapical tissue damage that induced by E.faecalis.

Anti-Inflammatory and Autonomic Effects of Electroacupuncture in a Rat Model of Diet-Induced Obesity

2018 ◽  
Vol 36 (2) ◽  
pp. 103-109 ◽  
Author(s):  
Xiaoyan Jie ◽  
Xu Li ◽  
Jian-Qing Song ◽  
Dan Wang ◽  
Jian-Hua Wang
Keyword(s):  
High Fat Diet ◽  
Low Frequency ◽  
Control Group ◽  
Vagal Activity ◽  
Diet Induced Obesity ◽  
Tnf Α ◽  
Before And After ◽  
Anti Inflammatory ◽  
Normal Food ◽  
Necrosis Factor

Objective To study the effect of electroacupuncture (EA) on the cholinergic anti-inflammatory pathway (CAP) by measurement of vagal activity in rats with high-fat diet (HFD)-induced obesity. Methods Diet-induced obesity (DIO) was induced in 30 rats by feeding them a HFD for 12 weeks. A further 10 rats fed normal food comprised the lean diet (LD) control group. DIO rats were further subdivided into three groups that received a HFD only (HFD group, n=10), a HFD plus electroacupuncture (HFD+EA group, n=10) or a HFD plus minimal acupuncture (HFD+MA group, n=10). EA and MA treatments were continued for 8 weeks. Heart rate variability (HRV) was used to measure the function of the autonomic nervous system before and after treatment. ELISA was used to determine acetylcholine (ACh) and tumour necrosis factor (TNF)-α levels in the serum. Real-time PCR was used to assess the mRNA expression of α7-subtype nicotinic acetylcholine cholinergic receptors (α7nAChRs) and TNF-α in the mesenteric white adipose tissues (MWAT). Results EA but not MA significantly reduced rats’ bodyweight. No difference was found in the low frequency (LF), high frequency (HF) and the balance between LF and HF (LF/HF) components of HRV before treatment. After the EA intervention, HF was elevated and LF/HF was reduced in the HFD+EA group comparedwith the HFD group. TNF-α in the serum and MWAT were increased in the HFD group, but were reduced in the HFD+EA group. Furthermore, EA promoted expression of α7nAChRs and ACh in the MWAT. There was no difference between the HFD and HFD+MA groups for any indices. Conclusions EA enhanced vagal activity, promoted ACh release and activated α7nAChRs in the MWAT, leading to inhibition of proinflammatory cytokine production.

Natural limonoids protect mice from alcohol-induced liver injury

2020 ◽  
Vol 31 (5) ◽  
Author(s):  
Abacuc Valansa ◽  
Borris Rosnay Tietcheu Galani ◽  
Pascal Dieudonne Djamen Chuisseu ◽  
Armelle Tontsa Tsamo ◽  
Vincent Brice Ayissi Owona ◽  
...  
Keyword(s):  
Liver Injury ◽  
Positive Control ◽  
Negative Control ◽  
Protective Effects ◽  
Tnf Α ◽  
Factor Α ◽  
Necrosis Factor ◽  
Different Doses ◽  
Better Than

AbstractBackgroundAlcoholic liver disease (ALD) is regarded as a global health problem with limited therapeutic options. Previous studies highlighted some anticancer, antiviral, and hepatoprotective activities of limonoids, but the effects of these compounds on ALD remain unknown. The present study aimed to evaluate the effect of some natural limonoids on ethanol-induced liver injury.MethodsThirty-five albino mice (Mus musculus) were administered with 40% ethanol in the presence or absence of the different limonoids [including three havanensin-type limonoids, TS1, TS3, Rubescin D isolated from an African medicinal plant, Trichilia rubescens Oliv. (Meliaceae), and one limonin], or silymarin at 50 mg/kg for 3 days. Thereafter, the effect of the most active compound was evaluated in a chronic model of ALD. For this purpose, 24 mice with each group consisting of six mice were administered orally with 40% ethanol and limonoid at different doses (50, 75, and 100 mg/kg) for 28 days. Finally, biochemical parameters such as alanine aminotransferase (ALT), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), triglyceride (TG), and tumor necrosis factor α (TNF-α) levels were quantified in liver homogenates.ResultsAll tested limonoids significantly (p < 0.01) reduced ALT levels relative to the negative control in the acute model. However, in comparison to other limonoids, limonin at 50 and 75 mg/kg significantly reduced TG, MDA, and TNF-α levels (1.8-fold); alleviated leukocyte infiltration in liver tissue; significantly increased the activity of SOD; and decreased those of CAT better than silymarin used as a positive control at 50 mg/kg.ConclusionsThese data suggest that limonin possesses protective effects on long-term alcohol poisoning partially due to antioxidant and anti-inflammatory mechanisms.

Disease Activity Improvement in Rheumatoid Arthritis Treated with Tumor Necrosis Factor-α Inhibitors Correlates with Increased Soluble Fas Levels

2014 ◽  
Vol 41 (10) ◽  
pp. 1961-1965 ◽  
Author(s):  
Eloisa Romano ◽  
Riccardo Terenzi ◽  
Mirko Manetti ◽  
Francesca Peruzzi ◽  
Ginevra Fiori ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Tumor Necrosis Factor ◽  
Disease Activity ◽  
Tumor Necrosis ◽  
Tumor Necrosis Factor Α ◽  
Soluble Fas ◽  
Tnf Α ◽  
Before And After ◽  
Factor Α ◽  
Necrosis Factor

Objective.Rheumatoid arthritis (RA) is characterized by chronic synovial inflammation and hyperplasia. Tumor necrosis factor-α (TNF-α) plays a pivotal role in RA by interfering with the Fas–Fas ligand (FasL) proapoptotic pathway. We investigated the circulating levels of soluble Fas (sFas) and soluble FasL (sFasL), and their possible correlation with disease activity and improvement after anti-TNF-α treatment in RA.Methods.Serum levels of sFas and sFasL were measured by quantitative ELISA in 52 patients with RA before and after 3 months of anti-TNF-α treatment (adalimumab, n = 32; infliximab, n = 20). Disease activity measures [Disease Activity Score at 28 joints-erythrocyte sedimentation rate (DAS28-ESR), C-reactive protein (CRP)] were recorded before and after treatment. Forty age-matched and sex-matched healthy subjects served as controls.Results.No significant differences in serum sFas levels were detected between anti-TNF-α-naive patients with RA and controls. After anti-TNF-α treatment, serum sFas levels significantly increased in patients with RA compared to both anti-TNF-α-naive patients and controls. Increased sFas levels inversely correlated with disease activity variables (DAS28-ESR: r = −0.739, CRP: r = −0.636, both p < 0.001). No significant differences in sFasL levels were detected in patients with RA before and after anti-TNF-α treatment.Conclusion.In RA, an increase in sFas levels closely correlates with improvement in disease activity induced by TNF-α inhibitors, suggesting their ability to modulate Fas-mediated synoviocyte apoptosis.

Mechanistic Evaluation of the Protective Effect of Carnosine on Acute Lung Injury in Sepsis Rats

Pharmacology ◽  
2017 ◽  
Vol 100 (5-6) ◽  
pp. 292-300 ◽  
Author(s):  
Chenliang Sun ◽  
Qinfeng Wu ◽  
Xuejie Zhang ◽  
Qianru He ◽  
Hongsheng Zhao
Keyword(s):  
Lipid Peroxidation ◽  
Therapeutic Potential ◽  
Albino Rats ◽  
Total Protein Content ◽  
Histopathological Analysis ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Bronchoalveolar Fluid ◽  
Factor Alpha ◽  
Necrosis Factor

This study analyzes the sepsis healing therapeutic potential of carnosine against experimentally sepsis-induced male albino rats. Carnosine in 2 different doses, 25 mg/kg and 50 mg/kg, were administered for 30 consecutive days. At the end of the treatment, lipid peroxidation, catalase, superoxide dismutase, glutathione peroxidase and myeloperoxidase activities were measured. Lungs weight and total protein content were determined in the bronchoalveolar fluid (BALF). Cytokines such as macrophage inhibitory factor (MIF), interleukin-8 (IL-8) and tumour necrosis factor-alpha (TNF-α) were determined in the BALF. In addition, the histopathological analysis was also carried out to understand the effect of carnosine in the cellular architecture. Carnosine treatment significantly renormalized the lipid peroxidation and other antioxidant enzymes. IL-β, TNF-α, and MIF were found to be reduced after carnosine treatment. After carnosine treatment, the intensity of sepsis was significantly reduced evidenced by histopathological analysis. In western blot analysis, carnosine treatment causes the upregulation of IκBα together with the downregulation of the expressions of p65 and p-IKKα/β (Ser 180/Ser 181).

scholarly journals Exercise and the cytokines-interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α): A review

2017 ◽  
Vol 1 (1) ◽  
pp. 3-8 ◽  
Author(s):  
Ambarish Vijayaraghava ◽  
Venkatesh Doreswamy
Keyword(s):  
Tumor Necrosis Factor ◽  
Interleukin 6 ◽  
Normative Data ◽  
Tumor Necrosis ◽  
Tumor Necrosis Factor Α ◽  
Plasma Samples ◽  
Tnf Α ◽  
Before And After ◽  
Factor Α ◽  
Necrosis Factor

Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were one of the first few cytokines to be discovered. The normative data for levels of cytokines IL-6 and TNF-α in particular and all other cytokines in general have not yet been established well. The normal levels for each of the cytokines vary from one race to another. Therefore, all studies need to be done in cases and controls belonging to the same race or same populations. The kits for cytokine assays are expensive and running the assays is laborious and time consuming. It is recommended that the serum/plasma samples are run in duplicates and triplicates to avoid error. Immunology and the field of cytokines is an area which has many domains unexplored. As yet, it is not clearly understood by what mechanisms and pathways each of the cytokines alter the levels of other cytokines. Exercise or physical activity is an intervention which can be administered easily and levels of cytokines measured before and after intervention in same individuals taking all the above mentioned factors into consideration. Hence it is imperative that we look into studies on exercise and cytokines to do further research in the field of cytokines.

scholarly journals CYCLOOXYGENASE MECHANISMS IN THE REGULATION OF RESPIRATORY EFFECTS OF TUMOR NECROSIS FACTOR

2019 ◽  
Vol 19 (1S) ◽  
pp. 17-17
Author(s):  
G A Danilova ◽  
A A Klinnikova ◽  
N P Aleksandrova
Keyword(s):  
Tumor Necrosis Factor ◽  
Tumor Necrosis ◽  
Inflammatory Cytokine ◽  
Ventilatory Response ◽  
Hypoxic Ventilatory Response ◽  
Respiratory Effects ◽  
Cox Inhibitor ◽  
Tnf Α ◽  
Before And After ◽  
Necrosis Factor

Introduction. It is known that the systemic level of the major pro-inflammatory cytokine increases in many respiratory diseases such as asthma, COPD and sleep apnea [1, 2]. The lung ventilation changes and the pathological types of breathing are typical in these diseases. By the reason, the research of the respiratory effects of cytokine is actual. The aim of this study was to compare the respiratory effects of tumor necrosis factor - α (TNF-α) before and after pretreatment with diclofenac, a nonspecific cyclooxygenase (COX) inhibitor. Materials and methods. Тhe experiments were performed in tracheostomized anaesthetized with urethane rats. A respiratory flow head connected to a pneumotachometer (AD Instruments ML141 Spirometer, Dunedin, New Zealand) was used to measure peak airflow and respiratory rate. The hypoxic ventilatory response was measured by using rebreathing with hypoxic gas mixture before and after the tail vein injection of TNF-α (10 μg/rat). In order to determine the role of the cyclooxygenase pathway in the ventilatory effects of TNF-α, introperitoneal administration of diclofenac, a nonspecific COX inhibitor, was used (0.5 mg/rat). Results and discussion. We have shown that the increase in level of TNF-α in blood increased the parameters of respiration such as minute ventilation (by 40%), tidal volume (by 18%), and the mean inspiratory flow (by 33%). The slope of the hypoxic ventilatory response decreased from 6.06 ± 0.91 to 3.48 ± 0.38 ml/min-1 mmHg-1 (by 40%) 40 min after administration of TNF-α (p < 0.05), the slope of tidal volume and mean inspiratory flow also decreased (by 27%) (p < 0.05). After pretreatment with diclofenac, the influence of TNF-α on breathing was dampened, as no significant changes were observed. Conclusion. We concluded that the elevation of inflammatory cytokine level in blood intensifies ventilation during the resting breathing that may be associated with increased central inspiratory activity. At the same time TNF-α reduces the chemoreflex sensitivity to hypoxia, thereby worsening the compensatory capabilities of the respiratory system. Thus, the results of our study suggest participation of inflammatory cytokines in mechanisms of central breathing control and chemoreception. Diclofenac pretreatment eliminated the respiratory effects of TNF-α. The data indicate that the ability of TNF-α to enhance basal ventilation and to reduce the ventilatory hypoxic response is mediated by the COX pathway.

scholarly journals Evaluation of tissue trauma and healing on the basis of tumour necrosis factor-alpha, interleukin-6, and C-reactive protein in peripheral blood during and after pyometra in bitches

2018 ◽  
Vol 74 (10) ◽  
pp. 658-664
Author(s):  
TUĞBA SEVAL FATMA TOYDEMIR KARABULUT ◽  
KIVILCIM SÖNMEZ
Keyword(s):  
Tumour Necrosis Factor ◽  
Tumour Necrosis ◽  
Tumour Necrosis Factor Alpha ◽  
C Reactive Protein ◽  
Necrosis Factor Alpha ◽  
Reactive Protein ◽  
Tnf Α ◽  
Before And After ◽  
Factor Alpha ◽  
Necrosis Factor

In this study, inflammation in the blood of bitches with pyometra (PG) was compared before and 15 days after ovariohysterectomy (OVH). The results were compared with those for control dioestrus bitches (CG) to reveal the evidence of inflammation in blood after a routine surgery. Inflammation was tracked by tumour necrosis factor-alpha (TNF-α), interleukine-6 (IL-6) and C-reactive protein (CRP), using immunocytochemistry (ICC) and immunofluorescence capture of blood cells in cell blocks. ICC is performed mainly during routine cytological examinations, whereas the use of cell blocks in blood examination is uncommon. Insofar as we know, this is the first study using cell block techniques on canine blood samples. Three commercially available antibodies against TNF-α, IL-6 and CRP, forming two panels, were evaluated. A standard streptoavidin-biotin complex technique was used for ICC. TNF-α and IL-6 labelling was scored for colour and intensity, and CRP for immunofluorescence capture. TNF-α and IL-6 colour and intensity scores differed significantly between the PG and CG groups, and were higher in PG before OVH (P&lt;0.01, P&lt;0.01, P&lt;0.001, P&lt;0.01, respectively). IL-6 intensity was significantly greater in PG 15 days after OVH (P&lt;0.05). CRP capture in PG was strong before OVH and high in both groups 15 days later. Low-dose anti-inflammatory agents or an anti-cytokine therapy may be useful in pyometra treatment in the future because these treatments may offer protection from systemic inflammatory response syndrome before and after OVH. .

scholarly journals The Membrane Proteinase 3 Expression on Neutrophils Was Downregulated After Treatment With Infliximab in Patients With Rheumatoid Arthritis

2008 ◽  
Vol 14 (2) ◽  
pp. 186-192 ◽  
Author(s):  
Takeshi Matsumoto ◽  
Toshihiro Kaneko ◽  
Masashi Seto ◽  
Hideo Wada ◽  
Toshihiko Kobayashi ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Resting State ◽  
Proteinase Inhibitor ◽  
Antibody Therapy ◽  
Proteinase 3 ◽  
Tnf Α ◽  
Before And After ◽  
Necrosis Factor ◽  
Stimulation Of

Proteinase 3 (PR3) expression on neutrophils was examined in rheumatoid arthritis (RA) patients before and after antitumor necrosis factor (TNF)-α therapy. Membrane PR3 expression from patients with either an infection or RA significantly increased. Membrane PR3 expression on neutrophils from RA patients treated with infliximab (anti-TNF-α antibody) therapy was less than in those without such treatment in a resting state, but the expression later increased after stimulation in vitro. Membrane PR3 expression increased because of the stimulation of TNFα, whereas it was significantly suppressed by plasma or α1-proteinase inhibitor. The condition of patients with RA improved after treatment with infliximab. Membrane PR3 expression on neutrophils in RA patients was downregulated by infliximab. As a result, PR3 might play an important role in the neutrophil-mediated inflammatory reaction in patients with either RA or an infection.

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