ELISA Results Research Articles

FSTL1 Can Be a Promising Target in TMJ Osteoarthritis via Regulating Chondrocyte Mitophagy and Apoptosis.

Previous studies have shown that follistatin-like protein 1 (FSTL1) is elevated in the synovial fluid of osteoarthritis and whether it is associated with disease development progress in cartilage degeneration is still unclear. The experiment was performed to explore the effect and mechanism of FSTL1 on chondrocyte degeneration and its further impaction in osteoarthritis as well as its treatment method. The patients who were diagnosed with temporomandibular joint (TMJ) disc displacement and osteoarthritis (OA) group was divided into 2 groups, anterior disc displacement (ADD) without bone resorption and ADD with bone resorption group according to the radiologic examination. The ELISA kit was used to determine the expression level of FSTL1 in patients TMJ environment. The function of FSTL1 in promoting chondrocyte degeneration was tested by quantitative reverse transcription polymerase chain reaction (Rt-qPCR) and western blot. The chondrocyte apoptosis and mitophagy were further test by flow cytometry and mitosox staining by upregulating and downregulating of FSTL1. In the end, the effectiveness of regulating FSTL1 in OA procedure was further validated by hematoxylin-eosin (HE), safranin O, and immunohistochemical (IHC) staining in vivo. There were 56 samples collected from the patients were included into this study. According to the ELISA results, FSTL1 expression levels of ADD without bone resorption groups were significantly lower than that in ADD with bone resorption group. Furthermore, the rate of cell apoptosis cells and the mitophagy procedure were highly activated when FSTL1 was upregulated. The morphology analysis of mitochondria showed significant changes when FSTL1 was highly upregulated in vitro. The in vivo and in vitro experiments showed that suppressing FSTL1 could alleviate the cartilage degeneration in TMJ OA progression. To sum up, upregulated expression level of FSTL1 in synovial fluid promoted the progression of TMJ OA by upregulating accelerating the chondrocyte apoptosis and mitophagy, and suppressing the FSTL1 in TMJ can rescue the OA progression. Therefore, it may be a promising result to consider the FSTL1 as a therapeutic target in the future.

Chemiluminescent Fe3O4@Nickel-Cobalt Double Hydroxide Magnetic Core-Shell Nanomaterial as an Analytical Interface for Label-Free CYFRA21-1 Immunosensing.

Cytokeratin 19 fragment (CYFRA21-1) is considered to be a potential marker for the diagnosis and of classification of lung cancer. It is highly desired to develop rapid and highly sensitive label-free chemiluminescence (CL) immunosensors for the detection of CYFRA21-1. In this work, magnetic core-shell nanocomposite Fe3O4@nickel-cobalt double hydroxide (Fe3O4@DH) was used for the first time as a carrier to synthesize N-(4-aminobutyl)-N-ethylisoluminol (ABEI) and gold and silver nanocluster (AuAgNC) bifunctionalized magnetic nanomaterials Fe3O4@DH/AuAgNCs-ABEI. The resulting Fe3O4@DH/AuAgNCs-ABEI demonstrated excellent CL performance, attributed to the exceptional catalytic capability of nickel-cobalt double hydroxide (NiCo-DH) and AuAgNCs, as well as the large specific surface area of NiCo-DH for AuAgNCs and ABEI enrichment. Moreover, Fe3O4@DH/AuAgNCs-ABEI exhibited superior stability compared to Fe3O4-based CL-functionalized magnetic nanomaterials, owing to the presence of NiCo-DH shells. On this basis, a label-free CL immunosensor using CYFRA21-1 antibody-modified Fe3O4@DH/AuAgNCs-ABEI as an analytical interface was constructed for the detection of CYFRA21-1 in the range from 1.0 × 10-13 to 1.0 × 10-8 g/mL. The detection limit of this CL immunosensor was 47.7 fg/mL, 4 orders of magnitude lower than existing CL methods. The CL immunosensor was able to accurately detect the concentration of CYFRA21-1 in serum samples, as evidenced by ELISA results. More importantly, it not only distinguished well between healthy persons and lung cancer patients (90.0% sensitivity and 90.0% specificity), but also effectively distinguished between patients with non-small cell lung cancer and small cell lung cancer (80.0% sensitivity and 86.7% specificity).

Lyme serology in non-endemic countries: Why do we request it and what do we find?

BackgroundThe primary objectives of our research were to analyze Lyme serology results from pediatric patients, identify the clinical reasons for ordering these tests, and assess the clinical relevance of the serology results in the context of Lyme disease.MethodsOur study, conducted at a reference pediatric hospital in a non-endemic region for Lyme disease, included all pediatric patients for whom Lyme serology was requested. ELISA and Western blot results were documented. Presenting complaints and findings at the time of admission were recorded. Diagnostic tests were categorized as first-step tests if conducted during the initial visit. Subsequent tests, sent due to the inability to establish a diagnosis based on the results of the first-line tests, were defined as second and third-step tests, respectively.Results219 patients, for whom Lyme serology had been requested for varying reasons, were included in our study. The most prevalent complaints and indications observed in the presentations of patients with serology requests were as follows: headache(24.7%), paresis/paresthesia(14.6%), and painful or blurred vision(13.2%). Serology was primarily requested in the presence of neurological(59.4%) and ocular symptoms(13.2%). 68% of the tests were requisitioned during the initial consultation. Nevertheless, it was discerned that no patient received a diagnosis of Lyme disease.ConclusionAccording to the guidelines, Lyme serology should only be performed when there is a realistic possibility of exposure to infected ticks, particularly in patients who have had untreated erythema migrans or those with a history of a tick bite presenting with unexplained joint or neurological symptoms. Our data, in line with these guidelines, suggests that unnecessary Lyme serology testing in non-endemic areas, where exposure is highly unlikely, may lead to false-positive results and unnecessary follow-up testing, as illustrated by the high rate of false-positive ELISAs in our cohort.

Unraveling immune checkpoint inhibitors effects on atherosclerosis: mechanisms and preventive approaches

Abstract Background Immune checkpoint inhibitors (ICIs) represent a novel and rapidly evolving field in cancer treatment. Despite increasing reports of accelerated atherosclerosis and ICI-related atherosclerotic cardiovascular (CV) events in clinical studies and case reports, there is a notable lack of data concerning the underlying mechanisms, appropriate monitoring, and potential interventions. Objectives To explore the role of ICIs in triggering and/or accelerating the atherosclerotic process, with a specific focus on the involvement of CD8+ T-cells. Additionally, we aimed to evaluate and compare the potential beneficial effects of statins and Proprotein Convertase Subtilisin/Kexin type 9 (PCSK9) inhibitors on atherosclerosis under ICIs treatment. Methods Thirty 8-week-old C57BL mice were divided into six groups as follows: (1) Control, normal diet; (2) Control, atherogenic diet; (3) Atherogenic diet + anti PD-1; (4) Atherogenic diet + statins + anti-PD1; (5) Atherogenic diet + PCSK9-i + anti PD-1; (6) Atherogenic diet + PCSK9 inhibitors. At 16 weeks, the carotid arteries and heart were removed for histological examination, and serum was withdrawn for blood work and further analysis. Results Anti-PD1 treatment resulted in extensive lymphocytic infiltration of the carotid intima, primarily constituted of CD8+ T cells. This infiltration was significantly attenuated when statins or PCSK-9 inhibitors were used in conjunction with anti-PD1 treatment. Serum analysis for CD4+ and CD8+ T cells by ELISA revealed no difference in CD4+ abundance between the groups. However, plasma CD8+ T cells were significantly increased with anti-PD1 treatment, and furthermore, when PCSK9 inhibitors were used in combination; but not when statins were used. Conclusion The induction of CD8+ T cell infiltration in the arterial carotid walls by anti-PD-1 was prevented with the use of either statins or PCSK9 inhibitors. In the plasma, while statins reduced the hyper-expression of CD8+ T cells observed under PD-1 treatment, PCSK9 inhibitors enhanced this expression. This may provide a first indication of the potential benefit of PCSK9 inhibitor treatment for both preventing arterial inflammation and enhancing the desired anti-tumor immune response under ICIs treatment. Figure legend: 1A. A cross-section of the carotid artery after 16 weeks of treatment, stained with H&E. Magnification is X100. 1B. A cross-section of the carotid artery after 16 weeks of treatment, stained with anti-CD8 Ab. Magnification is X100. 1C. quantitative CD8+ T cells staining in the carotid arteries. 1D. quantitative results of ELISA for plasma CD8+ T cells.

Comparison of triclosan and triclocarban in triggering immunotoxicity in larval zebrafish

As two efficient broad-spectrum sterilizing agents, triclosan (TCS) and triclocarban (TCC) are widely used, especially during the COVID-19 pandemic. The health risks caused by secondary pollution of TCS and TCC have aroused wide concern. Because of the similar mother nucleus structure and high lipophilicity, it remains unknown about the differences in the effect and mechanism of the toxicity (especially immunotoxicity) between TCS and TCC in organisms in the environment. In this study, we used zebrafish as a model to compare the immunotoxicity and mechanisms between the two pollutants at the same exposure concentration (0.6 µmol/L). The results showed that both TCS and TCC led to a hatching rate below 60% at the time point of 72 hours post fertilization (hpf) and the mortality rates of 40% and 50% at 120 hpf in larval zebrafish, respectively. The zebrafish exposed to TCS and TCC displayed malformations, such as shortened body, swimming sac closure, pericardial edema, yolk cyst deposition, and absorption disorder. Moreover, the developmental abnormalities caused by TCC were significantly severer than those caused by TCS. TCS exposure increased the proliferation rate of innate immune cells to 20% and decreased the number of mature T cells by 35%, while TCC exposure inhibited the differentiation of both innate immune cells and T cells, with the inhibition rates of 25% and 60%, respectively. The results of real-time quantitative PCR (RT-qPCR) and ELISA showed that TCS and TCC exposure up-regulated the expression levels of il-1β, il-6, and tnf-α, while il-10 and IgM exhibited opposite expression patterns. Additionally, both compounds slightly decreased C3 expression. The Pearson correlation analysis showed that the developmental toxicity induced by TCS and TCC had positive and negative correlations with the differentiation of immune cells, respectively. However, the toxicity induced by either TCS or TCC was positively correlated with the expression of pro-inflammatory cytokines. GO function and KEGG pathway enrichment analyses demonstrated that the target molecules of TCS and TCC were enriched in different signaling pathways, and the key network hub genes and the enriched regulatory pathways differed between TCS and TCC. The findings provide compelling evidence that TCS and TCC adopt different mechanisms in triggering immunotoxicity and offer a theoretical reference for the recognition, warning, and management of TCS and TCC-induced health risks.

Tinglu Yixin granule inhibited fibroblast-myofibroblast transdifferentiation to ameliorate myocardial fibrosis in diabetic mice

Ethnopharmacological relevanceMyocardial fibrosis is one of the pathological characteristics of advanced diabetic cardiomyopathy (DCM) and serves as the strong evidence of poor prognosis. Among them, the transdifferentiation of cardiac fibroblasts (CFs) may play a crucial role in the development of myocardial fibrosis in DCM. Tinglu Yixin granule (TLYXG) has been clinically used for many years and can significantly improve cardiac function of patients with DCM. However, the effect of TLYXG on myocardial fibrosis in DCM remains unknown, and the underlying mechanisms of its efficacy have yet to be fully understood. Aim of the studyThis study aimed to investigate the impact and underlying mechanism of TLYXG on myocardial fibrosis in diabetes mice. Materials and methodsThe bioactive compounds in TLYXG were identified using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). The potential mechanism of TLYXG in treating DCM was predicted using network pharmacology combined with molecular docking and protein-protein docking. The mice model of type 2 diabetes were established by intraperitoneal injection of streptozotocin (STZ) and the high-fat diet (HFD). Indicators of pancreatic islet function, lipids, oxidative stress, and inflammatory factors were tested using kits. Cardiac function was assessed in diabetic mice using echocardiography. Histologic staining was performed to evaluate myocardial hypertrophy and fibrosis. Mechanistically, the hypothesis was tested through rescue experiments. The expression levels of transient receptor potential channel 6 (TRPC6), transforming growth factor-β1 (TGF-β1), collagen I (COL-I) and alpha-smooth muscle actin (α-SMA), along with the mRNA and phosphorylation levels of SMAD family member 3 (Smad3) and protein 38 mitogen-activated protein kinase (p38 MAPK), were assessed using quantitative RT-qPCR, Western blot, immunohistochemistry, and immunofluorescence. Neonatal lactating mice were used to extract primary CFs for vitro experiments. Scratch and transwell assays were conducted to assess CFs migration and invasion abilities. Western blot and immunofluorescence were used to evaluate the expression levels of CFs transdifferentiation markers COL-I and α-SMA. ResultsA total of 168 active ingredients were detected in TLYXG based on UPLC-MS and databases. Network pharmacology indicated that TLYXG could improve DCM through inflammatory mediator regulation of TRP channels, TGF-beta signaling pathway, and MAPK signaling pathway. ELISA results showed that TLYXG could ameliorate metabolic levels, inflammation, and oxidative stress in diabetic mice. Echocardiography suggested that TLYXG improved cardiac systolic and diastolic dysfunction in diabetic mice. Histological analysis revealed that TLYXG alleviated myocardial fibrosis in diabetes mice. Additionally, molecular docking analysis indicated strong binding activity between the main active ingredients of TLYXG and TRPC6 of the TRP family. At the molecular level, TLYXG reduced the mRNA and protein expression levels of TRPC6 and TGF-β1 and inhibited the mRNA and phosphorylation levels of Smad3 and p38 MAPK. Furthermore, TLYXG inhibited CFs migration and invasion, and reduced the expression levels of the CFs transdifferentiation markers COL-I and α-SMA. ConclusionTLYXG inhibited the proliferation, migration, invasion and transdifferentiation of CFs by suppressing TGF-β1/Smad3/p38 MAPK signaling through down-regulation of TRPC6, thereby ameliorating myocardial fibrosis in diabetes mice.

Chidan Tuihuang granule modulates gut microbiota to influence NOD1/RIPK2 pathway in cholestatic liver injury recovery

BackgroundCholestatic liver injury (CLI), which occurs if bile acids are imbalanced and the liver becomes inflamed, is difficult to treat effectively ObjectiveWe investigated how the Chinese patent medicine Chidan Tuihuang granule (CDTH) ameliorates cholestatic liver injury with a focus on its effects on the NOD1/RIPK2 pathway and intestinal flora MethodsWe used an ANIT-induced SD rat model of CLI to evaluate the therapeutic effects of CDTH. The experimental design included control, model, UDCA (ursodeoxycholic acid) and CDTH treatment groups. UHPLC-Q-Orbitrap-HRMS was used to analyse the blood components of CDTH. The efficacy of CDTH was assessed by liver function tests, histopathological examination (HE and TUNEL staining), transmission electron microscopy, and ELISA to measure apoptosis and inflammatory markers. Mechanistic insights were obtained using transcriptomics and RT-qPCR, while alterations in the expression of key proteins were studied using western blotting, immunohistochemistry, and immunofluorescence. Furthermore, the impact of CDTH on the gut microbiota and its associated metabolite, meso-2,6-diaminopimelic acid (DAP), which is linked to NOD1 activation, was examined and confirmed through in vitro ResultsThe experimental results demonstrated a notable elevation in serum levels of AST, ALT, ALP, TBA, TBIL, and DBIL in the rats belonging to the model group, accompanied by the infiltration of inflammatory cells, hepatocyte degeneration, and necrosis in the liver tissue. CDTH administration significantly improved liver function and cholestasis indicators. Transmission electron microscopy and TUNEL staining revealed a marked reduction in liver cell apoptosis with CDTH treatment. ELISA results showed that CDTH effectively reduced inflammatory markers. Transcriptomic analysis showed that CDTH inhibited the NOD1/RIPK2 pathway, resulting in a significant decrease in the expression of NOD1, RIPK2 and associated genes in liver tissue. Gut microbiota analysis demonstrated that CDTH regulated intestinal flora structure, reducing the abundance of DAP-producing Gram-negative bacteria such as lactobacilli. In vitro experiments confirmed that CDTH enhanced cell viability by downregulating the DAP-mediated NOD1/RIPK2 signaling pathway secreted by intestinal bacteria ConclusionCDTH ameliorated liver damage in cholestatic rats by inhibiting the NOD1/RIPK2 signaling pathway through regulation of gut flora and downregulation of DAP metabolites.

Evaluation of a point-of-care multiplex immunochromatographic assay for the diagnosis of typhoid: results from a retrospective diagnostic accuracy study.

Currently available diagnostic tests for typhoid have several limitations, including low sensitivity and specificity. Dual Path Platform Typhoid assay is a multiplex rapid test that detects IgA antibodies to lipopolysaccharide and hemolysin E antigen. It is considered to have high sensitivity and specificity, and its results were found to be highly correlated with ELISA results. However, very few studies have been conducted to evaluate this test and limited information about the accuracy of this test is present. Hence, this study evaluated the new typhoid test.

Concordance Between Platelet Expression Assay (PEA) and Serotonin Release Assay (SRA) in Heparin-Induced Thrombocytopenia: Implications for Diagnostic Accuracy and Laboratory Test Utilization

Abstract Introduction The Serotonin Release Assay (SRA) serves as the gold standard functional assay for heparin-induced thrombocytopenia (HIT) diagnosis at our institution. The present turnaround time (TAT) for SRA results ranges from 4-5 days, occasionally extending beyond 8 days, given outsourcing to a reference laboratory. Recently, an alternative functional assay, the Platelet Expression Assay (PEA), has been introduced. The PEA detects platelet activation through p-selectin expression by flow cytometry, with a reported sensitivity of 100% and specificity of 95% for HIT. The PEA is also a send-out test, and thus has similar TAT and cost profiles as the SRA, but with potential to transition into an in-house assay in the future. Herein, we sought to determine the concordance of the PEA and SRA for the detection and diagnosis of HIT within our patient population. Methods A retrospective review was conducted on all cases in which a HIT ELISA (screening immunoassay) was performed by the Special Coagulation Laboratory during a one-year period. A 4Ts score was calculated if not already documented by chart review. In a subset analysis, specimens with a positive screening ELISA were concurrently tested by both SRA and PEA to compare performance. Results Of the 1341 cases, a 4Ts score was calculated and found to be less than 4 for 30-50% of cases. A total of 490 cases were sent out for SRA, of which 49 (10%) returned positive results. In a subset analysis of 20 samples from 14 patients, all with positive ELISAs, results revealed concordance between the SRA and PEA for all but one sample (19/20, 95%). Specifically, three successive samples from one patient had [negative, negative, positive] results for SRA, and [negative, positive, positive] results for PEA, respectively. This discordant result suggests a potential difference in sensitivity between the two assays. Further subset analysis focused on an additional 13 samples from 10 patients, all with SRA-positive results. Only 8 samples were concordant (8/13, 62%). One case showed a negative PEA and presumed false positive SRA, given the clinical history, while 4 samples showed likely true positive SRA results, given strength of ELISA results and 4Ts score, but were negative by PEA. Conclusions The lack of adherence to the 4Ts score algorithm to drive laboratory test utilization resulted in unnecessary ELISA and functional assay testing, potentially resulting in increased costs and false positive results. This result underscores the need for vigilance in assessing and refining diagnostic strategies. Additionally, our discordant findings in the subset analysis underscore the importance of evaluating the concordance and discordance between SRA and PEA results, providing insights into the diagnostic accuracy and potential limitations of each assay in identifying HIT.

Effects of pomegranate exocarp extract on H. pylori-induced pancreatic EMT: Molecular mechanisms and therapeutic potential

BackgroundPrevious studies have linked Helicobacter pylori infection with pancreatic diseases, including cancer. PurposeTo explore the influence of pomegranate exocarp extract (PEE) on epithelial-mesenchymal transition (EMT) in H. pylori-induced pancreatic rat tissue and to uncover the underlying molecular mechanisms. Study designTwenty-eight rats were divided into six groups: group 1 (negative control), group 2 (H. pylori-infected), group 3 (infected + PEE pretreatment), group 4 (infected + PEE treatment), group 5 (infected + metronidazole treatment), and group 6 (infected + metronidazole/PEE co-treatment). Methods and resultsThis study aimed to assess the effectiveness of pomegranate exocarp extract (PEE) in treating Helicobacter pylori infection and its associated pancreatic tissue changes in Wistar rats. The study involved Forty-eight male rats divided into six groups: H. pylori-infected control, PEE-preventive, PEE treatment, metronidazole treatment, PEE combined with metronidazole treatment, and negative control. The results showed a significant reduction in H. pylori concentration in the antrum in the PEE-treated groups (27.08 %) compared to that in the positive control group (p < 0.05). The group receiving the combined treatment exhibited the highest reduction (55.8 %) in H. pylori concentration (p < 0.005), with no significant difference observed between the PEE-preventive and metronidazole-treated groups. The ELISA results showed that the groups treated with PEE, PEE-preventive, and PEE combined with metronidazole experienced a significant increase in pancreatic E-cadherin levels by 47.7 %, 73.8 %, and 118.06 % respectively, and a substantial decrease in vimentin levels by 16.6 %, 31.6 %, and 43.5 % respectively, compared to the positive control group (p < 0.05). The results of the RT-qPCR analysis showed that the PEE treatment group, as well as the PEE preventive and PEE combined with metronidazole treatment groups, displayed significant downregulation of vimentin, sirtuin1, and lncRNA MALAT-1, and upregulation of E-cadherin compared to the positive control group. However, there was no significant difference between the PEE-preventive and metronidazole-treated groups (p < 0.05). Histopathological analysis showed considerable improvement in pancreatic tissue morphology in the PEE-treated groups. The inflammation score was significantly lower in these groups (p < 0.05), and the combined treatment group exhibited minimal signs of metaplasia and mononuclear cell infiltration. A computational study identified 54 human target genes of bioactive compounds in PEE. These findings shed light on the crucial interactions and pathways in treating pancreatic tumors. Additionally, GO enrichment and KEGG pathway analyses revealed significant pathways, such as the MAPK signaling and RTK pathway, enriched with genes targeted by PEE. Furthermore, evaluation of drug-likeness and ADME properties indicated that ellagic acid possesses drug-like properties and has a high potential for oral absorption. In conclusion, PEE has shown significant therapeutic potential in reducing H. pylori load and improving pancreatic tissue health while influencing key molecular markers and pathways associated with pancreatic tumors.

Immune infiltration related circular RNA, circGLIS2, facilitated progression of endometriosis through miR-4731-5p/IL-1β axis

Aberrantly expressed circRNAs affects various anti-tumour immune responses and immune cell regulation. However, the exact function of circGLIS2 on the pathogenesis of EMs remains unclear. In this study, we found that circGLIS2 was upregulated in EMs tissues and intimately related to clinicopathologic characteristics of EMs patients. Functionally, IHC and mouse model of EMs proved that circGLIS2 recruited immune cells infiltrated into ectopic endometrial microenvironment. RNA-seq, ELISA, RT-qPCR, and western blot results indicated that circGLIS2 influenced immune infiltration by regulating IL-1β. Besides, expression of circGLIS2 and IL-1β was also closely correlated in 284 human endometrium tissues. Mechanistically, RNA sequencing and intermolecular interaction prediction established “circGLIS2-miR-4731-5p-IL-1β” network. RNA pull down, RIP, dual luciferase reporter assay and expression regulation analysis confirmed the interaction among “circGLIS2-miR-4731-5p-IL-1β” axis. In summary, our findings demonstrated that circGLIS2 facilitated EMs progression by increasing immune cell infiltration via miR-4731-5p/IL-1β axis.

Investigating the relationship between blood metabolites and diabetic retinopathy using two-sample mendelian randomization and in vivo validation

We addressed fundamental questions about the influence of metabolites on the development of Diabetic retinopathy (DR), and explored the related pathological mechanism. Genome-wide association study (GWAS) database data for metabolites and DR were used to perform Mendelian randomization (MR) studies. The inverse variance weighting (IVW) was chosen as the primary analysis method. Sensitivity analysis was conducted using MR-PRESSO, leave-one-out and Cochran’s Q test. Confounding factors were eliminated to ensure robustness. We also conducted metabolic pathway analysis. In vivo experimental validation was conducted using Sprague Dawley rats. The serum metabolites of the DR group rats and normal group rats were examined to evaluate the MR results. The screen identified eighteen metabolites associated with DR risk, twelve of which were known components. Seven metabolites were positively correlated with DR risk, while five could reduce it. Eight metabolites associated with proliferative DR (PDR) risk were identified, four of which are known components. Three of these were positively associated with PDR risk and one metabolite reduced PDR risk. Additionally, two possible metabolic pathways involved in the biological mechanism of DR were identified. The ELISA results showed that the serum levels of isoleucine and 4-HPA were significantly increased in DR rats, while the level of inosine was decreased. This study offers novel insights into the biological mechanisms underlying DR. Metabolites that are causally linked to DR may serve as promising biomarkers and therapeutic targets.

Intraperitoneal MASP-1 Levels are Associated with Peritoneal Solute Transport Rate in Peritoneal Dialysis Patients: A Retrospective Cohort Study.

A major limitation of long-term peritoneal dialysis (PD) is peritoneal membrane dysfunction characterized by faster peritoneal solute-transport rate (PSTR). This study aimed to identify efficient complement factors in peritoneal effluents of continuous ambulatory peritoneal dialysis (CAPD) patients that can predict the PSTR. A multiplex suspension protein array was used to screened related complement pathways in overnight peritoneal effluents among 58 CAPD patients. Then the related complement factors in lectin and classical pathways in effluents were analyzed using ELISA kits among another cohort of 129 CAPD patients. Logistic regression modeling was fitted to predict the PSTR of PD patients. The multiplex suspension protein array showed complement factors including C2, C4b, C5, C5a, Factor D, Factor I, and MBL were detected in effluents of CAPD patients, and the effluent C2 Appearance rate (Ar) and C4b Ar levels were significantly correlated with D/P Cr and D/D0 glucose. The levels of effluents MASP-1 Ar, M-Ficolin Ar, C2 Ar and C4b Ar, which belong to the lectin pathway were also positively correlated with D/P Cr according the ELISA results and these parameters were expressed higher in the high and high-average (H/HA) groups according to the PET results. Moreover, effluent Masp-1 was independently associated with increased PSTR and adverseevents relatedperitonealtransportfunction failure. This study suggested that the lectin pathway may be involvedin local complement activation and peritoneal injury of PD patients, intraperitoneal level of Masp-1 was an independent predictor of increased PSTR in PD patients.

Activation of the AMPK/Nrf2 pathway ameliorates LPS-induced acute lung injury by inhibiting oxidative stress and reducing inflammation

BackgroundNumerous diseases-related acute lung injury (ALI) contributed to high mortality. Currently, the therapeutic effect of ALI was still poor. The detailed mechanism of ALI remained elusive and this study aimed to elucidate the mechanism of ALI.MethodThis study was performed to expose the molecular mechanisms of AMPK/Nrf2 pathway regulating oxidative stress in LPS-induced AMI mice. The mouse ALI model was established via intraperitoneal injection of LPS, then the lung tissue and blood samples were obtained, followed by injection with Dimethyl fumarate (DMF). Finally, Western blot, HE staining, injury score, lung wet/dry ratio, reactive oxygen species (ROS) and ELISA were used to elucidate the mechanism of AMPK/Nrf2 pathway in LPS -induced acute lung injury by mediating oxidative stress.ResultsThe lung tissue injury score was evaluated, showing higher scores in the model group compared to the AMPK activator and control groups. DCFH-DA indicated that LPS increased ROS production, while AMPK activator DMF reduced it, with the model group exhibiting higher ROS levels than the control and AMPK activator groups. The lung wet/dry ratio was also higher in the model group. Western blot analysis revealed LPS reduced AMPK and Nrf2 protein levels, but DMF reversed this effect. ELISA results showed elevated IL-6 and IL-1β levels in the model group compared to the AMPK activator and control groups.Conclusion:ConclusionActivating the AMPK/Nrf2 pathway can improve LPS-induced acute lung injury by down-regulation of the oxidative stress and corresponding inflammatory factor level.

Excessive Erythrophagocytosis Accounts for Systemic Inflammation in Chronic Kidney Disease.

Chronic kidney disease (CKD) is associated with persistent systemic inflammation. Reduced red blood cell (RBC) survival in patients with CKD has been identified for several decades. The purpose of this study is to explore whether excessive erythrophagocytosis exists and contributes to systemic inflammation in CKD. A CKD rat model was induced by 5/6 nephrectomy. Erythrocyte osmotic fragility was determined with hypotonic NaCl solutions. Erythrocyte deformability was evaluated by filterability. RBC cell death was quantified using fluorescence-activated cell sorting analyses of fluorescent annexin V-bound surface phosphatidylserine (PS). Erythrophagocytosis was evaluated in vivo and in vitro. RT-qPCR and immunohistochemistry were used to determine the inflammatory effects after erythrophagocytosis. Erythrocyte osmotic fragility and deformability progressively declined, and the percentage of PS-exposing RBCs progressively increased in CKD rats. Levels of erythrophagocytosis in vivo were evaluated by autologous injection of CFSE-labeled erythrocytes. In comparison with the control group, higher fluorescence intensity of CFSE was detected in the spleen homogenates of rats with CKD. In vitro, more of erythrocytes from 5/6Nx rats were phagocytosed by peritoneal macrophages in comparison to those from control rats. Compared with macrophages phagocytosed control erythrocytes, macrophages phagocytosed CKD erythrocytes exhibited higher mRNA levels of IL-6, CXCL-10, CXCL-11, iNOS, IL-1β, ICAM-1 and MCP-1. Compared with the control group, the red pulp of rats with CKD exhibited higher levels of p-NFκB, IL-6, iNOS and CXCL-10. ELISA results showed significantly increased plasma levels of both IL-6 and CXCL-10 in patients with long-term hemodialysis compared with those in healthy controls (2.30 ± 1.38 pg/mL vs 1.33 ± 0.65 pg/mL, P=0.01; 78.11 ± 27.34 pg/mL vs 37.45 ± 7.08 pg/mL, P=0.001). Our results indicated that excessive erythrophagocytosis may contribute to systemic inflammation in CKD.

In Vitro And In Silico Studies Of Lawsone On Inflammation-Induced Skin Cells For Development Of Skin Anti-Inflammatory Treatment

Lawsone, an active phytoconstituent of Lawsonia inermis sp., is proposed as a safer alternative for the current skin anti-inflammatory treatments that are mainly steroidal with unwanted side effects which does not conform to the concept of halalan toyyiban in medicine. This study aims to investigate the inhibitory mechanism of lawsone on pro-inflammatory cytokine, TNF (tumour necrosis factor)-α through in silico and in vitro methods. The inhibitory mechanism of lawsone were investigated via molecular docking and molecular dynamics which were performed via AutoDock Vina and GROMACS softwares, respectively. The results were then analysed via PyMol, Proteins Plus, RMSD and RMSF graphs. Subsequently, cytotoxicity of lawsone towards A431 skin epidermoid and 3T3 fibroblast cells, was determined via MTT assay. Lawsone was further tested on A431 cells stimulated with ethanol (200 mM) or hydrogen peroxide (250 µM) for 24 hours (acute) and 48 hours (chronic) to induce pro-inflammatory cytokine (TNF-α) release. Cell viabilities were determined via MTT assay and expression of TNF-α were measured via ELISA. In silico works predicted lawsone’s ability to bind to TNF-α with good binding affinity without disruption to the protein’s structure stability.Lawsone is proven to be safe towards non-pathological cells. Lawsone exhibited significant anti-inflammatory effect on inflammation induced A431 epidermoid cells. ELISA results were not as expected as compared to previous in silico study, but an anti-inflammatory pattern can be observed in the chronic ethanol-induced treatment groups. In conclusion, lawsone is shown to have potential to be developed as a halalan-toyyiban alternative for skin anti-inflammatory treatment.

Role the (IL1, IL6, and TNF) with Chronic Autoimmune Diseases in Mosul City / Iraq

Objective: Our study examined the relationship of the levels of cytokines IL1, 6, and TNF in three different immune diseases with age and Sex. Material and methods: The samples of the study were obtained from in Mosul city hospitals for the period from October 2023 to February 29, 2024. The study included 90 different medical cases suffering from immune diseases, 50 of whom were suffering from rheumatoid arthritis, 16 were suffering from erythematosus, and 24 were suffering from Celiac disease. The ages of the study participants were between 10 years and 65 years. Outcomes: Serum samples were examined for IL1, 6, and TNF using the ELISA technique. The results of ELISA revealed that Concentrations of Interleukins and TNF were low in all patients because the patients in the chronic phase of Autoimmune Diseases take treatment to reduce the cytokines which is hyperactive Therefore, cytokines concentrations appeared lower than standard concentrations. However, the results show a significant association between measuring IL-1 for a CD and measuring IL-6, TNF for an SLE among Autoimmune disease patients and their sex, and age. Conclusion: significance and impact of study: IL1, 6, and TNF are one of the most common cytokines on the globe. It has been connected directly or indirectly with various disorders, including autoimmune diseases. However, the link between IL1, 6 - TNF and Autoimmune Diseases is still controversial. Our results indicate that they play a possible role in the development of autoimmune diseases; however, more studies on a larger scale are required to confirm the results.

COL6A6 Peptide Vaccine Alleviates Atherosclerosis through Inducing Immune Response and Regulating Lipid Metabolism in Apoe−/− Mice

Atherosclerosis is an autoimmune disease characterized by lipid imbalances and chronic inflammation within blood vessels, with limited preventive and treatment options currently available. In this study, a vaccine prepared with COL6A6 peptide (named the Pep_A6 vaccine) was administered to immunize Apoe−/− mice, and the immune mechanism of the Pep_A6 vaccine against atherosclerosis was first investigated. The results of arterial oil red O staining demonstrated that the Pep_A6 vaccine significantly reduced the atherosclerotic plaque area in Apoe−/− mice fed with a high-fat diet for 20 weeks. A flow cytometry analysis revealed that the Pep_A6 vaccine inhibited Th1 cell differentiation and increased the proportion of Treg cells. Furthermore, there was a significant increase in Ly6Clow monocytes observed in the vaccinated group. The ELISA results showed that the Pep_A6 vaccine induced a significant expression of Pep_A6-specific antibody IgG and IgG1 in mouse serum. Additionally, we found that the Pep_A6 vaccine significantly decreased serum LDL-C content and regulated the expression of genes related to liver lipid metabolism. Together, our findings suggest that the Pep_A6 vaccine alleviates atherosclerosis by inducing a positive immune response and regulating lipid metabolism, providing new insights into potential prevention strategies for atherosclerosis as an innovative vaccine.

Skin Prick Tests and Enzyme-Linked Immunosorbent Assays among Allergic Patients Using Allergenic Local Pollen Extracts.

Allergic respiratory diseases are prevalent in the Philippines, with allergic rhinitis and asthma occurring at 20% and 8.7% of the population, respectively. The diagnosis of respiratory allergies is achieved by a combination of patient history and different screening tools, especially for the identification of the allergic triggers such as allergy skin prick test (SPT) and serum-specific IgE enzyme-linked immunosorbent assays (sIgE ELISA). The Philippines, being a tropical country, have a wide variety of plant species with potential to produce allergenic pollen grains. Knowledge of the sensitization profiles of Filipino allergic patients to our local pollen allergens is currently limited. The aim of this study is to determine the sensitization profile of patients with respiratory allergies (allergic rhinitis and/or asthma) through the allergy skin prick test (SPT) using allergenic local pollen extracts. It also aimed to determine if there is a positive agreement between the SPT and sIgE ELISA positivity rate and whether the results have relationship with the pollen purity and the protein content of the extracts. Pollen allergens were extracted from Amaranthus spinosus (pigweed), Mimosa pudica (makahiya), Tridax procumbens (wild daisy), Imperata cylindrica (cogon), Oryza sativa (rice), Pennisetum polystachion (foxtail grass), Sorghum halepense (Johnson grass), Albizia saman (acacia), Cocos nucifera (coconut), Leucaena leucocephala (ipil-ipil), and Mangifera indica (mango). SPT was performed at the Allergy Clinic of the University of the Philippines-Philippine General Hospital on patients with allergic rhinitis and/or bronchial asthma. Blood samples were collected from patients who developed wheal diameters of 3 mm or more than the negative control. Sera were tested against the same pollen extracts using ELISA. Of the one hundred sixty-five (165) patients who submitted for skin prick test, 129 showed positive SPT results to the pollen extracts. Weeds were the most sensitizing (51.9%-58.1%). Blood samples were collected from these patients and tested for sIgE ELISA and among them, 71 were positive in the sIgE ELISA. Highest sensitization rates in sIgE ELISA were found in coconut, pigweed, Johnson grass, and rice. The highest positive agreements or the proportion of patients with positive sIgE ELISA among those with positive SPT were in coconut, followed by Johnson grass, pigweed, and rice. Most of the pollen sensitized patients on SPT are polysensitized. SPT is a safe, simple, and rapid method for the diagnosis of IgE-mediated allergy. The lower number of positive patients in sIgE ELISA may be attributed to the low serum IgE levels and low quantities of effectual allergen components in extracts. Results of both SPT and ELISA must be correlated with a patient's clinical history, particularly the patient's exposures, and physical examination.

501 The effect of dietary camelina, flaxseed, and canola oil supplementation on skin fatty acid profile, inflammation and immune responses in healthy adult horses

Abstract As n-6 and n-3 fatty acids (FA) have pro- and anti-inflammatory effects, respectively, supplemental oils have the potential to impact inflammation, the immune response, and skin and coat health. This study aimed to evaluate the effects of camelina (CAM), in comparison to canola (OLA) and flaxseed (FLX) oil on the skin FA profile, inflammation, and the immune response in horses. Horses [n = 24; 14.2 ± 4.8 yr; 518 ± 63 kg body weight (BW)] were acclimated to sunflower oil over 4 wk. Groups were balanced by location, age, weight, sex, and breed and randomly assigned one of the three treatments (CAM, OLA, FLX). Horses were fed their assigned oil at a dose of 0.37 g/kg BW for 16 wk. Skin samples were collected on wk 16, and the skin FA composition was analyzed by gas chromatography. On wk 10 and 12, keyhole limpet hemocyanin (KLH) was administered intramuscularly to each horse to stimulate a systemic immune response. Blood was collected on wk 14 and 16 for the evaluation of plasma KLH immunoglobulin G (IgG) concentrations using a modified ELISA protocol and results were presented as optical density units. On wk 12, after the first KLH injection, a delayed-type hypersensitivity (DTH) test was performed by intradermally injecting KLH and the area of swelling was measured using the software ImageJ at 0.5-, 1-, 8-, 24-, 48-, 72-, and 96-h post-injection. Data were analyzed using the GLIMMIX procedure of SAS considering the effects of treatment, time, and their interaction as fixed, and the effects of horse and location as random. Repeated measures on horses were modelled using a spatial power covariance structure. Significance was declared at P ≤ 0.05 and when fixed effects were significant, means were separated using Tukey-Kramer adjustments. Immune and inflammatory responses were produced by the KLH injections; however, there were no differences among treatment groups regarding KLH antibody and DTH response (P &amp;gt; 0.44). The percentages of palmitic (P = 0.02) and stearic acid (P = 0.03) in the skin at wk 16 were greater in FLX as compared with CAM but neither differed from the OLA. The percentage of linoleic acid (P = 0.01) was greater in CAM than OLA but neither differed from FLX at wk 16. The percentage of all other FA was similar among treatment groups (P &amp;gt; 0.06). These results suggest that CAM is comparable to FLX and OLA regarding support of an immune and inflammatory response, and the oil FA composition was not reflected in the skin FA profile after feeding for 16 wk as hypothesized. In conclusion, at the dose provided, CAM supports a similar immune and inflammatory response to KLH as FLX and OLA; however, the lack of incorporation of FA from the diet into the skin should be further explored.