Endotoxin detection in end-stage kidney disease

Wong, Jonathan; Jeraj, Hassan; Viljoen, Adie; Farrington, Kenneth

doi:10.1136/jclinpath-2014-202622

Cited by 5 publications

(16 citation statements)

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“…We recently examined the performance of a kinetic turbidimetric LAL assay in HD patients and found that the assay was sensitive and precise for endotoxin detection in uraemic plasma[ 21 ], [ 22 ]. Using this assay, we investigated the influence of BG on endotoxin read-out measurements by measuring endotoxin in HD patients with and without the use of a BG-blocking buffer.…”

Section: Introductionmentioning

confidence: 99%

Is Endotoxemia in Stable Hemodialysis Patients an Artefact? Limitations of the Limulus Amebocyte Lysate Assay and Role of (1→3)-β-D Glucan

et al. 2016

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BackgroundElevated blood endotoxin levels are frequently reported in the dialysis population and are strongly linked with inflammation, a major predictor of mortality. Virtually all studies have employed the Limulus Amoebocyte Lysate (LAL) assay to detect endotoxin. However this assay is not endotoxin-specific and can be activated by (1→3)-β-glucan (BG), a component of fungal cell walls leading to false positive signals. Very few studies have taken account of this. We examined the influence of BG-based activation of the LAL assay on the detection of endotoxemia in this setting.MethodWe measured plasma endotoxin levels in 50 hemodialysis patients with and without the use of BG-blocking buffers. These buffers inhibit BG activation of the LAL assay to ensure that any signal detected is endotoxin-specific. Blood samples were measured for BG, interleukin-6 (IL-6), tumor necrosis factor-alfa (TNF-α) to examine the association between endotoxin signals, BG and inflammation.ResultsEndotoxin signals were detected in 50% of patients. On repeat measurement with a BG-blocking buffer, all detected endotoxin signals were extinguished. No patient had detectable endotoxemia. Plasma BG levels were significantly elevated in 58% of patients and were higher in those with detectable endotoxin signals using the LAL assay without BG-blocking buffers (78vs.54pg/mL;p<0.001). Endotoxin signal and BG levels did not correlate with levels of TNF-α or IL-6.ConclusionUse of the LAL assay for blood endotoxin detection in dialysis patients has its limitations due to high blood BG. Endotoxemia frequently reported in non-infected hemodialysis patients may be artefactual due to BG interference.

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Section: Introductionmentioning

confidence: 99%

Is Endotoxemia in Stable Hemodialysis Patients an Artefact? Limitations of the Limulus Amebocyte Lysate Assay and Role of (1→3)-β-D Glucan

et al. 2016

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“…Blood samples were collected peripherally using aseptic technique as previously described [17,18]. For haemodialysis patients, blood samples were collected pre-dialysis.…”

Section: Blood Sampling and Processingmentioning

confidence: 99%

“…Endotoxin was measured twice, once using the standard LAL assay [17] and then repeated with a BG-blocking agent which inhibits the factor G pathway preventing the LAL assay from false positive activation by BG that may be present in the sample [4].…”

Section: Endotoxin Measurementsmentioning

confidence: 99%

“…Endotoxin measurements were performed using the kinetic turbidimetric LAL assay (Endosafe KTA2, Charles River Laboratories) as previously described [17]. Plasma samples were diluted 1:10 with 0.1% Tween80 (Merck Chemicals) and heated to 70°C for 10 min to remove inhibitory factors present in plasma and cooled to room temperature prior to analysis.…”

Section: Endotoxin Assay Using Standard Lal Without Bg-blocking Buffermentioning

confidence: 99%

“…All study participants gave written informed consent. Ethical approval was granted by the NHS research ethics committee East of England -Cambridge South (reference [17] /EE/0019) and NHS research ethics committee London Hampstead (reference [17] /LO/0467). The study was conducted ethically in accordance to the Declaration of Helsinki.…”

Section: Fundingmentioning

confidence: 99%

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Beta-glucans in advanced CKD: role in endotoxaemia and inflammation

et al. 2020

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Background/aims: (1-3)-β-D glucans (BG) are cellular components of yeasts and fungi. Elevated blood levels may be an adjunct in diagnosing invasive fungal infection, though can be high in dialysis patients without fungaemia. BG can also induce false positive signals in endotoxin detection assays (Limulus Amoebocyte Lysate [LAL] assay). We explored the relationship between BG levels, renal impairment, endotoxaemia and inflammation. Methods: We measured serum BG levels, markers of inflammation and blood endotoxin levels in 20 controls, 20 with stages 1-3 chronic kidney disease (CKD), 20 with stages 4-5 CKD, 15 on peritoneal dialysis (PD) and 60 on haemodialysis (HD). Another 30 patients were studied before and after HD initiation. Results: BG levels increased with advancing CKD, being highest in HD patients, 22% of whom had elevated levels (> 80 pg/ml). Levels increased significantly following HD initiation. Levels also correlated positively with CRP, TNFα, IL-6 levels, independently of CKD stage. Blood endotoxin was detectable by LAL assays in 10-53% of the CKD cohort, being most prevalent in the HD group, and correlating positively with BG levels. Adding BG blocking agent to the assay reduced endotoxin detection confining it to only 5% of HD patients. Levels of inflammatory markers were higher in those with detectable endotoxin-whether false-or true positives. Conclusion: BG levels increased with decreasing renal function, being highest in dialysis patients. High BG levels were associated with false positive blood endotoxin signals, and with markers of inflammation, independently of CKD stage. The cause for high BG levels is unknown but could reflect increased gut permeability and altered mononuclear phagocytic system function.

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Potent Anti‐Inflammatory and Pro‐Resolving Effects of Anabasum in a Human Model of Self‐Resolving Acute Inflammation

Motwani

Bennett

Norris

et al. 2018

Clin Pharma and Therapeutics

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Anabasum is a synthetic analog of Δ8‐tetrahydrocannabinol (THC)‐11‐oic acid that in preclinical models of experimental inflammation exerts potent anti‐inflammatory actions with minimal central nervous system (CNS) cannabimimetic activity. Here we used a novel model of acute inflammation driven by i.d. UV‐killed E. coli in healthy humans and found that anabasum (5 mg) exerted a potent anti‐inflammatory effect equivalent to that of prednisolone in terms of inhibiting neutrophil infiltration, the hallmark of acute inflammation. These effects arose from the inhibition of the neutrophil chemoattractant LTB4, while the inhibition of antiphagocytic prostanoids (PGE2, TxB2, and PGF2α) resulted in enhanced clearance of inflammatory stimulus from the injected site. Anabasum at the higher dose of 20 mg possessed the additional properties of triggering the biosynthesis of specialized pro‐resolving lipid mediators including LXA4, LXB4, RvD1, and RvD3. Collectively, we demonstrate for the first time a striking anti‐inflammatory and pro‐resolution effects of a synthetic analog of THC in healthy humans.

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Endotoxin detection in end-stage kidney disease

Cited by 5 publications

References 32 publications

Is Endotoxemia in Stable Hemodialysis Patients an Artefact? Limitations of the Limulus Amebocyte Lysate Assay and Role of (1→3)-β-D Glucan

Is Endotoxemia in Stable Hemodialysis Patients an Artefact? Limitations of the Limulus Amebocyte Lysate Assay and Role of (1→3)-β-D Glucan

Beta-glucans in advanced CKD: role in endotoxaemia and inflammation

Potent Anti‐Inflammatory and Pro‐Resolving Effects of Anabasum in a Human Model of Self‐Resolving Acute Inflammation

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