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May 3, 2024 · Remote ischemic conditioning (RIC) has the potential to benefit graft function following kidney transplantation by reducing ischemia-reperfusion injury; however, the current clinical evidence is inconclusive.
- 10.1186/s12871-024-02549-y
- 2024
- BMC Anesthesiol. 2024; 24: 168.
- Context Patient Cohort and Ric Protocol
- Sample Preparation and Analysis by Mass Spectrometry
- Tissue Proteomics
- Plasma Proteomics
- Analysis by Mass Spectrometry
- Elisa Assays For SAA1 and CRP
- Measurements of S-Troponin and S-Myoglobin
- Proteomics Data Analysis
The CONTEXT trial concerned 225 included transplant recipients and evaluated effects of RIC, induced by episodes of obstruction to arterial flow in the leg on the opposite side of where the transplant was implanted using a tourniquet (4 × 5 min) on the thigh prior to kidney reperfusion . Trial registration: NCT01395719. Registered 14 July 2011, htt...
For this in-depth study to detect any underlying molecular changes due to RIC our identifier cohorts consisted of six pairs of recipients (Table 1). We have analysed tissue and plasma samples from these six kidney pairs (6 RIC and 6 control), at 2 and 4 time points, respectively. Pairs of kidneys from the same donor where the recipients were as sim...
Approximately 10 mg of kidney tissue (n = 6 per group, Table 1) was lysed in RIPA buffer (Pierce) using a bead beater homogenizer (Precellys). A BCA assay was performed and 50 µg of total protein from each sample was taken for Filter Aided Sample Preparation (FASP) digestion. In short, 30 kDa filters (Millipore) were equilibrated with 8 M urea buff...
One hundred microliter of plasma were precipitated by adding PEG 6000 to final concentration of 12% in order to deplete abundant plasma proteins. Depleted plasma protein concentrations (n = 6 per group, Table 1) were determined using a BCA assay (Pierce) and digested using a Thermo Scientific™ SMART Digest™ Kit. Briefly, 50 µg of protein was added ...
Liquid chromatography tandem mass spectrometry (LC–MS/MS) analysis was performed using a Dionex Ultimate 3000 nano-ultra high pressure reversed-phase chromatography system coupled on-line to a Q Exactive High Field (HF) mass spectrometer (Thermo Scientific) as described previously . In brief, samples were separated on an EASY-Spray PepMap RSLC C18 ...
To validate proteomic findings, ELISA was used to detect and quantify SAA1 (R&D Systems, SAA1 DuoSet) levels in plasma of patients from the exploratory as well as the validation cohort where samples from all time points were available (n = 143, Table 1). Plates were coated overnight with capture antibody at RT and blocked with Reagent Diluent for 1...
In plasma from the whole CONTEXT cohort , ELISA assays were used to quantify levels of S-troponin T (Roche, Troponin T-high sensitive) and S-myoglobin (ADVIA Centaur XPT Immunoassay System) in plasma at baseline (TD0), 90 min and post-operative day 1 (TD1) according to the manufacturer’s instructions. As data are mixed with some paired data (when k...
MS raw data was searched using MaxQuant software (v1.3.1). The search parameters used were as follows: trypsin with two missed cleavages allowed, oxidation (M) and deamidation (N, Q) were set as variable modifications and carbamidomethylation (C) as a fixed modification. The data was searched against human protein sequences using the UPR_homoSapien...
- Adam M. Thorne, Honglei Huang, Darragh P. O‘Brien, Marco Eijken, Marco Eijken, Nicoline Valentina Kr...
- 2020
Experimental and clinical studies have shown protection by remote ischemic conditioning (RIC). We hypothesized that recipient RIC before kidney graft reperfusion reduces the time to graft recovery.
Nov 2, 2020 · A randomised clinical study (CONTEXT) evaluated the effects of RIC compared to non-RIC controls in human kidney transplants. Methods: RIC was induced prior to kidney reperfusion by episodes of obstruction to arterial flow in the leg opposite the transplant using a tourniquet (4 × 5 min).
- Adam M. Thorne, Honglei Huang, Darragh P. O‘Brien, Marco Eijken, Marco Eijken, Nicoline Valentina Kr...
- 2020
Sep 29, 2021 · We used samples from the CONTEXT trial cohort that examined the effects of four cycles of RIC during kidney transplant surgery on kidney graft function. We aimed to evaluate if RIC affected the frequency of circulating myeloid (m)DCs and pDCs and the distinct subset of ILT3 + DCs as well as the expression of activation markers human leucocyte ...
- Marie B Nielsen, Marie B Nielsen, Kristian Ravlo, Kristian Ravlo, Marco Eijken, Nicoline V Krogstrup...
- 10.1111/cei.13658
- 2021
- Clin Exp Immunol. 2021 Nov; 206(2): 226-236.
This brief commentary aims to discuss the effects of RIC on clinical outcomes and proinflammatory cytokines in patients undergoing kidney transplantation.
Dec 1, 2020 · Using a porcine kidney transplant model, we aimed to decipher the subclinical molecular effects of a RIC regime, compared to non-RIC controls.
