Saiid Bina1, John Capacchione, Bayarsaikhan Munkhuu, Sheila Muldoon, Rolf Bünger. 1. 1Department of Anesthesiology, Uniformed Services University of the Health Sciences, Bethesda, MD. 2Department of Anatomy, Physiology, and Genetics, Uniformed Services University of the Health Sciences, Bethesda, MD.
Abstract
OBJECTIVE: Malignant hyperthermia is a pharmacogenetic disorder typically triggered by potent inhalation anesthetics and/or the depolarizing muscle relaxant succinylcholine in malignant hyperthermia-susceptible individuals. Since lymphocytes express the same Ca channel mutation found in malignant hyperthermia-susceptible muscle, we investigated agonist-induced adenosine formation in lymphocytes as an index of sarcoplasmic reticulum Ca-release-induced adenosine 5'-triphosphate turnover as a potential minimally invasive functional malignant hyperthermia assay. DESIGN: Application of lymphocytes for malignant hyperthermia diagnosis. SETTING: Hospitals and university laboratory. SUBJECTS: Malignant hyperthermia-susceptible patients (n = 13) and normal subjects (n = 11). INTERVENTIONS: Adenosine formation due to malignant hyperthermia-triggering agent halothane or the ryanodine receptor Ca channels agonist 4-chloro-m-cresol was compared in blood lymphocytes from malignant hyperthermia-susceptible patients and normal subjects. MEASUREMENTS AND MAIN RESULTS: Cai and adenosine were measured in fresh or immortalized blood lymphocytes incubated with 0-10 mM 4-chloro-m-cresol or 0-10.7 mM halothane. Cai levels were significantly higher in immortalized malignant hyperthermia-susceptible B cells treated with 0.75 mM 4-chloro-m-cresol relative to controls. Similarly, at 1 mM 4-chloro-m-cresol or 0.96 mM halothane, adenosine levels were significantly higher in malignant hyperthermia-susceptible lymphocytes or immortalized B cells relative to controls. Receiver-operating characteristic analyses showed areas under the 4-chloro-m-cresol receiver-operating characteristic curves near more than or equal to 0.96 (p ≈ 0.0001), suggesting that 4-chloro-m-cresol-induced adenosine could readily distinguish between malignant hyperthermia-susceptible and normal controls cells. CONCLUSIONS: Both 4-chloro-m-cresol and halothane caused adenosine accumulation in blood lymphocytes. Adenosine accumulation was markedly increased in malignant hyperthermia-susceptible lymphocytes compared with controls reflecting higher than normal adenosine 5'-triphosphate degradation in the malignant hyperthermia-susceptible cells. Although 4-chloro-m-cresol receiver-operating characteristic curves revealed that adenosine accumulation could readily distinguish between normal and malignant hyperthermia-susceptible lymphocytes, independent confirmation is required with a substantially larger number of enrolled subjects to correctly appreciate the clinical utility of the novel lymphocyte-adenosine protocol for malignant hyperthermia testing.
OBJECTIVE:Malignant hyperthermia is a pharmacogenetic disorder typically triggered by potent inhalation anesthetics and/or the depolarizing muscle relaxant succinylcholine in malignant hyperthermia-susceptible individuals. Since lymphocytes express the same Ca channel mutation found in malignant hyperthermia-susceptible muscle, we investigated agonist-induced adenosine formation in lymphocytes as an index of sarcoplasmic reticulum Ca-release-induced adenosine 5'-triphosphate turnover as a potential minimally invasive functional malignant hyperthermia assay. DESIGN: Application of lymphocytes for malignant hyperthermia diagnosis. SETTING: Hospitals and university laboratory. SUBJECTS:Malignant hyperthermia-susceptible patients (n = 13) and normal subjects (n = 11). INTERVENTIONS:Adenosine formation due to malignant hyperthermia-triggering agent halothane or the ryanodine receptor Ca channels agonist 4-chloro-m-cresol was compared in blood lymphocytes from malignant hyperthermia-susceptible patients and normal subjects. MEASUREMENTS AND MAIN RESULTS:Cai and adenosine were measured in fresh or immortalized blood lymphocytes incubated with 0-10 mM 4-chloro-m-cresol or 0-10.7 mM halothane. Cai levels were significantly higher in immortalized malignant hyperthermia-susceptible B cells treated with 0.75 mM 4-chloro-m-cresol relative to controls. Similarly, at 1 mM 4-chloro-m-cresol or 0.96 mM halothane, adenosine levels were significantly higher in malignant hyperthermia-susceptible lymphocytes or immortalized B cells relative to controls. Receiver-operating characteristic analyses showed areas under the 4-chloro-m-cresol receiver-operating characteristic curves near more than or equal to 0.96 (p ≈ 0.0001), suggesting that 4-chloro-m-cresol-induced adenosine could readily distinguish between malignant hyperthermia-susceptible and normal controls cells. CONCLUSIONS: Both 4-chloro-m-cresol and halothane caused adenosine accumulation in blood lymphocytes. Adenosine accumulation was markedly increased in malignant hyperthermia-susceptible lymphocytes compared with controls reflecting higher than normal adenosine 5'-triphosphate degradation in the malignant hyperthermia-susceptible cells. Although 4-chloro-m-cresol receiver-operating characteristic curves revealed that adenosine accumulation could readily distinguish between normal and malignant hyperthermia-susceptible lymphocytes, independent confirmation is required with a substantially larger number of enrolled subjects to correctly appreciate the clinical utility of the novel lymphocyte-adenosine protocol for malignant hyperthermia testing.
Authors: Yuri Hosokawa; Douglas J Casa; Henry Rosenberg; John F Capacchione; Emmanuel Sagui; Sheila Riazi; Luke N Belval; Patricia A Deuster; John F Jardine; Stavros A Kavouras; Elaine C Lee; Kevin C Miller; Sheila M Muldoon; Francis G O'Connor; Scott R Sailor; Nyamkhishig Sambuughin; Rebecca L Stearns; William M Adams; Robert A Huggins; Lesley W Vandermark Journal: J Athl Train Date: 2017-03-07 Impact factor: 2.860
Authors: Kerstin Hoppe; Guido Hack; Frank Lehmann-Horn; Karin Jurkat-Rott; Scott Wearing; Alberto Zullo; Antonella Carsana; Werner Klingler Journal: Sci Rep Date: 2016-09-20 Impact factor: 4.379