BACKGROUND: Cholesterol ester storage disease (CESD) and Wolman Disease (WD) are due to deficiency of lysosomal acid lipase (LAL). A new method is described for the measurement of LAL in dried blood spots (DBS) using Lalistat 2 an inhibitor of LAL. METHODS: LAL activity in DBS extracts was measured using the substrate 4-methylumbelliferyl palmitate. LAL activity was determined by measuring total lipase activity and lipase activity in the presence of Lalistat 2. The specificity of Lalistat 2 was investigated using human recombinant LAL (hrLAL) and human pancreatic lipase (hPL). RESULTS: Lalistat 2 inhibited hrLAL with 1% residual activity at 1 μM inhibitor but had no effect on hPL up to 10 μM. LAL activity in DBS samples obtained from normal controls (n=140) was 0.50-2.30 nmol/punch/h and in patients with CESD was <0.03 nmol/punch/h (n=11). Activity in carriers showed intermediate activity: 0.15-0.40 nmol/punch/h (n=15). CONCLUSIONS: Measurement of LAL using DBS is made difficult by the presence of other lipases in whole blood. Lalistat 2 is a specific inhibitor of LAL which allows the determination of LAL in DBS. Results show the method differentiates clearly between normal controls, carriers and affected cases.
BACKGROUND: Cholesterol ester storage disease (CESD) and Wolman Disease (WD) are due to deficiency of lysosomal acid lipase (LAL). A new method is described for the measurement of LAL in dried blood spots (DBS) using Lalistat 2 an inhibitor of LAL. METHODS: LAL activity in DBS extracts was measured using the substrate 4-methylumbelliferyl palmitate. LAL activity was determined by measuring total lipase activity and lipase activity in the presence of Lalistat 2. The specificity of Lalistat 2 was investigated using human recombinant LAL (hrLAL) and humanpancreatic lipase (hPL). RESULTS: Lalistat 2 inhibited hrLAL with 1% residual activity at 1 μM inhibitor but had no effect on hPL up to 10 μM. LAL activity in DBS samples obtained from normal controls (n=140) was 0.50-2.30 nmol/punch/h and in patients with CESD was <0.03 nmol/punch/h (n=11). Activity in carriers showed intermediate activity: 0.15-0.40 nmol/punch/h (n=15). CONCLUSIONS: Measurement of LAL using DBS is made difficult by the presence of other lipases in whole blood. Lalistat 2 is a specific inhibitor of LAL which allows the determination of LAL in DBS. Results show the method differentiates clearly between normal controls, carriers and affected cases.
Authors: Maidina Tuohetahuntila; Martijn R Molenaar; Bart Spee; Jos F Brouwers; Richard Wubbolts; Martin Houweling; Cong Yan; Hong Du; Brian C VanderVen; Arie B Vaandrager; J Bernd Helms Journal: J Biol Chem Date: 2017-06-14 Impact factor: 5.157
Authors: Yuritzi Santillán-Hernández; Enory Almanza-Miranda; Winnie W Xin; Kendrick Goss; Aurea Vera-Loaiza; María T Gorráez-de la Mora; Raul E Piña-Aguilar Journal: World J Gastroenterol Date: 2015-01-21 Impact factor: 5.742
Authors: Ariel Brautbar; Emili Leary; Kristen Rasmussen; Don P Wilson; Robert D Steiner; Salim Virani Journal: Curr Atheroscler Rep Date: 2015-04 Impact factor: 5.113
Authors: Stuart A Scott; Benny Liu; Irina Nazarenko; Suparna Martis; Julia Kozlitina; Yao Yang; Charina Ramirez; Yumi Kasai; Tommy Hyatt; Inga Peter; Robert J Desnick Journal: Hepatology Date: 2013-07-29 Impact factor: 17.425
Authors: Licia Polimeni; Daniele Pastori; Francesco Baratta; Giulia Tozzi; Marta Novo; Roberto Vicinanza; Giovanni Troisi; Gaetano Pannitteri; Fabrizio Ceci; Laura Scardella; Francesco Violi; Francesco Angelico; Maria Del Ben Journal: Intern Emerg Med Date: 2017-09-12 Impact factor: 3.397