The Experts below are selected from a list of 99 Experts worldwide ranked by ideXlab platform
Peter J. Meikle - One of the best experts on this subject based on the ideXlab platform.
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Urinary Lipid Profiling for the Identification of
2015Co-Authors: Fabry Hemizygotes, Maria Fuller, Tina Rozaklis, Phillip D. Whitfield, David Blacklock, John J. Hopwood, Peter C. Sharp, Peter J. MeikleAbstract:storage disorder resulting from a deficiency of the lysosomal hydrolase, -galactosidase, for which enzyme replacement therapy is now available. In this study, we aimed to identify Fabry heterozygotes not only for genetic counseling of families but because it is becom-ing increasingly obvious that many heterozygous (car-rier) females are symptomatic and should be considered for treatment. Methods: We measured 29 individual lipid species, including Ceramide, glucosylCeramide, lactosylceram-ide, and Ceramide Trihexoside, in urine samples from Fabry hemizygotes and heterozygotes and from control individuals by electrospray ionization tandem mass spectrometry. Individual analyte species and analyte ratios were analyzed for their ability to differentiate th
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Urinary Lipid Profiling for the Identification of
2014Co-Authors: Fabry Hemizygotes, Maria Fuller, Tina Rozaklis, Phillip D. Whitfield, David Blacklock, John J. Hopwood, Peter C. Sharp, Peter J. MeikleAbstract:storage disorder resulting from a deficiency of the lysosomal hydrolase, -galactosidase, for which enzyme replacement therapy is now available. In this study, we aimed to identify Fabry heterozygotes not only for genetic counseling of families but because it is becom-ing increasingly obvious that many heterozygous (car-rier) females are symptomatic and should be considered for treatment. Methods: We measured 29 individual lipid species, including Ceramide, glucosylCeramide, lactosylceram-ide, and Ceramide Trihexoside, in urine samples from Fabry hemizygotes and heterozygotes, and control indi-viduals by electrospray ionization tandem mass spec-trometry. Individual analyte species and analyte ratios were analyzed for their ability to differentiate the con
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Urinary Lipid Profiling for the Identification of Fabry Hemizygotes and Heterozygotes
Clinical chemistry, 2005Co-Authors: Maria Fuller, P. Sharp, Tina Rozaklis, Phillip D. Whitfield, David Blacklock, John J. Hopwood, Peter J. MeikleAbstract:Background: Fabry disease is an X-linked lysosomal storage disorder resulting from a deficiency of the lysosomal hydrolase, α-galactosidase, for which enzyme replacement therapy is now available. In this study, we aimed to identify Fabry heterozygotes not only for genetic counseling of families but because it is becoming increasingly obvious that many heterozygous (carrier) females are symptomatic and should be considered for treatment. Methods: We measured 29 individual lipid species, including Ceramide, glucosylCeramide, lactosylCeramide, and Ceramide Trihexoside, in urine samples from Fabry hemizygotes and heterozygotes and from control individuals by electrospray ionization tandem mass spectrometry. Individual analyte species and analyte ratios were analyzed for their ability to differentiate the control and patient groups. Results: The Fabry hemizygotes had increased concentrations of the substrate for the deficient enzyme, Ceramide Trihexoside, as well as lactosylCeramide and Ceramide, along with decreased concentrations of both glucosylCeramide and sphingomyelin. Ratios of these analytes improved differentiation between the control and Fabry groups, with the Fabry heterozygotes generally falling between the Fabry hemizygotes and the control group. Conclusions: These lipid profiles hold particular promise for the identification of Fabry individuals, may aid in the prediction of phenotype, and are potentially useful for the monitoring of therapy in patients receiving enzyme replacement.
Johannes M. F. G. Aerts - One of the best experts on this subject based on the ideXlab platform.
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HPLC for Simultaneous Quantification of Total Ceramide, GlucosylCeramide, and Ceramide Trihexoside Concentrations in Plasma
Clinical chemistry, 2007Co-Authors: Johanna E.m. Groener, Ben J. H. M. Poorthuis, Sijmen Kuiper, Mariette T.j. Helmond, Carla E. M. Hollak, Johannes M. F. G. AertsAbstract:Background: Simple, reproducible assays are needed for the quantification of sphingolipids, Ceramide (Cer), and sphingoid bases. We developed an HPLC method for simultaneous quantification of total plasma concentrations of Cer, glucosylCeramide (GlcCer), and Ceramide Trihexoside (CTH). Methods: After addition of sphinganine as internal calibrator, we extracted lipids from 50 L plasma. We deacylated Cer and glycosphingolipids by use of microwave-assisted hydrolysis in methanolic NaOH, followed by derivatization of the liberated amino-group with o-phthaldialdehyde. We separated the derivatized sphingoid bases and lysoglycosphingolipids by HPLC on a C18 reversed-phase column with a methanol/water mobile phase (88:12, vol/vol) and quantified them by use of a fluorescence detector at ex 340 nm and em 435 nm. Results: Optimal conditions in the Solids/Moisture System SAM-155 microwave oven (CEM Corp.) for the complete deacylation of Cer and neutral glycosphingolipids without decomposition were 60 min at 85% power, fan setting 7. Intra- and interassay CVs were
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hplc for simultaneous quantification of total Ceramide glucosylCeramide and Ceramide Trihexoside concentrations in plasma
Clinical Chemistry, 2007Co-Authors: Johanna E.m. Groener, Ben J. H. M. Poorthuis, Sijmen Kuiper, Mariette T.j. Helmond, Carla E. M. Hollak, Johannes M. F. G. AertsAbstract:Background: Simple, reproducible assays are needed for the quantification of sphingolipids, Ceramide (Cer), and sphingoid bases. We developed an HPLC method for simultaneous quantification of total plasma concentrations of Cer, glucosylCeramide (GlcCer), and Ceramide Trihexoside (CTH). Methods: After addition of sphinganine as internal calibrator, we extracted lipids from 50 μL plasma. We deacylated Cer and glycosphingolipids by use of microwave-assisted hydrolysis in methanolic NaOH, followed by derivatization of the liberated amino-group with o -phthaldialdehyde. We separated the derivatized sphingoid bases and lysoglycosphingolipids by HPLC on a C18 reversed-phase column with a methanol/water mobile phase (88:12, vol/vol) and quantified them by use of a fluorescence detector at λex 340 nm and λem 435 nm. Results: Optimal conditions in the Solids/Moisture System SAM-155 microwave oven (CEM Corp.) for the complete deacylation of Cer and neutral glycosphingolipids without decomposition were 60 min at 85% power, fan setting 7. Intra- and interassay CVs were <4% and <14%, respectively, and recovery rates were 87%–113%. The limit of quantification was 2 pmol (0.1 pmol on column), and the method was linear over the interval of 2–200 μL plasma. In samples from 40 healthy individuals, mean (SD) concentrations were 9.0 (2.3) μmol/L for Cer, 6.3 (1.9) μmol/L for GlcCer, and 1.7 (0.5) μmol/L for CTH. Plasma concentrations of GlcCer were higher in Gaucher disease patient samples and of CTH in Fabry disease patient samples. Conclusions: HPLC enables quantification of total Cer, GlcCer, and CTH in plasma and is useful for the follow-up of patients on therapy for Gaucher or Fabry disease.
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Trihexoside Concentrations in Plasma
2006Co-Authors: Johanna E.m. Groener, Ben J. H. M. Poorthuis, Sijmen Kuiper, Mariette T.j. Helmond, Carla E. M. Hollak, Johannes M. F. G. AertsAbstract:Background: Simple, reproducible assays are needed for the quantification of sphingolipids, Ceramide (Cer), and sphingoid bases. We developed an HPLC method for simultaneous quantification of total plasma concentrations of Cer, glucosylCeramide (GlcCer), and Ceramide Trihexoside (CTH). Methods: After addition of sphinganine as internal calibrator, we extracted lipids from 50 �L plasma. We deacylated Cer and glycosphingolipids by use of microwave-assisted hydrolysis in methanolic NaOH, followed by derivatization of the liberated amino-group with o-phthaldialdehyde. We separated the derivatized sphingoid bases and lysoglycosphingolipids by HPLC on a C18 reversed-phase column with a methanol/water mobile phase (88:12, vol/vol) and quantified them by use of a fluorescence detector at � ex 340 nm and � em 435 nm. Results: Optimal conditions in the Solids/Moisture System SAM-155 microwave oven (CEM Corp.) for the complete deacylation of Cer and neutral glycosphingolipids without decomposition were 60 min at 85% power, fan setting 7. Intra- and interassay CVs were <4 % and <14%, respectively, and recovery rates were 87%–113%. The limit of quantification was 2 pmol (0.1 pmol on column), and the method was linear over the interval of 2–200 �L plasma. In samples from 40 healthy individuals, mean (SD) concentrations were 9.0 (2.3
Kimiyo Raymond - One of the best experts on this subject based on the ideXlab platform.
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Multiplex assay for the tandem detection of Ceramide Trihexosides and sulfatides: Efficient first tier screening for Fabry, MLD, MSD, and Saposin B in urine
Molecular Genetics and Metabolism, 2019Co-Authors: Kim K. Nickander, Jean M. Lacey, Dimitar Gavrilov, Dietrich Matern, Devin Oglesbee, Piero Rinaldo, Silvia Tortorelli, Kimiyo RaymondAbstract:Fabry disease, metachromatic leukodystrophy (MLD), multiple sulfatase deficiency (MSD), saposin B deficiency (SAPB) and mucolipidosis II (MLII) are disorders caused by enzyme deficiencies leading to accumulation of Ceramide Trihexosides and/or sulfatides in tissues and increased excretion in urine. Ceramide Trihexosides and sulfatides are extracted in chloroform from urine using a simple extraction method (modified from Alharbi et al, 2016) and analyzed in both positive mode for Ceramide Trihexosides and negative mode for sulfatides using MALDI-TOF mass spectrometry. Validation included specimens from confirmed patients affected with Fabry disease (N=16 males N=11 females), MLD (N=23), MSD (N=7), SAPB (N=2) and MLII (N=5). Fabry disease has a specific Ceramide Trihexoside profile pattern without presence of sulfatides. Overlapping sulfatide profiles, without Ceramide Trihexosides, for MLD and MSD can be confirmed by glucosaminoglycan analysis. SAPB has both the Ceramide Trihexoside profile pattern similar to Fabry disease and the sulfatide profile pattern similar to MLD and MSD, but reduced peak intensities. Overlapping profiles to MLII, which can have both Ceramide Trihexosides and sulfatides as well, can be confirmed by oligosaccharide analysis. To further delineate profile patterns, peak ratios to respective internal standards in positive and negative modes (N=30) are analyzed using a custom-made automated post-analytical multivariate pattern recognition software, Collaborative Laboratory Integrated Reports (CLIR https://clir.mayo.edu). This software generates additional ratios (N=248) between the analyte/IS ratios to aid in reliable pattern recognition.
N. Sakuragawa - One of the best experts on this subject based on the ideXlab platform.
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In vitro study of encapsulation therapy for Fabry disease using genetically engineered CHO cell line.
Cell transplantation, 2002Co-Authors: Y. Naganawa, Hitoshi Sakuraba, K. Ohsugi, R. Kase, Isao Date, N. SakuragawaAbstract:Fabry disease is an X-linked recessive disorder caused by a deficiency of the lysosomal hydrolase alpha-galactosidase A (alpha-gal). The deficiency of this enzyme leads to the systemic deposition of Ceramide Trihexoside (CTH) in various tissues and organs. Enzyme replacement using IV doses of recombinant human alpha-gal produced in CHO cells or in human fibroblasts is currently being evaluated in clinical trials as a potential therapy for this disease. However, it requires lifelong therapy involving a large amount of purified alpha-gal. As a novel approach for treatment of Fabry disease we used polymer encapsulated Chinese hamster ovary (CHO) cells genetically modified to express alpha-gal. The secreted high levels of alpha-gal passed through the semipermeable polymeric membrane. Using coculture system with Fabry fibroblasts, the secreted enzyme was taken up in cells, resulting in reduced accumulation of CTH in Fabry fibroblasts. This in vitro study demonstrated that an encapsulated alpha-gal-secreting cell line can be used to treat Fabry mice by transplantation in vivo. Judging from the protection against immune rejection by a semipermeable synthetic membrane, this novel approach may be applied to treat patients with Fabry disease and other lysosomal storage diseases.
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Enzymatic corrections for cells derived from Fabry disease patients by a recombinant adenovirus vector
Journal of Human Genetics, 2000Co-Authors: K. Ohsugi, K. Kobayashi, K. Itoh, H. Sakuraba, N. SakuragawaAbstract:Fabry disease is an X-linked inherited metabolic disorder caused by a deficiency of α-galactosidase (α-gal), resulting in the accumulation of Ceramide Trihexoside (CTH) in body fluids and in many organs and tissues. We constructed a recombinant adenovirus with a human α-gal cDNA (AxCAG α-gal), and transfected this vector to skin fibroblasts from Fabry patients. Transfected cells expressed high amounts of α-gal in their cytoplasm, and a high level of α-gal activity was detected in the medium. The accumulated CTH in the fibroblasts disappeared 3 days after infection. The secreted α-gal also eliminated the accumulated CTH from uninfected patient's cells. The enzyme may be taken up through mannose-6-phosphate receptors, as the addition of mannose-6-phosphate to the medium completely inhibited the uptake of the enzyme. The infected cells continued to express α-gal for more than 10 days. These results suggest that AxCAG α-gal could be used as enzyme replacement gene therapy for Fabry disease.
Johanna E.m. Groener - One of the best experts on this subject based on the ideXlab platform.
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HPLC for Simultaneous Quantification of Total Ceramide, GlucosylCeramide, and Ceramide Trihexoside Concentrations in Plasma
Clinical chemistry, 2007Co-Authors: Johanna E.m. Groener, Ben J. H. M. Poorthuis, Sijmen Kuiper, Mariette T.j. Helmond, Carla E. M. Hollak, Johannes M. F. G. AertsAbstract:Background: Simple, reproducible assays are needed for the quantification of sphingolipids, Ceramide (Cer), and sphingoid bases. We developed an HPLC method for simultaneous quantification of total plasma concentrations of Cer, glucosylCeramide (GlcCer), and Ceramide Trihexoside (CTH). Methods: After addition of sphinganine as internal calibrator, we extracted lipids from 50 L plasma. We deacylated Cer and glycosphingolipids by use of microwave-assisted hydrolysis in methanolic NaOH, followed by derivatization of the liberated amino-group with o-phthaldialdehyde. We separated the derivatized sphingoid bases and lysoglycosphingolipids by HPLC on a C18 reversed-phase column with a methanol/water mobile phase (88:12, vol/vol) and quantified them by use of a fluorescence detector at ex 340 nm and em 435 nm. Results: Optimal conditions in the Solids/Moisture System SAM-155 microwave oven (CEM Corp.) for the complete deacylation of Cer and neutral glycosphingolipids without decomposition were 60 min at 85% power, fan setting 7. Intra- and interassay CVs were
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hplc for simultaneous quantification of total Ceramide glucosylCeramide and Ceramide Trihexoside concentrations in plasma
Clinical Chemistry, 2007Co-Authors: Johanna E.m. Groener, Ben J. H. M. Poorthuis, Sijmen Kuiper, Mariette T.j. Helmond, Carla E. M. Hollak, Johannes M. F. G. AertsAbstract:Background: Simple, reproducible assays are needed for the quantification of sphingolipids, Ceramide (Cer), and sphingoid bases. We developed an HPLC method for simultaneous quantification of total plasma concentrations of Cer, glucosylCeramide (GlcCer), and Ceramide Trihexoside (CTH). Methods: After addition of sphinganine as internal calibrator, we extracted lipids from 50 μL plasma. We deacylated Cer and glycosphingolipids by use of microwave-assisted hydrolysis in methanolic NaOH, followed by derivatization of the liberated amino-group with o -phthaldialdehyde. We separated the derivatized sphingoid bases and lysoglycosphingolipids by HPLC on a C18 reversed-phase column with a methanol/water mobile phase (88:12, vol/vol) and quantified them by use of a fluorescence detector at λex 340 nm and λem 435 nm. Results: Optimal conditions in the Solids/Moisture System SAM-155 microwave oven (CEM Corp.) for the complete deacylation of Cer and neutral glycosphingolipids without decomposition were 60 min at 85% power, fan setting 7. Intra- and interassay CVs were <4% and <14%, respectively, and recovery rates were 87%–113%. The limit of quantification was 2 pmol (0.1 pmol on column), and the method was linear over the interval of 2–200 μL plasma. In samples from 40 healthy individuals, mean (SD) concentrations were 9.0 (2.3) μmol/L for Cer, 6.3 (1.9) μmol/L for GlcCer, and 1.7 (0.5) μmol/L for CTH. Plasma concentrations of GlcCer were higher in Gaucher disease patient samples and of CTH in Fabry disease patient samples. Conclusions: HPLC enables quantification of total Cer, GlcCer, and CTH in plasma and is useful for the follow-up of patients on therapy for Gaucher or Fabry disease.
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Trihexoside Concentrations in Plasma
2006Co-Authors: Johanna E.m. Groener, Ben J. H. M. Poorthuis, Sijmen Kuiper, Mariette T.j. Helmond, Carla E. M. Hollak, Johannes M. F. G. AertsAbstract:Background: Simple, reproducible assays are needed for the quantification of sphingolipids, Ceramide (Cer), and sphingoid bases. We developed an HPLC method for simultaneous quantification of total plasma concentrations of Cer, glucosylCeramide (GlcCer), and Ceramide Trihexoside (CTH). Methods: After addition of sphinganine as internal calibrator, we extracted lipids from 50 �L plasma. We deacylated Cer and glycosphingolipids by use of microwave-assisted hydrolysis in methanolic NaOH, followed by derivatization of the liberated amino-group with o-phthaldialdehyde. We separated the derivatized sphingoid bases and lysoglycosphingolipids by HPLC on a C18 reversed-phase column with a methanol/water mobile phase (88:12, vol/vol) and quantified them by use of a fluorescence detector at � ex 340 nm and � em 435 nm. Results: Optimal conditions in the Solids/Moisture System SAM-155 microwave oven (CEM Corp.) for the complete deacylation of Cer and neutral glycosphingolipids without decomposition were 60 min at 85% power, fan setting 7. Intra- and interassay CVs were <4 % and <14%, respectively, and recovery rates were 87%–113%. The limit of quantification was 2 pmol (0.1 pmol on column), and the method was linear over the interval of 2–200 �L plasma. In samples from 40 healthy individuals, mean (SD) concentrations were 9.0 (2.3