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Tamotsu Kanzaki - One of the best experts on this subject based on the ideXlab platform.
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.ALPHA.-N-acetylgalactosaminidase Deficiency with Angiokeratoma Corporis Diffusum (Kanzaki Disease): Discovery and Development Thereafer-1986 through 2006-
Nishi Nihon Hifuka, 2006Co-Authors: Tamotsu KanzakiAbstract:A 48-year-old Japanese lady was examined in 1989. She was apparently healthy but showed numerous numbers of small petechiae-like angiokeratoma on her lower torso to upper thighs, axillae and beneath the breasts. Electron microscopic examination of the skin revealed largely dilated electron lucent lysosomes with fuzzy filamentous materials in vascular endothelial cells, fibroblasts, eccrine sweat gland cells and others. Urinary examination revealed unusual glycopeptides with GalNAc-Ser/Thr moieties. This Disease was reported as a novel lysosomal storage Disease with angiokeratoma corporis diffusum, crowned Kanzaki Disease (MIM#104170). Soon, this Disease was found to be caused by a deficit of α-N-acetylgalactosaminidase (α-NAGA, 4. 3. 2. 49) activity and a point mutation was found in the gene (R329W) encoding the enzyme. Another patient, 47-year-old Japanese woman, was found, and she also was apparently healthy, but had less angiokeratoma as compared to the first one. The gene mutation was found and the resultant mutant enzyme was R329Q. She excreted less amount of GalNAc-Ser in urine as compared to the first patient. These phenotypical differences between case 1 and 2 were estimated to be caused by the differences in the three-dimensional structures in mutated α-NAGAs (R329W v. s. R329Q). Schindler Disease also shows α-NAGA deficiency but shows very severe central nervous symptoms before the age of one. Electron microscopically electron-dence material deposited in lysosomes in Schindler Disease. Electron-dense material means that the substance are probably lipid- or lipoprotein-containg materials. This quiet contrasts to the findings in Kanzaki Disease. These evidences suggest that Kanzaki Disease is caused by a pure α-NAGA deficiency but Schindler Disease is probably caused by together α-NAGA deficiency with some other factors.
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Structural and immunocytochemical studies on α-N-acetylgalactosaminidase deficiency (Schindler/Kanzaki Disease)
Journal of Human Genetics, 2004Co-Authors: Hitoshi Sakuraba, Tomoko Fukushige, Fumiko Matsuzawa, Sei-ichi Aikawa, Masaharu Kotani, Hiroshi Nakada, Tamotsu KanzakiAbstract:α- N -Acetylgalactosaminidase (α-NAGA) deficiency (Schindler/Kanzaki Disease) is a clinically and pathologically heterogeneous genetic Disease with a wide spectrum including an early onset neuroaxonal dystrophy (Schindler Disease) and late onset angiokeratoma corporis diffusum (Kanzaki Disease). In α-NAGA deficiency, there are discrepancies between the genotype and phenotype, and also between urinary excretion products (sialyl glycoconjugates) and a theoretical accumulated material (Tn-antigen; Gal NAcα1- Ο -Ser/Thr) resulting from a defect in α-NAGA. As for the former issue, previously reported genetic, biochemical and pathological data raise the question whether or not E325K mutation found in Schindler Disease patients really leads to the severe phenotype of α-NAGA deficiency. The latter issue leads to the question of whether α-NAGA deficiency is associated with the basic pathogenesis of this Disease. To clarify the pathogenesis of this Disease, we performed structural and immunocytochemical studies. The structure of human α-NAGA deduced on homology modeling is composed of two domains, domain I, including the active site, and domain II. R329W/Q, identified in patients with Kanzaki Disease have been deduced to cause drastic changes at the interface between domains I and II. The structural change caused by E325K found in patients with Schindler Disease is localized on the N-terminal side of the tenth β-strand in domain II and is smaller than those caused by R329W/Q. Immunocytochemical analysis revealed that the main lysosomal accumulated material in cultured fibroblasts from patients with Kanzaki Disease is Tn-antigen. These data suggest that a prototype of α-NAGA deficiency in Kanzaki Disease and factors other than the defect of α-NAGA may contribute to severe neurological disorders, and Kanzaki Disease is thought to be caused by a single enzyme deficiency.
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Structural and immunocytochemical studies on α-N-acetylgalactosaminidase deficiency (Schindler/Kanzaki Disease)
Journal of Human Genetics, 2004Co-Authors: Hitoshi Sakuraba, Tomoko Fukushige, Fumiko Matsuzawa, Sei-ichi Aikawa, Masaharu Kotani, Hiroshi Nakada, Hirofumi Doi, Tamotsu KanzakiAbstract:α- N -Acetylgalactosaminidase (α-NAGA) deficiency (Schindler/Kanzaki Disease) is a clinically and pathologically heterogeneous genetic Disease with a wide spectrum including an early onset neuroaxonal dystrophy (Schindler Disease) and late onset angiokeratoma corporis diffusum (Kanzaki Disease). In α-NAGA deficiency, there are discrepancies between the genotype and phenotype, and also between urinary excretion products (sialyl glycoconjugates) and a theoretical accumulated material (Tn-antigen; Gal NAcα1- Ο -Ser/Thr) resulting from a defect in α-NAGA. As for the former issue, previously reported genetic, biochemical and pathological data raise the question whether or not E325K mutation found in Schindler Disease patients really leads to the severe phenotype of α-NAGA deficiency. The latter issue leads to the question of whether α-NAGA deficiency is associated with the basic pathogenesis of this Disease. To clarify the pathogenesis of this Disease, we performed structural and immunocytochemical studies. The structure of human α-NAGA deduced on homology modeling is composed of two domains, domain I, including the active site, and domain II. R329W/Q, identified in patients with Kanzaki Disease have been deduced to cause drastic changes at the interface between domains I and II. The structural change caused by E325K found in patients with Schindler Disease is localized on the N-terminal side of the tenth β-strand in domain II and is smaller than those caused by R329W/Q. Immunocytochemical analysis revealed that the main lysosomal accumulated material in cultured fibroblasts from patients with Kanzaki Disease is Tn-antigen. These data suggest that a prototype of α-NAGA deficiency in Kanzaki Disease and factors other than the defect of α-NAGA may contribute to severe neurological disorders, and Kanzaki Disease is thought to be caused by a single enzyme deficiency.
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structural and immunocytochemical studies on α n acetylgalactosaminidase deficiency Schindler kanzaki Disease
Journal of Human Genetics, 2004Co-Authors: Hitoshi Sakuraba, Tomoko Fukushige, Fumiko Matsuzawa, Sei-ichi Aikawa, Masaharu Kotani, Hiroshi Nakada, Hirofumi Doi, Tamotsu KanzakiAbstract:α-N-Acetylgalactosaminidase (α-NAGA) deficiency (Schindler/Kanzaki Disease) is a clinically and pathologically heterogeneous genetic Disease with a wide spectrum including an early onset neuroaxonal dystrophy (Schindler Disease) and late onset angiokeratoma corporis diffusum (Kanzaki Disease). In α-NAGA deficiency, there are discrepancies between the genotype and phenotype, and also between urinary excretion products (sialyl glycoconjugates) and a theoretical accumulated material (Tn-antigen; Gal NAcα1-Ο-Ser/Thr) resulting from a defect in α-NAGA. As for the former issue, previously reported genetic, biochemical and pathological data raise the question whether or not E325K mutation found in Schindler Disease patients really leads to the severe phenotype of α-NAGA deficiency. The latter issue leads to the question of whether α-NAGA deficiency is associated with the basic pathogenesis of this Disease. To clarify the pathogenesis of this Disease, we performed structural and immunocytochemical studies. The structure of human α-NAGA deduced on homology modeling is composed of two domains, domain I, including the active site, and domain II. R329W/Q, identified in patients with Kanzaki Disease have been deduced to cause drastic changes at the interface between domains I and II. The structural change caused by E325K found in patients with Schindler Disease is localized on the N-terminal side of the tenth β-strand in domain II and is smaller than those caused by R329W/Q. Immunocytochemical analysis revealed that the main lysosomal accumulated material in cultured fibroblasts from patients with Kanzaki Disease is Tn-antigen. These data suggest that a prototype of α-NAGA deficiency in Kanzaki Disease and factors other than the defect of α-NAGA may contribute to severe neurological disorders, and Kanzaki Disease is thought to be caused by a single enzyme deficiency.
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Improved Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometric Method for Identification of Amino Acid O-Glycosides in Patients with α-N-Acetylgalactosaminidase Deficiency
Clinical chemistry, 2002Co-Authors: Masahisa Horiuchi, Tomoko Fukushige, Takeyori Saheki, Tamotsu KanzakiAbstract:Thin-layer chromatography (TLC) has been widely applied to the diagnosis of lysosomal storage Diseases by analyzing abnormally excreted oligosaccharides in urine. However, TLC does not necessarily identify the excreted compounds accurately. Labeling the oligosaccharides with a tag, such as 8-aminonaphthalene-1,3,6-trisulfonic acid, allows easy identification of the excreted compounds by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) (1). However, this labeling is based on reductive amination of the reductive end of the oligosaccharides with the labeling compound, so abnormal amino acid O-glycosides with no reducing end cannot be identified. Indeed, patients with Kanzaki/Schindler Disease or aspartylglucosaminuria, who mainly excrete abnormal amino acid O-glycosides, can be identified only with difficulty by this method. In the present study, we successfully identified abnormal amino acid O-glycosides in urine from Kanzaki/Schindler Disease [α- N -acetylgalactosaminidase (α-NAGA) deficiency] patients (2), using partition chromatography and MALDI-TOF MS. The method involves three steps: purification on a solid-phase extraction column filled with cellulose;removal of sialic acids; and mass spectrometric analysis. This simple procedure reduces both the amount of urine and analysis time and precisely identifies excreted abnormal substances. For MS, 5- to 15-μL urine samples (equivalent to 5 μg of creatinine) were mixed with 250 pmol of raffinose as an internal standard. Urinary creatinine was measured by colorimetric analysis (3). The samples were then mixed with an ethanol–1-butanol solution (1:4 by volume) and applied to a column filled with …
Tomoko Fukushige - One of the best experts on this subject based on the ideXlab platform.
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Improved Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometric Method for Identifi- cation of Amino Acid O-Glycosides in Patients with -N-Acetylgalactosaminidase Deficiency, Masahisa Ho-
2015Co-Authors: Tomoko Fukushige, Takeyori SahekiAbstract:u.ac.jp) Thin-layer chromatography (TLC) has been widely ap-plied to the diagnosis of lysosomal storage Diseases by analyzing abnormally excreted oligosaccharides in urine. However, TLC does not necessarily identify the excreted compounds accurately. Labeling the oligosaccharides with a tag, such as 8-aminonaphthalene-1,3,6-trisulfonic acid, allows easy identification of the excreted com-pounds by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) (1). However, this labeling is based on reductive amination of the reductive end of the oligosaccharides with the labeling compound, so abnormal amino acid O-glycosides with no reducing end cannot be identified. Indeed, patients with Kanzaki/Schindler Disease or aspartylglucosaminuria
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Structural and immunocytochemical studies on α-N-acetylgalactosaminidase deficiency (Schindler/Kanzaki Disease)
Journal of Human Genetics, 2004Co-Authors: Hitoshi Sakuraba, Tomoko Fukushige, Fumiko Matsuzawa, Sei-ichi Aikawa, Masaharu Kotani, Hiroshi Nakada, Tamotsu KanzakiAbstract:α- N -Acetylgalactosaminidase (α-NAGA) deficiency (Schindler/Kanzaki Disease) is a clinically and pathologically heterogeneous genetic Disease with a wide spectrum including an early onset neuroaxonal dystrophy (Schindler Disease) and late onset angiokeratoma corporis diffusum (Kanzaki Disease). In α-NAGA deficiency, there are discrepancies between the genotype and phenotype, and also between urinary excretion products (sialyl glycoconjugates) and a theoretical accumulated material (Tn-antigen; Gal NAcα1- Ο -Ser/Thr) resulting from a defect in α-NAGA. As for the former issue, previously reported genetic, biochemical and pathological data raise the question whether or not E325K mutation found in Schindler Disease patients really leads to the severe phenotype of α-NAGA deficiency. The latter issue leads to the question of whether α-NAGA deficiency is associated with the basic pathogenesis of this Disease. To clarify the pathogenesis of this Disease, we performed structural and immunocytochemical studies. The structure of human α-NAGA deduced on homology modeling is composed of two domains, domain I, including the active site, and domain II. R329W/Q, identified in patients with Kanzaki Disease have been deduced to cause drastic changes at the interface between domains I and II. The structural change caused by E325K found in patients with Schindler Disease is localized on the N-terminal side of the tenth β-strand in domain II and is smaller than those caused by R329W/Q. Immunocytochemical analysis revealed that the main lysosomal accumulated material in cultured fibroblasts from patients with Kanzaki Disease is Tn-antigen. These data suggest that a prototype of α-NAGA deficiency in Kanzaki Disease and factors other than the defect of α-NAGA may contribute to severe neurological disorders, and Kanzaki Disease is thought to be caused by a single enzyme deficiency.
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Structural and immunocytochemical studies on α-N-acetylgalactosaminidase deficiency (Schindler/Kanzaki Disease)
Journal of Human Genetics, 2004Co-Authors: Hitoshi Sakuraba, Tomoko Fukushige, Fumiko Matsuzawa, Sei-ichi Aikawa, Masaharu Kotani, Hiroshi Nakada, Hirofumi Doi, Tamotsu KanzakiAbstract:α- N -Acetylgalactosaminidase (α-NAGA) deficiency (Schindler/Kanzaki Disease) is a clinically and pathologically heterogeneous genetic Disease with a wide spectrum including an early onset neuroaxonal dystrophy (Schindler Disease) and late onset angiokeratoma corporis diffusum (Kanzaki Disease). In α-NAGA deficiency, there are discrepancies between the genotype and phenotype, and also between urinary excretion products (sialyl glycoconjugates) and a theoretical accumulated material (Tn-antigen; Gal NAcα1- Ο -Ser/Thr) resulting from a defect in α-NAGA. As for the former issue, previously reported genetic, biochemical and pathological data raise the question whether or not E325K mutation found in Schindler Disease patients really leads to the severe phenotype of α-NAGA deficiency. The latter issue leads to the question of whether α-NAGA deficiency is associated with the basic pathogenesis of this Disease. To clarify the pathogenesis of this Disease, we performed structural and immunocytochemical studies. The structure of human α-NAGA deduced on homology modeling is composed of two domains, domain I, including the active site, and domain II. R329W/Q, identified in patients with Kanzaki Disease have been deduced to cause drastic changes at the interface between domains I and II. The structural change caused by E325K found in patients with Schindler Disease is localized on the N-terminal side of the tenth β-strand in domain II and is smaller than those caused by R329W/Q. Immunocytochemical analysis revealed that the main lysosomal accumulated material in cultured fibroblasts from patients with Kanzaki Disease is Tn-antigen. These data suggest that a prototype of α-NAGA deficiency in Kanzaki Disease and factors other than the defect of α-NAGA may contribute to severe neurological disorders, and Kanzaki Disease is thought to be caused by a single enzyme deficiency.
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structural and immunocytochemical studies on α n acetylgalactosaminidase deficiency Schindler kanzaki Disease
Journal of Human Genetics, 2004Co-Authors: Hitoshi Sakuraba, Tomoko Fukushige, Fumiko Matsuzawa, Sei-ichi Aikawa, Masaharu Kotani, Hiroshi Nakada, Hirofumi Doi, Tamotsu KanzakiAbstract:α-N-Acetylgalactosaminidase (α-NAGA) deficiency (Schindler/Kanzaki Disease) is a clinically and pathologically heterogeneous genetic Disease with a wide spectrum including an early onset neuroaxonal dystrophy (Schindler Disease) and late onset angiokeratoma corporis diffusum (Kanzaki Disease). In α-NAGA deficiency, there are discrepancies between the genotype and phenotype, and also between urinary excretion products (sialyl glycoconjugates) and a theoretical accumulated material (Tn-antigen; Gal NAcα1-Ο-Ser/Thr) resulting from a defect in α-NAGA. As for the former issue, previously reported genetic, biochemical and pathological data raise the question whether or not E325K mutation found in Schindler Disease patients really leads to the severe phenotype of α-NAGA deficiency. The latter issue leads to the question of whether α-NAGA deficiency is associated with the basic pathogenesis of this Disease. To clarify the pathogenesis of this Disease, we performed structural and immunocytochemical studies. The structure of human α-NAGA deduced on homology modeling is composed of two domains, domain I, including the active site, and domain II. R329W/Q, identified in patients with Kanzaki Disease have been deduced to cause drastic changes at the interface between domains I and II. The structural change caused by E325K found in patients with Schindler Disease is localized on the N-terminal side of the tenth β-strand in domain II and is smaller than those caused by R329W/Q. Immunocytochemical analysis revealed that the main lysosomal accumulated material in cultured fibroblasts from patients with Kanzaki Disease is Tn-antigen. These data suggest that a prototype of α-NAGA deficiency in Kanzaki Disease and factors other than the defect of α-NAGA may contribute to severe neurological disorders, and Kanzaki Disease is thought to be caused by a single enzyme deficiency.
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Improved Matrix-assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometric Method for Identification of Amino Acid O-Glycosides in Patients with α-N-Acetylgalactosaminidase Deficiency
Clinical chemistry, 2002Co-Authors: Masahisa Horiuchi, Tomoko Fukushige, Takeyori Saheki, Tamotsu KanzakiAbstract:Thin-layer chromatography (TLC) has been widely applied to the diagnosis of lysosomal storage Diseases by analyzing abnormally excreted oligosaccharides in urine. However, TLC does not necessarily identify the excreted compounds accurately. Labeling the oligosaccharides with a tag, such as 8-aminonaphthalene-1,3,6-trisulfonic acid, allows easy identification of the excreted compounds by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) (1). However, this labeling is based on reductive amination of the reductive end of the oligosaccharides with the labeling compound, so abnormal amino acid O-glycosides with no reducing end cannot be identified. Indeed, patients with Kanzaki/Schindler Disease or aspartylglucosaminuria, who mainly excrete abnormal amino acid O-glycosides, can be identified only with difficulty by this method. In the present study, we successfully identified abnormal amino acid O-glycosides in urine from Kanzaki/Schindler Disease [α- N -acetylgalactosaminidase (α-NAGA) deficiency] patients (2), using partition chromatography and MALDI-TOF MS. The method involves three steps: purification on a solid-phase extraction column filled with cellulose;removal of sialic acids; and mass spectrometric analysis. This simple procedure reduces both the amount of urine and analysis time and precisely identifies excreted abnormal substances. For MS, 5- to 15-μL urine samples (equivalent to 5 μg of creatinine) were mixed with 250 pmol of raffinose as an internal standard. Urinary creatinine was measured by colorimetric analysis (3). The samples were then mixed with an ethanol–1-butanol solution (1:4 by volume) and applied to a column filled with …
Roselyne Froissart - One of the best experts on this subject based on the ideXlab platform.
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Development of a new tandem mass spectrometry method for urine and amniotic fluid screening of oligosaccharidoses
Rapid Communications in Mass Spectrometry, 2017Co-Authors: M. Piraud, M. Pettazzoni, L. Menegaut, C. Caillaud, Y. Nadjar, Christine Vianey-saban, Roselyne FroissartAbstract:RATIONALE: The first step in the diagnosis of oligosaccharidoses is to evidence abnormal oligosaccharides excreted in urine, usually performed by the poorly sensitive but efficient thin layer chromatography (TLC) method. Developing a tandem mass spectrometry (MS/MS) technique could be of great interest to replace TLC. METHODS: Abnormal underivatized oligosaccharides have been recently studied using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, allowing the unambiguous identification of oligosaccharidoses. Based on this previous work, we developed an advantageous and efficient liquid chromatography (LC)/MS/MS method using a more common triple quadrupole tandem mass spectrometer for oligosaccharides analysis. RESULTS: Oligosaccharidoses (n = 97) and control (n = 240) urine samples were analysed. A specific pattern was obtained for each oligosaccharidosis using this method. In urine, it allows not only the identification of all the oligosaccharidoses previously identified by TLC (fucosidosis, alphamannosidosis, aspartylglucosaminuria, GM1 gangliosidosis, sialidosis, galactosialidosis and Schindler Disease), but also extends the field of diagnosis to mucolipidosis type II, Sandhoff Disease, and beta-mannosidosis. The same technique was applied to 16 amniotic fluid supernatants from oligosaccharidosis-affected foetuses (n = 16) compared with 37 unaffected. All the affected foetuses could be clearly identified: sialidosis (n = 3), galactosialidosis (n = 4), aspartylglucosaminuria (n = 1), mucolipidosis type II (n = 4) or GM1 gangliosidosis (n = 4). This technique can be applied to early prenatal diagnosis as well as to the oligosaccharidosis screening in the case of non-immune hydrops fetalis. CONCLUSIONS: The method is quick and easy to run, with an LC analysis time of 13 min per sample. The quantitative validation could not be obtained in the absence of a specific standard and of a labelled internal standard for each compound. Even if this LC/MS/MS method is only qualitative, it is very specific and much more sensitive than TLC. It allows the urinary screening of oligosaccharidoses, even mild or late-onset forms, and the screening of antenatal forms in amniotic fluid. Copyright (C) 2017 John Wiley & Sons, Ltd.
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Development of a new tandem mass spectrometry method for urine and amniotic fluid screening of oligosaccharidoses
Rapid communications in mass spectrometry : RCM, 2017Co-Authors: M. Piraud, M. Pettazzoni, L. Menegaut, C. Caillaud, Y. Nadjar, Christine Vianey-saban, Roselyne FroissartAbstract:Rationale The first step in the diagnosis of oligosaccharidoses is to evidence abnormal oligosaccharides excreted in urine, usually performed by the poorly sensitive but efficient thin layer chromatography (TLC) method. Developing a tandem mass spectrometry (MS/MS) technique could be of great interest to replace TLC. Methods Abnormal underivatized oligosaccharides have been recently studied using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, allowing the unambiguous identification of oligosaccharidoses. Based on this previous work, we developed an advantageous and efficient liquid chromatography (LC)/MS/MS method using a more common triple quadrupole tandem mass spectrometer for oligosaccharides analysis. Results Oligosaccharidoses (n = 97) and control (n = 240) urine samples were analysed. A specific pattern was obtained for each oligosaccharidosis using this method. In urine, it allows not only the identification of all the oligosaccharidoses previously identified by TLC (fucosidosis, alphamannosidosis, aspartylglucosaminuria, GM1 gangliosidosis, sialidosis, galactosialidosis and Schindler Disease), but also extends the field of diagnosis to mucolipidosis type II, Sandhoff Disease, and β-mannosidosis. The same technique was applied to 16 amniotic fluid supernatants from oligosaccharidosis-affected foetuses (n = 16) compared with 37 unaffected. All the affected foetuses could be clearly identified: sialidosis (n = 3), galactosialidosis (n = 4), aspartylglucosaminuria (n = 1), mucolipidosis type II (n = 4) or GM1 gangliosidosis (n = 4). This technique can be applied to early prenatal diagnosis as well as to the oligosaccharidosis screening in the case of non-immune hydrops fetalis. Conclusions The method is quick and easy to run, with an LC analysis time of 13 min per sample. The quantitative validation could not be obtained in the absence of a specific standard and of a labelled internal standard for each compound. Even if this LC/MS/MS method is only qualitative, it is very specific and much more sensitive than TLC. It allows the urinary screening of oligosaccharidoses, even mild or late-onset forms, and the screening of antenatal forms in amniotic fluid. Copyright © 2017 John Wiley & Sons, Ltd.
E. Rodriguez Diaz - One of the best experts on this subject based on the ideXlab platform.
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Mild phenotypic expression of α-N-acetylgalactosaminidase deficiency in two adult siblings
Journal of Inherited Metabolic Disease, 1994Co-Authors: A. Chabás, M. J. Coll, M. Aparicio, E. Rodriguez DiazAbstract:Two adult siblings with an α- N -acetylgalactosaminidase deficiency are described. The patients' major features are massive lymphoedema and angiokeratoma corporis diffusum. Neurological evaluation performed in one of the patients was considered within normal limits. Blood type is A positive in each case. Ultrastructural examination of skin revealed numerous vacuoles in endothelial cells and pericytes. Fibroblast activity of α- N -acetylgalactosaminidase was decreased to 0.6–2% of mean normal value. Chromatography of urinary oligosaccharides showed abnormal bands identical to those excreted by two infantile patients with Schindler Disease. The bands were identified as sialyloligosaccharides, and gas chromatography revealed the presence of N -acetylgalactosamine-rich compounds accounting for 30% of the total monosaccharide content of the oligosaccharide fraction. These findings confirm the heterogeneity of α- N -acetylgalactosaminidase deficiency and emphasize the need to consider this lysosomal storage Disease in the differential diagnosis of patients with angiokeratoma.
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Mild phenotypic expression of α- N -acetylgalactosaminidase deficiency in two adult siblings
Journal of inherited metabolic disease, 1994Co-Authors: A. Chabás, M. J. Coll, M. Aparicio, E. Rodriguez DiazAbstract:Two adult siblings with an α-N-acetylgalactosaminidase deficiency are described. The patients' major features are massive lymphoedema and angiokeratoma corporis diffusum. Neurological evaluation performed in one of the patients was considered within normal limits. Blood type is A positive in each case. Ultrastructural examination of skin revealed numerous vacuoles in endothelial cells and pericytes. Fibroblast activity of α-N-acetylgalactosaminidase was decreased to 0.6–2% of mean normal value. Chromatography of urinary oligosaccharides showed abnormal bands identical to those excreted by two infantile patients with Schindler Disease. The bands were identified as sialyloligosaccharides, and gas chromatography revealed the presence ofN-acetylgalactosamine-rich compounds accounting for 30% of the total monosaccharide content of the oligosaccharide fraction.
Alina D. Zamfir - One of the best experts on this subject based on the ideXlab platform.
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Application of ion mobility tandem mass spectrometry to compositional and structural analysis of glycopeptides extracted from the urine of a patient diagnosed with Schindler Disease
Rapid communications in mass spectrometry : RCM, 2015Co-Authors: Mirela Sarbu, Jasna Peter-katalinić, Feifei Zhu, David E. Clemmer, Alina D. ZamfirAbstract:Rationale Schindler Disease is caused by the deficient activity of α-N-acetylgalactosaminidase, which leads to an abnormal accumulation of O-glycopeptides in tissues and body fluids. In this work the Schindler condition is for the first time approached by ion mobility (IMS) tandem mass spectrometry (MS/MS), for determining urine glycopeptide fingerprints and discriminate isomeric structures. Methods IMS-MS experiments were conducted on a Synapt G2s mass spectrometer operating in negative ion mode. A glycopeptide mixture extracted from the urine of a patient suffering from Schindler Disease was dissolved in methanol and infused into the mass spectrometer by electrospray ionization using a syringe-pump system. MS/MS was performed by collision-induced dissociation (CID) at low energies, after mobility separation in the transfer cell. Data acquisition and processing were performed using MassLynx and Waters Driftscope software. Results IMS-MS data indicated that the attachment of one or two amino acids to the carbohydrate backbone has a minimal influence on the molecule conformation, which limits the discrimination of the free oligosaccharides from the glycosylated amino acids and dipeptides. The structural analysis by CID MS/MS in combination with IMS-MS of species exhibiting the same m/z but different configurations demonstrated for the first time the presence of positional isomers for some of the Schindler Disease biomarker candidates. Conclusions The IMS-MS and CID MS/MS platform was for the first time optimized and applied to Schindler Disease glycourinome. By this approach the separation and characterization of Neu5Ac positional isomers was possible. IMS CID MS/MS showed the ability to determine the type of the glycopeptide isomers from a series of possible candidates. Copyright © 2015 John Wiley & Sons, Ltd.
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Identification and structural characterization of novel O- and N-glycoforms in the urine of a Schindler Disease patient by Orbitrap mass spectrometry.
Journal of mass spectrometry : JMS, 2015Co-Authors: Mirela Sarbu, Jasna Peter-katalinić, Cristian V. A. Munteanu, Liana Dehelean, Andrei J. Petrescu, Alina D. ZamfirAbstract:Schindler Disease is an inherited metabolic disorder caused by the deficient activity of α-N-acetylgalactosaminidase enzyme. An accurate diagnosis requires, besides clinical examination, complex and costly biochemical and molecular genetic tests. In the last years, mass spectrometry (MS) based on nanofluidics and high-resolution instruments has become a successful alternative for Disease diagnosis based on the investigation of O-glycopeptides in patient urine. A complex mixture of glycoforms extracted from the urine of a 3-year-old patient was investigated by Orbitrap MS equipped with Nanospray Flex Ion Source in the negative ion mode. For structural characterization of several molecular species, collision-induced dissociation MS2–MS3 was carried out using collision energy values within 20–60 eV range. By our approach, 39 novel species associated to this condition were identified, among which O-glycopeptides, free O-glycans and one structure corresponding to an N-glycan never characterized in the context of Schindler Disease. The experiments conducted at a resolution of 60 000 allowed the discrimination and identification of a total number of 69 different species with an average mass accuracy of 9.87 ppm, an in-run reproducibility of almost 100%, an experiment-to-experiment and day-to-day reproducibility of about 95%. This study brings contributions in the diagnosis of Schindler Disease through the elucidation of potential biomarker species in urine. Our multistage MS results completed with 39 new glycoforms the inventory of potential biomarker structures associated to Schindler Disease. For the first time, an N-glycan was identified and structurally characterized in Schindler patient urine, which opens new research directions in the field. Copyright © 2015 John Wiley & Sons, Ltd.
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Automated chip-nanoelectrospray mass spectrometry for glycourinomics in Schindler Disease type I
Carbohydrate research, 2014Co-Authors: Mirela Sarbu, Adrian C. Robu, Jasna Peter-katalinić, Alina D. ZamfirAbstract:In this study an integrative mass spectrometry (MS) approach based on fully automated chip-nanoelectrospray quadrupole time-of-flight was optimized and applied for the discovery and structural characterization of O-glycopeptides in a fraction from the urine of a patient diagnosed with Schindler Disease type I. A mixture of O-glycopeptides extracted and purified from an age matched healthy subject served as the control. 49 glycoforms were discovered in the investigated urine fraction from Schindler Disease versus only 14 in control urine. Structures with relevant biological significance, previously not described, such as O-fucosylated tetrasaccharides and chains up to pentadecamers O-linked to serine, threonine, or threonine-proline were identified in the pathological urine and characterized by tandem MS (MS/MS). A number of 29 species discovered here, most of which with long chain glycans, were not previously reported as associated to this condition. All glycopeptides were detected in only 1 min analysis time, with a sample consumption situated in the femtomole range.
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Copper‐coated microsprayer interface for on‐line sheathless capillary electrophoresis electrospray mass spectrometry of carbohydrates
Journal of separation science, 2006Co-Authors: Alina D. Zamfir, Nicolae Dinca, Eugen Sisu, Jasna Peter-katalinićAbstract:A sturdy home-built sheathless CE/ESI-QTOF-MS system was developed and optimized for carbohydrate analysis. The interface and employed methodology provided a simple analytical solution to laborious CE/MS interfacing methods and to problems in characterization of complex carbohydrate mixtures that require high-resolution separation of the components. The CE/ESI interface, feasible in any MS laboratory, consists of a one-piece CE column having the CE terminus in-laboratory shaped as a microsprayer and coated with copper. The CE microsprayer was inserted into an in-house made stainless steel clenching device and the whole assembly was mounted onto a quadrupole TOF mass spectrometer. The analytical potential of the interface in terms of suitability, microsprayer performance, copper coat durability, ionization efficiency, spray stability, and sensitivity was tested first on a simple mixture of standard saccharides, which were separated, resolved, and detected with high separation efficiency. The approach was next assessed for the screening of a biological sample, a complex mixture of 0-glycosylated sialylated amino acids from urine of a patient suffering from Schindler Disease. Preliminary data allow this method to be considered as one of general applicability in structural glycobiology and glycomics and easy to be implemented for proteomic surveys as well.
