g(m1)-ganglioside and Sandhoff-Disease

Topics: Aspartylglucosaminuria; Carbohydrate Metabolism, Inborn Errors; Chromatography, Paper; Fucose; G(M1) Ganglioside; Gangliosidoses; Glucosidases; Glycoproteins; Humans; Lactose Intolerance; Mannose; Mucolipidoses; Mucopolysaccharidoses; Neuraminidase; Oligosaccharides; Sandhoff Disease

Topics: Adolescent; beta-Galactosidase; Brain Diseases, Metabolic; Child; Child, Preschool; Clinical Enzyme Tests; Female; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosidoses; Humans; Infant; Infant, Newborn; Isoenzymes; Mass Screening; Metabolism, Inborn Errors; Models, Genetic; Phenotype; Pregnancy; Prenatal Diagnosis; Sandhoff Disease; Tay-Sachs Disease

Gangliosides (GGs) are considered as diagnostic biomarkers and therapeutic targets and agents. The goal of this study was to develop a tandem mass spectrometry (MS/MS) method for the simultaneous measurement of both GM1 and GM2 gangliosides in human cerebrospinal fluid (CSF) samples in order to be able to determine their concentrations in patients with Tay-Sachs and Sandhoff disease and assess whether drugs or transplantation affect their concentrations.. An API-4000 tandem mass spectrometer equipped with TurboIonSpray source and Shimadzu HPLC system was employed to perform the analysis using isotope dilution with deuterium labeled internal standards. To a 1.5 mL conical plastic Eppendorf centrifuge tube, 40 microL of human CSF sample was added and mixed with 400 microL of internal standard solution for deproteinization. After centrifugation, 100 microL of supernatant was injected onto a C-18 column. After a 2.5 min wash, the switching valve was activated and the analytes were eluted from the column with a water/methanol gradient into the MS/MS system. Quantification by multiple reaction-monitoring (MRM) analysis was performed in the negative mode.. The within-day coefficients of variation were <3% for GM1 and <2% for GM2 and the between-day coefficients of variation were <5% for both GM1 and GM2 at all concentrations tested. Accuracy ranged between 98% and 102% for both analytes. Good linearity was also obtained within the concentration range of 10-200 ng/mL (6.5-129.3 nmol/L) for GM1 and 5-100 ng/mL (3.6-72.3 nmol/L) for GM2 (r> or =0.995).. A new simple, accurate, and fast isotope dilution tandem mass spectrometry method was developed for the simultaneous quantification of GM1 and GM2 gangliosides in a small amount of human CSF. Concentrations were measured in "normal" CSF and in CSF from patients with Tay-Sachs disease.

Topics: G(M1) Ganglioside; G(M2) Ganglioside; Humans; Indicator Dilution Techniques; Molecular Structure; Reproducibility of Results; Sandhoff Disease; Sensitivity and Specificity; Tandem Mass Spectrometry; Tay-Sachs Disease

GM1- and GM2-gangliosides were isolated from brain and radiolabelled. The labelled moieties were localized by hydrolysis with lysosomal enzymes, followed by thin-layer chromatography of the products. High-resolution loading tests with labelled gangliosides were developed and found to differentiate infantile and juvenile forms of GM1- and GM2-gangliosidoses as well as the identification of B, O and AB types of GM2-gangliosidosis.

Topics: Animals; Brain Chemistry; Chromatography, Thin Layer; Diagnosis, Differential; Fibroblasts; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosidoses; Gangliosidosis, GM1; Genetic Variation; Humans; Kinetics; Mice; Sandhoff Disease; Tay-Sachs Disease

Neurophysiological studies (EEG, ERG, VEP and BAEP) have been carried out on a total of fifty-four patients (fourty-five GM2 and nine GM1 gangliosidosis) at various stages of the disease process. In infantile GM2 gangliosidosis, the EEG was midly abnormal from an early age but by the age of one year there was a rapid and progressive deterioration. EEG changes in late onset GM2 gangliosidosis were very variable and unrelated to age or enzyme defect. In both Type 1 and Type 2 GM1 gangliosidosis there was a progressive deterioration of the EEG. Paroxysmal features were not prominent in any of the gangliosidoses, despite the occurrence of seizures. The ERG remained normal in both GM2 and GM1 patients. In the infantile GM2 patients there was progressive loss of the VEP between nine and fifteen months of age but the timing of VEP changes were more variable in all the other groups. Evidence of brainstem dysfunction was found in one of the two TSD patients tested. The combined neurophysiological features appear to be characteristic for each group of gangliosidosis and differ from other neurometabolic disorders of childhood.

Topics: Arousal; Brain; Child; Child, Preschool; Electroencephalography; Electroretinography; Evoked Potentials; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Humans; Infant; Sandhoff Disease; Synaptic Transmission; Tay-Sachs Disease

GM1 and GM2 gangliosidoses are progressive neurodegenerative diseases which accumulate intralysosomal gangliosides--and to a lesser extent oligosaccharides--chiefly in the central and peripheral nervous system owing to deficiencies of beta-galactosidase and hexosaminidases A or/and B, respectively. This intralysosomal "storage" in neuronal pericarya and their processes, and subsequent loss of such nerve cells provide the background for clinical symptoms of the central nervous system and the retina, while involvement of the peripheral nervous system and the visceral organs largely remains free of clinical findings. The morphological involvement of the latter organs is widespread though varying, thus allowing morphological investigations of lymphocytes, skin, or rectum for morphological diagnosis and as a screening procedure.

Topics: Astrocytes; Brain; Child; Dendrites; G(M1) Ganglioside; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Humans; Inclusion Bodies; Lysosomes; Microscopy, Electron; Nerve Degeneration; Neurons; Peripheral Nerves; Sandhoff Disease; Spinal Cord; Synaptic Membranes; Tay-Sachs Disease; Vacuoles

Topics: beta-Galactosidase; beta-N-Acetylhexosaminidases; Brain; Child; Fibroblasts; G(M1) Ganglioside; G(M2) Activator Protein; G(M2) Ganglioside; Gangliosides; Gangliosidoses; Glycoproteins; Hexosaminidases; Humans; Lysosomes; Mutation; Protein Deficiency; Proteins; Sandhoff Disease; Saposins; Sphingolipid Activator Proteins; Tay-Sachs Disease

Rectal mucosa biopsy specimens from patients with neuronal storage diseases were examined by electron microscopy. The diseases were Tay-Sachs disease, Sandhoff's disease, Niemann-Pick disease types B and C, late infantile metachromatic leukodystrophy, GM1 gangliosidosis type 1, beta-galactosidase-neuraminidase deficiency, I-cell disease, and mucopolysaccharidoses (Hunter's syndrome and Sanfilippo's syndrome type A). Unmyelinated nerve fibers, endothelial cells, fibroblasts, plasma cells, and histiocytes were seen in the specimens. Except for plasma cells, the results thus obtained for various cells were similar to those obtained from skin and conjunctival biopsy specimens, which have been already reported. There has been no report so far on ultrastructure of the plasma cell in these diseases. Storage materials, eg, dense bodies and membrane-bound vacuoles, were observed in the plasma cells in various storage diseases, with the exception of late infantile metachromatic leukodystrophy. Thus, electron microscopy of rectal mucosa is useful in making diagnoses and examining plasma cells in some neuronal storage diseases.

Topics: Adolescent; Axons; Biopsy; Brain Diseases, Metabolic; Child, Preschool; G(M1) Ganglioside; Humans; Infant; Intestinal Mucosa; Leukodystrophy, Metachromatic; Lymphocytes; Microscopy, Electron; Niemann-Pick Diseases; Plasma Cells; Rectum; Sandhoff Disease; Tay-Sachs Disease