chondroitin-sulfates and Mucopolysaccharidoses

Topics: beta-Galactosidase; Cells, Cultured; Child, Preschool; Chondroitin Sulfates; Female; Fucose; Gangliosidoses; Genetic Carrier Screening; Hexosyltransferases; Humans; Iduronidase; Liver; Lysosomes; Mannose; Mucolipidoses; Mucopolysaccharidoses; Mucopolysaccharidosis I; Mucopolysaccharidosis II; Mucopolysaccharidosis III; Mucopolysaccharidosis IV; Mucopolysaccharidosis VI

Mucopolysaccharidoses (MPSs) are complex lysosomal storage disorders that result in the accumulation of glycosaminoglycans (GAGs) in urine, blood, and tissues. Lysosomal enzymes responsible for GAG degradation are defective in MPSs. GAGs including chondroitin sulfate (CS), dermatan sulfate (DS), heparan sulfate (HS), and keratan sulfate (KS) are disease-specific biomarkers for MPSs. This article describes a stable isotope dilution-tandem mass spectrometric method for quantifying CS, DS, and HS in urine samples. The GAGs are methanolyzed to uronic or iduronic acid-N-acetylhexosamine or iduronic acid-N-sulfo-glucosamine dimers and mixed with internal standards derived from deuteriomethanolysis of GAG standards. Specific dimers derived from HS, DS, and CS are separated by ultra-performance liquid chromatography (UPLC) and analyzed by electrospray ionization tandem mass spectrometry (MS/MS) using selected reaction monitoring for each targeted GAG product and its corresponding internal standard. This UPLC-MS/MS GAG assay is useful for identifying patients with MPS types I, II, III, VI, and VII. © 2023 Wiley Periodicals LLC. Basic Protocol: Urinary GAG analysis by ESI-MS/MS Support Protocol 1: Prepare calibration samples Support Protocol 2: Preparation of stable isotope-labeled internal standards Support Protocol 3: Preparation of quality controls for GAG analysis in urine Support Protocol 4: Optimization of the methanolysis time Support Protocol 5: Measurement of the concentration of methanolic HCl.

Topics: Chondroitin Sulfates; Chromatography, Liquid; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Humans; Iduronic Acid; Isotopes; Mucopolysaccharidoses; Mucopolysaccharidosis I; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

Chondroitin 6 sulphate (C6S) is a glycosaminoglycan (GAG) whose accumulation is notable in mucopolysaccharidosis type IVA and VII. Flaxseed,. We assess the effect of total ethanol flaxseed extract (EFSE) in an animal model of C6S accumulation.. The study was performed in adult male Wistar rats (. EFSE showed significant potential for chelation. Its use for the treatment of GAG accumulation could be suggested and generalized to a larger study population.

Topics: Animals; Antioxidants; Chondroitin Sulfates; Flax; Glycosaminoglycans; Humans; Male; Mucopolysaccharidoses; Plant Extracts; Rats; Rats, Wistar

Eleven patients from Yakutia with a new lysosomal disease assumed then as mucopolysaccharidosis-plus syndrome (MPS-PS) were reported by Gurinova et al. in 2014. Up to now, a total number of 39 patients have been reported; in all of them, the c.1492C>T (p.Arg498Trp) variant of the VPS33A gene was detected. Here, we describe the first Polish MPS-PS patient with a novel homozygous c.599G>C (p.Arg200Pro) VPS33A variant presenting over 12 years of follow-up with some novel clinical features, including fetal ascites (resolved spontaneously), recurrent joint effusion and peripheral edemas, normal growth, and visceral obesity. Functional analyses revealed a slight presence of chondroitin sulphate (only) in urine glycosaminoglycan electrophoresis, presence of sialooligosaccharides in urine by thin-layer chromatography, and normal results of lysosomal enzymes activity and lysosphingolipids concentration in dried blood spot. The comparison with other MPS-PS described cases was also provided. The presented description of the natural history of MPS-PS in our patient may broaden the spectrum of phenotypes in this disease.

Topics: Chondroitin Sulfates; Glycosaminoglycans; Humans; Mucopolysaccharidoses; Mutation; Poland; Sphingolipids; Vesicular Transport Proteins

Mucopolysaccharidoses comprise a group of rare metabolic diseases, in which the lysosomal degradation of glycosaminoglycans (GAGs) is impaired due to genetically inherited defects of lysosomal enzymes involved in GAG catabolism. The resulting intralysosomal accumulation of GAG-derived metabolites consequently manifests in neurological symptoms and also peripheral abnormalities in various tissues like liver, kidney, spleen and bone. As each GAG consists of differently sulfated disaccharide units, it needs a specific, but also partly overlapping set of lysosomal enzymes to accomplish their complete degradation. Recently, we identified and characterized the lysosomal enzyme arylsulfatase K (Arsk) exhibiting glucuronate-2-sulfatase activity as needed for the degradation of heparan sulfate (HS), chondroitin sulfate (CS) and dermatan sulfate (DS). In the present study, we investigated the physiological relevance of Arsk by means of a constitutive Arsk knockout mouse model. A complete lack of glucuronate desulfation was demonstrated by a specific enzyme activity assay. Arsk-deficient mice show, in an organ-specific manner, a moderate accumulation of HS and CS metabolites characterized by 2-O-sulfated glucuronate moieties at their non-reducing ends. Pathophysiological studies reflect a rather mild phenotype including behavioral changes. Interestingly, no prominent lysosomal storage pathology like bone abnormalities were detected. Our results from the Arsk mouse model suggest a new although mild form of mucopolysacharidose (MPS), which we designate MPS type IIB.

Topics: Animals; Arylsulfatases; Chondroitin Sulfates; Enzyme Activation; Heparitin Sulfate; Mice; Mice, Knockout; Mucopolysaccharidoses

The Mucopolysaccharidoses (MPS) are group of inherited metabolic diseases caused by the deficiency of enzymes required to degrade glycosaminoglycans (GAGs) in the lysosomes. GAGs are sulfated polysaccharides involving repeating disaccharides, uronic acid and hexosamines including chondroitin sulfate (CS), dermatan sulfate (DS), heparan sulfate (HS) and keratan sulfate (KS). Hyaluronan is excluded in terms of being non-sulfated in the GAG family. Different types of mutations have been identified as the causative agent in all types of MPS. Herein, we planned to investigate the pathogenic mutations in different types of MPS including type I (IDUA gene), IIIA (SGSH) and IIIB (NAGLU) in the eight Iranian patients. Autozygosity mapping was performed to identify the potential pathogenic variants in these 8 patients indirectly with the clinical diagnosis of MPSs. so three panels of STR (Short Tandem Repeat) markres flanking IDUA, SGSH and NAGLU genes were selected for multiplex PCR amplification. Then in each family candidate gene was sequenced to identify the pathogenic mutation. Our study showed two novel mutations c.469 T > C and c.903C > G in the IDUA gene, four recurrent mutations: c.1A > C in IDUA, c.220C > T, c.1298G > A in SGSH gene and c.457G > A in the NAGLU gene. The c.1A > C in IDUA was the most common mutation in our study. In silico analysis were performed as well to predict the pathogenicity of the novel variants.

Topics: Adolescent; Child; Child, Preschool; Chondroitin Sulfates; Dermatan Sulfate; DNA Mutational Analysis; Female; Genetic Testing; Heparitin Sulfate; Humans; Infant; Keratan Sulfate; Male; Mucopolysaccharidoses; Multiplex Polymerase Chain Reaction; Mutation

Mucopolysaccharidoses (MPS) are complex storage disorders that result in the accumulation of glycosaminoglycans (GAGs) in urine, blood, brain and other tissues. Symptomatic patients are typically screened for MPS by analysis of GAG in urine. Current screening methods used in clinical laboratories are based on colorimetric assays that lack the sensitivity and specificity to reliably detect mild GAG elevations that occur in some patients with MPS. We have developed a straightforward, reliable method to quantify chondroitin sulfate (CS), dermatan sulfate (DS) and heparan sulfate (HS) in urine by stable isotope dilution tandem mass spectrometry. The GAGs were methanolyzed to uronic acid-N-acetylhexosamine or iduronic acid-N-glucosamine dimers and mixed with stable isotope labeled internal standards derived from deuteriomethanolysis of GAG standards. Specific dimers derived from HS, DS and CS were separated by ultra-performance liquid chromatography and analyzed by electrospray ionization tandem mass spectrometry using selected reaction monitoring for each targeted GAG product and its corresponding internal standard. The method was robust with a mean inaccuracy from 1 to 15%, imprecision below 11%, and a lower limit of quantification of 0.4mg/L for CS, DS and HS. We demonstrate that the method has the required sensitivity and specificity to discriminate patients with MPS III, MPS IVA and MPS VI from those with MPS I or MPS II and can detect mildly elevated GAG species relative to age-specific reference intervals. This assay may also be used for the monitoring of patients following therapeutic intervention. Patients with MPS IVB are, however, not detectable by this method.

Topics: Adolescent; Adult; Aged; Child; Child, Preschool; Chondroitin Sulfates; Chromatography, Liquid; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Humans; Infant; Middle Aged; Mucopolysaccharidoses; Mucopolysaccharidosis II; Mucopolysaccharidosis III; Mucopolysaccharidosis IV; Mucopolysaccharidosis VI; Radioisotope Dilution Technique; Reference Values; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Young Adult

The evaluation of plasmatic galactosaminoglycans, dermatan sulfate (DS) and chondroitin sulfate (CS) can be helpful in the early identification of MPS patients, also considering that primary storage of one type of GAG can lead to secondary accumulation of other lysosomal substrates. We explore the possibility to determine plasmatic DS and CS in numerous healthy pediatric (and sometimes adult) subjects depending on age and in patients affected by various forms of MPS. A highly sensitive HPLC separation and fluorescence detection was applied for plasma/serum DS and CS determination after a specific enzymatic treatment able to release their constituent disaccharides. DS and CS content decrease significantly with age in controls having high values in the first year (~8 μg/mL). A highly significant decrease was observed for 1-5-year-old (∼-33%) and 5-10-year-old (∼-65%) healthy subgroups. No further decrease was determined showing a stabilization after 5 years of age. MPS I Scheie and Hurler patients showed rather similar DS and CS content significantly higher than controls matched for age. Similarly, MPS II, III and IV subjects all presented significantly higher plasmatic DS and CS content compared to healthy subjects matched for age. The same trend was determined for the only patient affected by MPS VI. Plasmatic DS and CS analyzed by the present procedure may be a useful diagnostic and screening marker for various forms of MPS.

Topics: Adolescent; Adult; Aged; Child; Child, Preschool; Chondroitin Sulfates; Dermatan Sulfate; Female; Humans; Infant; Infant, Newborn; Male; Middle Aged; Mucopolysaccharidoses; Polysaccharides

Mucopolysaccharidoses (MPSs) are complex lysosomal storage disorders that result in the accumulation of glycosaminoglycans (GAGs) in urine, blood, and tissues. Lysosomal enzymes responsible for GAG degradation are defective in MPSs. GAGs including chondroitin sulfate (CS), dermatan sulfate (DS), heparan sulfate (HS), and keratan sulfate (KS) are disease-specific biomarkers for MPSs. This unit describes a stable isotope dilution-tandem mass spectrometric method for quantifying CS, DS, and HS in urine samples. The GAGs are methanolyzed to uronic or iduronic acid-N-acetylhexosamine or iduronic acid-N-sulfo-glucosamine dimers and mixed with internal standards derived from deuteriomethanolysis of GAG standards. Specific dimers derived from HS, DS, and CS are separated by ultra-performance liquid chromatography (UPLC) and analyzed by electrospray ionization tandem mass spectrometry (MS/MS) using selected reaction monitoring for each targeted GAG product and its corresponding internal standard. This new GAG assay is useful for identifying patients with MPS types I, II, III, VI, and VII.

Topics: Biomarkers; Chondroitin Sulfates; Chromatography, Liquid; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Humans; Keratan Sulfate; Mucopolysaccharidoses; Quality Control; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

MPS disorders result from a deficiency or absence of glycosaminoglycan (GAG) degrading enzymes leading to an imbalance between the synthesis and degradation of GAGs and their subsequent accumulation in a range of cells. The inhibition of GAG synthesis using small chemical inhibitors has been proposed as a novel therapeutic approach to treatment. Several inhibitors have been shown to decrease heparan sulphate GAG synthesis and in this study we evaluated a novel fluorinated analog of N-acetylglucosamine (2-acetamido-1,3,6-tri-O-acetyl-4-deoxy-4-fluoro-D-glucopyranose (F-GlcNAc)) and rhodamine B for their ability to also inhibit the synthesis of chondroitin/dermatan and keratan sulphate GAGs present in bovine cartilage. Both inhibitors decreased GAG synthesis in chondrocyte monolayer culture and in cartilage chip explant culture in a dose dependent manner. Both inhibitors decreased the size of newly synthesised proteoglycans and in the case of F-GlcNAc this was due to a decrease in newly synthesised GAG chain size. Rhodamine B, however, did not affect GAG chain size, while both inhibitors decreased the amount of chondroitin/dermatan and keratan sulphate GAG equally. The expression of genes responsible for the initiation and elongation of chondroitin/dermatan sulphate and keratan sulphate GAGs were downregulated in the presence of rhodamine B but not in the presence of F-GlcNAc. Thus the 2 inhibitors appear to have differing effects on GAG synthesis, with F-GlcNAc inhibiting the epimerisation of UDP-GlcNAc to UDP-GalNAc thus decreasing the availability of monosaccharides for addition to the growing GAG chain, whereas rhodamine B is more likely to reduce the number of GAG chains. Together with previous data these 2 inhibitors are capable of non-specific inhibition of GAG synthesis, reducing the production of chondroitin/dermatan sulphate, keratan sulphate and heparan sulphate GAGs. As such they would be applicable to therapy in a range of MPS disorders.

Topics: Acetylglucosamine; Animals; Cartilage; Cattle; Chondrocytes; Chondroitin Sulfates; Dermatan Sulfate; Gene Expression Profiling; Gene Expression Regulation; Glycosaminoglycans; Keratan Sulfate; Mucopolysaccharidoses; Proteoglycans; Rhodamines

The fact that mucopolysaccharidoses (MPSes) are now treatable, and that the earlier treatment is initiated the better, is an indication for neonatal screening. The most efficient approach seems likely to be a multi-tier procedure in which screening for urinary glycosaminoglycan (GAG) is followed by enzyme determinations in heelprick blood of newborns screening positive. Hitherto the method of choice for the determination of GAG has been the measurement of absorbance by a complex of GAG and 1,9-dimethylmethylene blue (DMB).. We evaluated a DMB method in which absorbance by DMB is measured following its addition to the eluate obtained from paper-borne newborn urine samples and is normalized relative to urinary creatinine. Calibration is performed with chondroitin-6-sulfate (Ch-6-S).. The limits of detection and quantification of GAG were 1.98 and 5.94 mg/dl, respectively. The within-run coefficients of variation (CVs) of the GAG/creatinine ratio for 25, 31, and 70 mg/dl solutions of Ch-6-S in urine were 21.8, 16.4, and 10.5%, respectively, and the corresponding between-run CVs were 25.0, 13.5, and 10.1%. Recovery from the urine spiked with 31 mg Ch-6-S/dl was 94.8%. Accuracy was also acceptable for all other GAGs except hyaluronic acid. For neonatal screening, the diagnostic threshold was tentatively established as 800 mg GAG/g creatinine, the 95th centile of samples from 903 infants aged 3-28 days, but the value of the GAG/creatinine ratio was negatively correlated with age. Application of the new method to samples from older individuals with and without MPS achieved 100% sensitivity and specificity when used with an age-dependent threshold taken from the literature on the original DMB method.. If used in the first tier of a multi-tier screening protocol, the proposed method would allow the detection of abnormal levels of all GAGs except hyaluronic acid.

Topics: Aging; Calibration; Chondroitin Sulfates; Creatinine; Dermatan Sulfate; Glycosaminoglycans; Heparin; Heparinoids; Humans; Hyaluronic Acid; Infant, Newborn; Methylene Blue; Mucopolysaccharidoses; Neonatal Screening; Paper; Reproducibility of Results; Sensitivity and Specificity

A direct dye-binding technique has been developed to measure total glycosaminoglycans in urine. Fifty or 100 microliters of urine was mixed with a solution of Azure A (10 mg/l) and Azure B (10 mg/l) and the resulting decrease in absorbance at 610 nm was measured. The standard curve with chondroitin sulfate C was linear from 1 to 20 micrograms/assay. The normal value for total urinary glycosaminoglycans in adults was 1.8 +/- 0.6 g/mol creatinine. Other normal values were age-dependent. The assay is inexpensive, simple, precise, sensitive and suitable for screening for the mucopolysaccharidoses in pediatric patients.

Topics: Adolescent; Adult; Azure Stains; Child; Child, Preschool; Chondroitin Sulfates; Glycosaminoglycans; Humans; Indicators and Reagents; Infant; Infant, Newborn; Mucopolysaccharidoses; Spectrophotometry, Ultraviolet

A 68-year-old man and a 66-year-old woman had diffuse corneal stromal deposits that stained with alcian blue and colloidal iron but did not react with periodic acid-Schiff stain and lipid stains. Similar deposits were found within postmortem sclera in one case, but not in other ocular or extraocular tissues. The abnormal material was sensitive to testicular hyaluronidase and chondroitinase. The material reacted with monoclonal antibody 9-A-2 after digestion by chondroitinase AC in one case and ABC in both cases, which is consistent with the identification of the glycosaminoglycans chondroitin 4-sulfate and dermatan sulfate. Electron microscopic examination of the cornea in both cases disclosed granular material in vacuoles dispersed extracellularly and, rarely, in keratocytes. Results of blood and skin fibroblast enzyme assays for clinically relevant mucopolysaccharidoses and mucolipidoses were normal in both patients, and there were no somatic abnormalities suggesting a storage disease.

Topics: Aged; Chondroitin Sulfates; Corneal Diseases; Corneal Stroma; Dermatan Sulfate; Female; Histocytochemistry; Humans; Male; Mucopolysaccharidoses; Pedigree; Scleral Diseases

The clinical, radiological and biochemical findings in a black girl with a rare, inherited mucopolysaccharide storage disease, Sanfilippo's syndrome (mucopolysaccharidosis (MPS) III) type C, are described. Practical points concerning the biochemical diagnosis of this condition, arising from unusual characteristics of the deficient enzyme acetyl CoA: alpha-glucosaminide N-acetyltransferase, are discussed. Because phenotypic manifestations of mucopolysaccharidosis are mild in all four types of Sanfilippo's syndrome and screening tests for mucopolysacchariduria in these patients may be negative, many cases may be passed unrecognized or simply labelled as cases of nonspecific mental retardation. It is suggested that Sanfilippo's syndrome is grossly underdiagnosed in the RSA and clinicians are urged to develop a greater awareness of the existence, and often subtle presentation, of the condition.

Topics: Acetyltransferases; Black or African American; Black People; Child, Preschool; Chondroitin Sulfates; Female; Heparitin Sulfate; Humans; Mucopolysaccharidoses; Mucopolysaccharidosis III; Radiography; South Africa

Topics: Child; Chondroitin Sulfates; Diabetes Mellitus; Glomerulonephritis; Glycosaminoglycans; Heparitin Sulfate; Humans; Mucopolysaccharidoses; Nephritis, Hereditary; Nephrotic Syndrome; Proteinuria

Topics: Animals; Cartilage; Cell Differentiation; Chondroitin Sulfates; Exostoses, Multiple Hereditary; Female; Genes, Recessive; Hydrocephalus; Kidney; Mice; Microscopy, Electron; Mucopolysaccharidoses; Pregnancy; Sternum

A sensitive method was developed for the separation and quantitation of four unsaturated disaccharides (delta Di-0S, delta Di-4S, delta Di-6S, and delta Di-diS) by high performance liquid chromatography. The unsaturated disaccharides were coupled with a fluorescent compound, 2-aminopyridine. Complete separation of the resulting pyridylamino derivatives was achieved on a column of muBondapak-C18 with 8 mM KH2PO4-Na2HPO4 (pH 6.0)/methanol (30/l, by volume) as a mobile phase. There was a linear relationship between the fluorescence emission (peak height), and the amount of each authentic disaccharide used for the coupling reaction. This method was applied to analyze commercially available chondroitin sulfates A and C, dermatan sulfate, and urinary glycosaminoglycans obtained from patients with mucopolysaccharidosis after digestion with chondroitinases. The data indicated that the present method is useful for the separation and quantitation of nmol-pmol levels of the unsaturated disaccharides produced from chondroitin sulfate isomers by chondroitinases and can be used for their structural characterization.

Topics: Aminopyridines; Animals; Cartilage; Chondroitin; Chondroitin Sulfates; Chondroitinases and Chondroitin Lyases; Chondroitinsulfatases; Chromatography, High Pressure Liquid; Disaccharides; Fluorescent Dyes; Glycosaminoglycans; Humans; Isomerism; Mucopolysaccharidoses; Sharks; Skin; Swine; Whales

Topics: Chondroitin Sulfates; Dermatan Sulfate; Electrophoresis, Cellulose Acetate; Glycosaminoglycans; Heparin; Humans; Keratan Sulfate; Mucopolysaccharidoses; Temperature

Topics: Chondroitin; Chondroitin Sulfates; Chondroitinases and Chondroitin Lyases; Chromatography, High Pressure Liquid; Dermatan Sulfate; Disaccharides; Glycosaminoglycans; Humans; Mucopolysaccharidoses

Topics: Child, Preschool; Chlorides; Chondroitin Sulfates; Dermatan Sulfate; Edetic Acid; Electrophoresis; Electrophoresis, Cellulose Acetate; Glycosaminoglycans; Heparin; Heparitin Sulfate; Humans; Hyaluronic Acid; Keratan Sulfate; Lithium; Lithium Chloride; Liver; Male; Mucopolysaccharidoses

The high-performance liquid chromatographic (HPLC) method for the determination of unsaturated sulfated disaccharides is a comprehensive and reliable method which expedites ensymatic studies of isomeric chondroitin sulfates. Responses for these unsaturated disaccharides derived from urinary chondroitin sulfates were linear from 100 ng to 10 micrograms injected and good quantitation was obtained for 25 microliters or less of samples placed on the column. This method which is more sensitive and accurate than methods now being used has considerable potential for the chemical diagnosis of patients with mucopolysaccharidoses and for the clarification of glycosaminoglycan structure. The isomeric chondroitin sulfates in urines from patients with mucopolysaccharidoses were studied by enzyme digestion with chondroitinases followed by HPLC determination of the sulfated unsaturated disaccharides produced. Evaluation by HPLC of the unsaturated 4-sulfated disaccharide produced by digestion of the urinary GAG with chondroitinases ABC and AC revealed rapidly and quantitatively the large amounts of dermatan sulfate present in Hurler, Hunter, and Maroteaux-Lamy urines. Chondroitin 4-sulfate predominated in Sanfilippo urinary isomeric chondroitin sulfates whereas chondroitin 6-sulfate and chondroitin 4-sulfate were shown to be present in nearly equal amounts in Morquio urine. An oversulfated chondroitin sulfate was detected in small amounts in some of these urines. This was demonstrated by the detection of an unsaturated disulfated disaccharide after digestion with chondroitinase ABC but not with chondroitinase AC.

Topics: Chondroitin; Chondroitin Sulfates; Chondroitinases and Chondroitin Lyases; Chromatography, High Pressure Liquid; Glycosaminoglycans; Humans; Isomerism; Mucopolysaccharidoses

A 14-year-old boy was found to excrete excessive amounts of acidic glycosaminoglycans which were predominantly chondroitin 4-sulfate and chondroitin 6-sulfate. Clinical features included dwarfism, mental retardation, coarse facies, deformities of the spine, hip joints and thorax, and granulations in leucocytes. The clinical and biochemical features found in this boy were compared with the known types of mucopolysaccharidosis and it has been concluded that this case is a new type of mucopolysacchariduria.

Topics: Adolescent; Chondroitin; Chondroitin Sulfates; Dwarfism; Glycosaminoglycans; Humans; Intellectual Disability; Male; Mucopolysaccharidoses

The patient is a north african female, fourth born child in a family with consanguinity. Facial dysmorphia, clubfeet, swollen extremities and heel borne ponctuate calcifications are observed soon after birth. beta glucuronidase activity is very low in serum, leukocytes and skin fibroblasts. At 18 months, gorwth and psychomotor development are normal. Flat facies and dorsolumbar cyphosis are striking. There is no clinical sign of storage disease, neither ocular or cytologic (blood, bone-marrow) abnormalities. Squeletal abnormalities are predominant on cervical and lumbar column and pelvis. Urinary excretion of chondroitin 4 sulfate and chondroitin 6 sulfate is increased. A 4 year old sister is affected. Facial dysmorphia, mild squeletal abnormalities are observed. Again, growth is normal and there is no symptom of storage disease. Enzymic expression of parent heterozygotism is marked in serum studies, but less marked in leukocytes and fibroblasts. The last two children are heterozygotes. At the time of a 5th pregnancy, enzymic activity studies of amniotic fluid and amniotic cells have shown that the foetus was an inaffected female. This child was normal at birth.

Topics: Abnormalities, Multiple; Child; Chondroitin Sulfates; Consanguinity; Female; Glucuronidase; Humans; Infant, Newborn; Mucopolysaccharidoses

[1-3H]Galactitol-6-sulfate, N- [1-3H]acetylgalactosaminitol-6-sulfate, N-[1-3H]acetylglucosaminitol-6-sulfate, N-acetylglucosamine-6-sulfate, and 6-sulfated tetrasaccharides from chondroitin-6-sulfate have been used for the measurement of 6-sulfatase activity of extracts of normal skin fibroblasts and of fibroblasts cultured from patients with genetic mucopolysaccharidoses. With these substrates, extracts of fibroblasts derived from Morquio patients lack or have greatly reduced activities for galactitol-6-sulfate, N-acetylgalactosaminitol-6-sulfate, and 6-sulfated tetrasaccharides but have normal activity for N-acetylglucosamine-6-sulfate and its alditol; those derived from a patient with a newly discovered mucopolysaccharidosis have greatly reduced activity for N-acetylglucosamine-6-sulfate and its alditol but normal activity for galactitol-6-sulfate, N-acetylgalactosaminitol-6-sulfate, and the 6-sulfated tetrasaccharides. These findings demonstrate the existence of two different hexosamine-6-sulfate sulfatases, specific for the glucose or galactose configuration of their substrates. Their respective deficiencies, causing inability to degrade keratan sulfate and heparan sulfate in one case and keratan sulfate and chondroitin-6-sulfate in the other, are responsible for different clinical phenotypes.

Topics: Acetylgalactosamine; Acetylglucosamine; Cells, Cultured; Child, Preschool; Chondroitin Sulfates; Chondroitinsulfatases; Fibroblasts; Galactitol; Heparitin Sulfate; Humans; Hydrogen-Ion Concentration; Keratan Sulfate; Male; Mucopolysaccharidoses; Mucopolysaccharidosis III; Mucopolysaccharidosis IV; Skin; Substrate Specificity; Sulfatases

A 17-year-old patient clinically manifesting gargoyle face, dwarfism, skeletal bone deformity, mild mental retardation and benign course was presented. Biochemically, increased urinary excretion of acidic glycosaminoglycans was confirmed and chondroitin 4-sulfate and chondroitin 6-sulfate were substantiated to be the major components of the acid mucopolysacchariduria. Light microscopically, variable numbers of foam cells were observed in the biopsy materials of the lymph nodes, liver and skin, as well as in the smears of bone marrow aspirates. In the liver, the parenchymal cells appeared vacuolated. Histochemically, accumulation of sulfated acid glycosaminoglycans was demonstrated in the cytoplasm of the foam cells proliferating in these tissues, as well as in the liver cells. Electron microscopically, all of these storage cells were found to contain numerous, membrane-bound, vacuolar inclusions filled with flocculent, finely reticulogranular materials of low electron density similar to those seen in the storage cells of Hurler, Hunter or Sanfilippo's syndrome. Empty vacuoles were often coexistent. Accordingly, this case should be termed "chondroitin 4- and 6-sulfate mucopolysaccharidosis", with emphasis on the possibility of a new type of genetic mucopolysaccharidosis.

Topics: Adolescent; Chondroitin; Chondroitin Sulfates; Humans; Liver; Lymph Nodes; Male; Mucopolysaccharidoses; Skin

The approach, identification of clinical phenotype followed by lysosomal enzyme assays in cell culture, used in the classification of the genetic mucopolysaccharidoses I-VI has been applied to the chondroitinsulphaturias. There was evidence of heterogeneity in the first 9 patients reported.

Topics: Cells, Cultured; Chondroitin; Chondroitin Sulfates; Genotype; Glucuronidase; Humans; Infant; Lysosomes; Male; Mucopolysaccharidoses; Phenotype

The basic defects of a number of genetic disorders affecting the connective tissue have been elucidiated during the last few years. A similar progress in understanding the pathobiochemistry of other dermatological diseases is expected. The present review summarizes our knowledge about structure, synthesis, degradation, macromolecular organization and possible function of glycosaminoglycans, which are an important constitutent of the ground substance in the skin. The difficulties of relating the clinical symptoms of diseases with the basic defects are examplified for the mucopolysaccharidoses.

Topics: Adolescent; Adult; Age Factors; Aged; Chemical Phenomena; Chemistry; Chondroitin Sulfates; Dermatan Sulfate; Enzyme Therapy; Female; Fibroblasts; Glycosaminoglycans; Heparitin Sulfate; Histocytochemistry; Humans; Hyaluronic Acid; Infant, Newborn; Middle Aged; Mucopolysaccharidoses; Pregnancy; Proteoglycans; Skin

The mucopolysaccharidoses are a group of genetic diseases characterized by storage of incompletely degraded glycosaminoglycans. Such storage causes marked distortion of many tissues with consequent severe somatic changes and mental retardation. Storage of glycosaminoglycans results from markedly diminished activity of specific hydrolases requisite for the normal degradation of glycosaminoglycans. The specific enzymic defects have been identified in nine different diseases. In some cases evidence has been obtained indicating the existence of additional allelic diseases based on the same enzyme. The knowledge obtained from these studies has made prenatal diagnosis possible and has led to the possibility that therapy may be undertaken utilizing enzyme replacement.

Topics: Acetylglucosaminidase; Alleles; Arylsulfatases; Chondroitin Sulfates; Dermatan Sulfate; Glucuronidase; Glycosaminoglycans; Heparitin Sulfate; History, 20th Century; Humans; Iduronidase; Mucopolysaccharidoses; Mucopolysaccharidosis I; Sulfatases

Topics: Adolescent; Child; Child, Preschool; Chondroitin Sulfates; Chondroitinases and Chondroitin Lyases; Chromatography, Paper; Dermatan Sulfate; Female; Glycosaminoglycans; Heparitin Sulfate; Humans; Infant; Male; Mucopolysaccharidoses