Excretion Of Acid Mucopolysaccharides Research Articles

Increased urinary excretion of acid mucopolysaccharides and glycopeptides in hypothyroidism following thyroid hormone therapy

Urinary excretion of acid mucopolysaccharides and glycopeptides in the urine of an untreated patient with cretinism were measured before and after thyroid hormone replacement. Urinary uronic acid and hexose excretion in the CPC-precipitated fraction increased four to ten times after thyroid hormone administration. The maximum excretion was observed after 2 months of thyroid replacement. The excreted acid mucopolysaccharides consisted of chondroitin sulphate A/C and minor quantities of heparan sulphate. Urinary excretion of glycopeptides, particularly small molecular glycopeptides rose also four to five-fold with thyroid hormone administration. These data suggest that thyroid hormone markedly influences the metabolism of acid mucopolysaccharides and glycoproteins. Possibly, the lack of thyroid hormone caused a decreased activity of various lysosomal glycosidases and sulphatases.

1232 BETA-GLUCURONIDASE DEFICIENCY: A MUCOPOLYSACCHARIDOSIS WITH CHONDROITIN SULFATURIA

Beta-glucuronidase deficiency is an autosomal recessive mucopolysaccharidosis with variable clinical manifestations similar to the other mucopolysaccharidoses. The urinary excretion of acid mucopolysaccharides (AMPS) has not been very clear, with reports suggesting the excretion of dermatan, heparan and chondroitin sulfates. These suggestions were based primarily on electrophoretic mobility. This report deals with the isolation and characterization of the urinary AMPS from three unrelated patients with β-glucuronidase deficiency. Urinary AMPS were isolated using gel chromatography with Sephadex G-25, followed by precipitation of the AMPS fraction with cetylpiridinium chloride. Dermatan sulfate was identified by the formation of a complex with CuSO4, the elution pattern from Dowex 1, X2Cl− and susceptibility to chondroitinases. Heparan sulfate was estimated by the determination of the N-sulfated hexosamine, a reaction specific for this compound. Chondroitin 4 and 6 sulfates were determined by the pattern of elution from Dowex 1 and digestibility with chondroitinase AC. Total AMPS isolated ranged from 18.4mg to 71.0mg/24h. The major fraction (>90%) was chondroitin 4/6 sulfate. Dermatan sulfate comprised <8% and heparan sulfate <2%. Gas chromatography/mass spectrometry of the uronic acids hydrolyzed from these fractions confirmed these findings. These studies indicate that chondroitin sulfaturia is a characteristic urinary finding of β-glucuronidase deficiency.

Abnormal urinary excretion of glycosaminoglycans in Albers-Schönberg disease

Abnormal urinary excretion of acid mucopolysaccharides in two children affected with Albers-Schönberg disease is described. The urine specimens were preliminarily checked for their acid glycosaminoglycan (AGAG) content by a screening test. The AGAGs were then evaluated by uronic acid determination using the carbazole method, and the ratio between high molecular mass and low molecular mass AGAGs was determined. Qualitative characterization before and after enzymatic digestion with chondroitinases AC (ChAC) and ABC (ChABC) of the total AGAGs contained in the specimen and of each AGAG separated with Dowex 1×2 was also performed by electrophoresis. The pattern of the urinary mucopolysaccharides in the two patients showed an abnormal quantity of heparan sulphate (HS) and a smaller quantity of dermatan sulphate (DS). Such abnormal urinary AGAGs excretion should confirm the assumption of some authors that malignant infantile osteopetrosis may be caused by a deficiency in the lysosomal enzymatic activity, or by some anomaly in the cell membrane function.

Neuraminidase studies in sialidosis.

In 1968, two patients were described in the literature who exhibited clinical features resembling Hurler disease, yet had a normal urinary excretion of acid mucopolysaccharides. Histological and ultrastructural studies of neuronal, visceral and mesenchymal tissues revealed the lysosomal storage of compounds thought to be acid mucopolysaccharides and glycolipids (BERARD et al., 1968; SPRANGER et al., 1968; FREITAG et al., 1971) Together with morphologically similar disorders, these patients were later classified as mucolipidosis I (SPRANGER and WIEDEMANN, 1970). More recent biochemical investigations showed an accumulation of sialic acid-containing compounds in cultured fibroblasts and in leukocytes of such patients (CANTZ et al., 1977; SPRANGER et al., 1977; KELLY and GRAETZ, 1977), and a massive urinary excretion of sialyl oligosaccharides, whose structures resembled the glycan portion of glycoproteins MICHALSKI et al., 1977). In fibroblasts, there was a profound deficiency of an “acid” (presumably lysosomal) neuraminidase towards substrates such as sialyllactose, fetuin, and methoxyphenyl neuraminic acid, whereas other lysosomal hydrolases were within the normal range (CANTZ et al., 1977; SPRANGER et al., 1977; KELLY and GRAETZ, 1977). From these studies it became evident that the metabolic defect in mucolipidosis I consists of an impaired catabolism of glycoproteinderived sialyl oligosaccharides due to the genetic deficiency of a neuraminidase, i.e., that the disease is a sialidosis.

Treatment of Mucopolysaccharidosis

The therapeutic effectiveness of leucocyte transfusion (LT) was compared with that of plasma infusion (PI) clinically by range of motion (ROM) of joints and biochemically from the standpoint of alpha-L-iduronidase activity and urinary excretion of acid mucopolysaccharides (AMPS) in 2 patients with Hurler's and Scheie's syndromes. Both syndromes are considered to be due to the lack of alpha-L-iduronidase activity, a congenital metabolic disorder. As a result, leukocyte transfusion surpasses plasma infusion with respect to enzyme content, the grade and duration of clinical improvement in the stiffness of joints. Clinical improvement in the stiffness of joints was correlated with the degradation of AMPS when the ratio of urinary AMPS fragments to the total large molecule AMPS has become 50% or more after the leucocyte transfusion and plasma infusion.

Familial Juvenile Neuronal Storage Disease

Two patients had an illness characterized by a positive family history, juvenile onset, macular cherry-red spots, myoclonus, generalized convulsions, and cerebellar ataxia. Neither had dementia, gargoyle facies, bone or joint deformities, or visceromegaly. Vacuolated lymphocytes were not seen in the peripheral blood or bone marrow. Specimens from the rectum and vermiform appendix showed Sudan black B-, Sudan III-, and PAS-positive granules within the neurons of the myenteric plexus. On electron microscopic examination, lysosome-like bodies, membranous cytoplasmic bodies, pleomorphic lamellated bodies, dense bodies, and lipofuscin-like bodies in the neurons were seen, with a suggestion of morphological transitional forms among them. Sialoglycopeptides, especially sialic acid, were increased in the urine, but excretion of acid mucopolysaccharides was normal. Assays of lysosomal enzymes in leucocytes showed normal enzymatic activity. On the basis of the clinical, biochemical, and histological results, we suggest that these two cases and four similar cases reported in the literature be classified differently from the previously described lipidoses, although it is not known whether these cases represent a new entity or merely a clinical variant of juvenile lipidosis.

Urinary excretion of acid mucopolysaccharides in schizophrenia

1. (1) The 24-hr urinary excretion of acid mucopolysaccharides measured as glucuronic acid was found to be significantly lower in male schizophrenics as compared to normal male subjects of a corresponding age group. 2. (2) No hyaluronic acid was detected in normal urine. About one-third of total AMPS was present as hyaluronic acid in schizophrenic urine. 3. (3) Hyaluronic acid in schizophrenic urine was identified by electrophoresis, ion-exchange chromatographic behavior, infrared spectroscopy and by the presence of glucosamine in its hydrolysate.

Beta-D-glucuronidase deficiency mucopolysaccharidosis

A two year old girl presented with mild facial dysmorphism reminiscent of Hurler disease, moderate mental retardation, and hepatosplenomegaly. Hercorneae were clear. Radiologically, there was moderate dysostosis multiplex. Peripheral granulocytes contained abundant, coarse Alder-Reilly granulations. There was an increased urinary excretion of acid mucopolysaccharides. Determination of lysosomal glycosidases showed a markedly decreased activity of β-D-glucuronidase in cultured fibroblasts, leudocytes, and serum. In the parents, in one grandmother and in one sister, β-D-glucuronidase activity in serum and leucocytes was decreased to between 30 and 50%. The disorder has been classified as mucopolysaccharidosis,type VII. Our results indicate that a reliable diagnosis of the homozygous and heterozygous deficient state can be made by determining β-D-glucuronidase in serum.

Mucopolysaccharidosis. VII. Beta-glucuronidase deficiency.

An 18-month-old female showed a markedly decreased activity of the lysosomal enzyme β-D-glucuronidase in cultured fibroblasts, leucocytes, and serum. In obligate heterozygotes, the serum and leucocyte β-D-glucuronidase activity was decreased to approximately 50% of the control values. Clinically, the patient had a mild facial dysmorphism reminiscent of Hurler disease, moderate mental retardation, and hepatosplenomegaly. The corneae were clear. Peripheral granulocytes contained abundant, coarse Alder-Reilly granulations. The urinary excretion of acid mucopolysaccharides was increased. The disorder has been classified as mucopolysaccharidosis, type VII. It is assumed that this new mucopolysaccharidosis is caused by the defective action of β-glucuronidase which results in the faulty degradation and intralysosomal storage of acid mucopolysaccharides.

Influence of fresh plasma infusions on patients with genetic mucopolysaccharidosis.

The pattern of urinary acid mucopoly-saccharide excretion before, during and after fresh plasma infusions to five patients with mucopolysaccharidosis has been studied using the cetylpyridinium chloride (CPC) precipitation methcd. In all of the patients (three cases with Hurler syndrome, one case with Scheie syndrome and one case with Morquio syndrome), the results obtained were summarized as follows: 1) The plasma infusions were followed by a transient decreased urinary excretion of acid mucopolysaccharides. 2) Furthermore, the infusions were followed by an increased excretion of the non-CPC-precipitates. 3) Ratios of acid mucopolysaccharides to non-CPC-precipitates showed the lowest values on the third or fourth day after plasma infusions. 4) It is suggested that plasma infusions did not modify the course of the patients with mucopolysaccharidosis.

Early diagnosis of hypermucopolysacchariduria

Urinary excretion of acid mucopolysaccharides was determined in seven children with Hurler's disease and in 1000 apparently normal children using a simple turbidity method. All the seven patients with Hurler's syndrome gave values of mucopolysaccharides which were considerably higher than those obtained in the control groups. In the screening test, 32 children with various diseases showed a marked increase in mucopolysaccharides excretion. It must be emphasized that one of those children —aged 1 month at the time of the analysis—was diagnosed later on as a new case of Hurler's syndrome.

Roentgen findings in mucolipidosis III (Pseudo-Hurler polydystrophy).

The skeletal changes seen in 6 patients with mucolipidosis III (pseudo-Hurler polydystrophy) are described: these include claw hands, flattening of the femoral epiphyses, platyspondyly, and flaring of the iliac wings. These changes are variable and not sufficiently characteristic to permit differentiation from certain mucopolysaccharidoses and other mucolipidoses on radiological grounds alone. Correlation with the clinical findings (particularly the onset and progression of symptoms, the degree of physical and mental retardation, and the normal excretion of acid mucopolysaccharides in the urine) is necessary for a specific diagnosis.

I-cell disease: A clinical picture

Eight patients are presented who have a particular type of “Hurler”-like bodyconfiguration, severe skeletal dysplasia, severe psychomotor retardation, and normal urinary excretion of acid mucopolysaccharides, constituting in the sum a new entity, here designated as “I-cell” disease (or mucolipidosis II). The condition is evident from birth, with gradual accentuation of clinical features, complicated by respiratory infections, and with a fatal outcome. Familial occurrence in two instances and parental consanguinity in one, make autosomal recessive mode of inheritance most likely. Fibroblasts of these patients show an abundance of special cytoplasmic inclusions (“I cells”), have numerous deficiencies in lysosomal hydrolases, and contain excessive amounts of lipids and mucopolysaccharides. Results obtained on analysis of liver and brain specimens have less notable abnormalities. The pathogenesis of this new syndrome remains unknown. It must be differentiated especially from the Hurler's disease, GM1-gangliosidosis type 1, and lipomucopolysaccharidosis. Eight patients are presented who have a particular type of “Hurler”-like bodyconfiguration, severe skeletal dysplasia, severe psychomotor retardation, and normal urinary excretion of acid mucopolysaccharides, constituting in the sum a new entity, here designated as “I-cell” disease (or mucolipidosis II). The condition is evident from birth, with gradual accentuation of clinical features, complicated by respiratory infections, and with a fatal outcome. Familial occurrence in two instances and parental consanguinity in one, make autosomal recessive mode of inheritance most likely. Fibroblasts of these patients show an abundance of special cytoplasmic inclusions (“I cells”), have numerous deficiencies in lysosomal hydrolases, and contain excessive amounts of lipids and mucopolysaccharides. Results obtained on analysis of liver and brain specimens have less notable abnormalities. The pathogenesis of this new syndrome remains unknown. It must be differentiated especially from the Hurler's disease, GM1-gangliosidosis type 1, and lipomucopolysaccharidosis.

Das Nagel-Patella-Syndrom

Report on a family with nail-patella syndrome and on its four cardinal symptoms, i.e. onychodystrophy, habitual patellar luxation, subluxation of the capitulum radii and iliac horns, and on other abnormalities of the fibrous tissue and osseous system. An autosomal-dominant inheritance and the combination to the ABO blood group system was found. An increased excretion of acid mucopolysaccharides and hydroxyproline in 24-hour urine samples could not be stated. But the excretion of proline was, in contrast to a control group, significantly increased. Renal insufficiency was by clinical investigations excluded.

Chemical and enzymic studies of a family with skeletal abnormalities associated with mental retardation

In continuation of our previous studies of a family suffering from dwarfism, skeletal abnormalities and mental retardation, the present communication represents a chemical and enzymic study of the abnormalities found in the daily excretion of acid mucopolysaccharides, and in the lysosomal enzymic activities of biological fluids and skin biopsies. The data obtained reveal only a slight increase in the daily excretion of acid mucopolysaccharides, but significant changes in the proportion between the different types of acid mucopolysaccharides. In this respect, especially significant was the proportionally increased urinary excretion of desulfated acid mucopolysaccharides. These changes were associated with a significant decrease in the activity of acid p- nitrophenyl-β- d- galactopyranosidase , but normal or increased activities of α- d-mannosidase, β- d-glucosaminidase, α- l-fucosidase, acid phosphatase and β- d-glucuronidase. The abnormalities are discussed in relation to the clinical findings and support the idea of a specific clinical entity of lysosomal origin.

Biochemical definition of the mucopolysaccharidoses.

The excretion pattern of urinary acid mucopolysaccharides was determined in 72 patients with different clinical types of mucopolysaccharidoses and mucolipidoses. Using a chromatographic fractionation method, characteristic excretion patterns were found in the six classical types of mucopolysaccharidoses. Patients with Hunter's disease excreted relatively more heparansulfate and less dermatansulfate than patients with Hurler's disease. In Sanfilippo's disease the excretion of heparansulfate only was increased. In Morquio's disease abnormal amounts of substances with characteristics of chondroitin-4(6)-sulfate were found in addition to keratansulfate. In one patient with Ullrich-Scheie's disease and six patients with Maroteaux's disease relatively large amounts of substances with characteristics of dermatansulfate were present. There was very little overlap between the excretion patterns of different types of mucopolysaccharidoses. In the types of mucolipidoses investigated, the urinary excretion of acid mucopolysaccharides was normal. The intrafamilial variability in eight pairs of related children was small. Our data suggest that the excretion pattern of urinary acid mucopolysaccharides, as determined by the Dowex Column Chromatography Method, is a valuable aid in the definition of the mucopolysaccharidoses.

Biochemical studies of otosclerosis: urinary acid mucopolysaccharides.

Twenty-four hour urine samples, collected from thirty-six otosclerotic and thirty-eight non-otosclerotic male patients, were assayed for hexuronic acid (an index of total acid mucopolysaccharide content). The urines were electrophoresed in order to identify the individual acid mucopolysaccharides. No differences in total acid mucopolysaccharide excretion were observed to occur between otosclerotic and non-otosclerotic patients. The individual mucopolysaccharides comprising the total fraction were identical in the two groups; and no abnormal components were found in any of the samples, otosclerotic or non-otosclerotic. Thus, it may be concluded that changes in the ground substance, demonstrated histochemically and postulated to occur in the pathogenesis of otosclerosis, are not reflected in the urinary excretion of acid mucopolysaccharides. This would indicate that these changes are confined to the otosclerotic focal areas and are not part of the generalized mesenchymopathy with which otosclerosis is believed to be associated.

Urinary acid mucopolysaccharide excretion in skin disease.

The urinary output of non-sulphated and sulphated acid mucopolysaccharides was measured in a group of psoriatic patients, a group of patients with other dermatoses, and a group of healthy controls. The total excretion of acid mucopolysaccharides was raised in both groups of patients, the increment being equally distributed between the non-sulphated and the sulphated fractions.

Effect of hypophysectomy on the excretion of acid mucopolysaccharides in rat urine.

The effect of hypophysectomy on the excretion of acid mucopolysaccharides in the urine has been investigated in rats. Acid mucopolysaccharides were determined in 24 hr specimens of urine collected from 10 individual rats during 15 consecutive days before and 8 days after hypophysectomy. The results expressed both as ng of uronic acid per 24 hr and μg of uronic acid per 24 hr per 100 g body weight showed that hypophysectomy caused a decrease in the levels of acid mucopolysaccharides in urine, which was statistically highly significant (p<0.001). (Endocrinology 83: 861, 1968)

The effect of simulated altitude (500mm Hg) on urine acid mucopolysaccharides excretion in hypophysectomized rats.

The influence of simulated altitude of 500 mm Hg for 5 hr on the daily excretion of urinary acid mucopolysaccharides in hypophysectomized rats was investigated. Hypophysectomy per se caused a decreased in the excretion of acid mucopolysaccharides. Under the influence of simulated altitude the rats showed non significantly diminished excretion of acid mucopolysaccharides on the day of the experiment. These results were in contrast to those obtained with normal rats.