[Prognostication and prophylaxis of the adhesions recurrence postoperatively in patients suffering acute adhesive peritoneal disease complicated by ileus].

The results of treatment of 92 patients, suffering an acute ileus, were analyzed. After urgent operative interventions for an acute ileus the recurrence have occurred in 19.6% patients. To reduce the operative intervention traumaticity the preference was given to local viscerolysis conduction. For the adhesions occurrence prophylaxis a barrier medicines were used, what have promoted the reduction of contents of a connective tissue metabolites, excluding oxyprolin, concentration of which have exceeded such in a control, what have guaranteed the risk lowering for postoperative adhesions occurrence.

Enzyme replacement therapy attenuates disease progression in two Japanese siblings with mucopolysaccharidosis type VI.

Mucopolysaccharidosis type VI (MPS VI) is a progressive, multisystem autosomal recessive lysosomal disorder resulting from deficient N-acetylgalactosamine-4-sulphatase (ASB) and the consequent accumulation of glycosaminoglycan (GAG). Preclinical and clinical studies had demonstrated clinical benefits of early initiation of systemic therapies in patients with MPS. In this case report, two siblings with MPS VI started enzyme replacement therapy (ERT) with weekly infusions of recombinant human ASB (Galsulfase) at 1mg/kg. Sibling 1 started ERT 5.6 years of age and Sibling 2 was 6 weeks old. The disease status in these two siblings prior to and for no less than 36 months of ERT was followed up and compared. The treatment was well tolerated by both siblings. During 36 months of ERT, symptoms typical of MPS VI including short stature, progressive dysmorphic facial features, hepatosplenomegaly, hearing impairment, corneal clouding, and dysostosis multiplex were largely absent in the younger sibling. Her cardiac functions and joint mobility were well preserved. On the other hand, her affected brother had typical MPS VI phenotypic features described above before commencing ERT at the equivalent age, of 3 years. There was significant improvement in the shoulder range of motion and hearing loss after 36 months of treatment and cardiac function was largely preserved. His skeletal deformity and short stature remained unchanged. The results showed that early ERT initiated at newborn is safe and effective in preventing or slowing down disease progression of MPS VI including bone deformities. These observations indicate that early diagnosis and treatment of MPS VI before development of an irreversible disease is critical for optimal clinical outcome.

Safety and efficacy of idursulfase in the treatment of mucopolysaccharidosis II (Hunter syndrome): a post-marketing study in Japan.

OBJECTIVES: Enzyme replacement therapy with idursulfase has been shown to improve somatic signs and symptoms of mucopolysaccharidosis type II (MPS II). Idursulfase is available in Japan (since 2007), based on the outcome of clinical trials conducted in the United States, but data from Japanese patients are limited. METHODS: This was a postmarketing study of Japanese MPS II patients treated with 0.5 mg/kg intravenous idursulfase weekly, conducted over a period of 8 years after initial administration. Assessments included the safety profile, survival rate, degree of clinical improvement, change in urinary uronic acid (UA) concentration, and 6-minute walk test (6MWT). RESULTS: The safety and efficacy analysis populations included 145 and 143 patients, respectively. The incidence of serious adverse events was 42.8% and the incidence of adverse drug reactions was 48.3%. The 7-year survival rate was 82.7%. Improvements in the clinical features of hepatosplenomegaly, skin, joint, and respiratory disorders were reported (per investigator's assessment). The mean change in urinary UA concentration was -128.39 mg/g creatinine, and that of 6MWT walking distance was +31.8 m. CONCLUSION: Long-term idursulfase treatment was well tolerated, and effective in improving clinical features, reducing urinary UA, and slowing disease progression in Japanese MPS II patients.

Increased urinary excretion of acidic mucopolysaccharides in exophthalmos.

The excretion of mucopolysaccharides normally found in urine (chondroitin, chondroitin sulfates A and C, keratosulfate, and heparitin sulfate) is increased approximately twofold in patients with progresive exophthalmos. A threefold elevation of total serum mucopolysaccharides is also found. These increases are unrelated to thyroid function.

Urinary glycosaminoglycan excretion as a biochemical marker in patients with bladder carcinoma.

Urinary glycosaminoglycan excretion was examined in 25 individuals with bladder cancer in comparison to glycosaminoglycan excretion by eight normal individuals. Urinary glycosaminoglycan was isolated by gel filtration and quantified as macromolecular uronate concentration. Electrophoresis in calcium acetate and densitometry of Alcian blue-stained electrophoretograms were used to separate and quantify the relative amounts of individual glycosaminoglycans. Elevated excretion of macromolecular uronate was noted in 53% of the cancer cases. The highest levels were found among individuals with metastatic disease. Three electrophoretic bands were always detected in the control and cancer groups: chondroitin sulfate, heparan sulfate (both confirmed by chemical and enzymatic degradation), and a third band (Band 1) of unknown composition. A fourth band, corresponding to dermatan sulfate, was seen in some high-grade metastatic tumors. Band 1 excretion was elevated in a significant fraction of all patients. Seven of 12 metastatic cases but only two of 13 localized cases showed increased heparan sulfate excretion. Diagnostic limits were drawn from the observed distributions of normals, and with these limits 92% of the cancer cases, including 12 of 12 metastatic cases, could be identified. The results strongly suggest noninvasive urinary glycosaminoglycan analysis may well provide a new biochemical approach for detecting and monitoring the pathogeneses of bladder cancer.

Total and single species of uronic acid-bearing glycosaminoglycans in urine of newborns of 2-3days of age for early diagnosis application.

BACKGROUND: Urine are easily accessible and relatively simple to process and uronic acid-bearing glycosaminoglycans (UA-GAGs) may serve as biomarkers for several diseases, like for mucopolysaccharidosis. METHODS: We report a study from a large cohort of healthy newborns of 2-3days to have a basic profile of total content of urinary UA-GAGs, their composition and structural signatures utilizing a rapid extractive method and sensitive separation of enzymatic released disaccharides by capillary electrophoresis-light induced fluorescence. Results were also compared with those obtained from normal adult subjects. RESULTS: A total of UA-GAGs content of ~35µg/mg creatinine was observed in 331 newborns versus 1.5µg/mg creatinine of adult urine composed of ~90% chondroitin sulfate (CS), ~7% heparan sulfate (HS) and ~3% hyaluronic acid (HA). No significant differences were observed with adults. Specific ratios between the main CS disaccharides were informative of a significant greater 4-sulfation and charge density for newborn compared to adults. The HS from newborn urine was mainly composed by the non-sulfated (~64%) and mono-sulfated (~28%) disaccharides. No significant differences were observed versus adult urine. CONCLUSIONS: The present method is able to measure changes in UA-GAG composition and their structure independently of the age of subjects and rapidly applicable to the newborn diagnosis without necessity to have creatinine levels. Moreover, modifications in charge density values as well as the presence of sulfate groups in specific positions may be indicative of altered conditions.

Serum and urine glycosaminoglycans in myeloid leukemia and myelodysplasia.

Urinary glycosaminoglycan excretion is increased in a variety of human diseases, including malignancy. We have measured serum and urine glycosaminoglycan levels by the carbazole method of uronic acid determination in patients with myeloid leukemia or myelodysplasia. Eleven patients were studied during active disease as well as eight in complete remission. Serum levels in patients with active disease did not differ significantly from 11 healthy volunteers with no hematological disease. In contrast, the median urine level for the patients with active disease was 7.6 mg uronate/g Creatinine (Creat) compared to 2.6 for controls (p less than 0.002). Interestingly, the eight patients in complete remission also had a significant increase in uronate excretion with a median of 7.3 (p less than 0.002). These results suggest that elevated urinary glycosaminoglycan levels in leukemia are not due to impaired ability of the liver to clear circulating glycosaminoglycans or overproduction by leukemic cells. The observed increase in glycosaminoglycan excretion may be due to altered bone marrow matrix metabolism that is often not reversed by the achievement of hematologic remission.

Urinary glycoconjugates in schizophrenic patients.

The level of glycoconjugates excreted in the urine of five schizophrenic patients was compared to that excreted by six controls. Urinary samples were fractionated by means of gel filtration and anion exchange chromatography to yield (i) fraction I consisting of basic, neutral, or slightly acidic glycopeptides and/or oligosaccharides, (ii) fraction II consisting of acidic glycopeptides and/or oligosaccharides, and (iii) fraction III consisting of glycosaminoglycans (acidic mucopolysaccharides). The hexose levels of fraction II (p less than 0.05) and uronate levels of fraction III (p less than 0.025) were significantly reduced in schizophrenic patients. The ratio of galactose/mannose in the glycoconjugates of fraction II was lower than normal in the urine from schizophrenic patients. Significantly (p less than 0.05) lower levels of rhamnose and higher levels of fucose were found in the glycoconjugates of fraction I from schizophrenic patients. Contrary to a previous report, we found no evidence for the presence of an abnormally elevated rhamnose-containing glycoprotein or glycoconjugate in fraction II. I appears that the pattern of metabolism of glycoproteins and glycosaminoglycans in schizophrenic patients deviates from the normal.

Urinary excretion of glycosaminoglycans in disseminated neoplasm.

Urinary glycosaminoglycan excretion was studied in 24 cases of disseminated neoplasm, 12 of which had unequivocal evidence of skeletal involvement. Urinary hydroxyproline, cetylpyridinium chloride (CPC)-precipitable uronic acid, and CPC-precipitable hexosamine were expressed as a ratio to urinary creatinine. Glycosaminoglycans contained in urine concentrated x 1000 by vacuum-dialysis were separated by electrophoresis on cellulose acetate and stained with alcian blue. Of the 12 cases with clear evidence of skeletal involvement, eight (66%) showed elevation of serum alkaline phosphatase, five (42%) showed elevation of urinary hydroxyproline, and three (25%) showed elevation of urinary uronic acid. It is concluded that urinary uronic acid is not a sensitive index of skeletal involvement in disseminated neoplasm. The most striking feature of the study was the identification of a well-defined fraction indist inguishable from hyaluronic acid in seven (58%) of the cases with evidence of skeletal involvement. Hyaluronic acid is not normally identifiable in adult human urine. The hyaluronic acid excretors showed more consistent biochemical evidence of bone disease (elevation of serum alkaline phosphatase and urinary hydroxyproline) than the non-excretors. The possibility that the urinary hyaluronic acid is derived from degradation of skeletal hyaluronic acid is discussed. An alternative explanation is that the hyaluronic acid is derived from neoplastic cells as part of a reversion of glycosaminoglycan synthesis to a more ;fetal' state, a glycosaminoglycan counterpart of the production of oncofetal antigens by neoplastic cells.

Excretion patterns of glycosaminoglycans and glycoproteins in normal human urine.

Glycosaminoglycans and glycoproteins in the urine of 100 healthy, active, human subjects were examined by cellulose acetate electrophoresis and salt gradient, ion-exchange, column chromatography. The cetylpyridinium chloride (CPC) turbidity and uronic acid:creatinine ratio was also studied. Fractions were identified by electrophoretic mobility, staining reactions, susceptibility to enzyme digestion, identification of amino- and neutral sugars, hexosamine, uronic acid, and sulphate assays, and optical rotation. The CPC turbidity is relatively high in childhood, falling to lower levels in adults, but rising again to relatively high levels in old age. The uronic acid: creatinine ratio is high in children, falling to a low level in adult life, and rising only slightly in old age. Three major electrophoretic fractions, corresponding with glycoprotein, heparan sulphates, and chondroitin sulphates, were identified in every urine sample. Hyaluronic acid was identified in some samples. A small amount of keratan sulphate was present in the ;heparan sulphate' fraction. Chondroitin sulphate excretion is high in children. Adults excrete relatively less chondroitin sulphate and more heparan sulphate. In old age, the proportion of glycoprotein increases. The excretion pattern in the first few days of life resembles that of the adult. It is stressed that extreme caution must be exercised in interpreting the urinary glycosaminoglycan pattern of a child.

Urinary excretion of glycosaminoglycans in the various forms of gargoylism.

The urinary excretion of glycosaminoglycans in 28 cases of gargoylism, embracing the Hurler, Hunter, Sanfilippo, Morquio, and Scheie syndromes (McKusick, 1966), has been examined using the cetylpyridinium chloride (CPC) turbidity test, the uronic acid/creatinine ratio, and the electrophoretic pattern of urine concentrates, as routine procedures. Ion-exchange column chromatographic techniques were also employed for the fractionation of glycosaminoglycans and aminosugars. Molecular weights were investigated by gel filtration and ultracentrifugation. The CPC turbidity test was positive in every case. The uronic acid/creatinine ratio provided a sensitive index of increased glycosaminoglycan excretion. Cases of the Hurler syndrome showed the highest, and cases of the Morquio and Scheie syndromes the lowest, ratios. A correlation was observed between the uronic acid/creatinine ratio and the clinical severity of the disease. Cellulose acetate electrophoresis differentiated clearly between the two major forms of gargoylism, the Hurler and Sanfilippo syndromes, but differentiation between the Hurler, Hunter, and Scheie syndromes was more difficult on electrophoretic data alone. Results obtained with cases diagnosed as the Morquio syndrome were disappointing. The existence of formes frustes of the Sanfilippo syndrome among the mentally subnormal is predicted. Errors caused by bacterial contamination of urine samples are emphasized. The atypical behaviour of urinary glycosaminoglycans in analytical procedures is discussed. Molecular weight studies suggested heterogeneity. The nature of the basic defect in gargoylism is discussed.

Urinary excretion of glycosaminoglycans and hydroxyproline in Paget's disease of bone, compared with neoplastic invasion of bone.

Urinary glycosaminoglycan and hydroxyproline excretion was studied in 11 patients with clear evidence of Paget's disease of bone. Urinary hydroxyproline, cetyl pyridinium chloride (CPC)-precipitable uronic acid and CPC-precipitable hexosamine were expressed as ratios to urinary creatinine. Urine samples were concentrated x 1000 by vacuum dialysis and the glycosaminoglycans examined by electrophoresis on cellulose acetate followed by staining with alcian blue. All the cases studied showed markedly raised hydroxyproline excretion, whereas the uronic acid excretion was normal or only slightly raised in 10 of the 11 cases studied. One patient who had a raised uronic acid and raised hydroxyproline concentration was shown to have osteosarcoma as a complication of Paget's disease. THE VERY HIGH HYDROXYPROLINE: creatinine ratio in all cases of Paget's disease (mean 241.8 mmol hydroxyproline/mol creatinine) contrasted sharply with the cases of disseminated neoplasm, where the ratio was either normal or slightly raised (mean 29.3 mmol hydroxyproline/mol creatinine). The ratio of hydroxyproline to CPC-precipitable uronic acid was also markedly raised in cases of Paget's disease (mean 77.3 mmol hydroxyproline/mmol uronic acid) and was lower in the neoplastic group (mean 14.1 mmol hydroxyproline/mmol uronic acid) but showed no advantage over the hydroxyproline: creatinine ratio in differentiating the two groups. THE URINARY HYDROXYPROLINE: creatinine ratio promises to be of value in differentiating between Paget's disease of bone and neoplastic invasion of bone. A marked rise in CPC-precipitable uronic acid excretion alone is more suggestive of neoplastic invasion of bone, and if associated with a marked increase in hydroxyproline excretion, it raises the possibility of neoplastic change in Paget's disease of bone. The results of this study also suggest that bone collagen, rather than bone tissue in general, is primarily affected in Paget's disease.

Dietary fibres ameliorate decreased synthesis of heparan sulphate in streptozotocin induced diabetic rats.

The role of dietary fibers in diabetes has been studied by several workers. Long term dietary treatment with increased amounts of fiber-rich low-glycaemic index natural foods improves blood glucose and reduces the number of hypoglycemic events in type I diabetic patients. On the other hand Rohrbach and Martin and Cohen and Surma described changes in the general and biochemical structure of renal tissues such as the glomerular basement membranes. One of these changes was the reduction and undersulfation of the glycoconjugate and glycosaminoglycan heparan sulfate, which plays an important role in renal structure and function. The purpose of the present study was to determine specific effects of two types of dietary fiber on the composition of kidney glycoconjugates in an animal model of diabetes type I. Streptozotocin-treated diabetic rats were fed either a control diet or diets containing 10% wheat bran (insoluble dietary fiber) or 5% guar gum (soluble dietary fiber). Effects of these fibers on glycaemic control and nephropathy were assessed using previously described methodologies. The effect of dietary fiber in the glycoconjugate composition of kidneys of control and diabetic animals was studied by estimating their total hexose content, sulfated glycosaminoglycans, hexosamines and uronic acids. The activities of enzymes that participate in the synthesis of saccharides and glycoconjugates (L-glutamine-fructose-6-phosphate aminotransferase) and their degradation (N-acetyl-beta-glucosaminidase and beta-glucuronidase) were also evaluated. Results indicated that both soluble and insoluble dietary fibers ameliorated a significant increase in the activity of GFAT. Heparan sulfate was also isolated and quantified. Results indicated that the renal content of heparan sulfate decreased in diabetic animals and that this decrement was ameliorated by the ingestion of both soluble and insoluble fiber in the diet.

Familial progressive myoclonic epilepsy. A clinical, genetical, biochemical and patho-anatomical study of a family with a 6-year follow-up.

Six siblings, including 4 cases of myoclonic epilepsy, their parents and 2 grandmothers were subjected to systematic investigation, and the patients were followed-up. The genetic studies revealed in the mother's family a patient with Lafora bodies demonstrated at autopsy. No chromosome abnormalities were found nor any linkage to the HLA system. The affected family members were characterized biochemically by an increased excretion of total glycosaminoglycans and/or an abnormal electrophoretic pattern of urinary glycosaminoglycans with an increased proportion of low-sulfated glycosaminoglycans. In the healthy family members this pattern of electrophoresis could also be demonstrated in the father and the paternal grandmother. Based on the biochemical results and the genetic studies it is suggested that the family members with progressive familial myoclonic epilepsy present a combination of at least 2 hereditary defects. The course of the disease has been relatively benign and treatment with sodium valproate, baclofen and clonazepam has shown quite satisfying results. In consequence of the biochemical findings combined treatment with A and E vitamins has been initiated.

Glycosaminoglycan excretion in osteogenesis imperfecta.

This study was carried out in order to determine whether osteogenesis imperfecta affected individuals showed a different pattern of glycosaminoglycan excretion in urine than do normals. Quantitative excretion was compared by three different methods, and the qualitative composition of the excreted glycosaminoglycan was compared by electrophoresis. No difference was noted in the amount of glycosaminoglycan excreted by normal or affected individuals as measured by macromolecular uronic acid, Alcian Blue bound by urinary constituents or amount of glycosaminoglycan actually isolated by partition. However, the affected individuals showed a statistically significant increase in Alcian Blue binding over macromolecular uronic acid not seen in the normal group, which may possibly indicate the excretion of a substance capable of binding Alcian Blue, but which is not glycosaminoglycan, by osteogenesis imperfecta affected individuals. No qualitative differences in glycosaminoglycan excretion between the two groups were detectable by electrophoresis.

Compositional changes of urinary acidic glycosaminoglycans in progressive systemic sclerosis.

The compositional changes of acidic glycosaminoglycans (AGAG) in the urine of progressive systemic sclerosis (PSS) patients were studied using chondroitinases and heparitinase in appropriate enzyme assays and by electrophoretic characterization. Daily urinary excretion of AGAG in patients with PSS was increased, when compared to normals. The proportion of urinary AGAG in PSS patients, which was undigested by chondroitinase-ABC (most probably representing heparan sulfates (HS)), increased significantly from the normal value. The substance was found to be mainly HS as determined by the electrophoretic pattern, thin-layer chromatographic analysis and by its susceptibility to heparitinase. After digestion of urinary chondroitin sulfate isomers with chondroitinases, the unsaturated disaccharides were mainly separated into 4-sulfated and 6-sulfated disaccharide units by paper chromatography. In all of the patients with PSS, the ratio of the unsaturated 4-sulfated disaccharide to the unsaturated disaccharide was higher than that in normal subjects. These observations indicate an abnormal turnover of AGAG in patients with PSS.

A method for the quantitative determination of urinary glycosaminoglycans.

A method is presented by which the urinary glycosaminoglycans can be isolated in a macromolecular fraction with complete recovery by preparative gel filtration. The urinary glycosaminoglycans can be quantitated by determinations of uronic acid and sulfaminohexose in the macromolecular fraction. Using untreated, random urine specimens of less than 5 ml volume, clinical mucopolysaccharidoses types I, II, and III could be readily distinguished.

Urinary excretion of glycosaminoglycans in psoriasis.

Urine from patients with generalised plaque psoriasis contained substantially more precipitable glycosaminoglycans (GAG) and uronic acid than the urine of healthy controls. The difference was not related to sex, age, renal function, the hospital environment, or to the presence of arthritis. Successful topical treatment with tar or dithranol, or PUVA therapy, did not affect the rate of GAG excretion. Cellulose acetate electrophoresis of the GAG from patients and controls showed similar patterns dominated by chondroitin sulphate. There was no evidence to favour the skin lesions as the source of the additional glycosaminoglycans and the findings are consistent with the concept of psoriasis as a general disease.

Glycohydrolases and lysosomal stability in adjuvant induced arthritis.

Adjuvant induced arthritis in rats was studied by the changes in serum and urinary protein-bound carbohydrate metabolites, changes in serum and tissue lysosomal glycohydrolases and lysosomal fragility. From the second week onwards the urinary excretion of hexosamine and uronic acid is increased. Serum levels of protein bound hexose, hexosamine, sialic acid and fucose are increased significantly in both the acute and chronic phases of the disease. There is no change in the total activity of lysosomal glycohydrolases, viz., beta-glucuronidase, beta-N-acetyl glucosaminidase and cathepsin D in the tissues of liver, kidney and spleen except that of liver enzymes in the chronic phase which are elevated significantly. The free activities of lysosomal glycohydrolases investigated, viz., beta-glucuronidase, beta-N-acetyl glucosaminidase, beta-galactosidase, alpha-mannosidase and cathepsin D are increased in liver and spleen in the acute phase. The free activities of beta-glucuronidase, beta-N-acetyl glucosaminidase and cathepsin D of kidney showed no change whereas those of beta-galactosidase and alpha-mannosidase are increased. In the chronic phase of the disease the free activities of all glycohydrolases are significantly increased in all tissues. Serum glycohydrolases are significantly increased in both acute and chronic phases. Studies on lysosomal preparations showed increased fragility of lysosomes derived from liver and kidney of arthritic rats in both phases of the disease.