[THE QUANTIFICATION OF SULPHATED GLYCOSAMINOGLYCANS IN RAT URINE IN EXPERIMENTAL HEMORRHAGIC CYSTITIS].

The paper presents the study of the excretion of sulfated glycosaminoglycans (GAG) in the urine of rats in experimental hemorrhagic cystitis induced by cyclophosphamide and treated with glycosaminoglycan replacement therapy. Rats were given intraperitoneal injections of cyclophosphamide at a dose of 100 mg per 1 kg body weight and subsequently treated with intragastric administration of the combined preparation of glycosaminoglycans containing glucosamine hydrochloride and chondroitin sulfate at a dose of 10 and 100 mg per 1 kg of body weight. Within 24 or 72 hours after cystitis induction there was a statistically significant increase in urinary GAG excretion. The study also found a decrease (from 1.34 to 1.22 mg/dL) in urinary GAG within 0 to 72 hours following induction of acute cystitis without glycosaminoglycan therapy. In the subchronic model of inflammation in the bladder, upon repeated administration of low doses of cyclophosphamide (50 mg/kg), decrease in urinary GAG within 0 to 72 hours (1,32±0,13 mg/dL) as well as increased excretion after 96 hours at a concentration of 2,29±0,13 mg/L after initiation cystitis were found.

Suppression of NF-κB p65 nuclear translocation and tumor necrosis factor-α by Pongamia pinnata seed extract in adjuvant-induced arthritis.

Pongamia pinnata is a plant known for its therapeutic usage in Indian traditional medicine. Despite the controversy regarding toxic flavonoid and erucic acid content, the seed of this plant is consumed in tribal medicine and its oil is used in Ayurveda to treat psoriasis and arthritis. This study explored the potential anti-arthritic effects of a P. pinnata seed (hexane) extract (PSE) at non-lethal doses in an adjuvant-induced arthritic rat model; possible mechanisms of any observed effects were also explored. After establishing the lethal doses arising from oral exposure to the extract, the material was administered per os daily at two doses (0.3 g/kg/day; 0.5 g/kg/day) to arthritic rats. Other rats received indomethacin or vehicle (control). Treatments were performed for a total of 14 days. One day after the final exposure, the rats were euthanized to permit harvest of various cells, blood, and tissues for analyses. Paw diameter and tissue myeloperoxidase activity in the paws were evaluated as indices for edema and neutrophil infiltration into the tissue. The severity of arthritis in the experimental rats was assessed via measures of urinary hydroxyproline (HP) and glucosamine, and of serum pro-inflammatory TNFα and anti-inflammatory IL-10. The extent of NF-κB p65 nuclear translocation in peritoneal macrophages harvested from naïve rats and then treated in vitro was also assessed. The results indicated that exposure to PSE significantly decreased paw diameter, tissue myeloperoxidase level, and levels of urinary HP and glucosamine, as well as of serum TNFα and IL-10 in adjuvant-injected (arthritic) rats. In vitro PSE treatment also resulted in a marked inhibition of NF-κB p65 nuclear translocation in primary cultures of peritoneal macrophages. Thus, PSE appears to be able to prevent experimental arthritis, in part, by helping to maintain the balance between pro- and anti-inflammatory cytokines and by inhibiting NF-κB activation.

Determination of urinary hexosamines for diagnosis of bladder pain syndrome.

INTRODUCTION AND HYPOTHESIS: Bladder pain syndrome (BPS) is a chronic disease characterized by urgency, bladder pain, and frequency, and urinary glycosaminoglycans are thought to reflect bladder epithelial deficiency in BPS. Sensitive and specific evaluation of total urinary glycosaminoglycans may be useful for the clinical diagnosis of BPS and its treatment. METHODS: A procedure for the simultaneous determination of glucosamine and galactosamine produced from urinary glycosaminoglycans has been performed in BPS patients and healthy subjects. RESULTS: The total content of urinary hexosamines in BPS patients significantly increased by ~130% with the increase in glucosamine greater than galactosamine. CONCLUSIONS: A significant increase in total hexosamines content and in particular in glucosamine belonging to urinary heparan sulfate was determined in BPS patients compared with controls. We propose HS and in particular its low-molecular mass fragments and glucosamine assay as useful markers for a biochemical diagnosis of BPS and for monitoring this syndrome.

A high molecular weight protein extract of Mastobranchus indicus (Mi-64) having antiarthritic activity in experimental animals.

Mi-64, a high molecular weight protein (130 kDa), obtained from the tissue homogenate of marine polychaete (Mastobranchus indicus) collected from the Indian Sunderban has antiarthritic activity in experimental animals. The FCA-induced arthritis model was developed in Wistar albino rats to evaluate the antiarthritic effects of Mi-64. After FCA induction, the rats were treated with Mi-64 (0.25 and 0.5 mg kg(-1) body weight) for 10 days. We have determined the paw/ankle swellings, urinary hydroxyproline and glucosamine, serum acid and alkaline phosphatases to assess the antiarthritic activity. The levels of interleukin-1 beta (IL-1ß), IL-6, cytokine-induced neutrophil chemoattractant-1 (CINC-1), tumor necrosis factor-alpha (TNF-α), and IL-10 were measured by ELISA. Results showed that Mi-64 significantly reduced paw/ankle swellings and restored the urinary hydroxyproline/glucosamine and serum phosphatases. Mi-64 significantly inhibited the overproduction of IL-1ß, IL-6, CINC-1, and TNF-α and augmented IL-10 production. The data suggest that Mi-64 produced significant antiarthritic effects that may be mediated by balancing the pro- and anti-inflammatory cytokines.

Selenium metabolites in human urine after ingestion of selenite, L-selenomethionine, or DL-selenomethionine: a quantitative case study by HPLC/ICPMS.

To obtain quantitative information on human metabolism of selenium, we have performed selenium speciation analysis by HPLC/ICPMS on samples of human urine from one volunteer over a 48-hour period after ingestion of selenium (1.0 mg) as sodium selenite, L-selenomethionine, or DL-selenomethionine. The three separate experiments were performed in duplicate. Normal background urine from the volunteer contained total selenium concentrations of 8-30 microg Se/L (n=22) but, depending on the chromatographic conditions, only about 30-70% could be quantified by HPLC/ICPMS. The major species in background urine were two selenosugars, namely methyl-2-acetamido-2-deoxy-1-seleno-beta-D-galactopyranoside (selenosugar 1) and its deacylated analog methyl-2-amino-2-deoxy-1-seleno-beta-D-galactopyranoside (selenosugar 3). Selenium was rapidly excreted after ingestion of the selenium compounds: the peak concentrations (approximately 250-400 microg Se/L, normalized concentrations) were recorded within 5-9 hours, and concentrations had returned to close to background levels within 48 hours, by which time 25-40% of the ingested selenium, depending on the species ingested, had been accounted for in the urine. In all experiments, the major metabolite was selenosugar 1, constituting either approximately 80% of the total selenium excreted over the first 24 hours after ingestion of selenite or L-selenomethionine or approximately 65% after ingestion of DL-selenomethionine. Selenite was not present at significant levels (<1 microg Se/L) in any of the samples; selenomethionine was present in only trace amounts (approximately 1 microg/L, equivalent to less than 0.5% of the total Se) following ingestion of L-selenomethionine, but it constituted about 20% of the excreted selenium (first 24 hours) after ingestion of DL-selenomethionine, presumably because the D form was not efficiently metabolized. Trimethylselenonium ion, a commonly reported urine metabolite, could not be detected (<1 microg/L) in the urine samples after ingestion of selenite or selenomethionine. Cytotoxicity studies on selenosugar 1 and its glucosamine isomer (selenosugar 2, methyl-2-acetamido-2-deoxy-1-seleno-beta-D-glucosopyranoside) were performed with HepG2 cells derived from human hepatocarcinoma, and these showed that both compounds had low toxicity (about 1000-fold less toxic than sodium selenite). The results support earlier studies showing that selenosugar 1 is the major urinary metabolite after increased selenium intake, and they suggest that previously accepted pathways for human metabolism of selenium involving trimethylselenonium ion as the excretionary end product may need to be re-evaluated.

Aspartylglucosaminuria: psychomotor retardation masquerading as a mucopolysaccharidosis.

A 5-year-old girl with coarse facies, visceromegaly, and vacuolated lymphocytes is presented as the first case of aspartylglucosaminuria diagnosed in this country. This metabolic defect in glycoprotein catabolism can be clinically confused with other storage diseases such as the mucopolysaccharidoses and mucolipidoses. It is not diagnosed by routine laboratory screening methods. Special studies are required to confirm the diagnosis, but a thin-layer chromatography method for screening urine is presented for use when the diagnosis is suspected. The developmental potential in this inborn error of metabolism is documented.

Aspartylglycosaminuria: a generalized storage disease. Morphological and histochemical studies.

Aspartylglycosaminuria (AGU) is a hereditary metabolic disorder characterized by slowly progressive mental deterioration from infancy, urinary excretion of large amounts of aspartylglycosamine, and decreased activity of the lysosomal enzyme aspartylglcosamine amido hydrolase in various body tissues and fluids. The nature and distribution of the morphological and histochemical alterations in AUG are described in the light of the first AGU patient investigated post mortem and brain and liver. Most nerve cells and hepatocytes contained large vacuoles without any histochemically demonstrable lipid or carbohydrate material. Ultrastructural studies revealed numerous electron-lucent vaculoles, limited by a single, membrane, in the cytoplasm of these cells. In addition to evenly disperesed finely granular or reticular material the vacuoles contained small electron-opaque "lipid" droplets and polymorphic membraneous or granular aggregates. Similar vacuoles were also seen in a number of other cell types, particularly in the kupffer cells and brain macrophages, as well as in the capillary pericytes. Biochemical studies suggest that the principal storage material consists of aspartylglycosamine itself; glycoasparagines of higher molecular weight are present as only minor components. Correlated morphological and biochemical studies thus definitely establish that AGU is a generalized storage disorder. The condition is apparently due to decreased activity of aspartylglycosamine amido hydrolase, with accumulation of products of flycoprotein carabolism in cytoplasmic vacuoles in both epithelial and mesenchymal cells.