SERUM ENZYMES OF LYSOSOMAL ORIGIN AS INDICATORS OF THE METABOLIC CONTROL IN NON-INSULIN-DEPENDENT DIABETICS

Several lysosomal enzymes (\u3b2-N-acetyl-D-glucosaminidase, \u3b2-D-glucuronidase, \u3b1-D-galactosidase, \u3b2-D-galactosidase, \u3b1-D-glucosidase, \u3b2-D-glucosidase, \u3b1-L-fucosidase and \u3b1-D-mannosidase) were determined in the serum of 54 non-insulin-dependent diabetics with different degrees of metabolic control and without complications and in 18 non-insulin-dependent diabetics with complications. The serum levels of \u3b2-N-acetyl-D-glucosaminidase, \u3b2-D-glucuronidase, \u3b1-D-galactosidase, and \u3b1-D-mannosidase were significantly (p<0.01) higher in the diabetics without complications. The levels of \u3b2-N-acetyl-D-glucosaminidase and \u3b2-D-glucuronidase were inversely proportional to the degree of metabolic control, in a statistically significant manner. Moreover the levels of these enzymes decreased to normal values during a 2-month period of controlled oral hypoglycemic drug-diet therapy resulting in metabolic compensation. The presence of complications was indicated by a further increase of serum \u3b2-N-acetyl-D-glucosaminidase and \u3b2-D-glucuronidase; however the portion of lysosomal enzyme activities due to complications remained unchanged after controlled therapy aimed at compensating the metabolism. The conclusion is drawn that in non-insulin-dependent diabetics, as already shown for insulin dependent-diabetics, serum lysosomal enzymes, especially \u3b2-N-acetyl-D-glucosaminidase and \u3b2-D-glucuronidase, are good intraindividual indicators of the metabolic control of the disease

AUTOMATED FLUOROMETRIC ASSAY PROCEDURE FOR GLUCOHYDROLASES USING A ROUTINE CENTRIFUGAL ANALYZER ASSAY OF ENZYMES OF LYSOSOMAL ORIGIN IN PLASMA .2

The manual fluorimetric procedure, considered as a reference method for the determination of N-acetyl-beta-D-glucosaminidase, beta-D-glucuronidase and beta-D-galactosidase in human plasma, was automated as a routine method, using the IL Monarch centrifugal analyser. Using a liquid standard with a known enzyme content, the automated assay correlated fairly well with the reference manual method (r values very close to 1). Its analytical imprecision was much lower than that of the manual method. The automated assay of N-acetyl-beta-D-glucosaminidase, beta-D-glucuronidase and beta-D-galactosidase gave coefficients of variation of 5.7-6.9, 3.6-5.0 and 3.8-4.2%, respectively, detection limits of 4, 2 and 1 mU/l plasma respectively, and linear responses of up to 73, 8.4 and 0.9 U/l of plasma respectively. Furthermore, the method required only small volumes of undiluted plasma (4-10 mu l). This method appears to be reliable, sensitive, simple enough for routine analyses and as cost effective as the most common routine serum enzyme assays

Homocysteine concentration in primary and systemic sclerosis associated Raynaud&apos;s phenomenon

OBJECTIVE: To investigate whether patients with systemic sclerosis (SSc) have raised homocysteine (Hcy) plasma levels, thought to be an independent risk factor for vascular disease, and to study the relationship between Hcy and endothelial damage, and between Hcy and methylene-tetrahydrofolate reductase (MTHFR) genotypes, and patients' vitamin nutritional status, which are among the more frequent causes of hyperhomocysteinemia. METHODS: We measured Hcy, von Willebrand factor (vWF), folic acid, and vitamin B12 plasma levels and analyzed the frequencies of MTHFR mutations in 30 patients with SSc and 12 patients with primary Raynaud's phenomenon (RP); 29 healthy subjects served as controls. RESULTS: Patients with SSc had higher Hcy and vWF concentrations than those with RP (p < 0.01 and p < 0.02, respectively) or controls (p < 0.02 and p < 0.0001, respectively). Folic acid and vitamin B12 were lower in SSc than in RP (p < 0.01 and p < 0.02, respectively) or controls (p < 0.05). MTHFR genotype did not influence Hcy, folate, or vitamin B12 concentrations, but patients homozygous for the mutant gene had higher vWF levels. CONCLUSION: Patients with SSc, but not those with RP, had significantly higher Hcy and vWF plasma levels. Nutritional rather than inherited factors seem to have a pathogenic role in SSc hyperhomocysteinemia

Homocysteine concentration in primary and systemic sclerosis associated Raynaud's phenomenon

OBJECTIVE: To investigate whether patients with systemic sclerosis (SSc) have raised homocysteine (Hcy) plasma levels, thought to be an independent risk factor for vascular disease, and to study the relationship between Hcy and endothelial damage, and between Hcy and methylene-tetrahydrofolate reductase (MTHFR) genotypes, and patients' vitamin nutritional status, which are among the more frequent causes of hyperhomocysteinemia. METHODS: We measured Hcy, von Willebrand factor (vWF), folic acid, and vitamin B12 plasma levels and analyzed the frequencies of MTHFR mutations in 30 patients with SSc and 12 patients with primary Raynaud's phenomenon (RP); 29 healthy subjects served as controls. RESULTS: Patients with SSc had higher Hcy and vWF concentrations than those with RP (p < 0.01 and p < 0.02, respectively) or controls (p < 0.02 and p < 0.0001, respectively). Folic acid and vitamin B12 were lower in SSc than in RP (p < 0.01 and p < 0.02, respectively) or controls (p < 0.05). MTHFR genotype did not influence Hcy, folate, or vitamin B12 concentrations, but patients homozygous for the mutant gene had higher vWF levels. CONCLUSION: Patients with SSc, but not those with RP, had significantly higher Hcy and vWF plasma levels. Nutritional rather than inherited factors seem to have a pathogenic role in SSc hyperhomocysteinemi