ABSTRACT
A method of identification and quantitative determination of baclofen in blood by HPLC with mass spectrometry detection has been developed. It is characterized by high sensitivity, specificity, linearity, accuracy, reproducibility, and a low detection for quantitative determination. The method has been used for diagnostics of acute baclofen poisoning in patients.
Subject(s)
Baclofen/blood , Blood Chemical Analysis/methods , Muscle Relaxants, Central/blood , Baclofen/poisoning , Case-Control Studies , Chromatography, High Pressure Liquid/methods , Humans , Mass Spectrometry/methods , Muscle Relaxants, Central/poisoning , Sensitivity and SpecificityABSTRACT
Excess of vitamin A induces decrease of neutral phospholipase A1 and A2 activity in rat testes homogenates on the 4th day, and increase of beta-galactosidase activity on the 8th day of treatment. It is suggested that phospholipase A activity decrease is of great importance in development of testicular disorders, caused by disbalance of vitamin A.
Subject(s)
Phospholipases/metabolism , Testis/enzymology , Vitamin A/toxicity , Animals , Lysosomes/enzymology , Male , Rats , Testis/drug effectsABSTRACT
The effect of 120- and 240-h starvation on rats hepatocytes ultrastructure and particularly the changes of the lysosomes were studied. Eelectronmicroscopically and cytochemically there have been observed diminution of the number of mitochondria and degranulation and vacuolzation of the ER. At the same time Golgi complex was hypertrophied and the number of lysosomes was much increased, mainly those of the autophagic type. Biochemically was shown, that the activity of some acid hydrolases (beta-glucosidase, alpha- and beta-galactosidases, beta-N-acetylglucosaminidase, beta-glucuronidase and arylsulphatases A and B) in the liver of starved rats was markedly expressed. The sedimentation properties of the lysosomes and the lysosomal membrane stability was damaged as well. The data received have been discussed in the light of the reconstructive role of lysosomes.
Subject(s)
Liver/enzymology , Lysosomes/enzymology , Starvation , Acetylglucosaminidase/metabolism , Acid Phosphatase/metabolism , Animals , Cerebroside-Sulfatase/metabolism , Chondro-4-Sulfatase/metabolism , Galactosidases/metabolism , Glucose-6-Phosphatase/metabolism , Glucosidases/metabolism , Glucuronidase/metabolism , Liver/ultrastructure , Lysosomes/ultrastructure , Male , Microsomes, Liver/enzymology , Microsomes, Liver/ultrastructure , Mitochondria, Liver/enzymology , Mitochondria, Liver/ultrastructure , Proteins/metabolism , RatsABSTRACT
Effect of ethanol on functional activity of isolated perfused rat liver was studied (rate of O2 utilization, absorption of bromosulpholeine from perfusate, bile formation); total activity and activity in supernatant of nine marker enzymes were also determined (malate dehydrogenase, beta-glucuronidase, arylsulphatases A and B, beta-galactosidase, beta-glucosidase, acetylesterase, glucoso-6-phosphatase, alanine aminotransferase and aspartate aminotransferase). Activity of the enzymes was simultaneously studied in perfusate. Ethanol (0.5%) caused distinct impairement in functional activity of isolated liver; rate of bile formation and absorption of bromosulpholeine from perfusate were primarily altered. Degree of impairements in functional activity of liver tissue correlated with the concentration of ethanol in perfusate. In analysis of correlation between the total activity of the enzymes in liver tissue and their activity in supernatants and perfusate it was shown that the concentration (1%) of ethanol used did not produce damaye effect on plasma membranes and membranes of subcellular structures of hepatocytes, but, within certain limits, it displayed a stabilizing effect.
Subject(s)
Ethanol/pharmacology , Liver/metabolism , Animals , Bile/metabolism , Cell Membrane/drug effects , Cell Membrane/metabolism , Cell Membrane/ultrastructure , In Vitro Techniques , Liver/drug effects , Oxygen Consumption/drug effects , Perfusion , RatsABSTRACT
Effect of different concentration of non-ionic detergents (Triton X-100, Triton X-305, BRIJ-35 and Triton WR-1339) on total and non-sedimentable activity of 8 rat liver lysosome enzymes (acid phosphatase, acid DNase, acid RNase, arylsulphatases A and B, beta-glucuronidase, beta-galactosidase, beta-glucosidase and beta-acetylglucosaminidase) was studied. Only Triton X-100 at the concentration of 0.1% (and higher) was found to release completely lysosome enzymes. Low concentrations of Triton X-100 (0.025-0.05%) were used to characterize the strength of enzyme binding: the level of releasing acid DNase, beta-galactosidase, beta-glucuronidase and acid phsophatase being considerably higher than that of other lysosome enzymes studied. On the basis of the data obtained a method is worked out, which is suitable for series studies of the stability of lysosome membranes under different physiological and pathological conditions. The essence of the method is the treatment of membrane particles with increasing concentrations of Triton X-100 (0.025; 0.05 AND 0.1%) AND THE SUCCESSIVE ESTIMATION OF NON-Sedimentable activity of marker enzymes. The method detected troubles in the stability of rat liver lysosome membranes under starvation, protein deficiency and aging.
