Enzymes of lysosomal origin in human plasma and serum: Assay conditions and parameters influencing the assay

Lombardo, A.; Caimi, Luigi; Marchesini, Sergio; Goi, G.; Tettamanti, Guido

doi:10.1016/0009-8981(80)90339-3

Cited by 56 publications

(33 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3), which clearly supports the hypothesis suggested by Lombardo et al (32). On the other hand, the partial correlation study which was carried out seems to indicate that no significant release of leukocyte ß-jV-acetylhexosaminidase to serum takes place during coagulation.…”

Section: Discussionsupporting

confidence: 88%

“…On the other hand, Suzuki et al (30) and O'Brien (31) point out that ß-N-acetylhexosaminidase would be inhibited by heparin, which could explain the lower activity obtained in plasma collected with this anticoagulant. However, according to Lombardo et al (32), heparin, citrate, oxalate or EDTA would not affect per se ß-JV-acetylhexosaminidase activity, as our results have also shown. Likewise, these authors find significantly higher ß-7V-acetylhexosaminidase activity in serum than in plasma, and suggest that the release of enzyme from the platelets could occur during coagulation.…”

Section: Discussionsupporting

confidence: 82%

See 1 more Smart Citation

Assay of Serum/Plasma β-N-Acetylhexosaminidase Isoenzymes by Heat Inactivation Using a Continuous Spectrophotometric Method Adapted to a Centrifugal Analyzer

Lf¹,

Jc²

1997

Clinical Chemistry and Laboratory Medicine

View full text Add to dashboard Cite

Summary: Activity of serum/plasma -N-acetylhexosaminidase (EC 3.2.1.52) was determined by means of a continuous spectrophotometric method using 3,3'-dichlorophenylsulphonphthaleinyl-N-acetyl--Z)-glucosaniinide as substrate, with very satisfactory results. Incubation of an undiluted aliquot (1 ml) of samples at 52 °C for 8 hours with an adjusted pH 5.5-6.0 provoked only the inactivation of isoenzyme A, thus allowing the evaluation of -JVacetylhexosaminidase isoenzyme composition. In 25 serum samples from control subjects and pregnant women, a good correlation between the percentage of isoenzyme B obtained by this procedure and the fluorimetric assay of O'Brien et al. (New Engl J Med 1970; 273:15-20) was found (r = 0.983, Syx = 1.51), with no statistically significant difference between the means (43.2 vs 42.8%). In 84 healthy adult subjects, an average value of 30.3% for the proportion of isoenzyme B was obtained, with an interval of 25.4-35.0%, in agreement with results reported by other authors.

show abstract

Section: Discussionsupporting

confidence: 88%

Section: Discussionsupporting

confidence: 82%

Assay of Serum/Plasma β-N-Acetylhexosaminidase Isoenzymes by Heat Inactivation Using a Continuous Spectrophotometric Method Adapted to a Centrifugal Analyzer

Lf¹,

Jc²

1997

Clinical Chemistry and Laboratory Medicine

View full text Add to dashboard Cite

show abstract

“…An important role for IL-6 has been previously implicated in a variety of other cellular responses such as the mediation of the acute phase reaction in hepatocytes (49,50) and the modulation of immunologic responses dependent on lymphocytes (51) functions. This report represents the first demonstration that IL-6 may induce intercellular adherence molecules on mesenchymal cells and thereby influence neutrophil functions.…”

Section: Discussionmentioning

confidence: 99%

Neutrophil adherence to isolated adult cardiac myocytes. Induction by cardiac lymph collected during ischemia and reperfusion.

Youker¹,

Smith²,

Anderson³

et al. 1992

J. Clin. Invest.

178

View full text Add to dashboard Cite

Canine neutrophils can be induced to adhere in vitro to isolated adult cardiac myocytes by stimulation of the neutrophils with chemotactic factors such as zymosan-activated serum (ZAS) only if the myocytes have been previously exposed to cytokines such as interleukin 1 (IL-1) or tumor necrosis factor-a. These cytokines induce synthesis and surface expression ofintercellular adhesion molecule-i (ICAM-1) on the myocyte, and neutrophil adhesion is almost entirely CD18 and ICAM-1 dependent. The present study examines cardiac-specific lymph collected from awake dogs during i-h coronary occlusion and 3 d of reperfusion for its ability to induce both ICAM-1 expression in cardiac myocytes, and neutrophil-myocyte adherence. Reperfusion lymph induced ICAM-1 expression in isolated myocytes, and myocyte adherence to ZAS-stimulated neutrophils that was completely inhibited by anti-CD18 and anti-ICAM-i monoclonal antibodies. This activity peaked at 90 min of reperfusion and persisted for up to 72 h. Preischemic lymph was not stimulatory. IL-1 appeared not to be a stimulating factor in lymph in that dilutions of lymph were found to inhibit the stimulatory effects of recombinant IL-1it. However, investigation of interleukin 6 (IL-6) revealed that recombinant IL-6 stimulated myocyte adhesiveness for ZAS-stimulated neutrophils (ED90 = 0.002 U/ml) and expression of ICAM-1 by isolated myocytes. IL-6 neutralizing antibody markedly reduced the ability of reperfusion lymph to stimulate adhesion and ICAM-1 expression, and estimates of levels of IL-6 in reperfusion lymph ranged from 0.035 to 0.14 U/ml. These results indicate that cytokines capable of promoting neutrophil-myocyte adhesion occur in extracellular fluid during reperfusion of ischemic myocardium, and that one of these cytokines is IL-6. Neutrophilmyocyte adhesion may be of pathogenic significance because it may enhance the cytotoxic activity of the neutrophil. (J. Clin. Invest. 1992. 89:602-609.) Key words: cardiac lymph * cytokine induction * intercellular adhesion molecule-i * interleukin 6 * ischemia-reperfusion injury Introduction Numerous studies indicate that neutrophils contribute to myocardial cell injury after ischemia particularly under conditions

show abstract

“…The activities of ß-galactosidase, total hexosaminidase and hexosaminidase A in leukocytes varied independently of the processing conditions. Lombardo et al (28) consider ß-galactosidase to be the most labile enzyme. They observed that ß-galactosidase and ß-glucuronidase are released to a great extent from leukocytes and platelets, leading to an increase in plasma activities during refrigerated storage of blood (7).…”

Section: Discussionmentioning

confidence: 99%

Effect of collection, transport, processing and storage of blood specimens on the activity of lysosomal enzymes in plasma and leukocytes

Burin

Dutra-Filho

Brum³

et al. 2000

Braz J Med Biol Res

View full text Add to dashboard Cite

This study was designed to evaluate the effect of different conditions of collection, transport and storage on the quality of blood samples from normal individuals in terms of the activity of the enzymes ß-glucuronidase, total hexosaminidase, hexosaminidase A, arylsulfatase A and ß-galactosidase. The enzyme activities were not affected by the different materials used for collection (plastic syringes or vacuum glass tubes). In the evaluation of different heparin concentrations (10% heparin, 5% heparin, and heparinized syringe) in the syringes, it was observed that higher doses resulted in an increase of at least 1-fold in the activities of ß-galactosidase, total hexosaminidase and hexosaminidase A in leukocytes, and ß-glucuronidase in plasma. When the effects of time and means of transportation were studied, samples that had been kept at room temperature showed higher deterioration with time (72 and 96 h) before processing, and in this case it was impossible to isolate leukocytes from most samples. Comparison of heparin and acid citrate-dextrose (ACD) as anticoagulants revealed that ß-glucuronidase and hexosaminidase activities in plasma reached levels near the lower normal limits when ACD was used. In conclusion, we observed that heparin should be used as the preferable anticoagulant when measuring these lysosomal enzyme activities, and we recommend that, when transport time is more than 24 h, samples should be shipped by air in a styrofoam box containing wet ice.

show abstract

Enzymes of lysosomal origin in human plasma and serum: Assay conditions and parameters influencing the assay

Cited by 56 publications

References 21 publications

Assay of Serum/Plasma β-N-Acetylhexosaminidase Isoenzymes by Heat Inactivation Using a Continuous Spectrophotometric Method Adapted to a Centrifugal Analyzer

Assay of Serum/Plasma β-N-Acetylhexosaminidase Isoenzymes by Heat Inactivation Using a Continuous Spectrophotometric Method Adapted to a Centrifugal Analyzer

Neutrophil adherence to isolated adult cardiac myocytes. Induction by cardiac lymph collected during ischemia and reperfusion.

Effect of collection, transport, processing and storage of blood specimens on the activity of lysosomal enzymes in plasma and leukocytes

Contact Info

Product

Resources

About