Platelets are stabilized in vivo by such mediators as nitric oxide, prostaglandin I2, prostacyclin, and endothelial ADPase
to prevent pathologic thrombus formation.
Platelets are continually exposed to a number of activating factors, such as collagen, thrombin, ADP, fibrinogen, vWF, and inhibitory factors such as prostacyclin ([PGI.sub.2]) and ADPase
In the absence of 10% DMSO, the adults worms homogenized presented an ADPase
activity of 76.49 [+ or -] 11.47 nmol Pi [mg.sup.-1] [min.sup.-1] and an ATPase activity of 55.80 [+ or -] 8.36 nmol Pi [mg.sup.-1] [min.sup.-1].
In the present work, we aimed to determine whether ecto-nucleotidases activities (ATPase, ADPase
, and AMPase) are present and measurable in the ultrasound-guided aspirated contents of ovarian cysts and whether these activities are altered in endometriotic cysts compared with simple cysts, affecting the inflammatory condition of endometriosis.
The reaction medium used to assay ADPase
activity contained 120 mM NaCl, 5.0 mM KCl, 60 mM glucose, 5 mM Ca[Cl.sub.2], and 50 mM Tris-HCl buffer, pH 7.5 in a final volume of 200 [micro]L.
Briefly, the single cell suspensions (5-10 x [10.sup.4] cells) were incubated for 45-60 min at 37[degrees]C in a final volume of 80 [micro]L RPMI-1640 in neutral pH supplemented with 4 mmol/L [beta]-glycerophosphate and the following tracer substrates: 500 [micro]M [[sup.3]H]ATP (ATPase), 500 [micro]M [[sup.3]H]ADP (ADPase
), 300 [micro]M [[sup.3]H]AMP (ecto-57-nucleotidase), 300 [micro]M [[sup.3]H]adenosine (adenosine deaminase), and 400 [micro]M [[sup.3]H]AMP plus 600 [micro]M [gamma]-phosphate-donating ATP (adenylate kinase).