• 1.

    Mackness MI, Arrol S, Durrington PN. Paraoxonase prevents accumulation of lipoperoxides on low-density lipoprotein. FEBS Lett 1991; 286:152154.

    • Search Google Scholar
    • Export Citation
  • 2.

    Marsillach J, Aragonès G, Beltrán R, et al. The measurement of the lactonase activity of paraoxonase-1 in the clinical evaluation of patients with chronic liver impairment. Clin Biochem 2009; 42:9198.

    • Search Google Scholar
    • Export Citation
  • 3.

    Ciftci H, Savas M, Yeni E, et al. Serum paraoxonase activity in patients with low glomerular filtration rates. Ren Fail 2010; 32:562565.

    • Search Google Scholar
    • Export Citation
  • 4.

    Dong JT. Prevalent mutations in prostate cancer. J Cell Biochem 2006; 97:433447.

  • 5.

    Dullaart RP, Vries R, Sluiter WJ, et al. High plasma C-reactive protein is related to low paraoxonase-I activity independently of high leptin and low adiponectin in type 2 diabetes mellitus. Clin Endocrinol 2009; 70:221226.

    • Search Google Scholar
    • Export Citation
  • 6.

    Mackness B, Mackness MI, Arrol S, et al. Serum paraoxonase 55 and 192 polymorphism and paraoxonase activity and concentration in non-insulin dependent diabetes mellitus. Atherosclerosis 1998; 139:341349.

    • Search Google Scholar
    • Export Citation
  • 7.

    Kotani K, Sakane N, Sano Y, et al. Changes on the physiological lactonase activity of serum paraoxonase 1 by a diet intervention for weight loss in healthy overweight and obese women. J Clin Biochem Nutr 2009; 45:329334.

    • Search Google Scholar
    • Export Citation
  • 8.

    Ferretti G, Bacchetti T, Moroni C, et al. Paraoxonase activity in high-density lipoproteins: a comparison between healthy and obese females. J Clin Endocrinol Metab 2005; 90:17281733.

    • Search Google Scholar
    • Export Citation
  • 9.

    Mackness M, Durrington P, Mackness B. Paraoxonase I activity, concentration and genotype in cardiovascular disease. Curr Opin Lipidol 2004; 15:399404.

    • Search Google Scholar
    • Export Citation
  • 10.

    Kujiraoka T, Oka T, Ishihara M, et al. A sandwich enzyme-linked immunosorbent assay for human serum paraoxonase concentration. J Lipid Res 2000; 41:13581363.

    • Search Google Scholar
    • Export Citation
  • 11.

    Himbergen TM, Roest M, Graaf J, et al. Indications that paraoxonase-1 contributes to plasma high density lipoprotein levels in familial hypercholesterolemia. J Lipid Res 2005; 46:445451.

    • Search Google Scholar
    • Export Citation
  • 12.

    Khersonsky O, Tawfik DS. Chromogenic and fluorogenic assays for the lactonase activity of serum paraoxonases. Chembiochem 2006; 7:4953.

    • Search Google Scholar
    • Export Citation
  • 13.

    Motta S, Letellier C, Ropert M, et al. Protecting effect of vitamin E supplementation on submaximal exercise-induced oxidative stress in sedentary dogs as assessed by erythrocyte membrane fluidity and paraoxonase-1 activity. Vet J 2009; 181:288295.

    • Search Google Scholar
    • Export Citation
  • 14.

    Erel O. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin Biochem 2004; 37:277285.

    • Search Google Scholar
    • Export Citation
  • 15.

    Haagen L, Brock A. A new automated method for phenotyping arylesterase (EC 3.1.1.2) based upon inhibition of enzymatic hydrolysis of 4-nitrophenyl acetate by phenyl acetate. Eur J Clin Chem Clin Biochem 1992; 30:391395.

    • Search Google Scholar
    • Export Citation
  • 16.

    Browne RW, Koury ST, Marion S, et al. Accuracy and biological variation of human serum paraoxonase 1 activity and polymorphism (Q192R) by kinetic enzyme assay. Clin Chem 2007; 53:310317.

    • Search Google Scholar
    • Export Citation
  • 17.

    Martinez-Subiela S, Ceron JJ. Effects of haemolysis, lipaemia, hyperbilirubinemia, and anticoagulants in canine C-reacive protein, serum amyloid A, and ceruloplasnib assays. Can Vet J 2005; 46:625629.

    • Search Google Scholar
    • Export Citation
  • 18.

    Draganov DI, Teiber JF, Speelman A, et al. Human paraoxonases (PON1, PON2, and PON3) are lactonases with overlapping and distinct substrate specificities. J Lipid Res 2005; 46:12391247.

    • Search Google Scholar
    • Export Citation
  • 19.

    Tor ER, Holstege DM, Galey FD. Determination of cholinesterase activity in brain and blood samples using a plate reader. J AOAC Int 1997; 5:13081313.

    • Search Google Scholar
    • Export Citation
  • 20.

    Huen K, Richter R, Furlong C, et al. Validation of PON1 enzyme activity assays for longitudinal studies. Clin Chim Acta 2009; 402:6774.

    • Search Google Scholar
    • Export Citation
  • 21.

    Dantoine TF, Debord J, Charmes JP, et al. Decrease of serum paraoxonase activity in chronic renal failure. J Am Soc Nephrol 1998; 9:20822088.

    • Search Google Scholar
    • Export Citation
  • 22.

    Jayakumari N, Thejaseebai G. High prevalence of low serum paraoxonase-1 in subjects with coronary artery disease. J Clin Biochem Nutr 2009; 45:278284.

    • Search Google Scholar
    • Export Citation
  • 23.

    Alleman AR. The effects of haemolysis and lipaemia on serum biochemical constituents. Vet Med 1990; 85:12721284.

  • 24.

    Parra S, Marsillach J, Aragones G, et al. Methodological constraints in interpreting serum paraoxonase-1 activity measurements: an example from a study in HIV-infected patients. Lipids Health Dis 2010; 9:3236.

    • Search Google Scholar
    • Export Citation
  • 25.

    Mackness MI, Mackness B, Durrington PN, et al. Paraoxonase: biochemistry, genetics and relationship to plasma lipoproteins. Curr Opin Lipidol 1996; 7:6976.

    • Search Google Scholar
    • Export Citation
  • 26.

    Mackness I. Why plasma should not be used to study paraoxonase? Atherosclerosis 1998; 136:195196.

  • 27.

    Ham TH, Grauel JA, Dunn RF, et al. Physical properties of red cells as related to effects in vivo. IV. Oxidant drugs producing abnormal intracellular concentration of haemoglobin (eccentrocytes) with a rigid-red-cell haemolytic syndrome. J Lab Clin Med 1973; 82:898910.

    • Search Google Scholar
    • Export Citation
  • 28.

    Chan TK, Chan WC, Weed RI. Erythrocyte hemighosts: a hallmark of severe oxidative injury in vivo. Br J Hematol 1982; 50:575582.

  • 29.

    Harvey JW, Rackear D. Experimental onion-induced haemolytic anemia in dogs. Vet Pathol 1985; 22:387392.

  • 30.

    Caldin M, Carli E, Furanello T, et al. A retrospective study of 60 cases of eccentrocytosis in the dog. Vet Clin Pathol 2005; 34:224231.

    • Search Google Scholar
    • Export Citation
  • 31.

    Harvey JW. Pathogenesis, laboratory diagnosis, and clinical implications of erythrocyte enzyme deficiencies in dogs, cats, and horses. Vet Clin Pathol 2006; 35:144156.

    • Search Google Scholar
    • Export Citation
  • 32.

    Pavlica Z, Petelin M, Nemec A, et al. Measurement of total antioxidant capacity in gingival crevicular fluid and serum in dogs with periodontal disease. Am J Vet Res 2004; 65:15841588.

    • Search Google Scholar
    • Export Citation
  • 33.

    Camkerten I, Sahin T, Borazan G, et al. Evaluation of blood oxidant/antioxidant balance in dogs with sarcoptic mange. Vet Parasitol 2009; 161:106109.

    • Search Google Scholar
    • Export Citation
  • 34.

    Selec S, Aslan M, Horoz M, et al. Oxidative status and serum PON1 activity in beta-thalassemia minor. Clin Biochem 2007; 40:287291.

  • 35.

    Ferré N, Camps J, Prats E, et al. Serum paraoxonase activity: a new additional test for the improved evaluation of chronic liver damage. Clin Chem 2002; 48:261268.

    • Search Google Scholar
    • Export Citation
  • 36.

    Novak F, Vavrova L, Kodydkova J, et al. Decreased paraoxonase activity in critically ill patients with sepsis. Clin Exp Med 2010; 10:2125.

    • Search Google Scholar
    • Export Citation

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Validation of spectrophotometric assays for serum paraoxonase type-1 measurement in dogs

Asta Tvarijonaviciute DVM1, Fernando Tecles DVM, PhD2, Marco Caldin DVM, PhD3, Silvia Tasca DVM4, and José Cerón DVM, PhD5
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  • 1 Department of Animal Medicine and Surgery, Veterinary School, University of Murcia, 30100 Murcia, Spain
  • | 2 Department of Animal Medicine and Surgery, Veterinary School, University of Murcia, 30100 Murcia, Spain
  • | 3 San Marco Veterinary Hospital, Padova, Italy.
  • | 4 San Marco Veterinary Hospital, Padova, Italy.
  • | 5 Department of Animal Medicine and Surgery, Veterinary School, University of Murcia, 30100 Murcia, Spain

Abstract

Objective—To evaluate and validate 3 spectrophotometric assays for measuring serum activity of paraoxonase type-1 (PON1), an enzyme associated with high-density lipoproteins, in dogs.

Animals—22 healthy adult dogs and 10 dogs with eccentrocytosis.

Procedures—2 methods were adapted for use in 96-well microplates with phenyl acetate and 5-thiobutyl butyrolactonase as substrates, and 1 was adapted for use in an automated analyzer with p-nitrophenyl acetate as substrate. Blood samples were collected from all dogs, serum was harvested, and serum PON1 activity was measured with each method.

Results—Imprecision was low for all 3 methods, with the exception of interassay imprecision for 5-thiobutyl butyrolactonase, and results were linear across serial sample dilutions. The 3 methods were able to detect low PON1 activity when EDTA was used for blood sample collection, yielded lower PON1 values in sick dogs with eccentrocytosis than in healthy dogs, and yielded highly correlated results.

Conclusions and Clinical Relevance—The methods described here may allow a wider use of PON1 activity as a biomarker of oxidative stress in dogs in clinical and research settings. Results of each method were robust and precise (with the exception of the interassay values for the lactonase method), and the methods were easy to set up in a laboratory.

Abstract

Objective—To evaluate and validate 3 spectrophotometric assays for measuring serum activity of paraoxonase type-1 (PON1), an enzyme associated with high-density lipoproteins, in dogs.

Animals—22 healthy adult dogs and 10 dogs with eccentrocytosis.

Procedures—2 methods were adapted for use in 96-well microplates with phenyl acetate and 5-thiobutyl butyrolactonase as substrates, and 1 was adapted for use in an automated analyzer with p-nitrophenyl acetate as substrate. Blood samples were collected from all dogs, serum was harvested, and serum PON1 activity was measured with each method.

Results—Imprecision was low for all 3 methods, with the exception of interassay imprecision for 5-thiobutyl butyrolactonase, and results were linear across serial sample dilutions. The 3 methods were able to detect low PON1 activity when EDTA was used for blood sample collection, yielded lower PON1 values in sick dogs with eccentrocytosis than in healthy dogs, and yielded highly correlated results.

Conclusions and Clinical Relevance—The methods described here may allow a wider use of PON1 activity as a biomarker of oxidative stress in dogs in clinical and research settings. Results of each method were robust and precise (with the exception of the interassay values for the lactonase method), and the methods were easy to set up in a laboratory.

Contributor Notes

Address correspondence to Dr. Cerón (jjceron@um.es).