Biochemical assays in lung homogenates

Artifacts caused by trapped blood after perfusion

Carroll E Cross, Tyrus T. Watanabe, Glen K. Hasegawa, Gary N. Goralnik, Karen E. Roertgen, Tokio Kaizu, Karen M. Reiser, Arnold B. Gorin, Jerold A Last

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

The amounts of trapped whole blood in well-perfused homogenates of rat lung (from rat lungs weighing approximately 1-1.5 g wet weight) were quantitatively determined by a method which utilizes the difference in hemoglobin (Hb) absorbance at 415 nm after reduction of HbO2 to deoxyHb by dithionite. By adding known amounts of whole blood to unfiltered crude lung homogenates, we constructed standard curves which allowed us to determine residual blood contents of perfused homogenates. Homogenates contained a mean of 20 ± 2.4 (SE) μl of blood/lung. These data were used to calculate the effects that contaminating whole blood might make upon measured values of various biochemical parameters in lung cytosols and homogenates. Depending on the specific parameter evaluated, blood contamination can contribute a major (e.g., catalase, β-N-acetylglucosaminidase and protein), a trivial (e.g., lysozyme, DNA) or an intermediate (e.g., superoxide dismutase, glutathione peroxidase) fraction of the total amount of a given component present in lung homogenate or cytosol. In studies in which the overall magnitudes of measured lung biochemical changes have been ascribed mechanistic significance (such as in lung damage, repair, protective, or adaptative processes) or in which lung enzyme specific activities are being expressed, the effects of blood contamination must be taken into account.

Original languageEnglish (US)
Pages (from-to)99-109
Number of pages11
JournalToxicology and Applied Pharmacology
Volume48
Issue number1 PART 1
DOIs
StatePublished - Mar 30 1979

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Artifacts
Assays
Blood
Perfusion
Lung
Rats
Contamination
Cytosol
Dithionite
Acetylglucosaminidase
Weighing
Muramidase
Glutathione Peroxidase
Catalase
Superoxide Dismutase
Hemoglobins
Repair
DNA
Enzymes
Weights and Measures

ASJC Scopus subject areas

  • Pharmacology
  • Toxicology

Cite this

Biochemical assays in lung homogenates : Artifacts caused by trapped blood after perfusion. / Cross, Carroll E; Watanabe, Tyrus T.; Hasegawa, Glen K.; Goralnik, Gary N.; Roertgen, Karen E.; Kaizu, Tokio; Reiser, Karen M.; Gorin, Arnold B.; Last, Jerold A.

In: Toxicology and Applied Pharmacology, Vol. 48, No. 1 PART 1, 30.03.1979, p. 99-109.

Research output: Contribution to journalArticle

Cross, CE, Watanabe, TT, Hasegawa, GK, Goralnik, GN, Roertgen, KE, Kaizu, T, Reiser, KM, Gorin, AB & Last, JA 1979, 'Biochemical assays in lung homogenates: Artifacts caused by trapped blood after perfusion', Toxicology and Applied Pharmacology, vol. 48, no. 1 PART 1, pp. 99-109. https://doi.org/10.1016/S0041-008X(79)80012-5
Cross, Carroll E ; Watanabe, Tyrus T. ; Hasegawa, Glen K. ; Goralnik, Gary N. ; Roertgen, Karen E. ; Kaizu, Tokio ; Reiser, Karen M. ; Gorin, Arnold B. ; Last, Jerold A. / Biochemical assays in lung homogenates : Artifacts caused by trapped blood after perfusion. In: Toxicology and Applied Pharmacology. 1979 ; Vol. 48, No. 1 PART 1. pp. 99-109.
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