Quantitative Histochemical and Cytochemical Assays

Noorden, C. J. F. Van; Gossrau, Reinhart

doi:10.1007/978-94-011-3094-3_6

“…nant hepatocytes was not so clear as in rat tumors, where strongly positive hepatocytes were found to coexist with completely negative cells (8). Considering that enzyme cytochemistry catalyzing the oxidation of NADPH by nitrobluetetrazolium can detect less than 15 pmol H2 m~r -~* s -~ in the section (25) and that the limit for the cytochemical detection of this enzyme has been estimated to be as low as 69 mIUlmg protein by comparative biochemical/cytochemical studies in rat hepatoma-derived cell lines (26), liver cells with less than 0.05 absorbance units could be reasonably assumed to be devoid of ALDH-3.…”

Section: Discussionmentioning

confidence: 99%

Cytochemical detection of a class 3 aldehyde dehydrogenase in human hepatocellular carcinoma

Chieco¹,

Stecca²,

Bolondi

³

et al. 1995

Liver

4

0

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The development of hepatocellular carcinoma in rodents treated with different chemical compounds is associated with the appearance in the cytosol of neoplastic liver cells of an unusual aldehyde dehydrogenase isozyme of class 3 (ALDH‐3) which is very active with aromatic aldehydes. This tumor‐associated isozyme is readily detected by enzyme cytochemistry using the substrate benzaldehyde with NADP as coenzyme. To determine whether human hepatocellular carcinomas express ALDH‐3, the activity of this isozyme was examined in frozen sections from 68 echo‐guided human liver biopsies. In 54 cases the guided biopsy was performed on one or more nodules suggestive for hepatocellular carcinoma found at ultrasonography within the liver parenchyma. The remaining 14 patients were affected by chronic active hepatitis or cirrhosis. An intense enzymatic activity was ascertained in 5 out of 36 hepatocellular carcinomas. In non‐neoplastic liver, in macroregenerative nodules and in metastatic adenocarcinomas enzymatic activity was not detectable. ALDH‐3‐positive tumors were typical hepatocellular carcinomas (histological grade II and III). These results suggest that ALDH‐3 is a phenotype associated with malignancy in human liver tumors.

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In situ kinetic measurements of d-amino acid oxidase in rat liver with respect to its substrate specificity

Frederiks

¹

,

Noorden

²

,

Marx

³

et al. 1993

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D-Amino acid oxidase activity was demonstrated in peroxisomes of rat liver using unfixed cryostat sections and a histochemical technique using cerium ions as capture reagent for hydrogen peroxide and diaminobenzidine, cobalt ions and exogenous hydrogen peroxide to visualize the final reaction product for light microscopical analysis. Cytophotometric analysis of liver sections revealed similar zero-order reaction velocities of D-amino acid oxidase with activity twice as high in periportal areas as in pericentral areas of liver lobuli when using either D-proline or D,L-thiazolidine-2-carboxylic acid as substrates. On the other hand, a 4-5 times higher KM value was found for D-proline than for D,L-thiazolidine-2-carboxylic acid. The KM values in periportal and pericentral areas were similar for each substrate. These findings support the suggestion that the physiological substrate for D-amino acid oxidase may be D,L-thiazolidine-2-carboxylic acid, the adduct of cysteamine and glyoxylic acid. D-Amino acid oxidase may play a role in vivo in the production of oxalate which may participate in metabolic control processes as an intracellular messenger molecule.

show abstract

In situ kinetic measurements ofd-amino acid oxidase in rat liver with respect to its substrate specificity

Frederiks

¹

,

Noorden

²

,

Marx

³

et al. 1993

View full text Add to dashboard Cite

D-Amino acid oxidase activity was demonstrated in peroxisomes of rat liver using unfixed cryostat sections and a histochemical technique using cerium ions as capture reagent for hydrogen peroxide and diaminobenzidine, cobalt ions and exogenous hydrogen peroxide to visualize the final reaction product for light microscopical analysis. Cytophotometric analysis of liver sections revealed similar zero-order reaction velocities of D-amino acid oxidase with activity twice as high in periportal areas as in pericentral areas of liver lobuli when using either D-proline or D,L-thiazolidine-2-carboxylic acid as substrates. On the other hand, a 4-5 times higher KM value was found for D-proline than for D,L-thiazolidine-2-carboxylic acid. The KM values in periportal and pericentral areas were similar for each substrate. These findings support the suggestion that the physiological substrate for D-amino acid oxidase may be D,L-thiazolidine-2-carboxylic acid, the adduct of cysteamine and glyoxylic acid. D-Amino acid oxidase may play a role in vivo in the production of oxalate which may participate in metabolic control processes as an intracellular messenger molecule.

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Quantitative Histochemical and Cytochemical Assays

Cited by 5 publications

References 79 publications

Cytochemical detection of a class 3 aldehyde dehydrogenase in human hepatocellular carcinoma

Cytochemical detection of a class 3 aldehyde dehydrogenase in human hepatocellular carcinoma

In situ kinetic measurements of d-amino acid oxidase in rat liver with respect to its substrate specificity

In situ kinetic measurements ofd-amino acid oxidase in rat liver with respect to its substrate specificity

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